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Sample records for pilocarpine

  1. Pilocarpine iontophoresis test

    DEFF Research Database (Denmark)

    Hjortskov, N; Jepsen, Leif; Nielsen, B

    1995-01-01

    The pilocarpine iontophoresis test (P-test) is used as a predictor of the capacity to produce sweat. Therefore, we studied the reproducibility of this test in 12 normal subjects on 10 consecutive days. Furthermore, we determined whether the P-test reflects whole-body and regional sweat secretion...... during exercise in the heat. Finally, we determined whether the P-test stimulates the eccrine sweat glands to maximal sweat secretion. Six growth hormone-deficient (GHD) patients who are known to have decreased sweating, and 11 healthy control subjects were studied. To induce maximal sweat secretion......, the patients exercised on a bicycle ergometer at a workload corresponding to 40% of their maximal aerobic power (VO2max). The 11 healthy subjects exercised at a workload of 150 W. All subjects exercised for 60 min in ambient air at 35 degrees C, with 50% relative humidity. The P-test showed a mean day...

  2. Pilocarpine iontophoresis test

    DEFF Research Database (Denmark)

    Hjortskov, N; Jepsen, Leif; Nielsen, B

    1995-01-01

    The pilocarpine iontophoresis test (P-test) is used as a predictor of the capacity to produce sweat. Therefore, we studied the reproducibility of this test in 12 normal subjects on 10 consecutive days. Furthermore, we determined whether the P-test reflects whole-body and regional sweat secretion...... during exercise in the heat. Finally, we determined whether the P-test stimulates the eccrine sweat glands to maximal sweat secretion. Six growth hormone-deficient (GHD) patients who are known to have decreased sweating, and 11 healthy control subjects were studied. To induce maximal sweat secretion......, the patients exercised on a bicycle ergometer at a workload corresponding to 40% of their maximal aerobic power (VO2max). The 11 healthy subjects exercised at a workload of 150 W. All subjects exercised for 60 min in ambient air at 35 degrees C, with 50% relative humidity. The P-test showed a mean day...

  3. [Electroanalytical chemistry study of pilocarpine].

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    Wang, N X; Chen, J M; Lu, R C; Zhang, X L; Deng, J Q

    1990-01-01

    The electrochemical behaviour of pilocarpine was investigated by differential pulse polarography and cyclic voltammetry. The optimum conditions for the determination of pilocarpine were as follows: The electrolyte was 0.4 mol/L HAc-NaAc(pH 4.0) buffer solution, Zn (II) 1.0 x 10(-3) mol/L; initial potential -0.80 V (vs Ag-AgCl), final potential -1.30 V (vs Ag-AgCl), and voltage sweep rate 5 mV/s. A fair reproducibility in the above procedure and a good relationship between peak current and pilocarpine concentration (4.0 x 10(-6)-4.0 x 10(-5) mol/L) were achieved. The detection limit was 8.0 x 10(-7) mol/L. This paper also reports that the application of cyclic voltammetry to study electrode process behavior has been established. The PVC membrane ion-selective electrode for pilocarpine also was studied. It is based on the use of tetraphenylborate pilocarpine ion pair complex as the active material. The PVC membrane electrode showed Nernstian response over the pilocarpine concentration range from 1.0 x 10(-2) to 3.0 x 10(-5) mol/L, with a slope of 36 mV/decade, the detection limit being 3.0 x 10(-6) mol/L. The electrode gave fast response and good reproducibility. This method provides a rapid and simple way to the determination of pilocarpine in pharmaceutical preparations.

  4. Pilocarpine

    Science.gov (United States)

    ... tell your doctor if you have asthma, acute iritis (uveitis; swelling and irritation inside the eye), or ... any products such as vitamins, minerals, or other dietary supplements. You should bring this list with you each ...

  5. Vibrational spectra of pilocarpine hydrochloride crystals

    Energy Technology Data Exchange (ETDEWEB)

    Bento, R.R.F. [Universidade Federal de Mato Grosso (UFMT), Cuiaba, MT (Brazil). Inst. de Fisica; Freire, P.T.C. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil). Dept. de Fisica]. E-mail: tarso@fisica.ufc.br; Teixeira, A.M.R.; Silva, J.H. [Universidade Regional do Cariri, Crato, CE (Brazil). Dept. Ciencias Fisicas e Biologicas; Lima Junior, J.A. [Universidade Estadual do Ceara (UECE), Limoeiro do Norte, CE (Brazil); Oliveira, M.C.F. de; Andrade-Neto, M. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil). Dept. de Quimica Organica e Inorganica; Romero, N.R. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil). Dept. de Farmacia; Pontes, F.M. [Universidade Estadual Paulista, Bauru, SP (Brazil). Faculdade de Ciencias

    2009-03-15

    Pilocarpine is a natural substance with potential application in the treatment of several diseases. In this work Fourier Transform (FT)-Raman spectrum and the Fourier Transform infra red (FT-IR) spectrum of pilocarpine hydrochloride C{sub 11} H{sub 17} N{sub 2} O{sup +}{sub 2} .Cl{sup -1} were investigated at 300 K. Vibrational wavenumber and wave vector have been predicted using density functional theory (B3LYP) calculations with the 6-31 G(d,p) basis set. A comparison with experiment allowed to assign most of the normal modes of the crystal. (author)

  6. Modulation of pilocarpine-induced seizures by cannabinoid receptor 1.

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    Rebecca L Kow

    Full Text Available Administration of the muscarinic agonist pilocarpine is commonly used to induce seizures in rodents for the study of epilepsy. Activation of muscarinic receptors has been previously shown to increase the production of endocannabinoids in the brain. Endocannabinoids act at the cannabinoid CB1 receptors to reduce neurotransmitter release and the severity of seizures in several models of epilepsy. In this study, we determined the effect of CB1 receptor activity on the induction in mice of seizures by pilocarpine. We found that decreased activation of the CB1 receptor, either through genetic deletion of the receptor or treatment with a CB1 antagonist, increased pilocarpine seizure severity without modifying seizure-induced cell proliferation and cell death. These results indicate that endocannabinoids act at the CB1 receptor to modulate the severity of pilocarpine-induced seizures. Administration of a CB1 agonist produced characteristic CB1-dependent behavioral responses, but did not affect pilocarpine seizure severity. A possible explanation for the lack of effect of CB1 agonist administration on pilocarpine seizures, despite the effects of CB1 antagonist administration and CB1 gene deletion, is that muscarinic receptor-stimulated endocannabinoid production is acting maximally at CB1 receptors to modulate sensitivity to pilocarpine seizures.

  7. Rebamipide and mosapride enhance pilocarpine-induced salivation

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    Kazuo Hike

    2009-08-01

    Full Text Available Background: During esophageal acid clearance, salivation plays an important role in defending the esophageal mucosa. Mosapride, an agent used in chronic, long-term therapy of gastro-esophageal reflux disease (GERD was regarded as mediating its efficacy through prokinetic properties. Rebamipide is also widely used as an anti-gastritis and anti-ulcer agent in GERD patients with chronic gastritis. The aim of this study is to investigate the effects of rebamipide, mosapride, and risperidone on the salivation induced by pilocarpine. Materials and Methods: The experiments were conducted on 4-week male SD rats (120-150g. The salivation was induced by intraperitoneally administrated pilocarpine and saliva was collected using preweighted small cotton balls inserted into the animal's mouth every 30 min for 180 min. Thirteen minutes before intraperitoneal administration of pilocarpine, rebamipide, mosapride, and risperidone were administered intraduodenally. Control rats were conducted by intraperitoneal administration of saline and intraduodenal administration of 0.5% methylcellulose solution. Rsults: The saliva weight at 0-30 min was significantly (p<0.01 increased after administration of pilocarpine, compared to control rats. An additional administration of mosapride and rebamipide increased the saliva weight at 0-30 min. The total volume of saliva for 150 min after administration of pilocarpine was the highest after preadministration of rebamipide, followed by mosapride, and risperidone. Conclusions: Increase in salivation produced by i.p. pilocarpine was enhanced by preadministration of rebamipide and mosapride.

  8. Lipoic acid effects on glutamate and taurine concentrations in rat hippocampus after pilocarpine-induced seizures

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    P S Santos

    2011-01-01

    Full Text Available Pilocarpine-induced seizures can be mediated by increases in oxidative stress and by cerebral amino acid changes. The present research suggests that antioxidant compounds may afford some level of neuroprotection against the neurotoxicity of seizures in cellular level. The objective of the present study was to evaluate the lipoic acid (LA effects in glutamate and taurine contents in rat hippocampus after pilocarpine-induced seizures. Wistar rats were treated intraperitoneally (i.p. with 0.9% saline (Control, pilocarpine (400 mg/kg, Pilocarpine, LA (10 mg/kg, LA, and the association of LA (10 mg/kg plus pilocarpine (400 mg/kg, that was injected 30 min before of administration of LA (LA plus pilocarpine. Animals were observed during 24 h. The amino acid concentrations were measured using high-performance liquid chromatograph (HPLC. In pilocarpine group, it was observed a significant increase in glutamate content (37% and a decrease in taurine level (18% in rat hippocampus, when compared to control group. Antioxidant pretreatment significantly reduced the glutamate level (28% and augmented taurine content (32% in rat hippocampus, when compared to pilocarpine group. Our findings strongly support amino acid changes in hippocampus during seizures induced by pilocarpine, and suggest that glutamate-induced brain damage plays a crucial role in pathogenic consequences of seizures, and imply that strong protective effect could be achieved using lipoic acid through the release or decrease in metabolization rate of taurine amino acid during seizures.

  9. Antioxidant mechanisms of iso-6-cassine in suppressing seizures induced by pilocarpine

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    Rivelilson Mendes de Freitas

    2011-06-01

    Full Text Available The aim of this study was to evaluate the in vitro antioxidant effects of 12-[(2R,5R,6R-5-hydroxy-6-methylpiperidin-2-yl]dodecan-2-one (iso-6-cassine; ISO and the anticonvulsant effects of ISO on pilocarpine-induced seizures in rats. Wistar rats were treated with 0.9% saline (i.p., control group, pilocarpine (400 mg/kg, i.p., pilocarpine group, and the association of ISO (1.0 mg/kg, i.p. plus pilocarpine (400 mg/kg, i.p., 30 min after administration of ISO (ISO plus pilocarpine group. After the treatments all groups were observed for 1h. The antioxidant effect of ISO on the pilocarpine model was assessed by determining the activity of glutathione peroxidase (GPx, glutathione-S-transferase (GST and catalase (CAT as well as the levels of reactive species (RS and lipid peroxidation (LP. In vitro, ISO (5 μM reduced RS and LP. ISO (1.0 mg/kg and abolished seizures and death induced by pilocarpine in rats. ISO protected against the increase in the RS and LP levels, GST activity as well as the inhibition of GPx activity caused by pilocarpine. In addition, ISO increased the catalase activity in hippocampus of seized rats. In conclusion, the dta suggest that ISO can present anticonvulsant and antioxidant properties in the pilocarpine model of seizures in rats.

  10. New insights from the use of pilocarpine and kainate models.

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    Leite, J P; Garcia-Cairasco, N; Cavalheiro, E A

    2002-06-01

    Local or systemic administration of pilocarpine and kainate in rodents leads to a pattern of repetitive limbic seizures and status epilepticus, which can last for several hours. A latent period follows status epilepticus and precedes a chronic phase, which is characterized by the occurrence of spontaneous limbic seizures. These distinct features, in a single animal preparation, of an acute damage induced by status epilepticus, a silent interval between injury and the onset of spontaneous seizures, and a chronic epileptic state have allowed antiepileptic drug (AED) studies with different purposes, (a) in the acute phase, identification of compounds with efficacy against refractory status epilepticus and/or neuroprotection against damage induced by sustained seizures; (b) in the latent period, identification of agents with a potential for preventing epileptogenesis and/or against seizure-induced long-term behavioral deficits and (c) in the chronic phase, testing drugs effective against partial and secondarily generalized seizures. Studies on pilocarpine and kainate models have pointed out that some AEDs or other compounds exert an antiepileptogenic effect. The analogy of the latent phase of pilocarpine and kainate models with the acquisition of amygdala kindling should encourage testing of drugs that have proved to suppress the evolution of amygdala kindling. Drug testing in the chronic phase should not address only the suppression of secondarily generalized motor seizures. Most of current tools used to quantify spontaneous seizure events need to be coupled to electrophysiology and more sophisticated systems for recording and analyzing behavior.

  11. A centrally mediated prolonged hypotension produced by oxotremorine or pilocarpine

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    Dage, R.C.

    1979-01-01

    1 Oxotremorine, methyloxotremorine, pilocarpine or arecoline were given intravenously to anaesthetized cats, dogs or rats, and intraperitoneally to conscious normotensive and spontaneously hypertensive rats, pretreated with doses of methylatropine that completely blocked peripheral muscarinic receptors, to ascertain their effects on blood pressure and heart rate. 2 Oxotremorine but not methyloxotremorine produced a prolonged hypotension in cats and dogs but not in rats. Heart rate was not changed. Pilocarpine, although less potent, produced an identical effect, whereas the effect of arecoline was short by comparison. The hypotensive effect of these drugs was reversed by atropine. 3 In dogs, oxotremorine produced a prolonged hypotension with no change in heart rate or cardiac output. 4 A decrease in spontaneous sympathetic nerve activity accompanied the hypotension in cats. Both effects were reversed by atropine but could be reinvoked by large doses of oxotremorine. 5 The oxotremorine-induced hypotension in cats was not altered by decerebration but was abolished by high cervical spinal section. 6 The results indicate that the prolonged hypotension elicited by oxotremorine is mediated by an action at muscarinic receptors in the brain stem resulting in a decrease in sympathetic nerve activity and peripheral resistance but not heart rate or cardiac output. PMID:760887

  12. The pilocarpine model of epilepsy: what have we learned?

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    Fulvio A. Scorza

    2009-09-01

    Full Text Available The systemic administration of a potent muscarinic agonist pilocarpine in rats promotes sequential behavioral and electrographic changes that can be divided into 3 distinct periods: (a an acute period that built up progressively into a limbic status epilepticus and that lasts 24 h, (b a silent period with a progressive normalization of EEG and behavior which varies from 4 to 44 days, and (c a chronic period with spontaneous recurrent seizures (SRSs. The main features of the SRSs observed during the long-term period resemble those of human complex partial seizures and recurs 2-3 times per week per animal. Therefore, the pilocarpine model of epilepsy is a valuable tool not only to study the pathogenesis of temporal lobe epilepsy in human condition, but also to evaluate potential antiepileptogenic drugs. This review concentrates on data from pilocarpine model of epilepsy.A administração sistêmica do potente agonista muscarínico pilocarpina em ratos promove alterações comportamentais e eletrográficas que podem ser divididas em três períodos distintos: (a período agudo o animal evolui progressivamente para o status epilepticus, que perdura por até 24h; (b período silencioso, caracterizado pela normalização progressiva do comportamento e do EEG e pode ter uma duração de 4 a 44 dias; período crônico, aparecimento de crises epilépticas espontâneas e recorrentes (SRSs. As características das SRSs observadas nos animas durante o período crônico são semelhantes às crises parciais complexas dos seres humanos e recorrem de 2-3 vezes por semana/animal. Além disso, o modelo de epilepsia induzido pela pilocarpina é válido não somente para se estudar a patogênese da epilepsia do lobo temporal em humanos como também para se testar a viabilidade de drogas antiepilépticas. Esse artigo de revisão aborda diversos aspectos do modelo de epilepsia induzido pela pilocarpina.

  13. Interictal discharges in the hippocampus of rats with long-term pilocarpine seizures.

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    Nagao, T; Avoli, M; Gloor, P

    1994-06-20

    Systemic administration of pilocarpine to adult rats induces an acute status epilepticus followed by spontaneous recurrent seizures after a 1-2-week silent period. We recorded field potentials in hippocampal slices obtained from rats with spontaneous recurrent seizures after pilocarpine-induced status. The frequency of the interictal discharges induced in these slices by 4-aminopyridine (4AP) was reduced and their duration was increased. Cutting the Schaffer collaterals caused interictal discharges in CA1 to disappear in normal rats and in rats 3 weeks after pilocarpine-induced status. However, 12 weeks after pilocarpine, these discharges remained in CA1 after such a cut but occurred at a lower frequency. These findings show that in rat hippocampi with a lesion similar to that of human Ammon's horn sclerosis some electrophysiological features of 4AP-induced interictal discharges are altered in comparison to those induced in normal hippocampi.

  14. Molecular Mechanism of Bovine Trabecular Meshwork Cells Apoptosis Induced by Dexamethasone and Protection by Pilocarpine

    Institute of Scientific and Technical Information of China (English)

    Yajuan Gu; Shujun Zeng; Pengxin Qiu; Yuping Wu; Dawei Peng; Guangmei Yan

    2005-01-01

    Purpose: To study the molecular mechanism of trabecular meshwork cells apoptosis induced by dexamethasone and the protection of pilocarpine.Methods: Determining mRNA expression with reverse transcription-polymerase chain reaction (RT-PCR), protein expression with Western blots and the percentage of apoptotic cells with fluorescent microscopy.Results: Dexamethasone up-regulated Fas proteins and affected Bax, caspase-8 and caspase-9 proteins in an action of first decrease then increase. Pre-treatment with pilocarpine decreased the four proteins expression, which were increased by dexamethasone. Pilocarpine self could decrease pro-apoptotic factors Bax, caspase-8 and caspase-9 proteins expression.Conclusion: Fas/FasL pathway participated in apoptotic process induced by dexamethasone in trabecular meshwork cells and the process was probably related with both caspase-8 and caspase-9 pathways. Pilocarpine protected the cells against apoptosis through down-regulating Fas, Bax, caspase-8 and caspase-9 proteins expression.

  15. Moxonidine and rilmenidine injected into the medial septal area reduces the salivation induced by pilocarpine.

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    Saad, Wilson Abrão; de Arruda Camargo, Luis Antonio; Simões, Silvio; Saad, William Abrão; Guarda, Renata Saad; Guarda, Ismael Francisco Mota Siqueira

    2004-05-31

    We determined the effects of moxonidine and rilmenidine 20 nmol (alpha(2)-adrenergic and imidazoline receptor agonists) injected into the medial septal area (MSA) on the pilocarpine-induced salivation, when injected intraperitoneally (i.p.), of male Holtzman rats weighing 250-300 g, with stainless-steel cannula implanted into the MSA. The rats were anesthetized with zoletil 50 mg kg(-1) b.wt. (tiletamine chloridrate 125.0 mg and zolazepan chloridrate 125.0 mg) into quadriceps muscle intramuscularly (IM), saliva was collected using pre-weighed small cotton balls inserted in the animal's mouth. The pre-treatment with moxonidine injected into the MSA reduced the salivation induced by pilocarpine (1 mg kg(-1)) injected i.p. (12+/-3 mg min(-1)) vs. control (99+/-9 mg min(-1)). The pre-treatment with rilmenidine 40 nmol also reduced the salivation induce by pilocarpine injected i.p. (20+/-5 mg min(-1)) vs. control (94+/-7 mg min(-1)). Idazoxan 40 nmol (imidazoline receptor antagonist) injected into the MSA previous to moxonidine and rilmenidine partially blocked the effect of moxonidine and totally blocked the rilmenidine effect in pilocarpine-induced salivation injected i.p. (60+/-8 and 95+/-10 mg min(-1), respectively). Yohimbine 40 nmol (alpha(2)-adrenergic receptor antagonist) injected into the MSA previously to moxonidine and rilmenidine partially blocked the moxonidine effect but produced no change on the rilmenidine effect on i.p. pilocarpine-induced salivation (70+/-6 and 24+/-6 mg min(-1), respectively). Injection of these alpha(2)-adrenergic and imidazoline agonists and antagonists agents i.p. produced no change on i.p. pilocarpine-induced salivation. These results show that central, but not peripheral, injection of alpha(2)-adrenergic and imidazoline agonists' agents inhibit pilocarpine-induced salivation. Idazoxan, an imidazoline receptor antagonist, totally inhibits the rilmenidine effect and partially inhibits the moxonidine effect on pilocarpine

  16. Pilocarpine-induced seizures in rodents--17 years on.

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    Turski, W A

    2000-01-01

    In 1983, we reported that intracerebral or systemic administration of cholinergic agents produced seizures and seizure-related brain damage in rodents. During the following 17 years, seizures induced by cholinomimetic drugs became a popular model of epilepsy. Seizures can by produced by cholinergic agonists acting directly at muscarinic receptors or by drugs enhancing cholinergic transmission due to the inhibition of acetylcholinesterase activity. Status epilepticus evoked by pilocarpine in rodents triggers long-lasting changes which can be described as: (I) acute-onset seizures lasting for several hours, (II) a silent period characterized by normalization of electroencephalographic patterns lasting for days, and (III) spontaneous recurrent seizures lasting for life. Therefore, seizures induced by cholinomimetics in rodents are of value for studies of basic mechanisms of epileptogenesis and action of antiepileptic drugs.

  17. Effect on epileptogenesis of carbamazepine treatment during the silent period of the pilocarpine model of epilepsy.

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    Capella, H M; Lemos, T

    2002-01-01

    This study addresses the question of epileptogenesis by investigating the effects of carbamazepine (CBZ) on the silent period of the pilocarpine model of epilepsy. Adult Wistar rats were subjected to status epilepticus (SE) induced by pilocarpine and treated with CBZ or saline, i.p, during 56 days. Latency for the first spontaneous seizure, incidence, frequency, and duration of seizures were monitored for seizures and the hippocampal damage. CBZ did not abort the epileptogenesis but minimized the expression of seizures and hippocampal damage.

  18. Does pilocarpine-induced epilepsy in adult rats require status epilepticus?

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    Graciela Navarro Mora

    Full Text Available Pilocarpine-induced seizures in rats provide a widely animal model of temporal lobe epilepsy. Some evidences reported in the literature suggest that at least 1 h of status epilepticus (SE is required to produce subsequent chronic phase, due to the SE-related acute neuronal damage. However, recent data seems to indicate that neuro-inflammation plays a crucial role in epileptogenesis, modulating secondarily a neuronal insult. For this reason, we decided to test the following hypotheses: a whether pilocarpine-injected rats that did not develop SE can exhibit long-term chronic spontaneous recurrent seizures (SRS and b whether acute neurodegeneration is mandatory to obtain chronic epilepsy. Therefore, we compared animals injected with the same dose of pilocarpine that developed or did not SE, and saline treated rats. We used telemetric acquisition of EEG as long-term monitoring system to evaluate the occurrence of seizures in non-SE pilocarpineinjected animals. Furthermore, histology and MRI analysis were applied in order to detect neuronal injury and neuropathological signs. Our observations indicate that non-SE rats exhibit SRS almost 8 (+/22 months after pilocarpine-injection, independently to the absence of initial acute neuronal injury. This is the first time reported that pilocarpine injected rats without developing SE, can experience SRS after a long latency period resembling human pathology. Thus, we strongly emphasize the important meaning of including these animals to model human epileptogenesis in pilocarpine induced epilepsy.

  19. Does Pilocarpine-Induced Epilepsy in Adult Rats Require Status epilepticus?

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    Navarro Mora, Graciela; Bramanti, Placido; Osculati, Francesco; Chakir, Asmaa; Nicolato, Elena; Marzola, Pasquina; Sbarbati, Andrea; Fabene, Paolo Francesco

    2009-01-01

    Pilocarpine-induced seizures in rats provide a widely animal model of temporal lobe epilepsy. Some evidences reported in the literature suggest that at least 1 h of status epilepticus (SE) is required to produce subsequent chronic phase, due to the SE-related acute neuronal damage. However, recent data seems to indicate that neuro-inflammation plays a crucial role in epileptogenesis, modulating secondarily a neuronal insult. For this reason, we decided to test the following hypotheses: a) whether pilocarpine-injected rats that did not develop SE can exhibit long-term chronic spontaneous recurrent seizures (SRS) and b) whether acute neurodegeneration is mandatory to obtain chronic epilepsy. Therefore, we compared animals injected with the same dose of pilocarpine that developed or did not SE, and saline treated rats. We used telemetric acquisition of EEG as long-term monitoring system to evaluate the occurrence of seizures in non-SE pilocarpineinjected animals. Furthermore, histology and MRI analysis were applied in order to detect neuronal injury and neuropathological signs. Our observations indicate that non-SE rats exhibit SRS almost 8 (+/22) months after pilocarpine-injection, independently to the absence of initial acute neuronal injury. This is the first time reported that pilocarpine injected rats without developing SE, can experience SRS after a long latency period resembling human pathology. Thus, we strongly emphasize the important meaning of including these animals to model human epileptogenesis in pilocarpine induced epilepsy. PMID:19503612

  20. Comparison between carbachol iontophoresis and intravenous pilocarpine stimulated accommodation in anesthetized rhesus monkeys.

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    Wendt, Mark; He, Lin; Glasser, Adrian

    2013-10-01

    Rhesus monkeys are an animal model for human accommodation and presbyopia and consistent and repeatable methods are needed to stimulate and measure accommodation in anesthetized rhesus monkeys. Accommodation has typically been pharmacologically stimulated with topical pilocarpine or carbachol iontophoresis. Intravenous (i.v.) pilocarpine has recently been shown to produce more natural, rapid and reproducible accommodative responses compared to topical pilocarpine. Here, i.v. pilocarpine was compared to carbachol iontophoresis stimulated accommodation. Experiments were performed under anaesthesia on five previously iridectomized monkeys aged 10-16 years. In three monkeys, accommodation was stimulated with carbachol iontophoresis in five successive experiments and refraction measured with a Hartinger coincidence refractometer. In separate experiments, accommodation was stimulated using a 5 mg/kg bolus of i.v. pilocarpine given over 30 s followed by a continuous infusion of 20 mg/kg/hr for 5.5 min in three successive experiments with the same monkeys as well as in single experiments with two additional monkeys. Refraction was measured continuously using photorefraction with baseline and accommodated refraction also measured with the Hartinger. In subsequent i.v. pilocarpine experiments with each monkey, accommodative changes in lens equatorial diameter were measured in real-time with video-image analysis. Maximum accommodation of three monkeys with carbachol iontophoresis (five repeats) was (mean ± SD; range) 14.0 ± 3.5; 9.9-20.3 D and with i.v. pilocarpine stimulation (three repeats) was 11.1 ± 1.1; 9.9-13.0 D. The average of the standard deviations of maximum accommodation from each monkey was 0.8 ± 0.3 D from carbachol iontophoresis and 0.3 ± 0.2 from i.v. pilocarpine. The average latency to the start of the response after carbachol iontophoresis was 2.5 ± 3.9; 0.0-12.0 min with a time constant of 12.7 ± 9.5; 2.3-29.2 min. The average

  1. Central muscarinic receptor subtypes involved in pilocarpine-induced salivation, hypertension and water intake.

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    Borella, T L; De Luca, L A; Colombari, D S A; Menani, J V

    2008-12-01

    Recent evidence has suggested that pilocarpine (ACh receptor agonist) injected peripherally may act centrally producing salivation and hypertension. In this study, we investigated the effects of specific M(1) (pirenzepine), M(2)/M(4) (methoctramine), M(1)/M(3) (4-DAMP) and M(4) (tropicamide) muscarinic receptor subtype antagonists injected into the lateral cerebral ventricle (LV) on salivation, water intake and pressor responses to peripheral pilocarpine. Male Holtzman rats with stainless steel cannulae implanted in the LV were used. Salivation was measured in rats anaesthetized with ketamine (100 mg per kg body weight) and arterial pressure was recorded in unanaesthetized rats. Salivation induced by i.p. pilocarpine (4 micromol per kg body weight) was reduced only by 4-DAMP (25-250 nmol) injected into the LV, not by pirenzepine, methoctramine or tropicamide at the dose of 500 nmol. Pirenzepine (0.1 and 1 nmol) and 4-DAMP (5 and 10 nmol) injected into the LV reduced i.p. pilocarpine-induced water intake, whereas metoctramine (50 nmol) produced nonspecific effects on ingestive behaviours. Injection of pirenzepine (100 nmol) or 4-DAMP (25 and 50 nmol) into the LV reduced i.v. pilocarpine-induced pressor responses. Tropicamide (500 nmol) injected into the LV had no effect on pilocarpine-induced salivation, pressor responses or water intake. The results suggest that central M(3) receptors are involved in peripheral pilocarpine-induced salivation and M(1) receptors in water intake and pressor responses. The involvement of M(3) receptors in water intake and pressor responses is not clear because 4-DAMP blocks both M(1) and M(3) receptors.

  2. Optimum dose range for the amelioration of tong term radiation-induced hyposalivation using prophylactic pilocarpine treatment

    NARCIS (Netherlands)

    Burlage, Fred Ronald; Roesink, Judith M.; Faber, Hette; Vissink, Arjan; Langendijk, Johannes A.; Kampinga, Harm H.; Coppes, Rob P.

    Background: To determine dose and time dependency of pilocarpine pre-treatment protection from late damage after unilateral irradiation of the rat parotid gland. Methods and materials: The right parotid gland of saline (1 mg/ml) or pilocarpine (4 mg/kg) pre-treated rats was irradiated with 10, 15

  3. Pilocarpine-induced seizure-like activity with increased BNDF and neuropeptide Y expression in organotypic hippocampal slice cultures

    DEFF Research Database (Denmark)

    Poulsen, Frantz Rom; Jahnsen, Henrik; Blaabjerg, Morten

    2002-01-01

    with the muscarinic receptor antagonist atropine (100 microM). Regardless of dose and exposure time, the pilocarpine treatment induced very limited neuronal cell death, recorded as cellular propidium iodide uptake. Cultures exposed to 5 mM pilocarpine for up to 7 days displayed increased BDNF expression when analyzed...

  4. Zinc chelation reduces hippocampal neurogenesis after pilocarpine-induced seizure.

    Directory of Open Access Journals (Sweden)

    Jin Hee Kim

    Full Text Available Several studies have shown that epileptic seizures increase hippocampal neurogenesis in the adult. However, the mechanism underlying increased neurogenesis after seizures remains largely unknown. Neurogenesis occurs in the subgranular zone (SGZ of the hippocampus in the adult brain, although an understanding of why it actively occurs in this region has remained elusive. A high level of vesicular zinc is localized in the presynaptic terminals of the SGZ. Previously, we demonstrated that a possible correlation may exist between synaptic zinc localization and high rates of neurogenesis in this area after hypoglycemia. Using a lithium-pilocarpine model, we tested our hypothesis that zinc plays a key role in modulating hippocampal neurogenesis after seizure. Then, we injected the zinc chelator, clioquinol (CQ, 30 mg/kg, into the intraperitoneal space to reduce brain zinc availability. Neuronal death was detected with Fluoro Jade-B and NeuN staining to determine whether CQ has neuroprotective effects after seizure. The total number of degenerating and live neurons was similar in vehicle and in CQ treated rats at 1 week after seizure. Neurogenesis was evaluated using BrdU, Ki67 and doublecortin (DCX immunostaining 1 week after seizure. The number of BrdU, Ki67 and DCX positive cell was increased after seizure. However, the number of BrdU, Ki67 and DCX positive cells was significantly decreased by CQ treatment. Intracellular zinc chelator, N,N,N0,N-Tetrakis (2-pyridylmethyl ethylenediamine (TPEN, also reduced seizure-induced neurogenesis in the hippocampus. The present study shows that zinc chelation does not prevent neurodegeneration but does reduce seizure-induced progenitor cell proliferation and neurogenesis. Therefore, this study suggests that zinc has an essential role for modulating hippocampal neurogenesis after seizure.

  5. Cyclooxygenase-2 inhibitor inhibits hippocampal synaptic reorganization in pilocarpine-induced status epilepticus rats

    Institute of Scientific and Technical Information of China (English)

    Hai-ju ZHANG; Ruo-peng SUN; Ge-fei LEI; Lu YANG; Chun-xi LIU

    2008-01-01

    Objective: To examine modulations caused by cyclooxygenase-2 (COX-2) inhibitors on altered microenvironments and overbalanced neurotransmitters in pilocarpine-induced epileptic status rats and to investigate possible mechanisms. Methods:Celecoxib (a COX-2 inhibitor) was administered 45 min prior to pilocarpine administration. The effects of COX-2 inhibitors on mIPSCs (miniature GABAergic inhibitory postsynaptic currents) of CA3 pyramidal cells in the hippocampus were recorded. Expressions of COX-2, c-Fos, newly generated neurons, and activated microgliosis wore analyzed by immunohistochemistry, and expressions of α-subunit of γ-amino butyric acid (GABAA) receptors and mitogen-activated protein kinase/extracellular sig-nal-regulated protein kinase (MAPK/ERK) activity were detected by Western blotting. Results: Pretreatment with celecoxib showed protection against pilocarpine-induced seizures. Celecoxib prevented microglia activation in the hilus and inhibited the abnormal neurogenesis and astrogliosis in the hippocampus by inhibiting MAPK/ERK activity and c-Fos transcription. Celecoxib also up-regulated the expression of GABAA receptors. NS-398 (N-2-cyclohexyloxy-4-nitrophenyl-methanesuifonamide), another COX-2 inhibitor, enhanced the frequency and decay time of mIPSCs. Conclusion: The COX-2 inhibitor celecoxib decreased neuronal excitability and prevented epileptogenesis in pilocarpine-induced status epilepticus rats. Celecoxib regulates synaptic reorganization by inhibiting astrogliosis and ectopic neurogenesis by attenuating MAPK/ERK signal activity, mediated by a GABAergic mechanism.

  6. Effects of isoflurane anesthesia and pilocarpine on rat parotid saliva flow

    DEFF Research Database (Denmark)

    Knudsen, Jacob Dronninglund; Nauntofte, Birgitte; Josipovic, M

    2011-01-01

    rats was 50% slower than that of the sham-irradiated rats. In conclusion, 1.5% isoflurane was found to be a good compromise between proper anesthesia and isoflurane-induced inhibition of saliva secretion. Pilocarpine induces saliva secretion in a dose-dependent matter, with supra-maximal stimulation...

  7. SDZ ENS 163 A NOVEL PILOCARPINE LIKE DRUG - PHARMACOLOGICAL INVITRO AND INVIVO PROFILE

    NARCIS (Netherlands)

    ENZ, A; BODDEKE, H; SAUTER, A; RUDIN, M; SHAPIRO, G

    1993-01-01

    The thiolactone analogue of pilocarpine, SDZ ENS 163, acts in vitro and in vivo as a partial agonist at M1/M3 and as an antagonist at M2 muscarinic receptors. In vitro, the properties of SDZ ENS 163 have been investigated in several functional models for muscarinic receptors: it is a full agonist at

  8. Anticonvulsive and antioxidant effects of curcumin on pilocarpine-induced seizures in rats

    Institute of Scientific and Technical Information of China (English)

    DU Peng; TANG Hai-yan; LI Xin; LIN Hao-jie; PENG Wei-feng; MA Yu; FAN Wei; WANG Xin

    2012-01-01

    Background Curcumin,an active ingredient of turmeric with antioxidant and anti-inflammatory properties has recently been reported to have anticonvulsant effects in several animal models of epilepsy.This study aimed to investigate the effects of curcumin on the pilocarpine rat model of status epilepticus.Methods The effect of intraperitoneal administration of curcumin (30,100,and 300 mg/kg) on pilocarpine-induced seizures in rats was tested.The correlation between seizure activity and hippocampal levels of nitric oxide synthase and free radicals was quantified.Whether curcumin treatment modulated these parameters was also investigated.Results Curcumin significantly increased seizure threshold at doses of 100 and 300 mg/kg.Rats with pilocarpineinduced seizures showed significantly elevated levels of malonaldehyde,nitric oxide synthase,and lactate dehydrogenase,but decreased levels of superoxide dismutase and glutathione compared with normal control rats.At doses of 100 and 300 mg/kg,curcumin reversed the effects of pilocarpine-indUced seizures on nitric oxide synthase,lactate dehydrogenase,glutathione,and superoxide dismutase.However,curcumin did not restore the elevated malonaldehyde levels.Conclusion Curcumin has anticonvulsant activity in the pilocarpine rat model of seizures,and that modulation of free radicals and nitric oxide synthase may be involved in this effect.

  9. Acute toxicity and anticonvulsant activity of liposomes containing nimodipine on pilocarpine-induced seizures in mice.

    Science.gov (United States)

    Moreno, Lina Clara Gayoso e Almendra Ibiapina; Cavalcanti, Isabella Macário Ferro; Satyal, Prabodh; Santos-Magalhães, Nereide Stela; Rolim, Hercília Maria Lins; Freitas, Rivelilson Mendes

    2015-01-12

    Nimodipine has been shown to have an inhibitory action on seizures and brain damage in rodents. However, the pharmaceutical applicability of this drug is limited by its low solubility in gastrointestinal fluids and high first-pass effect in the liver, which leads to low bioavailability. These difficulties can be overcome through the use of liposomes. The aim of the present study is to evaluate the toxicity and anticonvulsant activity of liposomes containing nimodipine (NMD-Lipo) on pilocarpine-induced seizures. NMD-Lipo was prepared using the lipid-film hydration method. Central nervous system toxicity of NMD-Lipo was assessed by Hippocratic screening. Systemic toxicity was evaluated by analyses of biochemical and hematological parameters and by observing possible signs of toxicity. The possible anticonvulsant activity was tested by the pilocarpine model. The administration of the NMD-Lipo at doses of 0.1, 1, and 10 mg/kg caused no toxicity in animals. Furthermore, NMD-Lipo prevented the installation of 100% of the pilocarpine-induced seizures and prevented the death of 100% of the mice treated with pilocarpine. These data shown that NMD-Lipo has an anticonvulsant activity significantly superior to free NMD, suggesting that the liposomes promoted a drug controlled release by improving its bioavailability and consequently increasing its pharmacological activity. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  10. Salivary gland-sparing prophylactic pilocarpine treatment has no effect on tumor regrowth after irradiation

    NARCIS (Netherlands)

    Licht, R; Kampinga, HH; Coppes, RP

    2002-01-01

    Radiotherapy, of head and neck cancer frequently damages the salivary glands. Prophylactic administration of the muscarinic receptor agonist pilocarpine reduces subsequent radiation damage to the salivary glands in rats, but its effects on tumor cell radiosensitivity and tumor regrowth after irradia

  11. Neuronal and glial pathological changes during epileptogenesis in the mouse pilocarpine model.

    Science.gov (United States)

    Borges, Karin; Gearing, Marla; McDermott, Dayna L; Smith, Amy B; Almonte, Antoine G; Wainer, Bruce H; Dingledine, Raymond

    2003-07-01

    The rodent pilocarpine model of epilepsy exhibits hippocampal sclerosis and spontaneous seizures and thus resembles human temporal lobe epilepsy. Use of the many available mouse mutants to study this epilepsy model would benefit from a detailed neuropathology study. To identify new features of epileptogenesis, we characterized glial and neuronal pathologies after pilocarpine-induced status epilepticus (SE) in CF1 and C57BL/6 mice focusing on the hippocampus. All CF1 mice showed spontaneous seizures by 17-27 days after SE. By 6 h there was virtually complete loss of hilar neurons, but the extent of pyramidal cell death varied considerably among mice. In the mossy fiber pathway, neuropeptide Y (NPY) was persistently upregulated beginning 1 day after SE; NPY immunoreactivity in the supragranular layer after 31 days indicated mossy fiber sprouting. beta2 microglobulin-positive activated microglia, normally absent in brains without SE, became abundant over 3-31 days in regions of neuronal loss, including the hippocampus and the amygdala. Astrogliosis developed after 10 days in damaged areas. Amyloid precursor protein immunoreactivity in the thalamus at 10 days suggested delayed axonal degeneration. The mortality after pilocarpine injection was very high in C57BL/6 mice from Jackson Laboratories but not those from Charles River, suggesting that mutant mice in the C57BL/6(JAX) strain will be difficult to study in the pilocarpine model, although their neuropathology was similar to CF1 mice. Major neuropathological changes not previously studied in the rodent pilocarpine model include widespread microglial activation, delayed thalamic axonal death, and persistent NPY upregulation in mossy fibers, together revealing extensive and persistent glial as well as neuronal pathology.

  12. Comparison of sweat rate during graded exercise and the local rate induced by pilocarpine

    Directory of Open Access Journals (Sweden)

    Vimieiro-Gomes A.C.

    2005-01-01

    Full Text Available Centrally stimulated sweat rate produced by graded exercise until exhaustion was compared to the local sweat rate induced by pilocarpine, often used as a sweating index for healthy individuals. Nine young male volunteers (22 ± 4 years were studied in temperate environment in two situations: at rest and during progressive exercise with 25 W increases every 2 min until exhaustion, on a cycle ergometer. In both situations, sweating was induced on the right forearm with 5 ml 0.5% pilocarpine hydrochloride applied by iontophoresis (1.5 mA, 5 min, with left forearm used as control. Local sweat rate was measured for 15 min at rest. During exercise, whole-body sweat rate was calculated from the body weight variation. Local sweat rate was measured from the time when heart rate reached 150 bpm until exhaustion and was collected using absorbent filter paper. Pharmacologically induced local sweat rate at rest (0.4 ± 0.2 mg cm-2 min-1 and mean exercise-induced whole-body sweat rate (0.4 ± 0.1 mg cm-2 min-1 were the same (P > 0.05 but were about five times smaller than local exercise-induced sweat rate (control = 2.1 ± 1.4; pilocarpine = 2.7 ± 1.2 mg cm-2 min-1, indicating different sudorific mechanisms. Both exercise-induced whole-body sweat rate (P < 0.05 and local sweat rate (P < 0.05 on control forearm correlated positively with pilocarpine-induced local sweat rate at rest. Assuming that exercise-induced sweating was a result of integrated physiological mechanisms, we suggest that local and whole-body sweat rate measured during graded exercise could be a better sweating index than pilocarpine.

  13. Protection of salivary function by concomitant pilocarpine during radiotherapy : A double-blind, randomized, placebo-controlled study

    NARCIS (Netherlands)

    Burlage, Fred R.; Roesink, Judith M.; Kampinga, Harm H.; Coppes, Rob P.; Terhaard, Chris; Langendijk, Johannes A.; van Luijk, Peter; Stokman, Monique A.; Vissink, Arjan

    2008-01-01

    Purpose: To investigate the effect of concomitant administration of pilocarpine during radiotherapy for head-and-neck squamous cell carcinoma (HNSCC) on postradiotherapy xerostomia. Methods and Materials: A prospective, double blind, placebo-controlled randomized trial including 170 patients with

  14. Structural comparison of cytochromes P450 2A6, 2A13, and 2E1 with pilocarpine

    Energy Technology Data Exchange (ETDEWEB)

    DeVore, Natasha M.; Meneely, Kathleen M.; Bart, Aaron G.; Stephens, Eva S.; Battaile, Kevin P.; Scott, Emily E. (Kansas); (HWMRI)

    2013-11-20

    Human xenobiotic-metabolizing cytochrome P450 (CYP) enzymes can each bind and monooxygenate a diverse set of substrates, including drugs, often producing a variety of metabolites. Additionally, a single ligand can interact with multiple CYP enzymes, but often the protein structural similarities and differences that mediate such overlapping selectivity are not well understood. Even though the CYP superfamily has a highly canonical global protein fold, there are large variations in the active site size, topology, and conformational flexibility. We have determined how a related set of three human CYP enzymes bind and interact with a common inhibitor, the muscarinic receptor agonist drug pilocarpine. Pilocarpine binds and inhibits the hepatic CYP2A6 and respiratory CYP2A13 enzymes much more efficiently than the hepatic CYP2E1 enzyme. To elucidate key residues involved in pilocarpine binding, crystal structures of CYP2A6 (2.4 {angstrom}), CYP2A13 (3.0 {angstrom}), CYP2E1 (2.35 {angstrom}), and the CYP2A6 mutant enzyme, CYP2A6 I208S/I300F/G301A/S369G (2.1 {angstrom}) have been determined with pilocarpine in the active site. In all four structures, pilocarpine coordinates to the heme iron, but comparisons reveal how individual residues lining the active sites of these three distinct human enzymes interact differently with the inhibitor pilocarpine.

  15. Evaluation of behavioral parameters and mortality in a model of temporal lobe epilepsy induced by intracerebroventricular pilocarpine administration.

    Science.gov (United States)

    Medina-Ceja, Laura; Pardo-Peña, Kenia; Ventura-Mejía, Consuelo

    2014-06-06

    The pilocarpine model of temporal lobe epilepsy (TLE) is a useful tool that is used to investigate the mechanisms underlying the generation and maintenance of seizures. Although this model has been modified significantly to reduce mortality and to promote the appearance of spontaneous recurrent seizures, to date, no detailed evaluation has been performed of the behavioral parameters and mortality in TLE induced by intracerebroventricular pilocarpine administration; therefore, this was the goal of the present study. A single dose of pilocarpine hydrochloride (2.4 mg in a total volume of 2 µl) was injected into the right lateral brain ventricle of rats; the convulsive behavior was rated using the Racine scale and the mortality was analyzed in these animals. We found that 30-90 min after animals received intracerebroventricular pilocarpine injections, 73% developed status epilepticus (SE) with an activity score of 4/5 on the Racine scale. Moreover, these seizures were associated with the propagation of epileptiform activity to different hippocampal regions. Of the animals that developed SE, spontaneous recurrent seizures were observed in 32.5% at different times after SE induction. A 35% mortality rate was observed, which included animals that died during pilocarpine injection and after SE induction. On the basis of these findings, and given the observed latency between the insult (SE induction by pilocarpine injection) and the manifestation of spontaneous recurrent seizures, we propose that this model is a useful tool for basic biomedical research of SE and TLE.

  16. Effect of duration of pilocarpine-induced status epilepticus on subsequent cognitive function in rats.

    Science.gov (United States)

    Balakrishnan, S; Nidhi, G; Pandhi, P

    2001-03-01

    The aim of this study was to determine the effect of the duration of pilocarpine-induced status epilepticus (SE) on subsequent cognitive function in rats. SE was induced by pilocarpine (320 mg/kg i.p.) and was terminated by injection of 1 mg/kg diazepam at 30, 60 and 90 min in 3 groups of 10 rats each. Cognitive function was tested by a passive avoidance task and was assessed at the baseline and on days 1, 7, 14 and 21 (post SE). It was found that cognitive function was disrupted on days 7, 14 and 21 post SE in rats who had SE for 60 and 90 min, whereas it was not affected in rats that had 30 min of SE. Hence, the duration of SE may affect future cognitive performance and mandates emergency treatment.

  17. Effects of isoflurane anesthesia and pilocarpine on rat parotid saliva flow

    DEFF Research Database (Denmark)

    Knudsen, Jacob Dronninglund; Nauntofte, Birgitte; Josipovic, M

    2011-01-01

    The purpose of this study was to investigate the effects of isoflurane on unstimulated and pilocarpine-stimulated parotid saliva secretion. Ten male Sprague-Dawley rats weighing 350-400 g were randomized into two groups, and the saliva flow rate and lag phase were measured at two doses...... of isoflurane in a crossover study design. Increasing the isoflurane concentration from 1% to 2% was associated with a 19% decrease in saliva secretion rate, and the lag to saliva secretion was increased by 155%. To clarify whether the effect of isoflurane (1.5%) on the parotid flow varied with stimulus...... rats was 50% slower than that of the sham-irradiated rats. In conclusion, 1.5% isoflurane was found to be a good compromise between proper anesthesia and isoflurane-induced inhibition of saliva secretion. Pilocarpine induces saliva secretion in a dose-dependent matter, with supra-maximal stimulation...

  18. Increased levels of acidic calponin during dendritic spine plasticity after pilocarpine-induced seizures.

    Science.gov (United States)

    Ferhat, Lotfi; Esclapez, Monique; Represa, Alfonso; Fattoum, Abdellatif; Shirao, Tomoaki; Ben-Ari, Yezekiel

    2003-01-01

    We have previously shown that, in HEK 293 cells, overexpression of acidic calponin, an actin-binding protein, induces remodeling of actin filaments, leading to a change in cell morphology. In addition, this protein is found in dendritic spines of adult hippocampal neurons. We hypothesized that this protein plays a role in regulating actin-based filaments during dendritic spine plasticity. To assess this hypothesis, the pilocarpine model of temporal lobe epilepsy was selected because an important reorganization of the glutamatergic network, which includes an aberrant sprouting of granule cell axons, neo-synaptogenesis, and dendritic spine remodeling, is well established in the dentate gyrus. This reorganization begins after the initial period of status epilepticus after pilocarpine injection, during the silent period when animals display a normal behavior, and reaches a plateau at the chronic stage when the animals have developed spontaneous recurrent seizures. Our data show that the intensity of immunolabeling for acidic calponin was clearly increased in the inner one-third of the molecular layer of the dentate gyrus, the site of mossy fiber sprouting, and neo-synaptogenesis, at 1 and 2 weeks after pilocarpine injection (silent period) when the reorganization was taking place. In contrast, in chronic pilocarpine-treated animals, when the reorganization was established, the levels of labeling for acidic calponin in the inner molecular layer were similar to those observed in control rats. In addition, double immunostaining studies suggested that the increase in acidic calponin levels occurred within the dendritic spines. Altogether, these results are consistent with an involvement of acidic calponin in dendritic spine plasticity.

  19. Salivary stimulation by prolonged release of pilocarpine using films in diabetic rats.

    Directory of Open Access Journals (Sweden)

    Jesús Rodríguez

    2015-04-01

    Full Text Available Introduction: The local use of prolonged drug delivery in the oral cavity provides many advantages, i.e., it increases pharmacologic actions in the desired local site, allows smaller doses and reduces adverse effects. Pilocarpine is a cholinergic drug approved by the FDA for treating glandular hypofunction; however, the adverse effects associated with it limit its use. Objective: To evaluate cytotoxicity of films in adherent fibroblasts and their ability to release pilocarpine in vivo for a prolonged time in the oral cavity of diabetic rats and its effect on salivary flow.Methods: Chitosan and Hydroxypropylmethylcellulose (Methocel K4MCR films were prepared in 1% acetic acid and pilocarpine was added under magnetic stirring. Cytotoxicity of films was evaluated in adherent fibroblasts HS27 and assessed by neutral red technique. The sialogogue effect of films was evaluated on the floor of the mouth of diabetic rats. Later, histopathological analysis was performed using hematoxylin and eosin and Masson’s trichrome stains. Results: Films were biocompatible and had 96% cell viability. It was possible to increase stimulation of salivary flow in diabetic rats (6.36±0.987mg/hr compared to the control group (0.5±0.06mg/hr. The histopathological analysis did not show inflammatory infiltrate in the area where films were placed. Conclusion: Films were biocompatible and had high cell viability. Also, they considerably increased salivary flow in diabetic rats, without triggering an inflammatory infiltrate in the area which indicates that it is a biocompatible product for sustained release and safe for pilocarpine administration.

  20. Cytidine 5'-diphosphocholine (CDP-choline) adversely effects on pilocarpine seizure-induced hippocampal neuronal death.

    Science.gov (United States)

    Kim, Jin Hee; Lee, Dong Won; Choi, Bo Young; Sohn, Min; Lee, Song Hee; Choi, Hui Chul; Song, Hong Ki; Suh, Sang Won

    2015-01-21

    Citicoline (CDP-choline; cytidine 5'-diphosphocholine) is an important intermediate in the biosynthesis of cell membrane phospholipids. Citicoline serves as a choline donor in the biosynthetic pathways of acetylcholine and neuronal membrane phospholipids, mainly phosphatidylcholine. The ability of citicoline to reverse neuronal injury has been tested in animal models of cerebral ischemia and clinical trials have been performed in stroke patients. However, no studies have examined the effect of citicoline on seizure-induced neuronal death. To clarify the potential therapeutic effects of citicoline on seizure-induced neuronal death, we used an animal model of pilocarpine-induced epilepsy. Temporal lobe epilepsy (TLE) was induced by intraperitoneal injection of pilocarpine (25mg/kg) in adult male rats. Citicoline (100 or 300 mg/kg) was injected into the intraperitoneal space two hours after seizure onset and a second injection was performed 24h after the seizure. Citicoline was injected once per day for one week after pilocarpine- or kainate-induced seizure. Neuronal injury and microglial activation were evaluated at 1 week post-seizure. Surprisingly, rather than offering protection, citicoline treatment actually enhanced seizure-induced neuronal death and microglial activation in the hippocampus compared to vehicle treated controls. Citicoline administration after seizure-induction increased immunoglobulin leakage via BBB disruption in the hippocampus compared with the vehicle-only group. To clarify if this adverse effect of citicoline is generalizable across alternative seizure models, we induced seizure by kainate injection (10mg/kg, i.p.) and then injected citicoline as in pilocarpine-induced seizure. We found that citicoline did not modulate kainate seizure-induced neuronal death, BBB disruption or microglial activation. These results suggest that citicoline may not have neuroprotective effects after seizure and that clinical application of citicoline after

  1. In vivo Pharmacological Evaluations of Pilocarpine-Loaded Antioxidant-Functionalized Biodegradable Thermogels in Glaucomatous Rabbits

    Science.gov (United States)

    Chou, Shih-Feng; Luo, Li-Jyuan; Lai, Jui-Yang

    2017-02-01

    To alleviate oxidative stress-induced ocular hypertension, grafting of antioxidant molecules to drug carriers enables a dual-function mechanism to effectively treat glaucomatous intraocular pressure (IOP) dysregulation. Providing potential application for intracameral administration of antiglaucoma medications, this study, for the first time, aims to examine in vivo pharmacological efficacy of pilocarpine-loaded antioxidant-functionalized biodegradable thermogels in glaucomatous rabbits. A series of gallic acid (GA)-grafted gelatin-g-poly(N-isopropylacrylamide) (GN) polymers were synthesized via redox reactions at 20–50 °C. Our results showed that raising redox radical initiation reaction temperature maximizes GA grafting level, antioxidant activity, and water content at 40 °C. Meanwhile, increase in overall hydrophilicity of GNGA carriers leads to fast polymer degradation and early pilocarpine depletion in vivo, which is disadvantageous to offer necessary pharmacological performance at prolonged time. By contrast, sustained therapeutic drug concentrations in aqueous humor can be achieved for long-term (i.e., 28 days) protection against corneal aberration and retinal injury after pilocarpine delivery using dual-function optimized carriers synthesized at 30 °C. The GA-functionalized injectable hydrogels are also found to contribute significantly to enhancement of retinal antioxidant defense system and preservation of histological structure and electrophysiological function, thereby supporting the benefits of drug-containing antioxidant biodegradable thermogels to prevent glaucoma development.

  2. Lovastatin decreases the synthesis of inflammatory mediators during epileptogenesis in the hippocampus of rats submitted to pilocarpine-induced epilepsy.

    Science.gov (United States)

    Gouveia, T L F; Scorza, F A; Iha, H A; Frangiotti, M I B; Perosa, S R; Cavalheiro, E A; Silva, J A; Feliciano, R S; de Almeida, A C; Naffah-Mazzacoratti, M G

    2014-07-01

    Statins may act on inflammatory responses, decreasing oxidative stress and also reducing brain inflammation in several brain disorders. Epileptogenesis is a process in which a healthy brain becomes abnormal and predisposed to generating spontaneous seizures. We previously reported that lovastatin could prevent neuroinflammation in pilocarpine-induced status epilepticus (SE). In this context, this study investigated the long-lasting effects of lovastatin on mRNA expression of proinflammatory cytokines (interleukin-1β, tumor necrosis factor α, interleukin-6) and the antiinflammatory cytokine IL-10 in the hippocampus during epileptogenesis by immunohistochemistry and real time polymerase chain reaction (RT-PCR) during the latent and chronic phases in the epilepsy model induced by pilocarpine in rats. For these purposes, four groups of rats were employed: saline (CONTROL), lovastatin (LOVA), pilocarpine (PILO), and pilocarpine plus lovastatin (PILO+LOVA). After pilocarpine injection (350mg/kg, i.p.), the rats were treated with 20mg/kg of lovastatin via an esophagic probe 2h after SE onset. All surviving rats were continuously treated during 15days, twice/day. The pilocarpine plus lovastatin group showed a significant decrease in the levels of IL-1β, TNF-α, and IL-6 during the latent phase and a decreased expression of IL-1β and TNF-α in the chronic phase when compared with the PILO group. Moreover, lovastatin treatment also induced an increased expression of the antiinflammatory cytokine, IL-10, in the PILO+LOVA group when compared with the PILO group in the chronic phase. Thus, our data suggest that lovastin may reduce excitotoxicity during epileptogenesis induced by pilocarpine by increasing the synthesis of IL-10 and decreasing proinflammatory cytokines in the hippocampus.

  3. Rhythmic patterns in the thoracic nerve cord of the stick insect induced by pilocarpine

    Science.gov (United States)

    BÜSchges; Schmitz; BÄSsler

    1995-01-01

    Bath application of the muscarinic agonist pilocarpine onto the deafferented stick insect thoracic nerve cord induced long-lasting rhythmic activity in leg motoneurones. Rhythmicity was induced at concentrations as low as 1x10(-4) mol l-1 pilocarpine. The most stable rhythms were reliably elicited at concentrations from 2x10(-3) mol l-1 to 5x10(-3) mol l-1. Rhythmicity could be completely abolished by application of atropine. The rhythm in antagonistic motoneurone pools of the three proximal leg joints, the subcoxal, the coxo-trochanteral (CT) and the femoro-tibial (FT), was strictly alternating. In the subcoxal motoneurones, the rhythm was characterised by the retractor burst duration being correlated with cycle period, whereas the protractor burst duration was almost independent of it. The cycle periods of the rhythms in the subcoxal and CT motoneurone pools were in a similar range for a given preparation. In contrast, the rhythm exhibited by motoneurones supplying the FT joint often had about half the duration. The pilocarpine-induced rhythm was generated independently in each hemiganglion. There was no strict intersegmental coupling, although the protractor motoneurone pools of the three thoracic ganglia tended to be active in phase. There was no stereotyped cycle-to-cycle coupling in the activities of the motoneurone pools of the subcoxal joint, the CT joint and the FT joint in an isolated mesothoracic ganglion. However, three distinct 'spontaneous, recurrent patterns' (SRPs) of motoneuronal activity were reliably generated. Within each pattern, there was strong coupling of the activity of the motoneurone pools. The SRPs resembled the motor output during step-phase transitions in walking: for example, the most often generated SRP (SRP1) was exclusively exhibited coincident with a burst of the fast depressor trochanteris motoneurone. During this burst, there was a switch from subcoxal protractor to retractor activity after a constant latency. The activity of

  4. Skin pretreatment with microneedles prior to pilocarpine iontophoresis increases sweat production.

    Science.gov (United States)

    Wing, David; Prausnitz, Mark R; Buono, Michael J

    2013-11-01

    Collection of sweat via pilocarpine iontophoresis is commonly used to diagnose cystic fibrosis (CF), with thousands of tests performed each day. The main source of resistance to the passage of pilocarpine ions to the sweat glands is the electrical resistance of the stratum corneum. It was hypothesized that pretreating the skin with 0·5 mm-long microneedles would significantly decrease this resistance, thus increasing pilocarpine's permeation into the skin. Improved permeation should result in significantly reduced time to sweat initiation, time to collection of a clinically meaningful amount of sweat, and increased total amount of sweat produced in 15 min. Subjects (n = 12) had two 5 cm(2) areas on the forearm measured, marked and randomized to experimental (microneedles + iontophoresis) or control (iontophoresis alone). Microneedle pretreatment was conducted using a 35-needle microneedle stamp in a manner that 20 applications completely covered the 5 cm(2) treatment area. This was repeated five times for a total of 100 applications. Both experimental and control sites were placed under iontophoresis (1·5 mA) for 5 min. Microneedle pretreatment significantly decreased mean skin resistance (260 ± 27 kΩ versus 160 ± 19 kΩ, P = 0·006), while significantly increasing mean sweat rate (0·76 ± 0·35 versus 0·54 ± 0·19 μl cm(2) min(-1) , P = 0·007). No significant difference was found concerning pain (P = 0·059), number of active sweat glands (P = 0·627) or the osmolality of the collected sweat (P = 0·636). The results of this study suggest that microneedle pretreatment prior to pilocarpine iontophoresis significantly increases sweat production. Such results have the potential to improve the methodology currently used to diagnose cystic fibrosis and, more broadly, to administer drugs via the skin.

  5. Glucose utilisation during status epilepticus in an epilepsy model induced by pilocarpine: a qualitative study

    Directory of Open Access Journals (Sweden)

    Scorza Fulvio Alexandre

    2002-01-01

    Full Text Available Status epilepticus (SE is a medical emergency and it is associated to brain damage. 2-deoxy-[14C] glucose (2-DG procedure has been used to measure the alterations in the functional activity of the brain induced by various pharmacological and toxicological agents. The aim of this study was to determine which changes occur in the seizure anatomic substrates during the SE induced by pilocarpine (PILO using [14C]-2 deoxyglucose functional mapping technique. Wistar male adult rats were submitted to SE PILO-induced for 6h and received [14C] 2-deoxyglucose injection via jugular vein 45 min before the 6th hour of SE. The control animals were submitted to all procedures but received saline and not pilocarpine. Brain sections were prepared and exposed X-ray film about seven days. The optical density of each region was obtained using a solid state digital analyser. The analysis revealed that 14C-2DG utilisation was pronounced in the SE rats on the areas corresponding to the hippocampal formation (+50.6%, caudate-putamen (+30.6%, frontoparietal cortex (+32.2%, amygdala (+31.7%, entorrinal cortex (+28.2%, thalamic nucleus (+93.5%, pre-tectal area (+50.1% and substantia nigra (+50.3% when compared to control. Our results suggest that the different activation levels of the distinct structures may be particularly important for understanding triggering and spreading mechanisms underlying epileptic activity during status epilepticus.

  6. Investigation of oxidative stress involvement in hippocampus in epilepsy model induced by pilocarpine.

    Science.gov (United States)

    Freitas, R M

    2009-10-25

    The relationship between free radical and scavenger enzymes has been found in the epileptic phenomena and reactive oxygen species have been implicated in seizure-induced neurodegeneration. Using the epilepsy model obtained by systemic administration of pilocarpine in rats, we investigated the lipid peroxidation, nitrite content, superoxide dismutase (SOD) and catalase activities in the hippocampus of rats during chronic period. The enzyme activities as well as the lipid peroxidation and nitrite concentrations were measured using spectrophotometric methods and the results compared to values obtained from saline-treated animals. The superoxide dismutase and catalase activities increased during the chronic phase. In addition, lipid peroxidation and nitrite levels increased in same period in the hippocampus of animals observed during spontaneous recurrent seizures. Previous studies showed that animals presenting seizures and submitted to 24h of status epilepticus showed normal levels of superoxide dismutase and increased in catalase activities as well as an increase in hippocampal lipid peroxidation and nitrite concentrations. These results show a direct evidence of lipid peroxidation and nitrite during seizure activity that could be responsible for neuronal damage in the hippocampus of rats, during the establishment of pilocarpine model of epilepsy.

  7. The expression of somatostatin receptors in the hippocampus of pilocarpine-induced rat epilepsy model.

    Science.gov (United States)

    Kwak, Sung-Eun; Kim, Ji-Eun; Choi, Hui-Chul; Song, Hong-Ki; Kim, Yeong-In; Jo, Seung-Mook; Kang, Tae-Cheon

    2008-01-01

    During the course of this study, we sought examine whether the expression of somatostatin receptors (SSTRs) is altered in the hippocampus following pilocarpine-induced status epilepticus (SE) in order to understand the role/function of SSTRs in the hippocampus after epileptogenic insults. SSTR1 and SSTR4 immunoreactivities were increased in the hippocampus at 1 week after SE. At 4 weeks after SE, SRIF1-family (SSTR 2A, SSTR2B, and SSTR5) immunoreactivity was increased only in neuropil. Both SSTR2A and 2B immunoreactivities were increased in CA2-3 pyramidal cells. However, SSTR3 and SSTR4 immunoreactivities were reduced in the CA1 pyramidal cells of epileptic rat due to neuronal loss. In addition, SSTR5 immunoreactivity was reduced in CA2 pyramidal cells and various interneurons. Both SSTR2B and SSTR4 immunoreactivities were increased within microglia following SE. Our findings suggest that increases in neuron-glial SSTR expressions may be closely related to the enhanced inhibition of the dentate gyrus and regulation of reactive microgliosis in the hippocampus of a pilocarpine model of temporal lobe epilepsy.

  8. Glucose utilisation during status epilepticus in an epilepsy model induced by pilocarpine: a qualitative study.

    Science.gov (United States)

    Scorza, Fulvio Alexandre; Arida, Ricardo Mario; Priel, Margareth Rose; Calderazzo, Lineu; Cavalheiro, Esper Abrão

    2002-06-01

    Status epilepticus (SE) is a medical emergency and it is associated to brain damage. 2-deoxy-[14C] glucose (2-DG) procedure has been used to measure the alterations in the functional activity of the brain induced by various pharmacological and toxicological agents. The aim of this study was to determine which changes occur in the seizure anatomic substrates during the SE induced by pilocarpine (PILO) using [14C]-2 deoxyglucose functional mapping technique. Wistar male adult rats were submitted to SE PILO-induced for 6h and received [14C] 2-deoxyglucose injection via jugular vein 45 min before the 6th hour of SE. The control animals were submitted to all procedures but received saline and not pilocarpine. Brain sections were prepared and exposed X-ray film about seven days. The optical density of each region was obtained using a solid state digital analyser. The analysis revealed that 14C-2DG utilisation was pronounced in the SE rats on the areas corresponding to the hippocampal formation (+50.6%), caudate-putamen (+30.6%), frontoparietal cortex (+32.2%), amygdala (+31.7%), entorrinal cortex (+28.2%), thalamic nucleus (+93.5%), pre-tectal area (+50.1%) and substantia nigra (+50.3%) when compared to control. Our results suggest that the different activation levels of the distinct structures may be particularly important for understanding triggering and spreading mechanisms underlying epileptic activity during status epilepticus.

  9. Progranulin promotes activation of microglia/macrophage after pilocarpine-induced status epilepticus.

    Science.gov (United States)

    Zhu, Shanshan; Tai, Chao; Petkau, Terri L; Zhang, Si; Liao, Chengyong; Dong, Zhifang; Wen, Wendy; Chang, Qing; Tian Wang, Yu; MacVicar, Brian A; Leavitt, Blair R; Jia, William; Cynader, Max S

    2013-09-12

    Progranulin (PGRN) haploinsufficiency accounts for up to 10% of frontotemporal lobe dementia. PGRN has also been implicated in neuroinflammation in acute and chronic neurological disorders. Here we report that both protein and mRNA levels of cortical and hippocampal PGRN are significantly enhanced following pilocarpine-induced status epilepticus. We also identify intense PGRN immunoreactivity that colocalizes with CD11b in seizure-induced animals, suggesting that PGRN elevation occurs primarily in activated microglia and macrophages. To test the role of PGRN in activation of microglia/macrophages, we apply recombinant PGRN protein directly into the hippocampal formation, and observe no change in the number of CD11b(+) microglia/macrophages in the dentate gyrus. However, with pilocarpine-induced status epilepticus, PGRN application significantly increases the number of CD11b(+) microglia/macrophages in the dentate gyrus, without affecting the extent of hilar cell death. In addition, the number of CD11b(+) microglia/macrophages induced by status epilepticus is not significantly different between PGRN knockout mice and wildtype. Our findings suggest that status epilepticus induces PGRN expression, and that PGRN potentiates but is not required for seizure-induced microglia/macrophage activation.

  10. Exposure to Mozart music reduces cognitive impairment in pilocarpine-induced status epilepticus rats.

    Science.gov (United States)

    Xing, Yingshou; Qin, Yi; Jing, Wei; Zhang, Yunxiang; Wang, Yanran; Guo, Daqing; Xia, Yang; Yao, Dezhong

    2016-02-01

    Patients with temporal lobe epilepsy (TLE) often display cognitive deficits. However, current epilepsy therapeutic interventions mainly aim at how to reduce the frequency and degree of epileptic seizures. Recovery of cognitive impairment is not attended enough, resulting in the lack of effective approaches in this respect. In the pilocarpine-induced temporal lobe epilepsy rat model, memory impairment has been classically reported. Here we evaluated spatial cognition changes at different epileptogenesis stages in rats of this model and explored the effects of long-term Mozart music exposure on the recovery of cognitive ability. Our results showed that pilocarpine rats suffered persisting cognitive impairment during epileptogenesis. Interestingly, we found that Mozart music exposure can significantly enhance cognitive ability in epileptic rats, and music intervention may be more effective for improving cognitive function during the early stages after Status epilepticus. These findings strongly suggest that Mozart music may help to promote the recovery of cognitive damage due to seizure activities, which provides a novel intervention strategy to diminish cognitive deficits in TLE patients.

  11. Protection of salivary function by concomitant pilocarpine during radiotherapy : A double-blind, randomized, placebo-controlled study

    NARCIS (Netherlands)

    Burlage, Fred R.; Roesink, Judith M.; Kampinga, Harm H.; Coppes, Rob P.; Terhaard, Chris; Langendijk, Johannes A.; van Luijk, Peter; Stokman, Monique A.; Vissink, Arjan

    2008-01-01

    Purpose: To investigate the effect of concomitant administration of pilocarpine during radiotherapy for head-and-neck squamous cell carcinoma (HNSCC) on postradiotherapy xerostomia. Methods and Materials: A prospective, double blind, placebo-controlled randomized trial including 170 patients with HN

  12. The contribution of the lateral posterior and anteroventral thalamic nuclei on spontaneous recurrent seizures in the pilocarpine model of epilepsy

    Directory of Open Access Journals (Sweden)

    Scorza Fulvio Alexandre

    2002-01-01

    Full Text Available The pilocarpine model of epilepsy in rats is characterised by the occurrence of spontaneous seizures (SRSs during the chronic period that recur 2-3 times per week during the whole animal life. In a previous study on brain metabolism during the chronic period of the pilocarpine model it was possible to observe that, among several brain structures, the lateral posterior thalamic nuclei (LP showed a strikingly increased metabolism. Some evidences suggest that the LP can participate in an inhibitory control system involved in the propagation of the seizures. The aim of the present study was to verify the role of LP in the expression and frequency of spontaneous seizures observed in the pilocarpine model. Ten adult male rats presenting SRSs were monitored for behavioural events by video system one month before and one month after LP ibotenic acid lesion. Another group of chronic epileptic rats (n=10 had the anteroventral thalamic nuclei (AV lesioned by ibotenic acid. After the surgical procedure, the animals were sacrified and the brains were processed for histological analysis by the Nissl method. The LP group seizure frequency was 3.1±1.9 before ibotenic acid injection and showed an increase (16.3±7.2 per week after LP lesion. No changes in SRSs frequency were observed in the AV group after ibotenic lesion in these nuclei. These results seem to suggest that LP play a role in the seizure circuitry inhibiting the expression of spontaneous seizures in the pilocarpine model.

  13. Expressional analysis of the astrocytic Kir4.1 channel in a pilocarpine-induced temporal lobe epilepsy model.

    Science.gov (United States)

    Nagao, Yuki; Harada, Yuya; Mukai, Takahiro; Shimizu, Saki; Okuda, Aoi; Fujimoto, Megumi; Ono, Asuka; Sakagami, Yoshihisa; Ohno, Yukihiro

    2013-01-01

    The inwardly rectifying potassium (Kir) channel Kir4.1 in brain astrocytes mediates spatial K(+) buffering and regulates neural activities. Recent studies have shown that loss-of-function mutations in the human gene KCNJ10 encoding Kir4.1 cause epileptic seizures, suggesting a close relationship between the Kir4.1 channel function and epileptogenesis. Here, we performed expressional analysis of Kir4.1 in a pilocarpine-induced rat model of temporal lobe epilepsy (TLE) to explore the role of Kir4.1 channels in modifying TLE epileptogenesis. Treatment of rats with pilocarpine (350 mg/kg, i.p.) induced acute status epilepticus, which subsequently caused spontaneous seizures 7-8 weeks after the pilocarpine treatment. Western blot analysis revealed that TLE rats (interictal condition) showed significantly higher levels of Kir4.1 than the control animals in the cerebral cortex, striatum, and hypothalamus. However, the expression of other Kir subunits, Kir5.1 and Kir2.1, remained unaltered. Immunohistochemical analysis illustrated that Kir4.1-immunoreactivity-positive astrocytes in the pilocarpine-induced TLE model were markedly increased in most of the brain regions examined, concomitant with an increase in the number of glial fibrillary acidic protein (GFAP)-positive astrocytes. In addition, Kir4.1 expression ratios relative to the number of astrocytes (Kir4.1-positive cells/GFAP-positive cells) were region-specifically elevated in the amygdala (i.e., medial and cortical amygdaloid nuclei) and sensory cortex. The present study demonstrated for the first time that the expression of astrocytic Kir4.1 channels was elevated in a pilocarpine-induced TLE model, especially in the amygdala, suggesting that astrocytic Kir4.1 channels play a role in modifying TLE epileptogenesis, possibly by acting as an inhibitory compensatory mechanism.

  14. Expressional analysis of the astrocytic Kir4.1 channel in a pilocarpine-induced temporal lobe epilepsy model

    Directory of Open Access Journals (Sweden)

    Yuki eNagao

    2013-07-01

    Full Text Available The inwardly-rectifying potassium (Kir channel Kir4.1 in brain astrocytes mediates spatial K+ buffering and regulates neural activities. Recent studies have shown that loss-of-function mutations in the human gene KCNJ10 encoding Kir4.1 cause epileptic seizures, suggesting a close relationship between the Kir4.1 channel function and epileptogenesis. Here, we performed expressional analysis of Kir4.1 in a pilocarpine-induced rat model of temporal lobe epilepsy (TLE to explore the role of Kir4.1 channels in modifying TLE epileptogenesis. Treatment of rats with pilocarpine (350 mg/kg, i.p. induced acute status epilepticus, which subsequently caused spontaneous seizures 7–8 weeks after the pilocarpine treatment. Western blot analysis revealed that TLE rats (interictal condition showed significantly higher levels of Kir4.1 than the control animals in the cerebral cortex, striatum and hypothalamus. However, the expression of other Kir subunits, Kir5.1 and Kir2.1, remained unaltered. Immunohistochemical analysis illustrated that Kir4.1-immunoreactivity-positive astrocytes in the pilocarpine-induced TLE model were markedly increased in most of the brain regions examined, concomitant with an increase in the number of glial fibrillary acidic protein (GFAP-positive astrocytes. In addition, Kir4.1 expression ratios relative to the number of astrocytes (Kir4.1-positive cells/GFAP-positive cells were region-specifically elevated in the amygdala (i.e., medial and cortical amygdaloid nuclei and sensory cortex. The present study demonstrated for the first time that the expression of astrocytic Kir4.1 channels was elevated in a pilocarpine-induced TLE model, especially in the amygdala, suggesting that astrocytic Kir4.1 channels play a role in modifying TLE epileptogenesis, possibly by acting as an inhibitory compensatory mechanism.

  15. ACUTE AND CHRONIC RESPONSES TO THE CONVULSANT PILOCARPINE IN DBA/2J AND A/J MICE

    Science.gov (United States)

    WINAWER, M. R.; MAKARENKO, N.; McCLOSKEY, D. P.; HINTZ, T. M.; NAIR, N.; PALMER, A. A.; SCHARFMAN, H. E.

    2009-01-01

    Characterizing the responses of different mouse strains to experimentally-induced seizures can provide clues to the genes that are responsible for seizure susceptibility, and factors that contribute to epilepsy. This approach is optimal when sequenced mouse strains are available. Therefore, we compared two sequenced strains, DBA/2J (DBA) and A/J. These strains were compared using the chemoconvulsant pilocarpine, because pilocarpine induces status epilepticus, a state of severe, prolonged seizures. In addition, pilocarpine-induced status is followed by changes in the brain that are associated with the pathophysiology of temporal lobe epilepsy (TLE). Therefore, pilocarpine can be used to address susceptibility to severe seizures, as well as genes that could be relevant to TLE. A/J mice had a higher incidence of status, but a longer latency to status than DBA mice. DBA mice exhibited more hippocampal pyramidal cell damage. DBA mice developed more ectopic granule cells in the hilus, a result of aberrant migration of granule cells born after status. DBA mice experienced sudden death in the weeks following status, while A/J mice exhibited the most sudden death in the initial hour after pilocarpine administration. The results support previous studies of strain differences based on responses to convulsants. They suggest caution in studies of seizure susceptibility that are based only on incidence or latency. In addition, the results provide new insight into the strain-specific characteristics of DBA and A/J mice. A/J mice provide a potential resource to examine the progression to status. The DBA mouse may be valuable to clarify genes regulating other seizure-associated phenomena, such as seizure-induced neurogenesis and sudden death. PMID:17904758

  16. Malnutrition in infancy as a susceptibility factor for temporal lobe epilepsy in adulthood induced by the pilocarpine experimental model.

    Science.gov (United States)

    Cabral, Francisco Romero; Priel, Margareth Rose; Silva Araujo, Bruno Henrique; Brito Torres, Laila; de Lima, Eliangela; Gurgel do Vale, Tiago; Pereira, Felipe; Alves de Amorim, Henrique; Abrão Cavalheiro, Esper; Amado Scerni, Débora; Naffah-Mazzacoratti, Maria da Graça

    2011-01-01

    Malnutrition during the earliest stages of life may result in innumerable brain problems. Moreover, this condition could increase the chances of developing neurological diseases, such as epilepsy. We analyzed the effects of early-life malnutrition on susceptibility to epileptic seizures induced by the pilocarpine model of epilepsy. Wistar rat pups were kept on a starvation regimen from day 1 to day 21 after birth. At day 60, 16 animals (8 = well-nourished; 8 = malnourished) were exposed to the pilocarpine experimental model of epilepsy. Age-matched well-nourished (n = 8) and malnourished (n = 8) rats were used as controls. Animals were video-monitored over 9 weeks. The following behavioral parameters were evaluated: first seizure threshold (acute period of the pilocarpine model); status epilepticus (SE) latency; first spontaneous seizure latency (silent period), and spontaneous seizure frequency during the chronic phase. The cell and mossy fiber sprouting (MFS) density were evaluated in the hippocampal formation. Our results showed that the malnourished animals required a lower pilocarpine dose in order to develop SE (200 mg/kg), lower latency to reach SE, less time for the first spontaneous seizure and higher seizure frequency, when compared to well-nourished pilocarpine rats. Histopathological findings revealed a significant cell density reduction in the CA1 region and intense MFS among the malnourished animals. Our data indicate that early malnutrition greatly influences susceptibility to seizures and behavioral manifestations in adult life. These findings suggest that malnutrition in infancy reduces the threshold for epilepsy and promotes alterations in the brain that persist into adult life. Copyright © 2011 S. Karger AG, Basel.

  17. Rhythmic patterns evoked in locust leg motor neurons by the muscarinic agonist pilocarpine.

    Science.gov (United States)

    Ryckebusch, S; Laurent, G

    1993-05-01

    1. When an isolated metathoracic ganglion of the locust was superfused with the muscarinic cholinergic agonist pilocarpine, rhythmic activity was induced in leg motor neurons. The frequency of this induced rhythm increased approximately linearly from 0 to 0.2 Hz with concentrations of pilocarpine from 10(-5) to 10(-4) M. Rhythmic activity evoked by pilocarpine could be completely and reversibly blocked by 3 x 10(-5) M atropine, but was unaffected by 10(-4) M d-tubocurarine. 2. For each hemiganglion, the observed rhythm was characterized by two main phases: a levator phase, during which the anterior coxal rotator, levators of the trochanter, flexors of the tibia, and common inhibitory motor neurons were active; and a depressor phase, during which depressors of the trochanter, extensors of the tibia, and depressors of the tarsus were active. Activity in depressors of the trochanter followed the activity of the levators of the trochanter with a short, constant interburst latency. Activity in the levator of the tarsus spanned both phases. 3. The levator phase was short compared with the period (0.5-2 s, or 10-20% of the period) and did not depend on the period. The interval between the end of a levator burst and the beginning of the following one thus increased with cycle period. The depressor phase was more variable, and was usually shorter than the interval between successive levator bursts. 4. Motor neurons in a same pool often received common discrete synaptic potentials (e.g., levators of trochanter or extensors of tibia), suggesting common drive during the rhythm. Coactive motor neurons on opposite sides (such as left trochanteral depressors and right trochanteral levators), however, did not share obvious common postsynaptic potentials. Depolarization of a pool of motor neurons during its phase of activity was generally accompanied by hyperpolarization of its antagonist(s) on the same side. 5. Rhythmic activity was generally evoked in both hemiganglia of the

  18. Different Effects of Topical Prazosin and Pilocarpine on Uveoscleral Outflow in Rabbit Eyes

    Institute of Scientific and Technical Information of China (English)

    Jianming Wang; Yanping Song; Baichao Ren; Naixue Sun; Yinggui Yu; Haitao Hu

    2003-01-01

    Purpose: To investigate the effects of topical prazosin and pilocarpine on uveoscleral outflow(Fu) in rabbits.Methods: Sixteen rabbits were randomly divided into the control group (5 rabbits, only topical application of normal saline in the right eye of each rabbit), Prazosin (PZ) treated group (6 rabbits, only 0.1% Prazosin eyedrop 0.1% in the right eye of each one) and Pilocarpine (PC) treated group (5 rabbits, 1% Pilocarpine eye drop in each right eye).Intraocular pressure (IOP)of bilateral eyes of each rabbit was measured before and 1 h after topical application of the eye drop.And the bilateral eyes were perfused with Fluorescein-isothiocyanate bovine serum albumin (FITC-BSA) as the tracer into the anterior chamber of each rabbit for 30 min at 90 min after topical treatment. Then the rabbits were killed for Fu measurement.Results: IOP of PZ-treated eyes decreased [ (0.71 +0.07)kPa ] in 1 hour after PZ application.IOP of PC-treated eyes decreased [(0.70+0.08)kPa] in 1 hour after PC application. The average value of Fu was (0.176+0.048)μl/min in control eyes. The average value of Fu in PZ-treated eyes was (0.339+0.018)μl/min. The average value of Fu in PC-treated eye was(0.123+0.022) μl/min.Conclusion: Both of PZ and PC can decrease IOP in rabbits following topical application.Topical PZ can increase Fu in rabbits and this is one of the mechanisms of PZ-induced IOP decrease.Topical PC can prevent Fu. PC decreases IOP not through uveoscleral pathway.This study demonstrates that uveoscleral pathway plays an important role in aqueous humor drainage. PZ may be a novel drug for decreasing IOP. FITC-BSA is an effective tracer for studying uveoscleral pathway.

  19. Enhanced proliferation of acinar and progenitor cells by prophylactic pilocarpine treatment underlies the observed amelioration of radiation injury to parotid glands

    NARCIS (Netherlands)

    Burlage, Fred R.; Faber, Hette; Kampinga, Harm H.; Langendijk, Johannes A.; Vissink, Arjan; Coppes, Rob P.

    2009-01-01

    Background: Administration of pilocarpine before irradiation can ameliorate radiation-induced hyposalivation. Indirect evidence Suggests that this effect may be mediated through induction of a compensatory response. In this study, this hypothesis is tested directly, by assessing the proliferation of

  20. The lesional and epileptogenic consequences of lithium-pilocarpine-induced status epilepticus are affected by previous exposure to isolated seizures: effects of amygdala kindling and maximal electroshocks.

    Science.gov (United States)

    André, V; Ferrandon, A; Marescaux, C; Nehlig, A

    2000-01-01

    In temporal lobe epilepsy, the occurrence of seizures seems to correlate with the presence of lesions underlying the establishment of a hyperexcitable circuit. However, in the lithium-pilocarpine model of epilepsy, neuronal damage occurs both in the structures belonging to the circuit of initiation and maintenance of the seizures (forebrain limbic system) as in the propagation areas (cortex and thalamus) and in the circuit of remote control of seizures (substantia nigra pars reticulata). To determine whether or not we could protect the brain from lesions and epileptogenesis induced by status epilepticus and identify cerebral structures involved in the genesis of epilepsy, we studied the effects of the chronic exposure to non-deleterious seizures, either focalized with secondary generalization (amygdala kindling, kindled-pilocarpine rats), or primary generalized (ear-clip electroshocks, electroshock-pilocarpine rats) on neuronal damage and epileptogenesis induced by lithium-pilocarpine status epilepticus. These animals were compared to rats subjected to status epilepticus but not pretreated with seizures (sham-kindled-pilocarpine or sham-electroshock-pilocarpine rats). Compared to sham-pilocarpine rats, neuronal damage was prevented in the limbic system of the kindled-pilocarpine rats, except in the hilus of the dentate gyrus and the entorhinal cortex, while it was enhanced in rats pretreated with electroshocks, mainly in the entorhinal and perirhinal cortices. Most sham-kindled- and sham-electroshock-pilocarpine rats (92-100%) developed recurrent seizures after a silent period of 40-54days. Likewise, all kindled-pilocarpine rats developed spontaneous seizures after the same latency as their sham controls, while only two of 10 electroshock-pilocarpine rats became epileptic after a delay of 106-151days. The present data show that the apparent antiepileptic properties of electroshocks correlate with extensive damage in midbrain cortical regions, which may prevent the

  1. Microglial ablation and lipopolysaccharide preconditioning affects pilocarpine-induced seizures in mice

    Energy Technology Data Exchange (ETDEWEB)

    Mirrione, M.M.; Mirrione, M.M.; Konomosa, D.K.; Ioradanis, G.; Dewey, S.L.; Agzzid, A.; Heppnerd, F.L.; Tsirka, St.E.

    2010-04-01

    Activated microglia have been associated with neurodegeneration in patients and in animal models of Temporal Lobe Epilepsy (TLE), however their precise functions as neurotoxic or neuroprotective is a topic of significant investigation. To explore this, we examined the effects of pilocarpine-induced seizures in transgenic mice where microglia/macrophages were conditionally ablated. We found that unilateral ablation of microglia from the dorsal hippocampus did not alter acute seizure sensitivity. However, when this procedure was coupled with lipopolysaccharide (LPS) preconditioning (1 mg/kg given 24 h prior to acute seizure), we observed a significant pro-convulsant phenomenon. This effect was associated with lower metabolic activation in the ipsilateral hippocampus during acute seizures, and could be attributed to activity in the mossy fiber pathway. These findings reveal that preconditioning with LPS 24 h prior to seizure induction may have a protective effect which is abolished by unilateral hippocampal microglia/macrophage ablation.

  2. Pilocarpine-induced status epilepticus alters hippocampal PKC expression in mice.

    Science.gov (United States)

    Liu, Jian Xin; Liu, Yong; Tang, Feng Ru

    2011-01-01

    We investigated the protein expression of different protein kinase C (PKC) isoforms (PKC-alpha, PKC-beta1, PKC-beta2, PKC-gamma, PKC-delta, PKC-epsilon, PKC-eta and PKC-zeta) in the hippocampus of normal control mice and progressive changes in PKC isoforms expression during and after pilocarpine induced status epilepticus (PISE). We showed the reduced expression of PKC-delta, PKC-eta and PKC-zeta in interneurons in the CA1 area and in the hilus of the dentate gyrus during or after PISE. Increased expression of PKC-alpha and PKC-beta1 was demonstrated in the stratum pyramidale of CA3 area, and PKC-epsilon was up-regulated in the stratum lucidum of the CA3 area during or after PISE. Our results suggest that hippocampal PKC isoforms may play different roles in seizure generation, and be targets for development of anti-convulsive drugs.

  3. Synchrotron FTIR micro-spectroscopy study of the rat hippocampal formation after pilocarpine-evoked seizures.

    Science.gov (United States)

    Chwiej, J; Dulinska, J; Janeczko, K; Dumas, P; Eichert, D; Dudala, J; Setkowicz, Z

    2010-10-01

    In the present work, synchrotron radiation Fourier transform infrared (SRFTIR) micro-spectroscopy and imaging were used for topographic and semi-quantitative biochemical analysis of rat brain tissue in cases of pilocarpine-induced epilepsy. The tissue samples were analyzed with a beam defined by small apertures and spatial resolution steps of 10 microm which allowed us to probe the selected cellular layers of hippocampal formation. Raster scanning of the samples has generated 2D chemical cartographies revealing the distribution of proteins, lipids and nucleic acids. Spectral analysis has shown changes in the saturation level of phospholipids and relative secondary structure of proteins. Special interest was put in the analysis of two areas of the hippocampal formation (sector 3 of the Ammon's horn, CA3 and dentate gyrus, DG) in which elemental abnormalities were observed during our previous studies. Statistically significant increase in the saturation level of phospholipids (increased ratio of the absorption intensities at around 2921 and 2958 cm(-1)) as well as conformational changes of proteins (beta-type structure discrepancies as shown by the increased ratio of the absorbance intensities at around 1631 and 1657 cm(-1) as well as the ratio of the absorbance at 1548 and 1657 cm(-1)) were detected in pyramidal cells of CA3 area as well as in the multiform and molecular layers of DG. The findings presented here suggest that abnormalities in the protein secondary structure and increases in the level of phospholipid saturation could be involved in mechanisms of neurodegenerative changes following the oxidative stress evoked in brain areas affected by pilocarpine-induced seizures.

  4. Assessment of the progressive nature of cell damage in the pilocarpine model of epilepsy

    Directory of Open Access Journals (Sweden)

    L. Covolan

    2006-07-01

    Full Text Available Pilocarpine-induced (320 mg/kg, ip status epilepticus (SE in adult (2-3 months male Wistar rats results in extensive neuronal damage in limbic structures. Here we investigated whether the induction of a second SE (N = 6 would generate damage and cell loss similar to that seen after a first SE (N = 9. Counts of silver-stained (indicative of cell damage cells, using the Gallyas argyrophil III method, revealed a markedly lower neuronal injury in animals submitted to re-induction of SE compared to rats exposed to a single episode of pilocarpine-induced SE. This effect could be explained as follows: 1 the first SE removes the vulnerable cells, leaving behind resistant cells that are not affected by the second SE; 2 the first SE confers increased resistance to the remaining cells, analogous to the process of ischemic tolerance. Counting of Nissl-stained cells was performed to differentiate between these alternative mechanisms. Our data indicate that different neuronal populations react differently to SE induction. For some brain areas most, if not all, of the vulnerable cells are lost after an initial insult leaving only relatively resistant cells and little space for further damage or cell loss. For some other brain areas, in contrast, our data support the hypothesis that surviving cells might be modified by the initial insult which would confer a sort of excitotoxic tolerance. As a consequence of both mechanisms, subsequent insults after an initial insult result in very little damage regardless of their intensity.

  5. Subthalamic and Cortical Local Field Potentials Associated with Pilocarpine-Induced Oral Tremor in the Rat.

    Science.gov (United States)

    Long, Lauren L; Podurgiel, Samantha J; Haque, Aileen F; Errante, Emily L; Chrobak, James J; Salamone, John D

    2016-01-01

    Tremulous jaw movements (TJMs) are rapid vertical deflections of the lower jaw that resemble chewing but are not directed at any particular stimulus. In rodents, TJMs are induced by neurochemical conditions that parallel those seen in human Parkinsonism, including neurotoxic or pharmacological depletion of striatal dopamine (DA), DA antagonism, and cholinomimetic administration. Moreover, TJMs in rodents can be attenuated by antiparkinsonian agents, including levodopa (L-DOPA), DA agonists, muscarinic antagonists, and adenosine A2A antagonists. In human Parkinsonian patients, exaggerated physiological synchrony is seen in the beta frequency band in various parts of the cortical/basal ganglia/thalamic circuitry, and activity in the tremor frequency range (3-7 Hz) also has been recorded. The present studies were undertaken to determine if tremor-related local field potential (LFP) activity could be recorded from motor cortex (M1) or subthalamic nucleus (STN) during the TJMs induced by the muscarinic agonist pilocarpine, which is a well-known tremorogenic agent. Pilocarpine induced a robust TJM response that was marked by rhythmic electromyographic (EMG) activity in the temporalis muscle. Compared to periods with no tremor activity, TJM epochs were characterized by increased LFP activity in the tremor frequency range in both neocortex and STN. Tremor activity was not associated with increased synchrony in the beta frequency band. These studies identified tremor-related LFP activity in parts of the cortical/basal ganglia circuitry that are involved in the pathophysiology of Parkinsonism. This research may ultimately lead to identification of the oscillatory neural mechanisms involved in the generation of tremulous activity, and promote development of novel treatments for tremor disorders.

  6. Effects of ketogenic diets on the occurrence of pilocarpine-induced status epilepticus of rats.

    Science.gov (United States)

    Gama, Iclea Rocha; Trindade-Filho, Euclides Marinho; Oliveira, Suzana Lima; Bueno, Nassib Bezerra; Melo, Isabelle Tenório; Cabral-Junior, Cyro Rego; Barros, Elenita M; Galvão, Jaqueline A; Pereira, Wanessa S; Ferreira, Raphaela C; Domingos, Bruna R; da Rocha Ataide, Terezinha

    2015-02-01

    Two sources of medium-chain triglycerides--triheptanoin with anaplerotic properties and coconut oil with antioxidant features--have emerged as promising therapeutic options for the management of pharmacoresistant epilepsy. We investigated the effects of ketogenic diets (KDs) containing coconut oil, triheptanoin, or soybean oil on pilocarpine-induced status epilepticus (SE) in rats. Twenty-four adult male Wistar rats were divided into 4 groups and fed a control diet (7% lipids) or a KD containing soybean oil, coconut oil, or triheptanoin (69.8% lipids). The ketogenic and control diets had a lipid:carbohydrate + protein ratio of 1:11.8 and 3.5:1, respectively. SE was induced in all rats 20 days after initiation of the dietary treatment, through the administration of pilocarpine (340 mg/kg; i.p.). The latency, frequency, duration, and severity of seizures before and during SE were observed with a camcorder. SE was aborted after 3 h with the application of diazepam (5 mg/kg; i.p.). The rats in the triheptanoin-based KD group needed to undergo a higher number of seizures to develop SE, as compared to the control group (P < 0.05). Total weight gain, intake, energy intake, and feed efficiency coefficient, prior to induction of SE, differed between groups (P < 0.05), where the triheptanoin-based KD group showed less weight gain than all other groups, less energy intake than the Control group and intermediate values of feed efficiency coefficient between Control and other KDs groups. Triheptanoin-based KD may have a neuroprotective effect on the establishment of SE in Wistar rats.

  7. Pilocarpine Ophthalmic

    Science.gov (United States)

    ... hands thoroughly with soap and water. Use a mirror or have someone else apply the gel. Remove ... not go away: blurred or dim vision stinging, burning, or discomfort in the eye itching or redness ...

  8. A phase III randomized, double-blind, placebo-controlled study of pilocarpine for vaginal dryness: North Central Cancer Treatment group study N04CA.

    Science.gov (United States)

    Loprinzi, Charles L; Balcueva, Ernie P; Liu, Heshan; Sloan, Jeff A; Kottschade, Lisa A; Stella, Philip J; Carlson, Mark D; Moore, Dennis F; Zon, Robin T; Levitt, Ralph; Jaslowski, Anthony J

    2011-01-01

    Vaginal dryness is a common problem for which effective and safe nonestrogenic treatments are needed. Based on preliminary promising data that pilocarpine attenuated vaginal dryness, the current trial was conducted. A double-blind, placebo-controlled, randomized trial design was used to compare pilocarpine, at target doses of 5 mg twice daily and 5 mg four times daily, with a placebo. Vaginal dryness was recorded by patient-completed questionnaires at baseline and weekly for 6 weeks after study initiation. The primary endpoint for this study was the area under the curve summary statistic composed of the longitudinal responses obtained at baseline and through the 6 weeks of treatment to a numerical analogue scale asking patients to rate their perceived amount of vaginal dryness. The primary analysis was carried out by a single t test using a two-side alternative to compare the collective pilocarpine treatment arms with the collective placebo arms. A total of 201 patients enrolled in this trial. The primary analysis, comparing vaginal dryness symptoms in the collective pilocarpine arms against the placebo arm, did not reveal any benefit for the pilocarpine treatment. This finding was confirmed by other secondary analyses. Toxicity evaluation revealed more nausea, sweating, rigors, and urinary frequency with the pilocarpine arms compared with the placebo arm.

  9. Pilocarpine-induced epilepsy alters the expression and daily variation of the nuclear receptor RORα in the hippocampus of rats.

    Science.gov (United States)

    Rocha, Anna Karynna Alves de Alencar; de Lima, Eliangela; do Amaral, Fernanda Gaspar; Peres, Rafael; Cipolla-Neto, José; Amado, Débora

    2016-02-01

    It is widely known that there is an increase in the inflammatory responses and oxidative stress in temporal lobe epilepsy (TLE). Further, the seizures follow a circadian rhythmicity. Retinoic acid receptor-related orphan receptor alpha (RORα) is related to anti-inflammatory and antioxidant enzyme expression and is part of the machinery of the biological clock and circadian rhythms. However, the participation of RORα in this neurological disorder has not been studied. The aim of this study was to evaluate the RORα mRNA and protein content profiles in the hippocampus of rats submitted to a pilocarpine-induced epilepsy model at different time points throughout the 24-h light-dark cycle analyzing the influence of the circadian rhythm in the expression pattern during the acute, silent, and chronic phases of the experimental model. Real-time PCR and immunohistochemistry results showed that RORα mRNA and protein expressions were globally reduced in both acute and silent phases of the pilocarpine model. However, 60days after the pilocarpine-induced status epilepticus (chronic phase), the mRNA expression was similar to the control except for the time point 3h after the lights were turned off, and no differences were found in immunohistochemistry. Our results indicate that the status epilepticus induced by pilocarpine is able to change the expression and daily variation of RORα in the rat hippocampal area during the acute and silent phases. These findings enhance our understanding of the circadian pattern present in seizures as well as facilitate strategies for the treatment of seizures.

  10. Pilocarpine protects cobalt chloride-induced apoptosis of RGC-5 cells: involvement of muscarinic receptors and HIF-1 alpha pathway.

    Science.gov (United States)

    Zhu, Xu; Zhou, Wei; Cui, Yongyao; Zhu, Liang; Li, Juan; Feng, Xuemei; Shao, Biyun; Qi, Hong; Zheng, Jun; Wang, Hao; Chen, Hongzhuan

    2010-04-01

    The retina is the most metabolically active tissue in the human body and hypoxia-induced retinal ganglion cell (RGC) death has been implicated in glaucomatous optic neuropathy. The aim of this study is to determine whether muscarinic receptor agonist pilocarpine, a classic antiglaucoma drug, possesses neuroprotection against cobalt chloride (CoCl(2))-mimetic hypoxia-induced apoptosis of rat retinal ganglion cells (RGC-5 cells) and its underlying mechanisms. Cell viability was determined by Cell Counting Kit-8 assay and apoptosis was examined by annexin V and mitochondrial membrane potential (MMP) assays. Expressions of hypoxia-induced factor-1 alpha (HIF-1 alpha), p53, and BNIP3 were investigated by quantitative real-time PCR and western blot analysis. After treatment of 200 microM CoCl(2) for 24 h, RGC-5 cells showed a marked decrease of cell viability by approximately 30%, increased apoptosis rate and obvious decline in MMP, which could largely be reversed by the pretreatment of 1 microM pilocarpine mainly via the activation of muscarinic receptors. Meanwhile, pretreatment of 1 microM pilocarpine could significantly prevent CoCl(2)-induced HIF-1 alpha translocation from cytoplasm to nucleus and down-regulate the expression of HIF-1 alpha, p53, and BNIP3. These studies demonstrated that pilocarpine had effective protection against hypoxia-induced apoptosis in RGCs via muscarinic receptors and HIF-1 alpha pathway. The findings suggest that HIF-1 alpha pathway as a "master switch" may be used as a therapeutic target in the cholinergic treatment of glaucoma.

  11. Cognitive functions after pilocarpine-induced status epilepticus: changes during silent period precede appearance of spontaneous recurrent seizures.

    Science.gov (United States)

    Hort, J; Broźek, G; Mares, P; Langmeier, M; Komárek, V

    1999-09-01

    To study the possible relation between spontaneous recurrent seizures (SRS) and the derangement of cognitive memory. Status epilepticus (SE) was induced in adult Long-Evans rats by pilocarpine (320 mg/kg, i.p.) and interrupted after 2 h by clonazepam (CZPs mg/kg, i.p.). In addition to the animals that were given pilocarpine and CZP (group P), two groups received ketamine (100 mg/kg, i.p.): the first group 15 minutes after SE onset (group K15), and the second immediately after the CZP (group K120). Control groups were formed from animals not treated with pilocarpine as well as animals that received pilocarpine but did not develop motor seizures. Spatial cognitive memory was tested in the Morris water maze. Testing was impossible for more than 6 days after SE in group P. Ketamine shortened this period for the two groups that received it. During the silent period, deteriorated cognitive memory progressively improved, but the performance of group P started to worsen before the appearance of SRS. Group K120 only expressed a tendency toward declining performance, whereas group K15 never developed SRS, and the behavior of these animals did not differ from that of the controls after the postseizure period was over. Histologically, massive hippocampal cell loss was seen in group P. Ketamine protected hippocampal cells in a time-dependent manner; group K15 did not exhibit any obvious necrosis in the hippocampus. There is no close relation between cognitive functions and the appearance of SRS, because ketamine, administered 120 min after the beginning of SE, prevented the derangment of cognitive functions but not the appearance of SRSs.

  12. Mitochondrial and nuclear changes in hippocampal neurons in a lithium-pilocarpine-induced status epilepticus rat model

    Institute of Scientific and Technical Information of China (English)

    Shuhai Tang; Li Zhang; Jianying Sun; Xiaojun Pan

    2009-01-01

    BACKGROUND: Mitochondrial damage plays a key role in neuronal damage.OBJECTIVE: To observe ultrastructural damage to mitochondria and nuclei, as well as caspase-3 expression, in hippocampal CA3 neurons of lithium-pilocarpine-induced status epilepticus rats.DESIGN, TIME AND SETTING: The neuropathological, randomized, controlled study was performed at the Animal Experimental Center, Shandong University, China in May 2008.MATERIALS: A total of 75 healthy, adult, male, Wistar rats were randomly assigned into model (n = 45) and control (n = 30) groups. Lithium-pilocarpine (Sigma, USA) was used in this study.METHODS: Rats in the model group were intraperitoneally injected with lithium chloride (3 mEq/kg),and 24 hours later with pilocarpine (45 mg/kg), to induce seizures for 2 hours. Rats in the control group were intraperitoneally infused with the same volume of saline. Rat hippocampal CA3 tissue was obtained at 3, 12, and 24 hours following status epilepticus.MAIN OUTCOME MEASURES: Neuronal changes were observed under an optical microscope. Ultrastructural changes in mitochondria and nuclei were observed using an electron microscope.caspase-3 mRNA levels were quantified by semiquantitative RT-PCR.RESULTS: After 3 hours of status epilepticus, mitochondria with swollen cristae and ruptured membranes were observed by electron microscopy. Nuclei with marginated chromatin were observed after 24 hours status epilepticus. RT-PCR results demonstrated increased caspase-3 expression at 12 hours, and significantly increased expression at 24 hours following termination of status epilepticus. This was in accordance with acidophilia occurrence, as indicated by hematoxylin-eosin staining, and time of ultrastructural damage to nuclei.CONCLUSION: In lithium-pilocarpine-induced status epilepticus rat models, ultrastructural damage to mitochondria in hippocampal neurons occurred during early stages, followed by increased caspase-3 expression and nuclear changes. These results suggested

  13. Glutamate binding is altered in hippocampus and cortex of Wistar rats after pilocarpine-induced Status Epilepticus.

    Science.gov (United States)

    Cunha, Alexandra Olimpio Siqueira; Mortari, Márcia Renata; Carolino, Ruither Oliveira Gomes; Coutinho-Netto, Joaquim; Dos Santos, Wagner Ferreira

    2007-08-31

    Several evidences have pointed to biochemical alterations in some brain structures after experimental Status Epilepticus (SE). Thus, the effects of pilocarpine-induced SE on the glutamate binding in the hippocampus and cortex of Wistar rats were evaluated. Groups of animals were submitted to a 3h SE induced by intrahippocampal microinjection of pilocarpine, which was interrupted by the administration of sodium thiopental. Two weeks later the animals were sacrificed and had their cerebral cortices and hippocampi removed in order to perform the binding experiments. The results show that the pilocarpine-induced SE provoked an increase in 2.5-fold in the B(max) values for glutamate binding in the cortex, but not in the hippocampus. Moreover, we observed a 4-fold increase for the Kd values in the hippocampus and a 2-fold increase in the cortex. These findings might indicate that the epileptogenesis involves alterations in the glutamate receptors that are not restricted to the limbic system. Moreover, changes in these receptors are not exclusively of number, but rather involve the affinity for their ligands.

  14. Remarkable increase in 14C-acetate uptake in an epilepsy model rat brain induced by lithium-pilocarpine.

    Science.gov (United States)

    Hosoi, Rie; Kitano, Daisuke; Momosaki, Sotaro; Kuse, Kenji; Gee, Antony; Inoue, Osamu

    2010-01-22

    The present study demonstrates changes in rat brain glial metabolism during the acute phase of epilepsy. Status epilepticus (SE) was induced using the lithium-pilocarpine model. Glial metabolism was measured with (14)C-acetate. Local cerebral blood flow and glucose metabolism were also measured using (14)C-N-isopropyl-p-iodoamphetamine (IMP) and (14)C-2-deoxyglucose (2DG), respectively. At the initiation of the seizure, (14)C-acetate uptake did not change significantly. However, a marked increase was observed 2 h after the pilocarpine injection in all brain regions studied. The increase of brain uptake was transient, and the maximum enhancement was seen at 2 h after the pilocarpine injection. The increase of (14)C-acetate uptake was almost to the same degree in all regions, whereas (14)C-IMP and (14)C-2DG uptakes showed a heterogeneous increase. In the case of (14)C-IMP, the highest increase was observed in the thalamus (280%), and a moderate increase (120 to 150%) was seen in the orbital cortex, cingulate cortex and pyriform cortex. (14)C-2DG uptake increased by 130 to 240% in most regions of the brain, however, an increase of only 40 and 20% was observed in the cerebellum and pons-medulla, respectively. These results demonstrated that glial energy metabolism was markedly enhanced during a prolonged seizure. To our knowledge, this study is the first observation showing large and widespread glial metabolic increases in the rat brain during status epilepticus.

  15. Replacement of asymmetric synaptic profiles in the molecular layer of dentate gyrus following cycloheximide in the pilocarpine model in rats.

    Directory of Open Access Journals (Sweden)

    Simone eBittencourt

    2015-11-01

    Full Text Available Mossy fiber sprouting is among the best-studied forms of post-lesional synaptic plasticity and is regarded by many as contributory to seizures in both humans and animal models of epilepsy. It is not known whether mossy fiber sprouting increases the number of synapses in the molecular layer or merely replaces lost contacts. Using the pilocarpine model of status epilepticus to induce mossy fiber sprouting, and cycloheximide to block this sprouting, we evaluated at the ultrastructural level the number and type of asymmetric synaptic contacts in the molecular layer of the dentate gyrus. As expected, whereas pilocarpine-treated rats had dense silver grain deposits in the inner molecular layer (reflecting mossy fiber sprouting, pilocarpine+cycloheximide-treated animals did not differ from controls. Both groups of treated rats (Pilo group and CHX+Pilo group had reduced density of asymmetric synaptic profiles (putative excitatory synaptic contacts, which was greater for cycloheximide-treated animals. For both treated groups the loss of excitatory synaptic contacts was even greater in the outer molecular layer than in the best studied inner molecular layer (in which mossy fiber sprouting occurs. These results indicate that mossy fiber sprouting tends to replace lost synaptic contacts rather than increase the absolute number of contacts. We speculate that the overall result is more consistent with restored rather than with increased excitability.

  16. In vivo evaluation of anticonvulsant and antioxidant effects of phenobarbital microemulsion for transdermal administration in pilocarpine seizure rat model.

    Science.gov (United States)

    Figueiredo, Kayo Alves; Medeiros, Shirlene Cesário; Neves, Jamilly Kelly Oliveira; da Silva, José Alexsandro; da Rocha Tomé, Adriana; Carvalho, André Luis Menezes; de Freitas, Rivelilson Mendes

    2015-04-01

    This study aimed to evaluate a microemulsion system (ME) containing phenobarbital in epilepsy model induced by pilocarpine in rats and to oxidative stress and histologic lesions in hippocampus. The microemulsion was applied to the shaved back of Wistar rats. The animals were divided into the following groups: control group (P400); ME50 40mg/kg, topically-t.p.; ME100, 40mg/kg, t.p.; EM50, 40mg/kg, t.p.; phenobarbital solution 40mg/kg (PS), oral. After 60min, behavioral changes were evaluated for 1h in the model of epileptical crisis induced by pilocarpine. Phenobarbital in microemulsion was able to increase the latency for status epilepticus (SE) (p<0.05), decrease the number of epileptical crisis (ME50: p<0.001; ME100: p<0.01) and decrease mortality rate by 80% compared to P400. In EM50 and PS groups, deaths were decreased by 53.3% and 100% respectively. The ME50 and ME100 groups were able to reduce oxidative stress in experimental animals when compared to the P400. The microemulsion was still capable of reducing neuronal damage in the hippocampal areas. The results of this study come in an innovative way, demonstrating the ability of transdermal ME50 and ME100 to reduce pilocarpine-induced epileptical crisis, oxidative stress, besides neuronal damages. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Dendritic morphology, synaptic transmission, and activity of mature granule cells born following pilocarpine-induced status epilepticus in the rat

    Directory of Open Access Journals (Sweden)

    Fei eGao

    2015-10-01

    Full Text Available To understand the potential role of enhanced hippocampal neurogenesis after pilocarpine-induced status epilepticus (SE in the development of epilepsy, we quantitatively analyzed the geometry of apical dendrites, synaptic transmission, and activation levels of normotopically distributed mature newborn granule cells in the rat.SE in male Sprague-Dawley rats lasting for more than 2 hours was induced by an intraperitoneal injection of pilocarpine. The complexity, spine density, miniature post-synaptic currents, and activity-regulated cytoskeleton-associated protein (Arc expression of granule cells born five days after SE were studied at least 10 weeks after CAG-GFP retroviral vector-mediated labeling.Mature granule cells born after SE had dendritic complexity similar to that of granule cells born naturally, but with denser mushroom-like spines in dendritic segments located in the outer molecular layer. Miniature inhibitory post-synaptic currents (mIPSCs were similar between the controls and rats subjected to SE; however, smaller miniature excitatory post-synaptic current (mEPSC amplitude with a trend toward less frequent was found in mature granule cells born after SE. After maturation, granule cells born after SE did not show denser Arc expression in the resting condition or after being activated by transient seizure activity than vicinal GFP-unlabeled granule cells.Thus our results suggest that normotopic granule cells born after pilocarpine-induced SE are no more active when mature than age-matched, naturally born granule cells.

  18. Effects of pilocarpine and kainate-induced seizures on N-methyl-d-aspartate receptor gene expression in the rat hippocampus

    Energy Technology Data Exchange (ETDEWEB)

    Przewlocka, B.; Labuz, D.; Machelska, H.; Przewlocki, R.; Turchan, J.; Lason, W. [Department of Molecular Neuropharmacology, Institute of Pharmacology, Polish Academy of Sciences, 12 Smetna Street, 31-343 Krakow (Poland)

    1997-04-14

    The effects of pilocarpine- and kainate-induced seizures on N-methyl-d-aspartate receptor subunit-1 messenger RNA and [{sup 3}H]dizocilpine maleate binding were studied in the rat hippocampal formation. Pilocarpine- but not kainate-induced seizures decreased N-methyl-d-aspartate receptor subunit-1 messenger RNA level in dentate gyrus at 24 and 72 h after drug injection. Both convulsants decreased the messenger RNA level in CA1 pyramidal cells at 24 and 72 h, the effects of kainate being more profound. Kainate also decreased the N-methyl-d-aspartate receptor subunit-1 messenger RNA level in CA3 region after 24 and 72 h, whereas pilocarpine decreased the messenger RNA level at 72 h only. At 3 h after kainate, but not pilocarpine, an increased binding of [{sup 3}H]dizocilpine maleate in several apical dendritic fields of pyramidal cells was found. Pilocarpine reduced the [{sup 3}H]dizocilpine maleate binding in stratum lucidum only at 3 and 24 h after the drug injection. Pilocarpine but not kainate induced prolonged decrease in N-methyl-d-aspartate receptor subunit-1 gene expression in dentate gyrus. However, at the latest time measured, kainate had the stronger effect in decreasing both messenger RNA N-methyl-d-aspartate receptor subunit-1 and [{sup 3}H]dizocilpine maleate binding in CA1 and CA3 hippocampal pyramidal cells. The latter changes corresponded, however, to neuronal loss and may reflect higher neurotoxic potency of kainate.These data point to some differences in hippocampal N-methyl-d-aspartate receptor regulation in pilocarpine and kainate models of limbic seizures. Moreover, our results suggest that the N-methyl-d-aspartate receptor subunit-1 messenger RNA level is more susceptible to limbic seizures than is [{sup 3}H]dizocilpine maleate binding in the rat hippocampal formation. (Copyright (c) 1997 Elsevier Science B.V., Amsterdam. All rights reserved.)

  19. Role of nitric oxide of the median preoptic nucleus (MnPO in the alterations of salivary flow, arterial pressure and heart rate induced by injection of pilocarpine into the MnPO and intraperitoneally

    Directory of Open Access Journals (Sweden)

    Wilson A. Saad

    2003-07-01

    Full Text Available We investigated the effect of L-NAME, a nitric oxide (NO inhibitor and sodium nitroprusside (SNP, an NO-donating agent, on pilocarpine-induced alterations in salivary flow, mean arterial blood pressure (MAP and heart rate (HR in rats. Male Holtzman rats (250-300 g were implanted with a stainless steel cannula directly into the median preoptic nucleus (MnPO. Pilocarpine (10, 20, 40, 80, 160 µg injected into the MnPO induced an increase in salivary secretion (P<0.01. Pilocarpine (1, 2, 4, 8, 16 mg/kg ip also increased salivary secretion (P<0.01. Injection of L-NAME (40 µg into the MnPO prior to pilocarpine (10, 20, 40, 80, 160 µg injected into the MnPO or ip (1, 2, 4, 8, 16 mg/kg increased salivary secretion (P<0.01. SNP (30 µg injected into the MnPO or ip prior to pilocarpine attenuated salivary secretion (P<0.01. Pilocarpine (40 µg injection into the MnPO increased MAP and decreased HR (P<0.01. Pilocarpine (4 mg/kg body weight ip produced a decrease in MAP and an increase in HR (P<0.01. Injection of L-NAME (40 µg into the MnPO prior to pilocarpine potentiated the increase in MAP and reduced HR (P<0.01. SNP (30 µg injected into the MnPO prior to pilocarpine attenuated (100% the effect of pilocarpine on MAP, with no effect on HR. Administration of L-NAME (40 µg into the MnPO potentiated the effect of pilocarpine injected ip. SNP (30 µg injected into the MnPO attenuated the effect of ip pilocarpine on MAP and HR. The present study suggests that in the rat MnPO 1 NO is important for the effects of pilocarpine on salivary flow, and 2 pilocarpine interferes with blood pressure and HR (side effects of pilocarpine, that is attenuated by NO.

  20. Gabapentin administration reduces reactive gliosis and neurodegeneration after pilocarpine-induced status epilepticus.

    Directory of Open Access Journals (Sweden)

    Alicia Raquel Rossi

    Full Text Available The lithium-pilocarpine model of epilepsy reproduces in rodents several features of human temporal lobe epilepsy, by inducing an acute status epilepticus (SE followed by a latency period. It has been proposed that the neuronal network reorganization that occurs during latency determines the subsequent appearance of spontaneous recurrent seizures. The aim of this study was to evaluate neuronal and glial responses during the latency period that follows SE. Given the potential role of astrocytes in the post-SE network reorganization, through the secretion of synaptogenic molecules such as thrombospondins, we also studied the effect of treatment with the α2δ1 thrombospondin receptor antagonist gabapentin. Adult male Wistar rats received 3 mEq/kg LiCl, and 20 h later 30 mg/kg pilocarpine. Once SE was achieved, seizures were stopped with 20 mg/kg diazepam. Animals then received 400 mg/kg/day gabapentin or saline for either 4 or 14 days. In vitro experiments were performed in dissociated mixed hippocampal cell culture exposed to glutamate, and subsequently treated with gabapentin or vehicle. During the latency period, the hippocampus and pyriform cortex of SE-animals presented a profuse reactive astrogliosis, with increased GFAP and nestin expression. Gliosis intensity was dependent on the Racine stage attained by the animals and peaked 15 days after SE. Microglia was also reactive after SE, and followed the same pattern. Neuronal degeneration was present in SE-animals, and also depended on the Racine stage and the SE duration. Polysialic-acid NCAM (PSA-NCAM expression was increased in hippocampal CA-1 and dentate gyrus of SE-animals. Gabapentin treatment was able to reduce reactive gliosis, decrease neuronal loss and normalize PSA-NCAM staining in hippocampal CA-1. In vitro, gabapentin treatment partially prevented the dendritic loss and reactive gliosis caused by glutamate excitotoxicity. Our results show that gabapentin treatment during the

  1. Expression and significance of GIT1 in hippocampus of lithium-pilocarpine-induced epileptic rats

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    Li-hua ZHENG

    2015-07-01

    Full Text Available Objective  To investigate the expression changes of G-protein-coupled receptor kinase-interacting protein 1 (GIT1 in lithium-pilocarpine-induced epileptic rat model and explore its role in the genesis and development of epilepsy.  Methods  The lithium-pilocarpine-induced model of status epilepticus (SE was established in 42 specific pathogen free (SPF male adult Wistar rats, and those rats were randomly divided into control group and 6 epilepsy groups (1, 3, 7, 14, 30, 60 d after SE. The expression of GIT1 mRNA was detected by fluroescent quantitative polymerase chain reaction (PCR, while the expression of GIT1 protein was examined by Western blotting and immunohistochemistry was applied to test the expression of CA1 region, dentate gyrus and parahippocampal cortex in rat hippocampus at different time points.  Results  GIT1 mRNA level rised in acute phase on 1st and 3rd day after SE (P = 0.012, 0.002, then increased continously in latency on 7th and 14th day (P = 0.003, 0.001, and reached the peak in chronic phase on 30th and 60th day (P = 0.000, for all. GIT1 protein expression rised in acute phase and increased continously in chronic phase, but there was no significant difference compared with control group (P > 0.05, for all. Then, it reached the peak in chronic phase (P = 0.000, for all. Until the 30th day, the GIT1 expression of CA1 region, dentate gyrus and parahippocampal cortex in the hippocampus of rats in 6 epilepsy groups was significantly higher than that of control group (P = 0.000, for all.  Conclusions  The up-regulated expression of GIT1 in the hippocampus of epileptic rat was probably involved in the formation process of chronic epilepsy by regulating cytoskeleton dynamic regrouping to influence excitatory neural networks. DOI: 10.3969/j.issn.1672-6731.2015.06.011

  2. Normal spatial and contextual learning for ketamine-treated rats in the pilocarpine epilepsy model.

    Science.gov (United States)

    McKay, B E; Persinger, M A

    2004-05-01

    Cognitive impairments frequently accompany epileptic disorders. Here, we examine two neuroprotective agents, the noncompetitive NMDA antagonist ketamine and the dopaminergic antagonist acepromazine, for their efficacy in attenuating cognitive impairments in the lithium-pilocarpine (LI-PILO) model of rat limbic epilepsy. Declarative-like cognitive behaviors were assessed in a Morris water maze task that consisted successively of spatial and nonspatial (cued platform) training. Whereas the ketamine-treated (Ket) LI-PILO rats performed equally in all respects to nonseized control rats for the spatial and nonspatial components of the water maze task, the acepromazine-treated (Ace) LI-PILO rats failed to demonstrate learning in either the hidden or cued platform variants of the task and did not demonstrate any place learning in the platform-removed probe trials. We further assessed nondeclarative (associative) cognitive behaviors with a standard contextual fear-conditioning protocol. LI-PILO rats treated with acepromazine failed to learn the Pavlovian relationship; Ket LI-PILO rats performed equivalently to nonseized controls. Cumulatively, these data suggest robust cognitive sparing for LI-PILO rats with pharmacological NMDA receptor antagonism following induction of status epilepticus (SE). This cognitive sparing occurs despite earlier findings that the mean amount of total brain damage with LI-PILO is equivalent for Ket and Ace rats.

  3. Anticonvulsant effect of argan oil on pilocarpine model induced status epilepticus in wistar rats.

    Science.gov (United States)

    Bahbiti, Youssef; Ammouri, Hammou; Berkiks, Inssaf; El Hessni, Aboubaker; Ouichou, Ali; Nakache, Redouan; Chakit, Miloud; Bikjdaouene, Leila; Mesfioui, Abdelhalem

    2016-09-12

    Argan oil (AO) is rich in oleic and linoleic acids, polyphenols, sterols, and tocopherols. This composition gives it numerous beneficial pharmacological effects such as hypolipemiant, hypotensive, and antiproliferative. Oxidative stress is a mechanism of cell death induced by seizures and status epilepticus (SE). This study aims at investigating AO effects on (i) latency to first seizure, seizure severity, weight loss, mortality rate, (ii) lipid peroxidation level, nitrite level, and catalase activity in the hippocampus after SE induced by pilocarpine (PC). Wistar rats (1-month old) were daily administered by oral gavage with AO (1 ml/100 g/day) or with NaCl 0.9% during 2 months before receiving PC (400 mg/kg). After the PC injection, all groups were observed for 24 h. The catalase activity, the lipid peroxidation, and nitrite concentrations were measured using spectrophotometric methods. AO pretreatment increased the latency to first seizures, decreased the weight loss, and reduced mortality rate after SE. AO pretreatment produces significant decrease of the lipid peroxidation and nitrite levels. On the contrary, AO increased the catalase activity in rat hippocampus after seizures. For the first time, our results suggest that AO pretreatment is capable of attenuating seizure severity and oxidative stress in the hippocampus of Wistar rats. This indicates that AO may exhibit a neuroprotection against the temporal lobe epilepsy. Further investigations are in progress to confirm this pharmacological property.

  4. Manganese-enhanced magnetic resonance imaging detects mossy fiber sprouting in the pilocarpine model of epilepsy

    Science.gov (United States)

    Malheiros, Jackeline M.; Polli, Roberson S.; Paiva, Fernando F.; Longo, Beatriz M.; Mello, Luiz E.; Silva, Afonso C.; Tannús, Alberto; Covolan, Luciene

    2012-01-01

    Summary Purpose Mossy fiber sprouting (MFS) is a frequent finding following status epilepticus (SE). The present study aimed to test the feasibility of using manganese-enhanced magnetic resonance imaging (MEMRI) to detect MFS in the chronic phase of the well-established pilocarpine (Pilo) rat model of temporal lobe epilepsy (TLE). Methods To modulate MFS, cycloheximide (CHX), a protein synthesis inhibitor, was co-administered with Pilo in a sub-group of animals. In vivo MEMRI was performed 3 months after induction of SE and compared to the neo-Timm histological labeling of zinc mossy fiber terminals in the dentate gyrus (DG). Key findings Chronically epileptic rats displaying MFS as detected by neo-Timm histology had a hyperintense MEMRI signal in the DG, while chronically epileptic animals that did not display MFS had minimal MEMRI signal enhancement compared to non-epileptic control animals. A strong correlation (r = 0.81, P<0.001) was found between MEMRI signal enhancement and MFS. Significance This study shows that MEMRI is an attractive non-invasive method to detect mossy fiber sprouting in vivo and can be used as an evaluation tool in testing therapeutic approaches to manage chronic epilepsy. PMID:22642664

  5. Serotonin Depletion Does not Modify the Short-Term Brain Hypometabolism and Hippocampal Neurodegeneration Induced by the Lithium-Pilocarpine Model of Status Epilepticus in Rats.

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    García-García, Luis; Shiha, Ahmed Anis; Bascuñana, Pablo; de Cristóbal, Javier; Fernández de la Rosa, Rubén; Delgado, Mercedes; Pozo, Miguel A

    2016-05-01

    It has been reported that fluoxetine, a selective serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitor, has neuroprotective properties in the lithium-pilocarpine model of status epilepticus (SE) in rats. The aim of the present study was to investigate the effect of 5-HT depletion by short-term administration of p-chlorophenylalanine (PCPA), a specific tryptophan hydroxylase inhibitor, on the brain hypometabolism and neurodegeneration induced in the acute phase of this SE model. Our results show that 5-HT depletion did modify neither the brain basal metabolic activity nor the lithium-pilocarpine-induced hypometabolism when evaluated 3 days after the insult. In addition, hippocampal neurodegeneration and astrogliosis triggered by lithium-pilocarpine were not exacerbated by PCPA treatment. These findings point out that in the early latent phase of epileptogenesis, non-5-HT-mediated actions may contribute, at least in some extent, to the neuroprotective effects of fluoxetine in this model of SE.

  6. Subacute administration of fluoxetine prevents short-term brain hypometabolism and reduces brain damage markers induced by the lithium-pilocarpine model of epilepsy in rats.

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    Shiha, Ahmed Anis; de Cristóbal, Javier; Delgado, Mercedes; Fernández de la Rosa, Rubén; Bascuñana, Pablo; Pozo, Miguel A; García-García, Luis

    2015-02-01

    The role of serotonin (5-hydroxytryptamine; 5-HT) in epileptogenesis still remains controversial. In this regard, it has been reported that serotonergic drugs can alter epileptogenesis in opposite ways. The main objective of this work was to investigate the effect of the selective 5-HT selective reuptake inhibitor (SSRI) fluoxetine administered subacutely (10mg/kg/day×7 days) on the eventual metabolic impairment induced by the lithium-pilocarpine model of epilepsy in rats. In vivo 2-deoxy-2-[(18)F]fluoro-d-glucose ([(18)F] FDG) positron emission tomography (PET) was performed to assess the brain glucose metabolic activity on days 3 and 30 after the insult. In addition, at the end of the experiment (day 33), several histochemical and neurochemical assessments were performed for checking the neuronal functioning and integrity. Three days after the insult, a marked reduction of [(18)F] FDG uptake (about 30% according to the brain region) was found in all brain areas studied. When evaluated on day 30, although a hypometabolism tendency was observed, no statistically significant reduction was present in any region analyzed. In addition, lithium-pilocarpine administration was associated with medium-term hippocampal and cortical damage, since it induced neurodegeneration, glial activation and augmented caspase-9 expression. Regarding the effect of fluoxetine, subacute treatment with this SSRI did not significantly reduce the mortality rate observed after pilocarpine-induced seizures. However, fluoxetine did prevent not only the short-term metabolic impairment, but also the aforementioned signs of neuronal damage in surviving animals to lithium-pilocarpine protocol. Finally, fluoxetine increased the density of GABAA receptor both at the level of the dentate gyrus and CA1-CA2 regions in pilocarpine-treated animals. Overall, our data suggest a protective role for fluoxetine against pilocarpine-induced brain damage. Moreover, this action may be associated with an increase of

  7. N-methyl-D-aspartate receptor NR2B subunit involved in depression-like behaviours in lithium chloride-pilocarpine chronic rat epilepsy model.

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    Peng, Wei-Feng; Ding, Jing; Li, Xin; Fan, Fan; Zhang, Qian-Qian; Wang, Xin

    2016-01-01

    Depression is a common comorbidity in patients with epilepsy with unclear mechanisms. This study is to explore the role of glutamate N-methyl-D-aspartate (NMDA) receptor NR1, NR2A and NR2B subunits in epilepsy-associated depression. Lithium chloride (Licl)-pilocarpine chronic rat epilepsy model was established and rats were divided into epilepsy with depression (EWD) and epilepsy without depression (EWND) subgroups based on forced swim test. Expression of NMDA receptor NR1, NR2A and NR2B subunits was measured by western blot and immunofluorescence methods. The immobility time (IMT) was significantly greater in Licl-pilocarpine model group than in Control group, which was also greater in EWD group than in EWND group. No differences of spontaneous recurrent seizure (SRS) counts over two weeks and latency were found between EWD and EWND groups. The number of NeuN positive cells was significantly less in Licl-pilocarpine model group than in Control group, but had no difference between EWD and EWND groups. The ratios of phosphorylated NR1 (p-NR1)/NR1 and p-NR2B/NR2B were significantly greater in the hippocampus in EWD group than in EWND group. Moreover, the expression of p-NR1 and p-NR2B in the CA1 subfield of hippocampus were both greater in Licl-pilocarpine model group than Control group. Selective blockage of NR2B subunit with ifenprodil could alleviate depression-like behaviours of Licl-pilocarpine rat epilepsy model. In conclusion, glutamate NMDA receptor NR2B subunit was involved in promoting depression-like behaviours in the Licl-pilocarpine chronic rat epilepsy model and might be a target for treating epilepsy-associated depression.

  8. Altered expression and localization of hippocampal A-type potassium channel subunits in the pilocarpine-induced model of temporal lobe epilepsy.

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    Monaghan, M M; Menegola, M; Vacher, H; Rhodes, K J; Trimmer, J S

    2008-10-15

    Altered ion channel expression and/or function may contribute to the development of certain human epilepsies. In rats, systemic administration of pilocarpine induces a model of human temporal lobe epilepsy, wherein a brief period of status epilepticus (SE) triggers development of spontaneous recurrent seizures that appear after a latency of 2-3 weeks. Here we investigate changes in expression of A-type voltage-gated potassium (Kv) channels, which control neuronal excitability and regulate action potential propagation and neurotransmitter release, in the pilocarpine model of epilepsy. Using immunohistochemistry, we examined the expression of component subunits of somatodendritic (Kv4.2, Kv4.3, KChIPl and KChIP2) and axonal (Kv1.4) A-type Kv channels in hippocampi of pilocarpine-treated rats that entered SE. We found that Kv4.2, Kv4.3 and KChIP2 staining in the molecular layer of the dentate gyrus changes from being uniformly distributed across the molecular layer to concentrated in just the outer two-thirds. We also observed a loss of KChIP1 immunoreactive interneurons, and a reduction of Kv4.2 and KChIP2 staining in stratum radiatum of CA1. These changes begin to appear 1 week after pilocarpine treatment and persist or are enhanced at 4 and 12 weeks. As such, these changes in Kv channel distribution parallel the acquisition of recurrent spontaneous seizures as observed in this model. We also found temporal changes in Kv1.4 immunoreactivity matching those in Timm's stain, being expanded in stratum lucidum of CA3 and in the inner third of the dentate molecular layer. Among pilocarpine-treated rats, changes were only observed in those that entered SE. These changes in A-type Kv channel expression may contribute to hyperexcitability of dendrites in the associated hippocampal circuits as observed in previous studies of the effects of pilocarpine-induced SE.

  9. Glycyrrhizin ameliorates oxidative stress and inflammation in hippocampus and olfactory bulb in lithium/pilocarpine-induced status epilepticus in rats.

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    González-Reyes, Susana; Santillán-Cigales, Juan Jair; Jiménez-Osorio, Angélica Saraí; Pedraza-Chaverri, José; Guevara-Guzmán, Rosalinda

    2016-10-01

    Glycyrrhizin (GL) is a triterpene present in the roots and rhizomes of Glycyrrhiza glabra that has anti-inflammatory, hepatoprotective and neuroprotective effects. Recently, it was demonstrated that GL produced neuroprotective effects on the postischemic brain as well as on the kainic acid injury model in rats. In addition to this, GL also prevented excitotoxic effects on primary cultures. The aims of the present study were to evaluate GL scavenging properties and to investigate GL's effect on oxidative stress and inflammation in the lithium/pilocarpine-induced seizure model in two cerebral regions, hippocampus and olfactory bulb, at acute time intervals (3 or 24h) after status epilepticus (SE). Fluorometric methods showed that GL scavenged three reactive oxygen species: hydrogen peroxide, peroxyl radicals and superoxide anions. In contrast, GL was unable to scavenge peroxynitrite, hydroxyl radicals, singlet oxygen and 2,2-diphenil-1-picrylhydrazyl (DPPH) radicals suggesting that GL is a weak scavenger. Additionally, administration of GL (50mg/kg, i.p.) 30min before pilocarpine administration significantly suppressed oxidative stress. Moreover, malondialdehyde levels were diminished and glutathione levels were maintained at control values in both cerebral regions at 3 and 24 after SE. At 24h after SE, glutathione S-transferase and superoxide dismutase activity increased in the hippocampus, while both glutathione reductase and glutathione peroxidase activity were unchanged in the olfactory bulb at that time. In addition, GL suppressed the induction of the proinflammatory cytokines interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in both cerebral regions evaluated. These results suggest that GL confers protection against pilocarpine damage via antioxidant and anti-inflammatory effects.

  10. Circuit mechanisms of seizures in the pilocarpine model of chronic epilepsy: cell loss and mossy fiber sprouting.

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    Mello, L E; Cavalheiro, E A; Tan, A M; Kupfer, W R; Pretorius, J K; Babb, T L; Finch, D M

    1993-01-01

    We used the pilocarpine model of chronic spontaneous recurrent seizures to evaluate the time course of supragranular dentate sprouting and to assess the relation between several changes that occur in epileptic tissue with different behavioral manifestations of this experimental model of temporal lobe epilepsy. Pilocarpine-induced status epilepticus (SE) invariably led to cell loss in the hilus of the dentate gyrus (DG) and to spontaneous recurrent seizures. Cell loss was often also noted in the DG and in hippocampal subfields CA1 and CA3. The seizures began to appear at a mean of 15 days after SE induction (silent period), recurred at variable frequencies for each animal, and lasted for as long as the animals were allowed to survive (325 days). The granule cell layer of the DG was dispersed in epileptic animals, and neo-Timm stains showed supra- and intragranular mossy fiber sprouting. Supragranular mossy fiber sprouting and dentate granule cell dispersion began to appear early after SE (as early as 4 and 9 days, respectively) and reached a plateau by 100 days. Animals with a greater degree of cell loss in hippocampal field CA3 showed later onset of chronic epilepsy (r = 0.83, p seizure spread. These results demonstrate that the pilocarpine model of chronic seizures replicates several of the features of human temporal lobe epilepsy (hippocampal cell loss, supra- and intragranular mossy fiber sprouting, dentate granule cell dispersion, spontaneous recurrent seizures) and that it may be a useful model for studying this human condition. The results also suggest that even though a certain amount of cell loss in specific areas may be essential for chronic seizures to occur, excessive cell loss may hinder epileptogenesis.

  11. Persistent Hyperactivity of Hippocampal Dentate Interneurons after a Silent Period in the Rat Pilocarpine Model of Epilepsy

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    Xiaochen eWang

    2016-04-01

    Full Text Available Profile of GABAergic interneuron activity after pilocarpine-induced status epilepticus (SE was examined in the rat hippocampal dentate gyrus by analyzing immediate early gene expression and recording spontaneous firing at near resting membrane potential.SE for exact 2 hours or more than 2 hours was induced in the male Sprague-Dawley rats by an intraperitoneal injection of pilocarpine. Expression of immediate early genes was examined at 1 hour, 1 week, 2 weeks or more than 10 weeks after SE. For animals to be examined at 1 hour after SE, SE lasted for exact 2 hours was terminated by an intraperitoneal injection of diazepam. Spontaneous firing at near the resting membrane potential was recorded in interneurons located along the border between the granule cell layer and the hilus more than 10 weeks after SE. Results showed that both c-fos and activity-regulated cytoskeleton associated protein (Arc in hilar GABAergic interneurons were up-regulated after SE in a biphasic manner; they were increased at 1 hour and more than 2 weeks, but not at 1 week after SE. Ten weeks after SE, nearly 60% of hilar GABAergic cells expressed c-fos. With the exception of calretinin (CR-positive cells, percentages of hilar neuronal nitric oxide synthase (nNOS-, neuropeptide Y (NPY-, parvalbumin (PV-, and somatostatin (SOM-positive cells with c-fos expression are significantly higher than those of controls more than 10 weeks after SE. Without the resting membrane potential to be more depolarizing and changed threshold potential level in SE-induced rats, cell-attached recording revealed that nearly 90% of hilar interneurons fired spontaneously at near the resting membrane potential while only 22% of the same cell population did so in the controls.In conclusion, pilocarpine-induced SE eventually leads to a state in which surviving dentate GABAergic interneurons become hyperactive with a subtype-dependent manner; this implies that a fragile balance between excitation and

  12. Differential patterns of synaptotagmin7 mRNA expression in rats with kainate- and pilocarpine-induced seizures.

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    Gordana Glavan

    Full Text Available Previous studies in rat models of neurodegenerative disorders have shown disregulation of striatal synaptotagmin7 mRNA. Here we explored the expression of synaptotagmin7 mRNA in the brains of rats with seizures triggered by the glutamatergic agonist kainate (10 mg/kg or by the muscarinic agonist pilocarpine (30 mg/kg in LiCl (3 mEq/kg pre-treated (24 h rats, in a time-course experiment (30 min-1 day. After kainate-induced seizures, synaptotagmin7 mRNA levels were transiently and uniformly increased throughout the dorsal and ventral striatum (accumbens at 8 and 12 h, but not at 24 h, followed at 24 h by somewhat variable upregulation within different parts of the cerebral cortex, amigdala and thalamic nuclei, the hippocampus and the lateral septum. By contrast, after LiCl/pilocarpine-induced seizures, there was a more prolonged increase of striatal Synaptotagmin7 mRNA levels (at 8, 12 and 24 h, but only in the ventromedial striatum, while in some other of the aforementioned brain regions there was a decline to below the basal levels. After systemic post-treatment with muscarinic antagonist scopolamine in a dose of 2 mg/kg the seizures were either extinguished or attenuated. In scopolamine post-treated animals with extinguished seizures the striatal synaptotagmin7 mRNA levels (at 12 h after the onset of seizures were not different from the levels in control animals without seizures, while in rats with attenuated seizures, the upregulation closely resembled kainate seizures-like pattern of striatal upregulation. In the dose of 1 mg/kg, scopolamine did not significantly affect the progression of pilocarpine-induced seizures or pilocarpine seizures-like pattern of striatal upregulation of synaptotagmin7 mRNA. In control experiments, equivalent doses of scopolamine per se did not affect the expression of synaptotagmin7 mRNA. We conclude that here described differential time course and pattern of synaptotagmin7 mRNA expression imply regional

  13. Dual mechanisms of rapid expression of anxiety-related behavior in pilocarpine-treated epileptic mice.

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    Otsuka, Shintaro; Ohkido, Taro; Itakura, Makoto; Watanabe, Shigeru; Yamamori, Saori; Iida, Yuuki; Saito, Masanori; Miyaoka, Hitoshi; Takahashi, Masami

    2016-07-01

    A mouse model of epilepsy was generated by inducing status epilepticus (SE) for either 1.5 or 4.5h with pilocarpine to study anxiety-related behaviors, changes in the electroencephalogram of the cerebral cortex and hippocampus, and expression of hippocampal proteins. The viability and rate of success of SE induction were high in C57BL/6N mice but not in C57BL/6J mice. C57BL/6N mice were immotile during the first 2days after SE; however, by the third day, most mice were recovered and exhibited strong anxiety-related behaviors in response to the light/dark preference test and open field test. There was a striking difference in the temporal appearance of anxiety-related behavior between the two SE durations: 1.5h SE mice exhibited strong anxiety-related behavior 3days after SE that gradually attenuated over the next few weeks, whereas 4.5h SE mice exhibited strong anxiety-related behavior 3days after SE that persisted even at nearly 1year after SE. Mice receiving both SE durations exhibited generalized seizures (GS) after SE; however, there was a marked difference in the timing and duration of GS appearance. Mice in the 4.5h SE group exhibited spontaneous GS from 4days to at least 96days after SE. In contrast, mice in the 1.5h SE group exhibited GS only within the first several days after SE; however, epileptic spike clusters continuously appeared in the cerebral cortex and hippocampus for up to twelve days after SE. Among the hippocampal proteins tested, only brain derived-neurotrophic factor (BDNF) exhibited altered expression in parallel with anxiety-related behavior. These results showed the possibility that BDNF expression in the hippocampus might cause anxiety-related behavior in adulthood.

  14. Down-regulation of BK channel expression in the pilocarpine model of temporal lobe epilepsy.

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    Pacheco Otalora, Luis F; Hernandez, Eder F; Arshadmansab, Massoud F; Francisco, Sebastian; Willis, Michael; Ermolinsky, Boris; Zarei, Masoud; Knaus, Hans-Guenther; Garrido-Sanabria, Emilio R

    2008-03-20

    In the hippocampus, BK channels are preferentially localized in presynaptic glutamatergic terminals including mossy fibers where they are thought to play an important role regulating excessive glutamate release during hyperactive states. Large conductance calcium-activated potassium channels (BK, MaxiK, Slo) have recently been implicated in the pathogenesis of genetic epilepsy. However, the role of BK channels in acquired mesial temporal lobe epilepsy (MTLE) remains unknown. Here we used immunohistochemistry, laser scanning confocal microscopy (LSCM), Western immunoblotting and RT-PCR to investigate the expression pattern of the alpha-pore-forming subunit of BK channels in the hippocampus and cortex of chronically epileptic rats obtained by the pilocarpine model of MTLE. All epileptic rats experiencing recurrent spontaneous seizures exhibited a significant down-regulation of BK channel immunostaining in the mossy fibers at the hilus and stratum lucidum of the CA3 area. Quantitative analysis of immunofluorescence signals by LSCM revealed a significant 47% reduction in BK channel immunofluorescent signals in epileptic rats when compared to age-matched non-epileptic control rats. These data correlate with a similar reduction in BK channel protein levels and transcripts in the cortex and hippocampus. Our data indicate a seizure-related down-regulation of BK channels in chronically epileptic rats. Further functional assays are necessary to determine whether altered BK channel expression is an acquired channelopathy or a compensatory mechanism affecting the network excitability in MTLE. Moreover, seizure-mediated BK down-regulation may disturb neuronal excitability and presynaptic control at glutamatergic terminals triggering exaggerated glutamate release and seizures.

  15. Dexamethasone exacerbates cerebral edema and brain injury following lithium-pilocarpine induced status epilepticus.

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    Duffy, B A; Chun, K P; Ma, D; Lythgoe, M F; Scott, R C

    2014-03-01

    Anti-inflammatory therapies are the current most plausible drug candidates for anti-epileptogenesis and neuroprotection following prolonged seizures. Given that vasogenic edema is widely considered to be detrimental for outcome following status epilepticus, the anti-inflammatory agent dexamethasone is sometimes used in clinic for alleviating cerebral edema. In this study we perform longitudinal magnetic resonance imaging in order to assess the contribution of dexamethasone on cerebral edema and subsequent neuroprotection following status epilepticus. Lithium-pilocarpine was used to induce status epilepticus in rats. Following status epilepticus, rats were either post-treated with saline or with dexamethasone sodium phosphate (10mg/kg or 2mg/kg). Brain edema was assessed by means of magnetic resonance imaging (T2 relaxometry) and hippocampal volumetry was used as a marker of neuronal injury. T2 relaxometry was performed prior to, 48 h and 96 h following status epilepticus. Volume measurements were performed between 18 and 21 days after status epilepticus. Unexpectedly, cerebral edema was worse in rats that were treated with dexamethasone compared to controls. Furthermore, dexamethasone treated rats had lower hippocampal volumes compared to controls 3 weeks after the initial insult. The T2 measurements at 2 days and 4 days in the hippocampus correlated with hippocampal volumes at 3 weeks. Finally, the mortality rate in the first week following status epilepticus increased from 14% in untreated rats to 33% and 46% in rats treated with 2mg/kg and 10mg/kg dexamethasone respectively. These findings suggest that dexamethasone can exacerbate the acute cerebral edema and brain injury associated with status epilepticus.

  16. Pyrrolidine dithiocarbamate protects the piriform cortex in the pilocarpine status epilepticus model.

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    Soerensen, Jonna; Pekcec, Anton; Fuest, Christina; Nickel, Astrid; Potschka, Heidrun

    2009-12-01

    Pyrrolidine dithiocarbamate (PDTC) has a dual mechanism of action as an antioxidant and an inhibitor of the transcription factor kappa-beta. Both, production of reactive oxygen species as well as activation of NF-kappaB have been implicated in severe neuronal damage in different sub-regions of the hippocampus as well as in the surrounding cortices. The effect of PDTC on status epilepticus-associated cell loss in the hippocampus and piriform cortex was evaluated in the rat fractionated pilocarpine model. Treatment with 150 mg/kg PDTC before and following status epilepticus significantly increased the mortality rate to 100%. Administration of 50 mg/kg PDTC (low-dose) did not exert major effects on the development of a status epilepticus or the mortality rate. In vehicle-treated rats, status epilepticus caused pronounced neuronal damage in the piriform cortex comprising both pyramidal cells and interneurons. Low-dose PDTC treatment almost completely protected from lesions in the piriform cortex. A significant decrease in neuronal density of the hippocampal hilar formation was identified in vehicle- and PDTC-treated rats following status epilepticus. In conclusion, the NF-kappaB inhibitor and antioxidant PDTC protected the piriform cortex, whereas it did not affect hilar neuronal loss. These data might indicate that the generation of reactive oxygen species and activation of NF-kappaB plays a more central role in seizure-associated neuronal damage in the temporal cortex as compared to the hippocampal hilus. However, future investigations are necessary to exactly analyze the biochemical mechanisms by which PDTC exerted its beneficial effects in the piriform cortex.

  17. Antiepileptogenic and Neuroprotective Effects of Pergularia daemia on Pilocarpine Model of Epilepsy

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    Antoine K. Kandeda

    2017-06-01

    Full Text Available In this study, we investigated antiepileptogenic and neuroprotective effects of the aqueous extract of Pergularia daemia roots (PDR using in vivo and in vitro experimental models. In in vivo studies, status epilepticus caused by pilocarpine injection triggers epileptogenesis which evolves during about 1–2 weeks. After 2 h of status epilepticus, mice were treated during the epileptogenesis period for 7 days with sodium valproate and vitamin C (standards which demonstrated to alter epileptogenesis, or Pergularia daemia. The animals were then, 1 week after status epilepticus, challenged with acute pentylenetetrazole (PTZ administration to test behaviorally the susceptibility to a convulsant agent of animals treated or not with the plan extract. Memory was assessed after PTZ administration in the elevated plus maze and T-maze paradigms at 24 and 48 h. Antioxidant and acetylcholinesterase activities were determined in the hippocampus after sacrifice, in vitro studies were conducted using embryonic rat primary cortical cultures exposed to L-glutamate. Cell survival rate was measured and apoptotic and necrotic cell death determined. The results showed that chronic oral administration of PDR significantly and dose-dependently increased the latency to myoclonic jerks, clonic seizures and generalized tonic–clonic seizures, and the seizure score. In addition, PDR at all doses (from 4.9 to 49 mg/kg significantly decreased the initial and retention transfer latencies in the elevated plus maze. Interestingly PDR at the same doses significantly increased the time spent and the number of entries in T-maze novel arm. PDR significantly increased the activities of acetylcholinesterase and antioxidant enzymes superoxide dismutase, catalase, and total glutathione and proteins, and decreased malondialdehyde level. Furthermore, PDR increased viability rate of primary cortical neurons after L-glutamate-induced excitotoxicity, in a dose dependent manner. Altogether

  18. Brain-derived neurotrophic factor superinduction parallels anti-epileptic--neuroprotective treatment in the pilocarpine epilepsy model.

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    Biagini, G; Avoli, M; Marcinkiewicz, J; Marcinkiewicz, M

    2001-03-01

    Antiepileptic drugs provide neuroprotection in several animal models of brain damage, including those induced by status epilepticus (SE). The mechanisms involved in this action are unknown, but neurotrophic factors such as brain-derived neurotrophic factor (BDNF) may play a role. In this study we investigated the changes in BDNF levels in rats in which SE had been induced by pilocarpine injection (400 mg/kg i.p.) and continued for several hours (unprotected group). In other animals (protected groups), SE was suppressed after 30 min by intraperitoneal injection of either diazepam (10 mg/kg) + pentobarbital (30 mg/kg) or paraldehyde (0.3 mg/kg). In diazepam + pentobarbital-treated rats the hippocampal damage caused by SE was significantly lower (p rats treated with diazepam + pentobarbital. In contrast, a decrease of BDNF immunoreactivity occurred in the unprotected group. In conclusion, these results show that neuroprotection induced by anti-epileptic drugs in pilocarpine-treated rats is accompanied by strong potentiation of BDNF synthesis in brain regions involved in SE.

  19. Selective loss and axonal sprouting of GABAergic interneurons in the sclerotic hippocampus induced by LiCl-pilocarpine.

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    Long, Lili; Xiao, Bo; Feng, Li; Yi, Fang; Li, Guoliang; Li, Shuyu; Mutasem, M Abuhamed; Chen, Si; Bi, Fangfang; Li, Yi

    2011-02-01

    In this study, we performed immunohistochemistry for somatostatin (SS), neuropeptide Y (NPY), and parvalbumin (PV) in LiCl-pilocarpine-treated rats to observe quantitative changes and axonal sprouting of GABAergic interneurons in the hippocampus, especially in the sclerotic hippocampus. Fluoro-Jade B (FJB) was performed to detect the specific degeneration of GABAergic interneurons. Compared with age-matched control rats, there were fewer SS/NPY/PV-immunoreactive (IR) interneurons in the hilus of the sclerotic hippocampus in pilocarpine-treated rats; hilar dentritic inhibitory interneurons were most vulnerable. FJB stain revealed degeneration was evident at 2 months after status epilepticus. Some SS-IR and NPY-IR interneurons were also stained for FJB, but there was no evidence of degeneration of PV-IR interneurons. Axonal sprouting of GABAergic interneurons was present in the hippocampus of epileptic rats, and a dramatic increase of SS-IR fibers was observed throughout all layers of CA1 region in the sclerotic hippocampus. These results confirm selective loss and degeneration of a specific subset of GABAergic interneurons in specific subfields of the hippocampus. Axonal sprouting of inhibitory GABAergic interneurons, especially numerous increase of SS-IR neutrophils within CA1 region of the sclerotic hippocampus, may constitute the aberrant inhibitory circum and play a significant role in the generation and compensation of temporal lobe epilepsy.

  20. Protective but not anticonvulsant effects of ghrelin and JMV-1843 in the pilocarpine model of Status epilepticus.

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    Chiara Lucchi

    Full Text Available In models of status epilepticus ghrelin displays neuroprotective effects mediated by the growth hormone secretagogue-receptor 1a (GHS-R1a. This activity may be explained by anticonvulsant properties that, however, are controversial. We further investigated neuroprotection and the effects on seizures by comparing ghrelin with a more effective GHS-R1a agonist, JMV-1843. Rats were treated either with ghrelin, JMV-1843 or saline 10 min before pilocarpine, which was used to induce status epilepticus. Status epilepticus, developed in all rats, was attenuated by diazepam. No differences were observed among the various groups in the characteristics of pilocarpine-induced seizures. In saline group the area of lesion, characterized by lack of glial fibrillary acidic protein immunoreactivity, was of 0.45 ± 0.07 mm(2 in the hippocampal stratum lacunosum-moleculare, and was accompanied by upregulation of laminin immunostaining, and by increased endothelin-1 expression. Both ghrelin (P<0.05 and JMV-1843 (P<0.01 were able to reduce the area of loss in glial fibrillary acidic protein immunostaining. In addition, JMV-1843 counteracted (P<0.05 the changes in laminin and endothelin-1 expression, both increased in ghrelin-treated rats. JMV-1843 was able to ameliorate neuronal survival in the hilus of dentate gyrus and medial entorhinal cortex layer III (P<0.05 vs saline and ghrelin groups. These results demonstrate diverse protective effects of growth hormone secretagogues in rats exposed to status epilepticus.

  1. Protective but Not Anticonvulsant Effects of Ghrelin and JMV-1843 in the Pilocarpine Model of Status epilepticus

    Science.gov (United States)

    Lucchi, Chiara; Curia, Giulia; Vinet, Jonathan; Gualtieri, Fabio; Bresciani, Elena; Locatelli, Vittorio; Torsello, Antonio; Biagini, Giuseppe

    2013-01-01

    In models of status epilepticus ghrelin displays neuroprotective effects mediated by the growth hormone secretagogue-receptor 1a (GHS-R1a). This activity may be explained by anticonvulsant properties that, however, are controversial. We further investigated neuroprotection and the effects on seizures by comparing ghrelin with a more effective GHS-R1a agonist, JMV-1843. Rats were treated either with ghrelin, JMV-1843 or saline 10 min before pilocarpine, which was used to induce status epilepticus. Status epilepticus, developed in all rats, was attenuated by diazepam. No differences were observed among the various groups in the characteristics of pilocarpine-induced seizures. In saline group the area of lesion, characterized by lack of glial fibrillary acidic protein immunoreactivity, was of 0.45±0.07 mm2 in the hippocampal stratum lacunosum-moleculare, and was accompanied by upregulation of laminin immunostaining, and by increased endothelin-1 expression. Both ghrelin (P<0.05) and JMV-1843 (P<0.01) were able to reduce the area of loss in glial fibrillary acidic protein immunostaining. In addition, JMV-1843 counteracted (P<0.05) the changes in laminin and endothelin-1 expression, both increased in ghrelin-treated rats. JMV-1843 was able to ameliorate neuronal survival in the hilus of dentate gyrus and medial entorhinal cortex layer III (P<0.05 vs saline and ghrelin groups). These results demonstrate diverse protective effects of growth hormone secretagogues in rats exposed to status epilepticus. PMID:24015271

  2. Differential expression of brain-derived neurotrophic factor transcripts after pilocarpine-induced seizure-like activity is related to mode of Ca2+ entry.

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    Poulsen, F R; Lauterborn, J; Zimmer, J; Gall, C M

    2004-01-01

    Activity-dependent brain-derived neurotrophic factor (BDNF) expression is Ca2+-dependent, yet little is known about the Ca2+ channel contributions that might direct selective expression of the multiple BDNF transcripts. Here, effects of pilocarpine-induced seizure activity on total BDNF expression and on the individual sensitivity of BDNF transcripts to glutamate receptor and Ca2+ channel blockers were evaluated using hippocampal slice cultures and in situ hybridization of transcript-specific cRNA probes directed against mRNAs for the four 5' exons (I-IV) of the BDNF gene. mRNAs for nerve growth factor (NGF) and tyrosine kinase B (trkB) also were studied. Pilocarpine (5 mM) induced a dose- and time-dependent increase in total BDNF (exon V) mRNA expression in the dentate granule cells and CA3-CA1 pyramidal cells with maximal effects at 6 and 24 h, respectively. Increases were blocked by co-treatment with the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX: 25 microM) and the N-methyl-d-aspartic acid receptor antagonist 2-amino-5-phosphonovaleric acid (APV; 25 microM), whereas the L-type voltage sensitive Ca2+ channel blocker nifedipine (20 microM) was without detectable effect. Maximal NGF and trkB mRNA expression was induced by pilocarpine at 4 and 12 h, respectively. For the individual BDNF transcripts, APV blocked pilocarpine-induced increases in transcript II, whereas nifedipine blocked increases in transcripts I and III. Transcript IV levels were not altered by treatment. These results indicate that transcript II makes the greatest contribution to pilocarpine effects on total BDNF mRNA content in this model and provides evidence for regional and Ca2+ channel-specific differences in activity-dependent regulation of the different BDNF transcripts in hippocampus.

  3. Anticonvulsant effect of time-restricted feeding in a pilocarpine-induced seizure model: Metabolic and epigenetic implications.

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    Jorge eLandgrave-Gómez

    2016-01-01

    Full Text Available A new generation of antiepileptic drugs has emerged; however, one-third of epilepsy patients do not properly respond to pharmacological treatments. The purpose of the present study was to investigate whether time-restricted feeding has an anticonvulsant effect and whether this restrictive diet promotes changes in energy metabolism and epigenetic modifications in a pilocarpine-induced seizure model. To resolve our hypothesis, one group of rats had free access to food and water ad libitum (AL and a second group underwent a time-restricted feeding (TRF schedule. We used the lithium-pilocarpine model to induce status epilepticus (SE, and behavioral seizure monitoring was analyzed. Additionally, an electroencephalography (EEG recording was performed to verify the effect of TRF on cortical electrical activity after a pilocarpine injection. For biochemical analysis, animals were sacrificed 24 hours after SE and hippocampal homogenates were used to evaluate the proteins related to metabolism and chromatin structure. Our results showed that TRF had an anticonvulsant effect as measured by the prolonged latency of forelimb clonus seizure, a decrease in the seizure severity score and fewer animals reaching SE. Additionally, the power of the late phase EEG recordings in the AL group was significantly higher than the TRF group. Moreover, we found that TRF is capable of inducing alterations in signaling pathways that regulate energy metabolism, including an increase in the phosphorylation of AMP dependent kinase (AMPK and a decrease in the phosphorylation of Akt kinase. Furthermore, we found that TRF was able to significantly increase the beta hydroxybutyrate (β-HB concentration, an endogenous inhibitor of histone deacetylases (HDACs. Finally, we found a significant decrease in HDAC activity as well as an increase in acetylation on histone 3 (H3 in hippocampal homogenates from the TRF group. These findings suggest that alterations in energy metabolism and the

  4. Electroshocks delay seizures and subsequent epileptogenesis but do not prevent neuronal damage in the lithium-pilocarpine model of epilepsy.

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    André, V; Ferrandon, A; Marescaux, C; Nehlig, A

    2000-11-01

    Electroconvulsive therapy, which is used to treat refractory major depression in humans increases seizure threshold and decreases seizure duration. Moreover, the expression of brain derived neurotrophic factor induced by electroshocks (ECS) might protect hippocampal cells from death in patients suffering from depression. As temporal lobe epilepsy is linked to neuronal damage in the hippocampus, we tested the effect of repeated ECS on subsequent status epilepticus (SE) induced by lithium-pilocarpine and leading to cell death and temporal epilepsy in the rat. Eleven maximal ECS were applied via ear-clips to adult rats. The last one was applied 2 days before the induction of SE by lithium-pilocarpine. The rats were electroencephalographically recorded to study the SE characteristics. The rats treated with ECS before pilocarpine (ECS-pilo) developed partial limbic (score 2) and propagated seizures (score 5) with a longer latency than the rats that underwent SE alone (sham-pilo). Despite this delay in the initiation and propagation of the seizures, the same number of ECS- and sham-pilo rats developed SE with a similar characteristic pattern. The expression of c-Fos protein was down-regulated by repeated ECS in the amygdala and the cortex. In ECS-pilo rats, c-Fos expression was decreased in the piriform and entorhinal cortex and increased in the hilus of the dentate gyrus. Neuronal damage was identical in the forebrain areas of both groups, while it was worsened by ECS treatment in the substantia nigra pars reticulata, entorhinal and perirhinal cortices compared to sham-pilo rats. Finally, while 11 out of the 12 sham-pilo rats developed spontaneous recurrent seizures after a silent period of 40+/-27 days, only two out of the 10 ECS-pilo rats became epileptic, but after a prolonged latency of 106 and 151 days. One ECS-pilo rat developed electrographic infraclinical seizures and seven did not exhibit any seizures. Thus, the extensive neuronal damage occurring in the

  5. Antiepileptic and neuroprotective effects of human umbilical cord blood mononuclear cells in a pilocarpine-induced epilepsy model.

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    Costa-Ferro, Zaquer Suzana Munhoz; de Borba Cunha, Fernanda; de Freitas Souza, Bruno Solano; Leal, Marcos Maurício Tosta; da Silva, Adelson Alves; de Bellis Kühn, Telma Ingrid Borges; Forte, Andresa; Sekiya, Eliseo Joji; Soares, Milena Botelho Pereira; Dos Santos, Ricardo Ribeiro

    2014-03-01

    Status epilepticus (SE) is a condition of persistent seizure that leads to brain damage and, frequently, to the establishment of chronic epilepsy. Cord blood is an important source of adult stem cells for the treatment of neurological disorders. The present study aimed to evaluate the effects of human umbilical cord blood mononuclear cells (HUCBC) transplanted into rats after induction of SE by the administration of lithium and pilocarpine chloride. Transplantation of HUCBC into epileptic rats protected against neuronal loss in the hippocampal subfields CA1, CA3 and in the hilus of the dentate gyrus, up to 300 days after SE induction. Moreover, transplanted rats had reduced frequency and duration of spontaneous recurrent seizures (SRS) 15, 120 and 300 days after the SE. Our study shows that HUCBC provide prominent antiepileptic and neuroprotective effects in the experimental model of epilepsy and reinforces that early interventions can protect the brain against the establishment of epilepsy.

  6. Indomethacin can downregulate the levels of inflammatory mediators in the hippocampus of rats submitted to pilocarpine-induced status epilepticus

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    Michele Juliane Vieira

    2014-09-01

    Full Text Available OBJECTIVE: Refractory status epilepticus is one of the most life-threatening neurological emergencies and is characterized by high morbidity and mortality. Additionally, the use of anti-inflammatory drugs during this period is very controversial. Thus, this study has been designed to analyze the effect of a low dose of indomethacin (a COX inhibitor on the expression of inflammatory molecules. METHOD: The hippocampus of rats submitted to pilocarpine-induced long-lasting status epilepticus was analyzed to determine the expression of inflammatory molecules with RT-PCR and immunohistochemistry. RESULTS: Compared with controls, reduced levels of the kinin B2 receptors IL1β and TNFα were found in the hippocampus of rats submitted to long-lasting status epilepticus and treated with indomethacin. CONCLUSIONS: These data show that low doses of indomethacin could be employed to minimize inflammation during long-lasting status epilepticus.

  7. Altered hippocampal myelinated fiber integrity in a lithium-pilocarpine model of temporal lobe epilepsy: a histopathological and stereological investigation.

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    Ye, Yuanzhen; Xiong, Jiajia; Hu, Jun; Kong, Min; Cheng, Li; Chen, Hengsheng; Li, Tingsong; Jiang, Li

    2013-07-19

    The damage of white matter, primarily myelinated fibers, in the central nervous system (CNS) of temporal lobe epilepsy (TLE) patients has been recently reported. However, limited data exist addressing the types of changes that occur to myelinated fibers inside the hippocampus as a result of TLE. The current study was designed to examine this issue in a lithium-pilocarpine rat model. Investigated by electroencephalography (EEG), Gallyas silver staining, immunohistochemistry, western blotting, transmission electron microscopy, and stereological methods, the results showed that hippocampal myelinated fibers of the epilepsy group were degenerated with significantly less myelin basic protein (MBP) expression relative to those of control group rats. Stereological analysis revealed that the total volumes of hippocampal formation, myelinated fibers, and myelin sheaths in the hippocampus of epilepsy group rats were decreased by 20.43%, 49.16%, and 52.60%, respectively. In addition, epilepsy group rats showed significantly greater mean diameters of myelinated fibers and axons, whereas the mean thickness of myelin sheaths was less, especially for small axons with diameters from 0.1 to 0.8µm, compared to control group rats. Finally, the total length of the myelinated fibers in the hippocampus of epilepsy group rats was significantly decreased by 56.92%, compared to that of the control group, with the decreased length most prominent for myelinated fibers with diameters from 0.4 to 0.8µm. This study is the first to provide experimental evidence that the integrity of hippocampal myelinated fibers is negatively affected by inducing epileptic seizures with pilocarpine, which may contribute to the abnormal propagation of epileptic discharge.

  8. Synchrotron radiation Fourier-transform infrared and Raman microspectroscopy study showing an increased frequency of creatine inclusions in the rat hippocampal formation following pilocarpine-induced seizures

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    Dulinska, J.; Setkowicz, Z.; Janeczko, K.; C. SANDT; Dumas, P.; Uram, L.; Gzielo-Jurek, K.; Chwiej, J.

    2011-01-01

    In the present work, synchrotron radiation Fourier-transform infrared (SRFTIR) and Raman microspectroscopies were used to evaluate a possible role of creatine in the pathogenesis and progress of pilocarpine-evoked seizures and seizure-induced neurodegenerative changes in the rat hippocampal tissue. The main goal of this study was to identify creatine deposits within the examined brain area, to analyze their frequency in epileptic animals and naive controls and to examine correlations between ...

  9. In vivo imaging of mGluR5 changes during epileptogenesis using [11C]ABP688 PET in pilocarpine-induced epilepsy rat model.

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    Hongyoon Choi

    Full Text Available INTRODUCTION: Metabotropic glutamate receptor 5 (mGluR5 that regulates glutamatergic neurotransmission contributes to pathophysiology of epilepsy. In this study, we monitored the changes of mGluR5 in vivo using [11C]ABP688 PET during the epileptogenesis in a pilocarpine-induced epilepsy rat model. METHODS: In vivo mGluR5 images were acquired using [11C]ABP688 microPET/CT in pilocarpine-induced chronic epilepsy rat models and controls. We also acquired microPET/CT at acute, subacute as well as chronic periods after status epilepticus. Non-displaceable binding potential (BPND of [11C]ABP688 was calculated using simplified reference tissue model in a voxel-based manner. mGluR5 BPND of the rat brains of epilepsy models and controls were compared. RESULTS: Status epilepticus developed after pilocarpine administration and was followed by recurrent spontaneous seizures for more than 3 weeks. In chronic epilepsy rat model, BPND in hippocampus and amygdala was reduced on a voxel-based analysis. Temporal changes of mGluR5 BPND was also found. In acute period after status epilepticus, mGluR5 BPND was reduced in the whole brain. BPND of caudate-putamen was restored in subacute period, while BPND of the rest of the brain was still lower. In chronic period, global BPND was normalized except in hippocampus and amygdala. CONCLUSIONS: In vivo imaging of mGluR5 using [11C]ABP688 microPET/CT could successfully reveal the regional changes of mGluR5 binding potential of the rat brain in a pilocarpine-induced epilepsy model. The temporal and spatial changes in mGluR5 availability suggest [11C]ABP688 PET imaging in epilepsy provide abnormal glutamatergic network during epileptogenesis.

  10. Modulation of seizures and synaptic plasticity by adenosinergic receptors in an experimental model of temporal lobe epilepsy induced by pilocarpine in rats.

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    Vianna, Eduardo Paulo Morowsky; Ferreira, Alice Teixeira; Doná, Flávia; Cavalheiro, Esper Abrão; da Silva Fernandes, Maria José

    2005-01-01

    Adenosine is a major negative neuromodulator of synaptic activity in the central nervous system and can exert anticonvulsant and neuroprotective effects in many experimental models of epilepsy. Extracellular adenosine can be formed by a membrane-anchored enzyme ecto-5'-nucleotidase. The purposes of this study were to characterize the role of adenosine receptors in modulating status epilepticus (SE) induced by pilocarpine and evaluate its neuroprotective action. Ecto-5'-nucleotidase activity was studied during the different phases of pilocarpine-induced epilepsy in rats. Adult rats were pretreated with different adenosinergic agents to evaluate the latency and incidence of SE induced by pilocarpine in rats. The neuroprotective effect also was evaluated. A proconvulsant effect was observed with DPCPX and DMPX that reduced the latency of SE in almost all rats. Pretreatment with the MRS 1220 did not alter the incidence of SE but reduced the latency to develop SE. An anticonvulsant and neuroprotective effect was detected with R-PIA. Rats pretreated with R-PIA had a decreased number of apoptotic cells in the hippocampus, whereas pretreatment with DPCPX did not modify the hippocampal damage. An intensification of neuronal death was observed in the dentate gyrus and CA3 when rats were pretreated with DMPX. MRS-1220 did not modify the number of apoptotic cells in the hippocampus. An increase in the ecto-5 -nucleotidase staining was detected in the hippocampus during silent and chronic phases. The present data show that adenosine released during pilocarpine-induced SE via A1-receptor stimulation can exhibit neuroprotective and anticonvulsant roles. Similar effects could also be inferred with A2a and A3 adenosinergic agents, but further experiments are necessary to confirm their roles. Ecto-5 -nucleotidase activity during silent and chronic phases might have a role in blocking spontaneous seizures by production of inhibitory neuromodulator adenosine, besides taking part in

  11. A Long-Term Treatment with Arachidonyl-2′-Chloroethylamide Combined with Valproate Increases Neurogenesis in a Mouse Pilocarpine Model of Epilepsy

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    Marta Andres-Mach

    2017-04-01

    Full Text Available Rational polytherapy in the treatment of refractory epilepsy has been the main therapeutic modality for several years. In treatment with two or more antiepileptic drugs (AEDs, it is of particular importance that AEDs be selected based on their high anticonvulsant properties, minimal side effects, and impact on the formation of new neurons. The aim of the study was to conduct an in vivo evaluation of the relationship between treatments with synthetic cannabinoid arachidonyl-2′-chloroethylamide (ACEA alone or in combination with valproic acid (VPA and hippocampal neurogenesis in a mouse pilocarpine model of epilepsy. All studies were performed on adolescent male CB57/BL mice with using the following drugs: VPA (10 mg/kg, ACEA (10 mg/kg, phenylmethylsulfonyl fluoride (PMSF—a substance protecting ACEA against degradation by fatty acid hydrolase, 30 mg/kg, pilocarpine (PILO, a single dose of 290 mg/kg and methylscopolamine (30 min before PILO to stop peripheral cholinergic effects of pilocarpine, 1 mg/kg. We evaluated the process of neurogenesis after a 10-day treatment with ACEA and VPA, alone and in combination. We observed a decrease of neurogenesis in the PILO control group as compared to the healthy control mice. Furthermore, ACEA + PMSF alone and in combination with VPA significantly increased neurogenesis compared to the PILO control group. In contrast, VPA 10-day treatment had no impact on the level of neurons in comparison to the PILO control group. The combination of ACEA, PMSF and VPA considerably stimulated the process of creating new cells, particularly neurons, while chronic administration of VPA itself had no influence on neurogenesis in the mouse pilocarpine model of epilepsy. The obtained results enabled an in vivo evaluation of neurogenesis after treatment with antiepileptic drugs in an experimental model of epilepsy.

  12. Loss of interneurons innervating pyramidal cell dendrites and axon initial segments in the CA1 region of the hippocampus following pilocarpine-induced seizures.

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    Dinocourt, Celine; Petanjek, Zdravko; Freund, Tamas F; Ben-Ari, Yezekiel; Esclapez, Monique

    2003-05-12

    In the pilocarpine model of chronic limbic seizures, vulnerability of GABAergic interneurons to excitotoxic damage has been reported in the hippocampal CA1 region. However, little is known about the specific types of interneurons that degenerate in this region. In order to characterize these interneurons, we performed quantitative analyses of the different populations of GABAergic neurons labeled for their peptide or calcium-binding protein content. Our data demonstrate that the decrease in the number of GAD mRNA-containing neurons in the stratum oriens of CA1 in pilocarpine-treated rats involved two subpopulations of GABAergic interneurons: interneurons labeled for somatostatin only (O-LM and bistratified cells) and interneurons labeled for parvalbumin only (basket and axo-axonic cells). Stratum oriens interneurons labeled for somatostatin/calbindin or somatostatin/parvalbumin were preserved. The decrease in number of somatostatin- and parvalbumin-containing neurons was observed as early as 72 hours after the sustained seizures induced by pilocarpine injection. Many degenerating cell bodies in the stratum oriens and degenerating axon terminals in the stratum lacunosum-moleculare were observed at 1 and 2 weeks after injection. In addition, the synaptic coverage of the axon initial segment of CA1 pyramidal cells was significantly decreased in pilocarpine-treated animals. These results indicate that the loss of somatostatin-containing neurons corresponds preferentially to the degeneration of interneurons with an axon projecting to stratum lacunosum-moleculare (O-LM cells) and suggest that the death of these neurons is mainly responsible for the deficit of dendritic inhibition reported in this region. We demonstrate that the loss of parvalbumin-containing neurons corresponds to the death of axo-axonic cells, suggesting that perisomatic inhibition and mechanisms controlling action potential generation are also impaired in this model.

  13. Loss of interneurons innervating pyramidal cell dendrites and axon initial segments in the CA1 region of the hippocampus following pilocarpine-induced seizures.

    OpenAIRE

    Dinocourt, Celine; Petanjek, Zdravko; Freund, Tamas,; Ben-Ari, Yezekiel; Esclapez, Monique

    2003-01-01

    International audience; In the pilocarpine model of chronic limbic seizures, vulnerability of GABAergic interneurons to excitotoxic damage has been reported in the hippocampal CA1 region. However, little is known about the specific types of interneurons that degenerate in this region. In order to characterize these interneurons, we performed quantitative analyses of the different populations of GABAergic neurons labeled for their peptide or calcium-binding protein content. Our data demonstrat...

  14. Coenzyme Q10 enhances the anticonvulsant effect of phenytoin in pilocarpine-induced seizures in rats and ameliorates phenytoin-induced cognitive impairment and oxidative stress.

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    Tawfik, Mona K

    2011-12-01

    Conventional antiepileptic drugs fail to adequately control seizures and predispose to cognitive impairment and oxidative stress with chronic usage in a significant proportion of patients with epilepsy. Coenzyme Q10 (CoQ10), an antioxidant compound, exhibits a wide range of therapeutic effects that are attributed to its potent antioxidant capacity. To evaluate the neuroprotective effects of CoQ10 in rats against the observed oxidative stress during seizures induced by pilocarpine, and to study its interactions with the conventional antiepileptic drug phenytoin, two experiments were performed. Experiment 1 was conducted to test the effect of phenytoin, CoQ10, or both on seizure severity and oxidative markers in the pilocarpine model of epilepsy. Experiment 2 was conducted to test the effect of 2 weeks of chronic treatment with phenytoin, CoQ10, or both on oxidative markers and behavioral tests in rats. Overall, CoQ10 reduced the severity of pilocarpine-induced seizures and the severity of oxidative stress. Moreover, it potentiated the antiepileptic effects afforded by phenytoin treatment, with the potential safety and efficacy in ameliorating oxidative stress and cognitive impairment caused by chronic phenytoin therapy. Our findings strongly suggest that CoQ10 can be considered a safe and effective adjuvant to phenytoin therapy in epilepsy both to ameliorate seizure severity and to protect against seizure-induced oxidative damage by reducing the cognitive impairment and oxidative stress associated with chronic use of phenytoin.

  15. Hippocampal gene expression analysis using the ORESTES methodology shows that homer 1a mRNA is upregulated in the acute period of the pilocarpine epilepsy model.

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    Avedissian, Marcelo; Longo, Beatriz M; Jaqueta, Carolina B; Schnabel, Beatriz; Paiva, Paulo B; Mello, Luiz Eugênio A M; Briones, Marcelo R S

    2007-01-01

    In the study of temporal lobe epilepsy (TLE) the characterization of genes expressed in the hippocampus is of central importance for understanding their roles in epileptogenic mechanisms. Although several large-scale studies on TLE gene expression have been reported, precise assignment of individual genes associated with this syndrome is still debatable. Here we investigated differentially expressed genes by comparison of mRNAs from normal and epileptic rat hippocampus in the pilocarpine model of epilepsy. For this we used a powerful EST sequencing methodology, ORESTES (Open Reading frame Expressed Sequence Tags), which generates sequence datasets enriched for mRNAs open reading frames (ORFs) rather than simple 5' and 3' ends of mRNAs. Analysis of our sequences shows that ORESTES readily enables the identification of epilepsy associated ORFs. PFAM analysis of protein motifs present in our ORESTES epilepsy database revealed diverse important protein family domains, such as cytoskeletal, cell signaling and protein kinase domains, which could be involved in processes underlying epileptogenesis. More importantly, we show that the expression of homer 1a, known to be coupled to mGluR and NMDA synaptic transmission, is associated with pilocarpine induced status epilepticus (SE). The combined use of the pilocarpine model of epilepsy with the ORESTES technique can significantly contribute to the identification of specific genes and proteins related to TLE. This is the first study applying a large-scale method for rapid shotgun sequencing directed to ORFs in epilepsy research.

  16. Anticonvulsant effect of Rhynchophylline involved in the inhibition of persistent sodium current and NMDA receptor current in the pilocarpine rat model of temporal lobe epilepsy.

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    Shao, Hui; Yang, Yang; Mi, Ze; Zhu, Guang-Xi; Qi, Ai-Ping; Ji, Wei-Gang; Zhu, Zhi-Ru

    2016-11-19

    Rhynchophylline (RIN) is a significant active component isolated from the Chinese herbal medicine Uncaria rhynchophylla. Several studies have demonstrated that RIN has a significant anticonvulsant effect in many types of epilepsy models in vivo. However, the mechanisms of the anticonvulsant effect remain elusive. Using combined methods of behavioral testing, immunofluorescence and electrophysiological recordings, we characterized the anticonvulsant effect of RIN in a pilocarpine-induced status epilepticus (SE) rat model of temporal lobe epilepsy (TLE) and investigated the underlying cellular mechanisms. In one set of experiments, rats received RIN treatment prior to pilocarpine injection. In a second set of experiments, rats received RIN treatment following the onset of stage 3 seizures. Pretreatment and posttreatment with RIN effectively reduced the seizure severity in the acute phase of TLE. Furthermore, RIN protected medial entorhinal cortex (mEC) layer III neurons from neuronal death and terminated spontaneous epileptiform discharge of mEC layer II neurons in SE-experienced rats. Whole-cell voltage-clamp recordings indicated that RIN inhibited neuronal hyperexcitability via inhibition of the persistent sodium current (INaP) and NMDA receptor current. Immunofluorescence experiments also demonstrated that RIN rectified the pilocarpine-induced upregulation of Nav1.6 and NR2B protein expression. In conclusion, our results identified RIN as an anticonvulsant agent that inhibited ictal discharge via INap and NMDA receptor current inhibition.

  17. Propofol effectively inhibits lithium-pilocarpine-induced status epilepticus in rats via downregulation of N-methyl-D-aspartate receptor 2B subunit expression

    Institute of Scientific and Technical Information of China (English)

    Henglin Wang; Zhuoqiang Wang; Weidong Mi; Cong Zhao; Yanqin Liu; Yongan Wang; Haipeng Sun

    2012-01-01

    Status epilepticus was induced via intraperitoneal injection of lithium-pilocarpine. The inhibitory ef-fects of propofol on status epilepticus in rats were judged based on observation of behavior, elec-troencephalography and 24-hour survival rate. Propofol (12.5-100 mg/kg) improved status epilep-ticus in a dose-dependent manner, and significantly reduced the number of deaths within 24 hours of lithium-pilocarpine injection. Western blot results showed that, 24 hours after induction of status epilepticus, the levels of N-methyl-D-aspartate receptor 2A and 2B subunits were significantly in-creased in rat cerebral cortex and hippocampus. Propofol at 50 mg/kg significantly suppressed the increase in N-methyl-D-aspartate receptor 2B subunit levels, but not the increase in N-methyl-D-aspartate receptor 2A subunit levels. The results suggest that propofol can effectively inhibit status epilepticus induced by lithium-pilocarpine. This effect may be associated with down-regulation of N-methyl-D-aspartate receptor 2B subunit expression after seizures.

  18. Variations in elemental compositions of rat hippocampal formation between acute and latent phases of pilocarpine-induced epilepsy: an X-ray fluorescence microscopy study.

    Science.gov (United States)

    Chwiej, J; Dulinska, J; Janeczko, K; Appel, K; Setkowicz, Z

    2012-06-01

    There is growing experimental evidence that tracing the elements involved in brain hyperexcitability, excitotoxicity, and/or subsequent neurodegeneration could be a valuable source of data on the molecular mechanisms triggering or promoting further development of epilepsy. The most frequently used experimental model of the temporal lobe epilepsy observed in clinical practice is the one based on pilocarpine-induced seizures. In the frame of this study, the elemental anomalies occurring for the rat hippocampal tissue in acute and silent periods after injection of pilocarpine in rats were compared. X-ray fluorescence microscopy was applied for the topographic and quantitative elemental analysis. The differences in the levels of elements such as P, S, K, Ca, Fe, Cu, and Zn between the rats 3 days (SE72) and 6 h (SE6) after pilocarpine injection as well as naive controls were examined. Comparison of SE72 and control groups showed, for specific areas of the hippocampal formation, lower levels of P, K, Cu, and Zn, and an increase in Ca accumulation. These results as well as further analysis of the differences between the SE72 and SE6 groups confirmed that seizure-induced excitotoxicity as well as mossy fiber sprouting are the mechanisms involved in the neurodegenerative processes which may finally lead to spontaneous seizures in the chronic period of the pilocarpine model. Moreover, in the light of the results obtained, Cu seems to play a very important role in the pathogenesis of epilepsy in this animal model. For all areas analyzed, the levels of this element recorded in the latent period were not only lower than those for controls but were even lower than the levels found in the acute period. The decreased hippocampal accumulation of Cu in the phase of behavior and EEG stabilization, a possible inhibitory effect of this element on excitatory amino acid receptors, and enhanced seizure susceptibility in Menkes disease (an inherited Cu transport disorder leading to Cu

  19. Altered MT1 and MT2 melatonin receptors expression in the hippocampus of pilocarpine-induced epileptic rats.

    Science.gov (United States)

    Rocha, Anna Karynna Alves de Alencar; de Lima, Eliangela; Amaral, Fernanda; Peres, Rafael; Cipolla-Neto, José; Amado, Débora

    2017-06-01

    Clinical and experimental findings show that melatonin may be used as an adjuvant to the treatment of epilepsy-related complications by alleviates sleep disturbances, circadian alterations and attenuates seizures alone or in combination with AEDs. In addition, it has been observed that there is a circadian component on seizures, which cause changes in circadian system and in melatonin production. Nevertheless, the dynamic changes of the melatoninergic system, especially with regard to its membrane receptors (MT1 and MT2) in the natural course of TLE remain largely unknown. The aim of this study was to evaluate the 24-hour profile of MT1 and MT2 mRNA and protein expression in the hippocampus of rats submitted to the pilocarpine-induced epilepsy model analyzing the influence of the circadian rhythm in the expression pattern during the acute, silent, and chronic phases. Melatonin receptor MT1 and MT2 mRNA expression levels were increased in the hippocampus of rats few hours after SE, with MT1 returning to normal levels and MT2 reducing during the silent phase. During the chronic phase, mRNA expression levels of both receptors return to levels close to control, however, presenting a different daily profile, showing that there is a circadian change during the chronic phase. Also, during the acute and silent phase it was possible to verify MT1 label only in CA2 hippocampal region with an increased expression only in the dark period of the acute phase. The MT2 receptor was present in all hippocampal regions, however, it was reduced in the acute phase and it was found in astrocytes. In chronic animals, there is a reduction in the presence of both receptors especially in regions where there is a typical damage derived from epilepsy. Therefore, we conclude that SE induced by pilocarpine is able to change melatonin receptor MT1 and MT2 protein and mRNA expression levels in the hippocampus of rats few hours after SE as well as in silent and chronic phases. Copyright © 2017

  20. Pilocarpine-induced seizures trigger differential regulation of microRNA-stability related genes in rat hippocampal neurons

    Science.gov (United States)

    Kinjo, Erika R.; Higa, Guilherme S. V.; Santos, Bianca A.; de Sousa, Erica; Damico, Marcio V.; Walter, Lais T.; Morya, Edgard; Valle, Angela C.; Britto, Luiz R. G.; Kihara, Alexandre H.

    2016-01-01

    Epileptogenesis in the temporal lobe elicits regulation of gene expression and protein translation, leading to reorganization of neuronal networks. In this process, miRNAs were described as being regulated in a cell-specific manner, although mechanistics of miRNAs activity are poorly understood. The specificity of miRNAs on their target genes depends on their intracellular concentration, reflecting the balance of biosynthesis and degradation. Herein, we confirmed that pilocarpine application promptly (<30 min) induces status epilepticus (SE) as revealed by changes in rat electrocorticogram particularly in fast-beta range (21–30 Hz). SE simultaneously upregulated XRN2 and downregulated PAPD4 gene expression in the hippocampus, two genes related to miRNA degradation and stability, respectively. Moreover, SE decreased the number of XRN2-positive cells in the hilus, while reduced the number of PAPD4-positive cells in CA1. XRN2 and PAPD4 levels did not change in calretinin- and CamKII-positive cells, although it was possible to determine that PAPD4, but not XRN2, was upregulated in parvalbumin-positive cells, revealing that SE induction unbalances the accumulation of these functional-opposed proteins in inhibitory interneurons that directly innervate distinct domains of pyramidal cells. Therefore, we were able to disclose a possible mechanism underlying the differential regulation of miRNAs in specific neurons during epileptogenesis. PMID:26869208

  1. Role of the Mitochondrial Calcium Uniporter in Rat Hippocampal Neuronal Death After Pilocarpine-Induced Status Epilepticus.

    Science.gov (United States)

    Wang, Cui; Xie, Nanchang; Wang, Yunlong; Li, Yulin; Ge, Xinjie; Wang, Menglu

    2015-08-01

    The mitochondrial calcium uniporter (MCU) is reportedly involved in oxidative stress, apoptosis, and many neurological diseases. However, the role of the MCU in epilepsy remains unknown. In this study, we found that the MCU inhibitor Ru360 significantly attenuated neuronal death and exerted an anti-apoptotic effect on rat hippocampal neurons after pilocarpine-induced status epilepticus (SE), while the MCU activator spermine increased seizure-induced neuronal death and apoptosis. In addition, Ru360 decreased the level of seizure-induced reactive oxygen species (ROS) in mitochondria isolated from rat hippocampi. Moreover, Ru360 restored the altered mitochondrial membrane potential and cytochrome c (CytC) release in epileptic hippocampi. However, spermine treatment exerted an opposite effect on seizure-induced ROS production and mitochondrial membrane potential alteration and CytC release compared with Ru360 treatment. Altogether, the findings of this study suggest that MCU inhibition exerts a neuroprotective effect on seizure-induced brain injury possibly through the mitochondria/ROS/CytC pathway.

  2. Extreme obesity in female rats following prepuberal induction of lithium-pilocarpine seizures and a single injection of acepromazine.

    Science.gov (United States)

    St-Pierre, L S; Persinger, M A

    2005-11-01

    Seizures were induced in female Wistar albino rats at either 35 or 55 days of age with a single systemic injection of lithium (3 mEq/kg) and pilocarpine (30 mg/kg); the rats were then treated with the atypical neuroleptic acepromazine (25 mg/kg). These rats manifested progressive weight gain for the rest of their lives. The effect was conspicuous by casual observation 6 weeks after treatment and occurred primarily in those rats that later developed spontaneous seizures. After 1 year, the rats were obese (>1000 g). Such weight gains, associated with almost three times the serum triglyceride levels, were not observed in male rats and have not been observed in hundreds of female rats that received this treatment as adults. Single postseizure injections of ketamine rather than acepromazine did not produce this obesity; the weights of these rats were similar to those of normal littermates. These results indicate that a single injection of a neuroleptic during limbic seizures before puberty can produce neuronal alterations that contribute to a lifetime of obesity.

  3. Toll-like receptor 3 deficiency decreases epileptogenesis in a pilocarpine model of SE-induced epilepsy in mice.

    Science.gov (United States)

    Gross, Adi; Benninger, Felix; Madar, Ravit; Illouz, Tomer; Griffioen, Kathleen; Steiner, Israel; Offen, Daniel; Okun, Eitan

    2017-04-01

    Epilepsy affects 60 million people worldwide. Despite the development of antiepileptic drugs, up to 35% of patients are drug refractory with uncontrollable seizures. Toll-like receptors (TLRs) are central components of the nonspecific innate inflammatory response. Because TLR3 was recently implicated in neuronal plasticity, we hypothesized that it may contribute to the development of epilepsy after status epilepticus (SE). To test the involvement of TLR3 in epileptogenesis, we used the pilocarpine model for SE in TLR3-deficient mice and their respective wild-type controls. In this model, a single SE event leads to spontaneous recurrent seizures (SRS). Two weeks after SE, mice were implanted with wireless electroencephalography (EEG) transmitters for up to 1 month. The impact of TLR3 deficiency on SE was assessed using separate cohorts of mice regarding EEG activity, seizure progression, hippocampal microglial distribution, and expression of the proinflammatory cytokines tumor necrosis factor (TNF)α and interferon (IFN)β. Our data indicate that TLR3 deficiency reduced SRS, microglial activation, and the levels of the proinflammatory cytokines TNFα and IFNβ, and increased survival following SE. This study reveals novel insights into the pathophysiology of epilepsy and the contribution of TLR3 to disease progression. Our results identify the TLR3 pathway as a potential future therapeutic target in SE. Wiley Periodicals, Inc. © 2017 International League Against Epilepsy.

  4. Chronic deficit in the expression of voltage-gated potassium channel Kv3.4 subunit in the hippocampus of pilocarpine-treated epileptic rats.

    Science.gov (United States)

    Pacheco Otalora, Luis F; Skinner, Frank; Oliveira, Mauro S; Farrell, Bianca; Arshadmansab, Massoud F; Pandari, Tarun; Garcia, Ileana; Robles, Leslie; Rosas, Gerardo; Mello, Carlos F; Ermolinsky, Boris S; Garrido-Sanabria, Emilio R

    2011-01-12

    Voltage gated K(+) channels (Kv) are a highly diverse group of channels critical in determining neuronal excitability. Deficits of Kv channel subunit expression and function have been implicated in the pathogenesis of epilepsy. In this study, we investigate whether the expression of the specific subunit Kv3.4 is affected during epileptogenesis following pilocarpine-induced status epilepticus. For this purpose, we used immunohistochemistry, Western blotting assays and comparative analysis of gene expression using TaqMan-based probes and delta-delta cycle threshold (ΔΔCT) method of quantitative real-time polymerase chain reaction (qPCR) technique in samples obtained from age-matched control and epileptic rats. A marked down-regulation of Kv3.4 immunoreactivity was detected in the stratum lucidum and hilus of dentate gyrus in areas corresponding to the mossy fiber system of chronically epileptic rats. Correspondingly, a 20% reduction of Kv3.4 protein levels was detected in the hippocampus of chronic epileptic rats. Real-time quantitative PCR analysis of gene expression revealed that a significant 33% reduction of transcripts for Kv3.4 (gene Kcnc4) occurred after 1 month of pilocarpine-induced status epilepticus and persisted during the chronic phase of the model. These data indicate a reduced expression of Kv3.4 channels at protein and transcript levels in the epileptic hippocampus. Down-regulation of Kv3.4 in mossy fibers may contribute to enhanced presynaptic excitability leading to recurrent seizures in the pilocarpine model of temporal lobe epilepsy.

  5. Cellular hybridization for BDNF, trkB, and NGF mRNAs and BDNF-immunoreactivity in rat forebrain after pilocarpine-induced status epilepticus.

    Science.gov (United States)

    Schmidt-Kastner, R; Humpel, C; Wetmore, C; Olson, L

    1996-01-01

    The messenger RNAs (mRNAs) for the neurotrophins, brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF), are upregulated during epileptic seizure activity, as visualized by in situ hybridization techniques. Neurotrophins might be protective against excitotoxic cell stress, and the upregulation during seizures might provide such cell protection. In this study, a high dose of pilocarpine (300 mg/kg) was used to induce long-lasting, limbic motor status epilepticus and a selective pattern of brain damage. The regulation of BDNF, trkB, and NGF mRNA was studied by in situ hybridization at 1, 3, 6, and 24 h after induction of limbic motor status epilepticus. BDNF immunoreactivity was examined with an anti-peptide antibody and the neuropathological process studied in parallel. BDNF mRNA increased in hippocampus, neocortex, piriform cortex, striatum, and thalamus with a maximum at 3-6 h. Hybridization levels increased earlier in the resistant granule and CA1 cells as compared to the vulnerable CA3 neurons. BDNF immunoreactivity was elevated in dentate gyrus at 3-6 h. trkB mRNA increased in the entire hippocampus. NGF mRNA in hippocampus appeared in dentate gyrus at 3-6 h and declined in hilar neurons at 6-24 h. Cell damage was found in the CA3 area, entire basal cortex, and layers II/III of neocortex. Endogenous neurotrophins are upregulated during status epilepticus caused by pilocarpine, which is related to the coupling between neuronal excitation and trophic factor expression. This upregulation of neurotrophic factors may serve endogenous protective effects; however, the excessive levels of neuronal hyperexcitation resulting from pilocarpine seizures lead to cell damage which cannot be prevented by endogenous neurotrophins.

  6. Alterations of hippocampal GAbaergic system contribute to development of spontaneous recurrent seizures in the rat lithium-pilocarpine model of temporal lobe epilepsy.

    Science.gov (United States)

    André, V; Marescaux, C; Nehlig, A; Fritschy, J M

    2001-01-01

    Reorganization of excitatory and inhibitory circuits in the hippocampal formation following seizure-induced neuronal loss has been proposed to underlie the development of chronic seizures in temporal lobe epilepsy (TLE). Here, we investigated whether specific morphological alterations of the GABAergic system can be related to the onset of spontaneous recurrent seizures (SRS) in the rat lithium-pilocarpine model of TLE. Immunohistochemical staining for markers of interneurons and their projections, including parvalbumin (PV), calretinin (CR), calbindin (CB), glutamic acid decarboxylase (GAD), and type 1 GABA transporter (GAT1), was performed in brain sections of rats treated with lithium-pilocarpine and sacrificed after 24 h, during the silent phase (6 and 12 days), or after the onset of SRS (10-18 days after treatment). Semiquantitative analysis revealed a selective loss of interneurons in the stratum oriens of CA1, associated with a reduction of GAT1 staining in the stratum radiatum and stratum oriens. In contrast, interneurons in CA3 were largely preserved, although GAT1 staining was also reduced. These changes occurred within 6 days after treatment and were therefore insufficient to cause SRS. In the dentate gyrus, extensive cell loss occurred in the hilus. The pericellular innervation of granule cells by PV-positive axons was markedly reduced, although the loss of PV-interneurons was only partial. Most strikingly, the density of GABAergic axons, positive for both GAD and GAT1, was dramatically increased in the inner molecular layer. This change emerged during the silent period, but was most marked in animals with SRS. Finally, supernumerary CB-positive neurons were detected in the hilus, selectively in rats with SRS. These findings suggest that alterations of GABAergic circuits occur early after lithium-pilocarpine-induced status epilepticus and contribute to epileptogenesis. In particular, the reorganization of GABAergic axons in the dentate gyrus might

  7. Expression pattern of NMDA receptors reveals antiepileptic potential of apigenin 8-C-glucoside and chlorogenic acid in pilocarpine induced epileptic mice.

    Science.gov (United States)

    Aseervatham, G Smilin Bell; Suryakala, U; Doulethunisha; Sundaram, S; Bose, P Chandra; Sivasudha, T

    2016-08-01

    The present study was aimed to evaluate the effect of apigenin 8-C-glucoside (Vitexin) and chlorogenic acid on epileptic mice induced by pilocarpine and explored its possible mechanisms. Intraperitonial administration of pilocarpine (85mg/kg) induced seizure in mice was assessed by behavior observations, which is significantly (p>0.05) reduced by apigenin 8-C-glucoside (AP8CG) (10mg/kg) and chlorogenic acid (CA) (5mg/kg), similar to diazepam. Seizure was accompanied by an imbalance in the levels of Gamma-aminobutyric acid (GABA) and glutamate in the pilocarpine administered group. Moreover, convulsion along with reduced acetylcholinesterase, increased monoamine oxidase and oxidative stress was observed in epileptic mice brain. AP8CG and CA significantly restored back to normal levels even at lower doses. Further, increased lipid peroxidation and nitrite content was also significantly attenuated by AP8CG and CA. However, CA was found to be more effective when compared to AP8CG. In addition, the mRNA expression of N-methyl-d-aspartate receptor (NMDAR), mGluR1 and mGlu5 was significantly (P≤0.05) inhibited by AP8CG and CA in a lower dose. The mRNA expression of GRIK1 did not differ significantly in any of the group and showed a similar pattern of expression. Our result shows that AP8CG and CA selectively inhibit NMDAR, mGluR1 and mGlu5 expression. Modification in the provoked NMDAR calcium response coupled with neuronal death. Hence, these findings underline that the polyphenolics, AP8CG and CA have exerted antiepileptic and neuroprotective activity by suppressing glutamate receptors.

  8. The spiny rat Proechimys guyannensis as model of resistance to epilepsy: chemical characterization of hippocampal cell populations and pilocarpine-induced changes.

    Science.gov (United States)

    Fabene, P F; Correia, L; Carvalho, R A; Cavalheiro, E A; Bentivoglio, M

    2001-01-01

    At variance with pilocarpine-induced epilepsy in the laboratory rat, pilocarpine administration to the tropical rodent Proechimys guyannensis (casiragua) elicited an acute seizure that did not develop in long-lasting status epilepticus and was not followed by spontaneous seizures up to 30 days, when the hippocampus was investigated in treated and control animals. Nissl staining revealed in Proechimys a highly developed hippocampus, with thick hippocampal commissures and continuity of the rostral dentate gyri at the midline. Immunohistochemistry was used to study calbindin, parvalbumin, calretinin, GABA, glutamic acid decarboxylase, and nitric oxide synthase expression. The latter was also investigated with NADPH-diaphorase histochemistry. Cell counts and densitometric evaluation with image analysis were performed. Differences, such as low calbindin immunoreactivity confined to some pyramidal cells, were found in the normal Proechimys hippocampus compared to the laboratory rat. In pilocarpine-treated casiraguas, stereological cell counts in Nissl-stained sections did not reveal significant neuronal loss in hippocampal subfields, where the examined markers exhibited instead striking changes. Calbindin was induced in pyramidal and granule cells and interneuron subsets. The number of parvalbumin- or nitric oxide synthase-containing interneurons and their staining intensity were significantly increased. Glutamic acid decarboxylase(67)-immunoreactive interneurons increased markedly in the hilus and decreased in the CA1 pyramidal layer. The number and staining intensity of calretinin-immunoreactive pyramidal cells and interneurons were significantly reduced. These findings provide the first description of the Proechimys hippocampus and reveal marked long-term variations in protein expression after an epileptic insult, which could reflect adaptive changes in functional hippocampal circuits implicated in resistance to limbic epilepsy.

  9. Antagomirs targeting microRNA-134 increase hippocampal pyramidal neuron spine volume in vivo and protect against pilocarpine-induced status epilepticus.

    Science.gov (United States)

    Jimenez-Mateos, Eva M; Engel, Tobias; Merino-Serrais, Paula; Fernaud-Espinosa, Isabel; Rodriguez-Alvarez, Natalia; Reynolds, James; Reschke, Cristina R; Conroy, Ronan M; McKiernan, Ross C; deFelipe, Javier; Henshall, David C

    2015-07-01

    Emerging data support roles for microRNA (miRNA) in the pathogenesis of various neurologic disorders including epilepsy. MicroRNA-134 (miR-134) is enriched in dendrites of hippocampal neurons, where it negatively regulates spine volume. Recent work identified upregulation of miR-134 in experimental and human epilepsy. Targeting miR-134 in vivo using antagomirs had potent anticonvulsant effects against kainic acid-induced seizures and was associated with a reduction in dendritic spine number. In the present study, we measured dendritic spine volume in mice injected with miR-134-targeting antagomirs and tested effects of the antagomirs on status epilepticus triggered by the cholinergic agonist pilocarpine. Morphometric analysis of over 6,400 dendritic spines in Lucifer yellow-injected CA3 pyramidal neurons revealed increased spine volume in mice given antagomirs compared to controls that received a scrambled sequence. Treatment of mice with miR-134 antagomirs did not alter performance in a behavioral test (novel object location). Status epilepticus induced by pilocarpine was associated with upregulation of miR-134 within the hippocampus of mice. Pretreatment of mice with miR-134 antagomirs reduced the proportion of animals that developed status epilepticus following pilocarpine and increased animal survival. In antagomir-treated mice that did develop status epilepticus, seizure onset was delayed and total seizure power was reduced. These studies provide in vivo evidence that miR-134 regulates spine volume in the hippocampus and validation of the seizure-suppressive effects of miR-134 antagomirs in a model with a different triggering mechanism, indicating broad conservation of anticonvulsant effects.

  10. Validation of suitable reference genes for expression studies in different pilocarpine-induced models of mesial temporal lobe epilepsy.

    Directory of Open Access Journals (Sweden)

    Thalita Ewellyn Batista Sales Marques

    Full Text Available It is well recognized that the reference gene in a RT-qPCR should be properly validated to ensure that gene expression is unaffected by the experimental condition. We investigated eight potential reference genes in two different pilocarpine PILO-models of mesial temporal lobe epilepsy (MTLE performing a stability expression analysis using geNorm, NormFinder and BestKepeer softwares. Then, as a validation strategy, we conducted a relative expression analysis of the Gfap gene. Our results indicate that in the systemic PILO-model Actb, Gapdh, Rplp1, Tubb2a and Polr1a mRNAs were highly stable in hippocampus of rats from all experimental and control groups, whereas Gusb revealed to be the most variable one. In fact, we observed that using Gusb for normalization, the relative mRNA levels of the Gfap gene differed from those obtained with stable genes. On the contrary, in the intrahippocampal PILO-model, all softwares included Gusb as a stable gene, whereas B2m was indicated as the worst candidate gene. The results obtained for the other reference genes were comparable to those observed for the systemic Pilo-model. The validation of these data by the analysis of the relative expression of Gfap showed that the upregulation of the Gfap gene in the hippocampus of rats sacrificed 24 hours after status epilepticus (SE was undetected only when B2m was used as the normalizer. These findings emphasize that a gene that is stable in one pathology model may not be stable in a different experimental condition related to the same pathology and therefore, the choice of reference genes depends on study design.

  11. Lovastatin modulates glycogen synthase kinase-3β pathway and inhibits mossy fiber sprouting after pilocarpine-induced status epilepticus.

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    Chun-Yao Lee

    Full Text Available This study was undertaken to assay the effect of lovastatin on the glycogen synthase kinase-3 beta (GSK-3β and collapsin responsive mediator protein-2 (CRMP-2 signaling pathway and mossy fiber sprouting (MFS in epileptic rats. MFS in the dentate gyrus (DG is an important feature of temporal lobe epilepsy (TLE and is highly related to the severity and the frequency of spontaneous recurrent seizures. However, the molecular mechanism of MFS is mostly unknown. GSK-3β and CRMP-2 are the genes responsible for axonal growth and neuronal polarity in the hippocampus, therefore this pathway is a potential target to investigate MFS. Pilocarpine-induced status epilepticus animal model was taken as our researching material. Western blot, histological and electrophysiological techniques were used as the studying tools. The results showed that the expression level of GSK-3β and CRMP-2 were elevated after seizure induction, and the administration of lovastatin reversed this effect and significantly reduced the extent of MFS in both DG and CA3 region in the hippocampus. The alteration of expression level of GSK-3β and CRMP-2 after seizure induction proposes that GSK-3β and CRMP-2 are crucial for MFS and epiletogenesis. The fact that lovastatin reversed the expression level of GSK-3β and CRMP-2 indicated that GSK-3β and CRMP-2 are possible to be a novel mechanism of lovatstain to suppress MFS and revealed a new therapeutic target and researching direction for studying the mechanism of MFS and epileptogenesis.

  12. Decreased expression of proteins involved in energy metabolism in the hippocampal granular layer of rats submitted to the pilocarpine epilepsy model.

    Science.gov (United States)

    Araujo, Bruno; Torres, Laila; Stein, Mariana; Cabral, Francisco Romero; Herai, Roberto; Okamoto, Oswaldo; Cavalheiro, Esper

    2014-02-21

    Long-term structural and functional changes in the hippocampus have been identified as the primary physiopathological basis for temporal lobe epilepsy. These changes include reactive gliosis and granule cell axonal sprouting within the dentate gyrus. The intimate mechanisms of these changes are beginning to be revealed. Here, we show the possibility of using laser capture microdissection (LCM) to isolate the dentate granular cell layer of Wistar rats submitted to the pilocarpine model of epilepsy. Using two-dimensional gel electrophoresis (2-D PAGE) and mass spectrometry for laser-captured cells, we identified molecular events that could be altered as part of the epileptic pathogenic process. According to our results, eight proteins related to energy metabolism were differentially expressed between both the control and pilocarpine-treated animals. These results provide, for the first time, new molecular insights into the altered protein profile of the epileptogenic dentate gyrus and can contribute to a better understanding of the phenomena involved in the genesis and maintenance of the epileptic state.

  13. Growth-associated phosphoprotein expression is increased in the supragranular regions of the dentate gyrus following pilocarpine-induced seizures in rats.

    Science.gov (United States)

    Naffah-Mazzacoratti, M G; Funke, M G; Sanabria, E R; Cavalheiro, E A

    1999-01-01

    Neuroplasticity has been investigated considering the neuronal growth-associated phosphoprotein as a marker of neuronal adaptive capabilities. In the present work, studying the hippocampal reorganization observed in the epilepsy model induced by pilocarpine, we carried out quantitative western blotting associated with immunohistochemistry to determine the distribution of growth-associated phosphoprotein in the hippocampus of rats in acute, silent and chronic periods of this epilepsy model. The fibers and punctate elements from the inner molecular layer of the dentate gyrus were strongly immunostained in animals killed 5 h after status epilepticus, compared with the same region in control animals. Rats presenting partial seizures showed no alterations in the immunostaining pattern compared with saline-treated animals. The hippocampal dentate gyrus of animals during the seizure-free period and presenting spontaneous recurrent seizures was also characterized by strong growth-associated phosphoprotein immunostaining of fibers and punctate elements in the inner molecular layer, contrasting with the control group. As determined by western blotting analysis, growth-associated phosphoprotein levels increased following status epilepticus and remained elevated at the later time-points, both during the silent period and during the period of chronic recurring seizures. Pilocarpine-treated animals, which did not develop status epilepticus, showed no change in growth-associated phosphoprotein levels, indicating that status epilepticus is important to induce growth-associated phosphoprotein overexpression. The measurement of this overexpression could represent one of the early signals of hippocampal reorganization due to status epilepticus-induced damage.

  14. Cannabidiol Post-Treatment Alleviates Rat Epileptic-Related Behaviors and Activates Hippocampal Cell Autophagy Pathway Along with Antioxidant Defense in Chronic Phase of Pilocarpine-Induced Seizure.

    Science.gov (United States)

    Hosseinzadeh, Mahshid; Nikseresht, Sara; Khodagholi, Fariba; Naderi, Nima; Maghsoudi, Nader

    2016-04-01

    Abnormal and sometimes severe behavioral and molecular symptoms are usually observed in epileptic humans and animals. To address this issue, we examined the behavioral and molecular aspects of seizure evoked by pilocarpine. Autophagy can promote both cell survival and death, but there are controversial reports about the neuroprotective or neurodegenerative effects of autophagy in seizure. Cannabidiol has anticonvulsant properties in some animal models when used as a pretreatment. In this study, we investigated alteration of seizure scores, autophagy pathway proteins, and antioxidant status in hippocampal cells during the chronic phase of pilocarpine-induced epilepsy after treatment with cannabidiol. Cannabidiol (100 ng, intracerebroventricular injection) delayed the chronic phase of epilepsy. Single administration of cannabidiol during the chronic phase of seizure significantly diminished seizure scores such as mouth clonus, head nodding, monolateral and bilateral forelimb clonus and increased the activity of catalase enzyme and reduced glutathione content. Such a protective effect in the behavioral scores of epileptic rats was also observed after repeated administrations of cannabidiol at the onset of the silent phase. Moreover, the amount of Atg7, conjugation of Atg5/12, Atg12, and LC3II/LC3I ratio increased significantly in epileptic rats treated with repeated injections of cannabidiol. In short, our results suggest that post-treatment of Cannabidiol could enhance the induction of autophagy pathway and antioxidant defense in the chronic phase of epilepsy, which could be considered as the protective mechanisms of cannabidiol in a temporal lobe epilepsy model.

  15. Differential expression of brain-derived neurotrophic factor transcripts after pilocarpine-induced seizure-like activity is related to mode of Ca2+ entry

    DEFF Research Database (Denmark)

    Poulsen, F R; Lauterborn, J; Zimmer, J;

    2004-01-01

    ) and tyrosine kinase B (trkB) also were studied. Pilocarpine (5 mM) induced a dose- and time-dependent increase in total BDNF (exon V) mRNA expression in the dentate granule cells and CA3-CA1 pyramidal cells with maximal effects at 6 and 24 h, respectively. Increases were blocked by co-treatment with the alpha......Activity-dependent brain-derived neurotrophic factor (BDNF) expression is Ca2+-dependent, yet little is known about the Ca2+ channel contributions that might direct selective expression of the multiple BDNF transcripts. Here, effects of pilocarpine-induced seizure activity on total BDNF expression...... and on the individual sensitivity of BDNF transcripts to glutamate receptor and Ca2+ channel blockers were evaluated using hippocampal slice cultures and in situ hybridization of transcript-specific cRNA probes directed against mRNAs for the four 5' exons (I-IV) of the BDNF gene. mRNAs for nerve growth factor (NGF...

  16. Mice deficient for the extracellular matrix glycoprotein tenascin-r show physiological and structural hallmarks of increased hippocampal excitability, but no increased susceptibility to seizures in the pilocarpine model of epilepsy.

    Science.gov (United States)

    Brenneke, F; Bukalo, O; Dityatev, A; Lie, A A

    2004-01-01

    Recognition molecules provide important cues for neuronal survival, axonal fasciculation, axonal pathfinding, synaptogenesis, synaptic plasticity, and regeneration. Our previous studies revealed a link between perisomatic inhibition and the extracellular matrix glycoprotein tenascin-R (TN-R). Therefore, we here studied neuronal excitability and epileptic susceptibility in mice constitutively deficient in TN-R. In vitro analysis of populational spikes in hippocampal slices of TN-R-deficient mice revealed a significant increase in multiple spikes in the CA1 region, as compared with wild-type mice. This difference between genotypes was only partially reduced after blockade of GABA(A) receptors with picrotoxin, indicating a deficit in GABAergic inhibition and an increase in intrinsic excitability of CA1 pyramidal cells in TN-R-deficient mice. Using a battery of immunohistochemical markers and histological stainings, we were able to identify two abnormalities in the hippocampus of TN-R-deficient mice possibly related to increased excitability: the high number of glial fibrillary acidic protein-positive astrocytes and low number of calretinin-positive interneurons in the CA1 and CA3 regions. In order to test whether the revealed abnormalities give rise to increased susceptibility to seizures in TN-R-deficient mice, we used the pilocarpine model of epilepsy. No genotype-specific differences were found with regard to the time-course of pilocarpine-induced and spontaneous seizures, neuronal cell loss, aberrant sprouting and distribution of synaptic and inhibitory interneuron markers. However, pilocarpine-induced astrogliosis and reduction in calretinin-positive interneurons were less pronounced in TN-R mutants, thereby resulting in an occlusion of effects induced by TN-R deficiency and pilocarpine. Thus, TN-R-deficient mutants show several electrophysiological and morphological hallmarks of increased neuronal excitability, which, however, do not give rise to more

  17. Transcranial focal electrical stimulation reduces the convulsive expression and amino acid release in the hippocampus during pilocarpine-induced status epilepticus in rats.

    Science.gov (United States)

    Santana-Gómez, César E; Alcántara-González, David; Luna-Munguía, Hiram; Bañuelos-Cabrera, Ivette; Magdaleno-Madrigal, Víctor; Fernández-Mas, Rodrigo; Besio, Walter; Rocha, Luisa

    2015-08-01

    The aim of the present study was to evaluate the effects of transcranial focal electrical stimulation (TFS) on γ-aminobutyric acid (GABA) and glutamate release in the hippocampus under basal conditions and during pilocarpine-induced status epilepticus (SE). Animals were previously implanted with a guide cannula attached to a bipolar electrode into the right ventral hippocampus and a concentric ring electrode placed on the skull surface. The first microdialysis experiment was designed to determine, under basal conditions, the effects of TFS (300 Hz, 200 μs biphasic square pulses, for 30 min) on afterdischarge threshold (ADT) and the release of GABA and glutamate in the hippocampus. The results obtained indicate that at low current intensities (Status Epilepticus".

  18. Evaluation of potential gender-related differences in behavioral and cognitive alterations following pilocarpine-induced status epilepticus in C57BL/6 mice.

    Science.gov (United States)

    Oliveira, Clarissa Vasconcelos de; Grigoletto, Jéssica; Funck, Vinícius Rafael; Ribeiro, Leandro Rodrigo; Royes, Luiz Fernando Freire; Fighera, Michele Rechia; Furian, Ana Flávia; Oliveira, Mauro Schneider

    2015-05-01

    Together with pharmacoresistant seizures, the quality of life of temporal lobe epilepsy (TLE) patients is negatively impacted by behavioral comorbidities including but not limited to depression, anxiety and cognitive deficits. The pilocarpine model of TLE has been widely used to study characteristics of human TLE, including behavioral comorbidities. Since the outcomes of pilocarpine-induced TLE might vary depending on several experimental factors, we sought to investigate potential gender-related differences regarding selected behavioral alterations in C57BL6 mice. We found that epileptic mice, independent of gender, displayed increased anxiety-like behavior in the open-field test. In the object recognition test, epileptic mice, regardless of gender, showed a decreased recognition index at 24 (but not at 4) hours after training. On the other hand, no significant differences were found regarding mice learning and memory performance in the Barnes maze paradigm. Motor coordination and balance as assessed by the beam walk and rotarod tests were not impaired in epileptic mice of both genders. However, female mice, independent of epilepsy, performed the beam walk and rotarod tasks better than their male counterparts. We also found that only male epileptic mice displayed disturbed behavior in the forced swim test, but the mice of both genders displayed anhedonia-like behavior in the taste preference test. Lastly, we found that the extent of hilar cell loss is similar in both genders. In summary, both genders can be successfully employed to study behavioral comorbidities of TLE; however, taking the potential gender differences into account may help choose the more appropriated gender for a given task, which may be of value for the minimization of the number of animals used during the experiments.

  19. Berberine exerts an anticonvulsant effect and ameliorates memory impairment and oxidative stress in a pilocarpine-induced epilepsy model in the rat

    Directory of Open Access Journals (Sweden)

    Gao F

    2014-11-01

    Full Text Available Fei Gao,1,* Ying Gao,2,* Yang-feng Liu,3 Li Wang,4 Ya-jun Li1 1Department of Neurology, First Affiliated Hospital of Xi’an Medical University, Xi’an, People’s Republic of China; 2Department of Radiotherapy Oncology, First Affiliated Hospital of Medical College of Xi’an Jiaotong University, Xi’an, People’s Republic of China; 3Department of Neurology, People’s Liberation Army No. 451 Hospital, Xi’an, People’s Republic of China; 4Department of Scientific Research, First Affiliated Hospital of Xi’an Medical University, Xi’an, People’s Republic of China *These authors contributed equally to this work Abstract: Though new antiepileptic drugs are emerging, approximately a third of epileptic patients still suffer from recurrent convulsions and cognitive dysfunction. Therefore, we tested whether berberine (Ber, a vegetable drug, has an anticonvulsant property and attenuates memory impairment in a pilocarpine (Pilo-induced epilepsy model in rats. The rats were injected with 400 mg/kg Pilo to induce convulsions, and Ber 25, 50, and 100 mg/kg were administrated by the intragastric route once daily 7 days before Pilo injection until the experiment was over. Convulsions were observed after Pilo injection. For the rats that developed status epilepticus (SE, malondialdehyde, glutathione levels, superoxide dismutase, and catalase activity in the hippocampus were measured 24 hours after SE. The rats received the Morris water-maze test 2 weeks after SE, and then were killed for fluoro-jade B staining to detect the degenerating neurons. We found Ber delayed latency to the first seizure and the time to develop SE in a dose-dependent manner. Malondialdehyde levels were decreased, while glutathione and catalase activity were strengthened in Ber-injected SE rats. In the Morris water-maze test, Ber decreased escape latency compared to saline-treated SE rats. Additionally, Ber reduced the number of fluoro-jade B-positive cells in the

  20. The Predictability of Preoperative Pilocarpine-Induced Lens Shift on the Outcomes of Accommodating Intraocular Lenses Implanted in Senile Cataract Patients

    Science.gov (United States)

    Li, Jin; Chen, Qi; Lin, Zhibo; Leng, Lin; Huang, Fang

    2016-01-01

    Purpose. To evaluate the predictability of lens shift induced by pilocarpine (LSPilo) on the outcomes of accommodating intraocular lens (Acc-IOL) implantation. Methods. Twenty-four eyes of 24 senile cataract patients who underwent phacoemulsification and Acc-IOL implantation were enrolled. LSPilo was evaluated with anterior segment optical coherence tomography (AS-OCT). At 3 months postoperatively, the best corrected distance visual acuities (BCDVA), distance-corrected near visual acuities (DCNVA), and subjective and objective accommodations were measured. IOL shifts under accommodation stimulus (IOLSAcc) were evaluated with AS-OCT. Results. The mean LSPilo was 112.29 ± 30.72 µm. LSPilo was not associated with any preoperative parameters. The mean IOLSAcc was 130.46 ± 42.71 µm. The mean subjective and objective accommodation were 1.54 ± 0.39 D and 1.27 ± 0.41 D, respectively. The mean postoperative BCDVA and DCNVA (log MAR value) were 0.22 ± 0.11 and 0.24 ± 0.12, respectively. LSPilo positively correlated with IOLSAcc (r = 0.541; P = 0.006), subjective accommodation (r = 0.412; P = 0.022), and objective accommodation (r = 0.466; P = 0.045), respectively. Conclusion. LSPilo is an independent preoperative parameter associated with the postoperative Acc-IOL mobility and pseudophakic accommodation. It may offer valuable information for ophthalmologists in determining the suitable candidates for Acc-IOL implantation. PMID:27516899

  1. Pyrrolidine dithiocarbamate (PDTC) inhibits the overexpression of MCP-1 and attenuates microglial activation in the hippocampus of a pilocarpine-induced status epilepticus rat model.

    Science.gov (United States)

    Lv, Rilang; Xu, Xiaoyun; Luo, Zheng; Shen, Nan; Wang, Feng; Zhao, Yongbo

    2014-01-01

    The aim of this study was to investigate the effects of pyrrolidine dithiocarbamate (PDTC) on MCP-1 expression and microglial activation in the hippocampus of a rat model of pilocarpine (PILO)-induced status epilepticus (SE). Moreover, seizure susceptibility, frequency and severity as well as brain damage were analyzed and changes in behavior were recorded. Chemokine MCP-1 expression and microglial activation were detected by immunohistochemistry (IHC). Fluoro-Jade C (FJC) and NeuN staining were used for the evaluation of tissue damage. Our results showed that although SE resulted in the upregulation of MCP-1 and microglial activation in the rat hippocampus 24 h after seizure onset, pretreatment with PDTC significantly inhibited the MCP-1 overexpression and attenuated the microglial activation. These effects were accompanied by neurodegenerative amelioration. To the best of our knowledge, these findings indicated for the first time that the activation of the nuclear factor-κB (NF-κB) pathway may contribute to MCP-1 upregulation and microglial activation in the context of epilepsy. PDTC was also shown to exert anticonvulsant activity and to have a neuroprotective effect on the hippocampal CA1 and CA3 regions, potentially through attenuating microglial activation.

  2. One hour of pilocarpine-induced status epilepticus is sufficient to develop chronic epilepsy in mice, and is associated with mossy fiber sprouting but not neuronal death

    Institute of Scientific and Technical Information of China (English)

    Ling-Lin Chen; Hang-Feng Feng; Xue-Xia Mao; Qing Ye; Ling-Hui Zeng

    2013-01-01

    Determining the minimal duration of status epilepticus (SE) that leads to the development of subsequent spontaneous seizures (i.e.,epilepsy) is important,because it provides a critical time-window for seizure intervention and epilepsy prevention.In the present study,male ICR (Imprinting Control Region) mice were injected with pilocarpine to induce acute seizures.SE was terminated by diazepam at 10 min,30 min,1 h,2 h and 4 h after seizure onset.Spontaneous seizures occurred in the 1,2 and 4 h SE groups,and the seizure frequency increased with the prolongation of SE.Similarly,the Morris water maze revealed that the escape latency was significantly increased and the number of target quadrant crossings was markedly decreased in the 1,2 and 4 h SE groups.Robust mossy fiber sprouting was observed in these groups,but not in the 10 or 30 min group.In contrast,Fluoro-Jade B staining revealed significant cell death only in the 4 h SE group.The incidence and frequency of spontaneous seizures were correlated with Timm score (P =0.004) and escape latency (P =0.004).These data suggest that SE longer than one hour results in spontaneous motor seizures and memory deficits,and spontaneous seizures are likely associated with robust mossy fiber sprouting but not neuronal death.

  3. Antagonist targeting microRNA-155 protects against lithium-pilocarpine-induced status epilepticus in C57BL/6 mice by activating brain-derived neurotrophic factor

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    Zhengxu eCai

    2016-05-01

    Full Text Available Epilepsy is a severe brain disorder affecting numerous patients. Recently, it is inferred that modulation of microRNA-155 (miR-155 could serve as a promising treatment of mesial temporal lobe epilepsy (MTLE. In the current study, the therapeutic potential of miR-155 antagonist against TLE was evaluated and the underlying mechanism involved in this regulation was explored. TLE model was induced by lithium-pilocarpine method. The effect of miR-155 antagonist on epilepticus symptoms of TLE mice was assessed using Racine classification and electroencephalogram (EEG recordings. The expression of brain-derived neurotrophic factor (BDNF and its association with miR-155 were also assessed with a series of experiments. Our results showed that level of miR-155 was significantly up-regulated after induction of TLE model. Based on the results of EEG and behavior analyses, seizures in mice were alleviated by miR-155 antagonist. Moreover, administration of miR-155 antagonist also significantly increased the level of BDNF. The results of dual luciferase assay and western blotting showed that miR-155 antagonist exerted its action on status epilepticus by directly regulating the activity of BDNF. Taken all the information together, our results demonstrated that miR-155 antagonist might firstly induce the expression of BDNF, which then contributed to the alleviation of epilepsy in the current study.

  4. Hippocampal distribution of IL-1β and IL-1RI following lithium-pilocarpine-induced status epilepticus in the developing rat

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    Dulce-Mariely Álvarez-Croda

    2016-01-01

    Full Text Available The contribution of Interleukin-1β (IL-1β to neuronal injury induced by status epilepticus (SE in the immature brain remains unclear. The goal of this study was to determine the hippocampal expression of IL-1β and its type 1 receptor (IL-1RI following SE induced by the lithium-pilocarpine model in fourteen-days-old rat pups; control animals were given an equal volume of saline instead of the convulsant. IL-1β and IL-1RI mRNA hippocampal levels were assessed by qRT-PCR 6 and 24 h after SE or control conditions. IL-1β and IL-1RI expression was detected in the dorsal hippocampus by immunohistochemical procedures; Fluoro-Jade B staining was carried out in parallel sections in order to detect neuronal cell death. IL-1β mRNA expression was increased 6 h following SE, but not at 24 h; however IL-1RI mRNA expression was unaffected when comparing with the control group. IL-1β and IL-1RI immunoreactivity was not detected in control animals. IL-1β and IL-1RI were expressed in the CA1 pyramidal layer, the dentate gyrus granular layer and the hilus 6 h after SE, whereas injured cells were detected 24 h following seizures. Early expression of IL-1β and IL-1RI in the hippocampus could be associated with SE-induced neuronal cell death mechanisms in the developing rat.

  5. 应用Fluoro-Jade C评价匹罗卡品模型中新大脑皮质神经变性%Assessment of neurodegeneration in neocortex of pilocarpine-treated mice by Fluoro-Jade C staining

    Institute of Scientific and Technical Information of China (English)

    王莲; 魏玲; 付莉

    2012-01-01

    目的 应用Fluoro-Jade C(FJC)在小鼠匹罗卡品癫痫模型中探测新大脑皮质结构中神经元的变性情况.方法 雄性昆明种小鼠10只(对照组5只,匹罗卡品处理组5只).对照组给予阿托品、生理盐水腹腔注射;匹罗卡品处理组给予阿托品、匹罗卡品腹腔注射诱发癫痫持续状态.在癫痫持续状态后12 h,通过经心灌注固定处死匹罗卡品处理组小鼠.对照组小鼠处死时间及方法同匹罗卡品处理组.在各新大脑皮质水平切制冠状切片,行FJC染色,在荧光显微镜下,观察FJC阳性细胞的形态和在新大脑皮质中的整体分布情况.结果 在匹罗卡品处理组,许多新大脑皮质出现呈亮黄绿色荧光的FJC阳性细胞,而对照组未见.结论 在小鼠匹罗卡品癫痫模型中,运用FJC染色技术在新大脑皮质中显示发生了大量神经元变性,有利于更好地理解颞叶癫痫的长期病理变化和自发反复发作的癫痫机制.%Objective To detect degenerative neurons that may occur in the neocortex of pilocarpine - treated mice by Fluoro -Jade C (FJC). Methods Ten male adult mice (Kunming strain) were divided into pilocarpine - treated group (n =5) and control goup (n=5). The control mice were injected intraperitoneally with atropine and saline. The pilocarpine - treated mice were injected intraperitoneally with atropine and pilocarpine to induce status epilepticus ( SE). Pilocarpine - treated mice were sacrificed at 12 h after SE. The control mice were also sacrificed at the same time. Brain coronal sections in the neocortex levels were made and stained by FJC staining. Finally, FJC - stained brain sections were observed under an epifluorescence microscope to identify the shape and distribution of the FJC positive cells in the neocortex. Results In pilocarpine - treated group, the FJC - positive staining cells showed a bright green color in the somas and fine processes with neuronal profiles. Numerous degenerative cells could

  6. Regulation of the spatial code for BDNF mRNA isoforms in the rat hippocampus following pilocarpine-treatment: a systematic analysis using laser microdissection and quantitative real-time PCR.

    Science.gov (United States)

    Baj, Gabriele; Del Turco, Domenico; Schlaudraff, Jessica; Torelli, Lucio; Deller, Thomas; Tongiorgi, Enrico

    2013-05-01

    Brain-derived neurotrophic factor (BDNF) is essential for neuronal survival, differentiation, and plasticity and is one of those genes that generate multiple mRNAs with different alternatively spliced 5'UTRs. The functional significance of many BDNF transcripts, each producing the same protein, is emerging. On the basis of the analysis of the four most abundant brain BDNF transcripts, we recently proposed the "spatial code hypothesis of BDNF splice variants" according to which the BDNF transcripts, through their differential subcellular localization in soma or dendrites, represent a mechanism to synthesize the protein at distinct locations and produce local effects. In this study, using laser microdissection of hippocampal laminae and reverse transcription-quantitative real-time PCR (RT-qPCR), we analyzed all known BDNF mRNA variants at resting conditions or following 3 h pilocarpine-induced status epilepticus. In untreated rats, we found dendritic enrichment of BDNF transcripts encoding exons 6 and 7 in CA1; exons 1, 6, and 9a in CA3; and exons 5, 6, 7, and 8 in DG. Considering the low abundance of the other transcripts, exon 6 was the main transcript in dendrites under resting conditions. Pilocarpine treatment induced an increase of BDNF transcripts encoding exons 4 and 6 in all dendritic laminae and, additionally, of exon 2 in CA1 stratum radiatum and exons 2, 3, 9a in DG molecular layer while the other transcripts were decreased in dendrites, suggesting restriction to the soma. These results support the hypothesis of a spatial code to differentially regulate BDNF in the somatic or dendritic compartment under conditions of pilocarpine-induced status epilepticus and, furthermore, highlight the existence of subfield-specific differences.

  7. PTEN在氯化锂-匹罗卡品癫痫模型中的表达研究%Expression of PTEN in the lithium -pilocarpine model of epilepsy

    Institute of Scientific and Technical Information of China (English)

    吕耀东; 王学峰

    2013-01-01

    目的:检测第10号染色体同源丢失性磷酸酶张力蛋白基因(phosphatase and tensin homolog deleted on chromosome ten ,PTEN)在癫痫模型大鼠中的表达情况,初步探讨其在癫痫发病中的作用。方法:成年雄性SD 大鼠腹腔注射氯化锂-匹罗卡品(lithium -pilocarpine )构建癫痫模型,分别在癫痫发作后不同时间点(1 d、3 d、7 d、14 d、30 d)提取海马组织,利用荧光定量PCR和western blot分别测定PTEN mRNA和蛋白质的表达。结果:荧光定量PCR和western blot显示,PTEN mRNA 和蛋白质在癫痫发作后1 d的表达显著降低(P<0.05),到30 d 时仍维持在较低的水平(P<0.05)。结论:PTEN在氯化锂-匹罗卡品致痫大鼠海马组织中表达的降低,提示PTEN可能参与了癫痫的发生发展过程。%Objective :To investigate the expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN) in a rat model of epilepsy and its potential role in epilepsy .Methods Adult male SD rats were intraperitoneally injected with lithium -pilocarpine for the epilepsy model .And at different time points (1 d、3 d、7 d、14 d、30 d) after seizures ,the expression of PTEN at mRNA and protein was detected in the hippocampus using real -time PCR and western blot respectively .Results :The expression of PTEN at mRNA and protein was significantly decreased at 1 d (P<0 .05) ,and remained at a lower level by 30 d (P<0 .05) after pilocarpine -induced seizures .Conclusion The expression of PTEN was markedly decreased in the hippocampus in the rat lithium -pilocarpine model of epilepsy ,indicating that PTEN may be involved in epileptogenesis .

  8. Biodegradable in situ gelling delivery systems containing pilocarpine as new antiglaucoma formulations: effect of a mercaptoacetic acid/N-isopropylacrylamide molar ratio

    Directory of Open Access Journals (Sweden)

    Lai JY

    2013-10-01

    Full Text Available Jui-Yang Lai Institute of Biochemical and Biomedical Engineering, Biomedical Engineering Research Center, Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan Abstract: Ocular drug delivery is one of the most commonly used treatment modalities in the management of glaucoma. We have recently proposed the use of gelatin and poly(N-isopropylacrylamide (PNIPAAm graft copolymers as biodegradable in situ forming delivery systems for the intracameral administration of antiglaucoma medications. In this study, we further investigated the influence of carrier characteristics on drug delivery performance. The carboxyl-terminated PNIPAAm samples with different molecular weights were synthesized by varying the molar ratio of mercaptoacetic acid (MAA/N-isopropylacrylamide (NIPAAm from 0.05 to 1.25, and were determined by end-group titration. The preparation of gelatin-g-PNIPAAm (GN copolymers from these thermoresponsive polymers was achieved using carbodiimide chemistry. Our results showed that the carboxylic end-capped PNIPAAm of high molecular weight may lead to the lower thermal phase transition temperature and slower degradation rate of GN vehicles than its low molecular weight counterparts. With a decreasing MAA/NIPAAm molar ratio, the drug encapsulation efficiency of copolymers was increased due to fast temperature-triggered capture of pilocarpine nitrate. The degradation of the gelatin network could greatly affect the drug release profiles. All of the GN copolymeric carriers demonstrated good corneal endothelial cell and tissue compatibility. It is concluded that different types of GN-based delivery systems exhibit noticeably distinct intraocular pressure-lowering effect and miosis action, thereby reflecting the potential value of a MAA/NIPAAm molar ratio in the development of new antiglaucoma formulations. Keywords: gelatin, poly(N-isopropylacrylamide, glaucoma, chain transfer agent, ocular drug delivery

  9. Berberine exerts an anticonvulsant effect and ameliorates memory impairment and oxidative stress in a pilocarpine-induced epilepsy model in the rat.

    Science.gov (United States)

    Gao, Fei; Gao, Ying; Liu, Yang-Feng; Wang, Li; Li, Ya-Jun

    2014-01-01

    Though new antiepileptic drugs are emerging, approximately a third of epileptic patients still suffer from recurrent convulsions and cognitive dysfunction. Therefore, we tested whether berberine (Ber), a vegetable drug, has an anticonvulsant property and attenuates memory impairment in a pilocarpine (Pilo)-induced epilepsy model in rats. The rats were injected with 400 mg/kg Pilo to induce convulsions, and Ber 25, 50, and 100 mg/kg were administrated by the intragastric route once daily 7 days before Pilo injection until the experiment was over. Convulsions were observed after Pilo injection. For the rats that developed status epilepticus (SE), malondialdehyde, glutathione levels, superoxide dismutase, and catalase activity in the hippocampus were measured 24 hours after SE. The rats received the Morris water-maze test 2 weeks after SE, and then were killed for fluoro-jade B staining to detect the degenerating neurons. We found Ber delayed latency to the first seizure and the time to develop SE in a dose-dependent manner. Malondialdehyde levels were decreased, while glutathione and catalase activity were strengthened in Ber-injected SE rats. In the Morris water-maze test, Ber decreased escape latency compared to saline-treated SE rats. Additionally, Ber reduced the number of fluoro-jade B-positive cells in the hippocampal CA1 region. Our data suggest that Ber exerts anticonvulsant and neuroprotective effects on Pilo-induced epilepsy in rats. Simultaneously, Ber attenuates memory impairment. The beneficial effect may be partly due to mitigation of the oxidative stress burden.

  10. Identification of endogenous reference genes for the analysis of microRNA expression in the hippocampus of the pilocarpine-induced model of mesial temporal lobe epilepsy.

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    Mykaella Andrade de Araújo

    Full Text Available Real-time quantitative RT-PCR (qPCR is one of the most powerful techniques for analyzing miRNA expression because of its sensitivity and specificity. However, in this type of analysis, a suitable normalizer is required to ensure that gene expression is unaffected by the experimental condition. To the best of our knowledge, there are no reported studies that performed a detailed identification and validation of suitable reference genes for miRNA qPCR during the epileptogenic process. Here, using a pilocarpine (PILO model of mesial temporal lobe epilepsy (MTLE, we investigated five potential reference genes, performing a stability expression analysis using geNorm and NormFinder softwares. As a validation strategy, we used each one of the candidate reference genes to measure PILO-induced changes in microRNA-146a levels, a gene whose expression pattern variation in the PILO injected model is known. Our results indicated U6SnRNA and SnoRNA as the most stable candidate reference genes. By geNorm analysis, the normalization factor should preferably contain at least two of the best candidate reference genes (snoRNA and U6SnRNA. In fact, when normalized using the best combination of reference genes, microRNA-146a transcripts were found to be significantly increased in chronic stage, which is consistent with the pattern reported in different models. Conversely, when reference genes were individually employed for normalization, we failed to detect up-regulation of the microRNA-146a gene in the hippocampus of epileptic rats. The data presented here support that the combination of snoRNA and U6SnRNA was the minimum necessary for an accurate normalization of gene expression at the different stages of epileptogenesis that we tested.

  11. Progressive metabolic changes underlying the chronic reorganization of brain circuits during the silent phase of the lithium-pilocarpine model of epilepsy in the immature and adult Rat.

    Science.gov (United States)

    Dubé, C; Boyet, S; Marescaux, C; Nehlig, A

    2000-03-01

    The lithium-pilocarpine (Li-Pilo) model of epilepsy reproduces most of the features of human temporal lobe epilepsy. In the present study, we explored the correlation between metabolic changes, neuronal damage, and epileptogenesis during the silent phase following status epilepticus (SE) induced by Li-Pilo in 10- (P10) and 21-day-old (P21) and adult rats. Cerebral metabolic rates for glucose (CMR(glcs)) were measured at 14 and 60 days after SE by the 2-[(14)C]deoxyglucose method and neurodegeneration was assessed by the silver staining and cresyl violet techniques. In P10 rats, there was no damage and no metabolic consequences at any time after SE. In P21 rats, metabolic decreases were recorded at 14 days after SE, mainly in damaged forebrain regions. Conversely at 60 days after SE, P21 rats exhibited metabolic increases in both forebrain-damaged and brain-stem-intact areas. Finally, in adult rats studied at 14 days after SE, CMR(glcs) decreased in damaged forebrain areas involved in the circuitry of spontaneous seizures and increased in nondamaged brain-stem areas involved in the remote control of epilepsy. The increase in CMR(glcs) in damaged forebrain areas of P21 rats at 60 days after SE may reflect the genesis of a new circuitry underlying the occurrence of spontaneous seizures. The metabolic increase recorded in nondamaged brain-stem areas of P21 and adult rats occurs in regions involved in the remote control of seizures and might underlie a process of protection against the occurrence of seizures. Copyright 2000 Academic Press.

  12. Quantitative longitudinal imaging of activated microglia as a marker of inflammation in the pilocarpine rat model of epilepsy using [(11)C]-( R)-PK11195 PET and MRI.

    Science.gov (United States)

    Yankam Njiwa, J; Costes, N; Bouillot, C; Bouvard, S; Fieux, S; Becker, G; Levigoureux, E; Kocevar, G; Stamile, C; Langlois, J B; Bolbos, R; Bonnet, C; Bezin, L; Zimmer, L; Hammers, A

    2017-04-01

    Inflammation may play a role in the development of epilepsy after brain insults. [(11)C]-( R)-PK11195 binds to TSPO, expressed by activated microglia. We quantified [(11)C]-( R)-PK11195 binding during epileptogenesis after pilocarpine-induced status epilepticus (SE), a model of temporal lobe epilepsy. Nine male rats were studied thrice (D0-1, D0 + 6, D0 + 35, D0 = SE induction). In the same session, 7T T2-weighted images and DTI for mean diffusivity (MD) and fractional anisotropy (FA) maps were acquired, followed by dynamic PET/CT. On D0 + 35, femoral arterial blood was sampled for rat-specific metabolite-corrected arterial plasma input functions (AIFs). In multiple MR-derived ROIs, we assessed four kinetic models (two with AIFs; two using a reference region), standard uptake values (SUVs), and a model with a mean AIF. All models showed large (up to two-fold) and significant TSPO binding increases in regions expected to be affected, and comparatively little change in the brainstem, at D0 + 6. Some individuals showed increases at D0 + 35. AIF models yielded more consistent increases at D0 + 6. FA values were decreased at D0 + 6 and had recovered by D0 + 35. MD was increased at D0 + 6 and more so at D0 + 35. [(11)C]-( R)-PK11195 PET binding and MR biomarker changes could be detected with only nine rats, highlighting the potential of longitudinal imaging studies.

  13. Differential paired-pulse responses between the CA1 region and the dentate gyrus are related to altered CLC-2 immunoreactivity in the pilocarpine-induced rat epilepsy model.

    Science.gov (United States)

    Kwak, Sung-Eun; Kim, Ji-Eun; Kim, Duk-Soo; Won, Moo Ho; Lee, Hong Jin; Choi, Soo-Young; Kwon, Oh-Shin; Kim, Jin-Sang; Kang, Tae-Cheon

    2006-10-18

    The epileptic hippocampus shows differential paired-pulse responses between the dentate gyrus and the CA1 region. However, little data are available to explain this phenomenon. In the present study, we identified the relationship between regional differences of paired-pulse response and voltage gated Cl(-) channel 2 (CLC-2)/vesicular GABA transport (VGAT) expression in a pilocarpine-induced rat model. During epileptogenic periods, paired-pulse inhibitions in the dentate gyrus and the CA1 region were markedly reduced. After recurrent seizure onset, paired-pulse inhibition in the dentate gyrus was markedly enhanced, while that in the CA1 region more reduced. Unlike VGAT, CLC-2 immunoreactivity was markedly reduced in the hippocampus during epileptogenic periods and was re-enhanced only in the dentate gyrus after recurrent seizure onset. Linear regression analysis showed an inverse proportional relationship between alterations in CLC-2 immunoreactivity and changes in normalized population spike amplitude ratio within the CA1 region and the dentate gyrus. Therefore, our findings suggest that the regionally specific alterations in CLC-2 immunoreactivity after SE may determine the properties of paired-pulse responses in the hippocampus of the pilocarpine-induced rat epilepsy model.

  14. Scorpion ethanol extract and valproic acid effects on hippocampal glial fibrillary acidic protein expression in a rat model of chronic-kindling epilepsy induced by lithium chloride-pilocarpine

    Institute of Scientific and Technical Information of China (English)

    Yi Liang; Hongbin Sun; Liang Yu; Baoming He; Yan Xie

    2012-01-01

    The present study analyzed the effects of ethanol extracts of scorpion on epilepsy prevention and hippocampal expression of glial fibrillary acidic protein in a lithium chloride-pilocarpine epileptic rat model. Results were subsequently compared with valproic acid. Results showed gradually-increased hippocampal glial fibrillary acidic protein expression following model establishment; glial fibrillary acidic protein mRNA expression was significantly increased at 3 days, reached a peak at 7 days, and then gradually decreased thereafter. Ethanol extracts of scorpion doses of 580 and 1 160 mg/kg, as well as 120 mg/kg valproic acid, led to a decreased number of glial fibrillary acidic protein-positive cells and glial fibrillary acidic protein mRNA expression, as well as decreased seizure grades and frequency of spontaneously recurrent seizures. The effects of 1 160 mg/kg ethanol extracts of scorpion were equal to those of 120 mg/kg valproic acid. These results suggested that the anti-epileptic effect of ethanol extracts of scorpion were associated with decreased hippocampal glial fibrillary acidic protein expression in a rat model of lithium chloride-pilocarpine induced epilepsy.

  15. Relationship between neuronal loss and interictal glucose metabolism during the chronic phase of the lithium-pilocarpine model of epilepsy in the immature and adult rat.

    Science.gov (United States)

    Dubé, C; Boyet, S; Marescaux, C; Nehlig, A

    2001-02-01

    The lithium-pilocarpine (Li-Pilo) model of epilepsy reproduces most of the features of human temporal lobe epilepsy. After having studied the metabolic changes occurring during the silent phase, in the present study, we explored the relationship between interictal metabolic changes and neuronal loss during the chronic phase following status epilepticus (SE) induced by Li-Pilo in 10-day-old (P10), 21-day-old (P21), and adult rats. Rats were observed and their EEG was recorded to detect the occurrence of spontaneous recurrent seizures (SRS). Local cerebral glucose utilization was measured during the interictal period of the chronic phase, between 2 and 7 months after SE, by the [(14)C]2-deoxyglucose method in rats subjected to SE at P10, P21, or as adults. Neuronal damage was assessed by cell counting on adjacent cresyl violet stained sections. When SE was induced at P10, rats did not become epileptic, did not develop lesions and cerebral glucose utilization was in the normal range 7 months later. When SE was induced in adult rats, they all became epileptic after a mean duration of 25 days and developed lesions in the forebrain limbic areas, which were hypometabolic during the interictal period of the chronic phase, 2 months after SE. When SE was induced in P21 rats, 24% developed SRS, and in 43% seizures could be triggered (TS) by handling, after a mean delay of 74 days in both cases. The remaining 33% did not become epileptic (NS). The three groups of P21 rats developed quite comparable lesions mainly in the hilus of the dentate gyrus, lateral thalamus, and entorhinal cortex; at 6 months after SE, the forebrain was hypometabolic in NS and TS rats while it was normo- to slightly hypermetabolic in SRS rats. These data show that interictal metabolic changes are age-dependent. Moreover, there is no obvious correlation, in this model, between interictal hypometabolism and neuronal loss, as reported previously in human temporal lobe epilepsy. Copyright 2000 Academic

  16. Acupuncture-Like Transcutaneous Electrical Nerve Stimulation Versus Pilocarpine in Treating Radiation-Induced Xerostomia: Results of RTOG 0537 Phase 3 Study

    Energy Technology Data Exchange (ETDEWEB)

    Wong, Raimond K.W., E-mail: wongrai@hhsc.ca [McMaster University, Juravinski Cancer Centre, Hamilton, Ontario (Canada); Deshmukh, Snehal [NRG Oncology Statistics and Data Management Center, Philadelphia, Pennsylvania (United States); Wyatt, Gwen [Michigan State University, East Lansing, Michigan (United States); Sagar, Stephen [McMaster University, Juravinski Cancer Centre, Hamilton, Ontario (Canada); Singh, Anurag K. [Roswell Park Cancer Institute, Buffalo, New York (United States); Sultanem, Khalil [McGill University, Montreal, Quebec (Canada); Nguyen-Tân, Phuc F. [Centre Hospitalier de l' Université de Montréal-Hôpital Notre-Dame, Montreal, Quebec (Canada); Yom, Sue S. [University of California San Francisco, San Francisco, California (United States); Cardinale, Joseph [Yale-New Haven Hospital Saint Raphael Campus, New Haven, Connecticut (United States); Yao, Min [University Hospitals of Cleveland, Cleveland, Ohio (United States); Hodson, Ian [McMaster University, Juravinski Cancer Centre, Hamilton, Ontario (Canada); Matthiesen, Chance L. [University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma (United States); Suh, John [Cleveland Clinic Foundation, Cleveland, Ohio (United States); Thakrar, Harish [John H. Stroger, Jr. Hospital of Cook County MB-CCOP, Chicago, Illinois (United States); Pugh, Stephanie L. [NRG Oncology Statistics and Data Management Center, Philadelphia, Pennsylvania (United States); Berk, Lawrence [University of South Florida H. Lee Moffitt Cancer Center, Tampa, Florida (United States)

    2015-06-01

    Purpose and Objectives: This report presents the analysis of the RTOG 0537 multicenter randomized study that compared acupuncture-like transcutaneous stimulation (ALTENS) with pilocarpine (PC) for relieving radiation-induced xerostomia. Methods and Materials: Eligible patients were randomized to twice-weekly 20-minute ALTENS sessions for 24 sessions during 12 weeks or PC (5 mg 3 times daily for 12 weeks). The primary endpoint was the change in the University of Michigan Xerostomia-Related Quality of Life Scale (XeQOLS) scores from baseline to 9 months from randomization (MFR). Secondary endpoints included basal and citric acid primed whole salivary production (WSP), ratios of positive responders (defined as patients with ≥20% reduction in overall radiation-induced xerostomia symptom burden), and the presence of adverse events based on the Common Terminology Criteria for Adverse Events version 3. An intention-to-treat analysis was conducted. Results: One hundred forty-eight patients were randomized. Only 96 patients completed the required XeQOLS and were evaluable at 9 MFR (representing merely 68.6% statistical power). Seventy-six patients were evaluable at 15 MFR. The median change in the overall XeQOLS in ALTENS and PC groups at 9 and 15 MFR were −0.53 and −0.27 (P=.45) and −0.6 and −0.47 (P=.21). The corresponding percentages of positive responders were 81% and 72% (P=.34) and 83% and 63% (P=.04). Changes in WSP were not significantly different between the groups. Grade 3 or less adverse events, mostly consisting of grade 1, developed in 20.8% of patients in the ALTENS group and in 61.6% of the PC group. Conclusions: The observed effect size was smaller than hypothesized, and statistical power was limited because only 96 of the recruited 148 patients were evaluable. The primary endpoint—the change in radiation-induced xerostomia symptom burden at 9 MFR—was not significantly different between the ALTENS and PC groups. There was significantly less

  17. Terapia combinada con timolol/dorzolamida versus timolol/pilocarpina en el glaucoma primario de ángulo abierto Combined therapy with timolol and dorzolamide vs timolol and pilocarpine used in primary open-angle glaucoma

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    Frank García González

    2006-06-01

    Full Text Available El propósito de este trabajo fue evaluar la eficacia de la terapia combinada, timolol/dorzolamida, en comparación con timolol/pilocarpina. Se empleó tratamiento médico en 38 pacientes con glaucoma primario de ángulo abierto, a los que se les colocó en forma aleatoria timolol 0,5 %/dorzolamida 2 % (n. 19 o timolol 0,5 % y pilocarpina 2 % (n. 19 y posteriormente se analizaron los descensos de presión intraocular, efectividad durante cuatro semanas, efectos adversos locales y sistémicos. En el grupo de pacientes tratados con timolol/dorzolamida la presión intraocular media inicial (sin tratamiento descendió de 22,84 ± 1,77 mm Hg hasta 18,24 ± 1,84 mm Hg a las cuatro semanas de tratamiento, p. 0,01 (reducción de 4,60 mm Hg 22,14 %. En el grupo de pacientes tratados con timolol/pilocarpina la presión intraocular media inicial (sin tratamiento descendió de 23, 06 ± 1,29 mm Hg hasta 19,07 ± 1,23 mm Hg a la cuarta semana de tratamiento, p. 0,01 (reducción de 3,95 mm Hg /17,31 %, no se observaron diferencias significativas (p > 0,05 entre ambos tratamientos y fueron igualmente eficaces para reducir la presión intraocular. La calidad de vida de los pacientes que recibieron la dorzolamida como tratamiento coadyuvante fue superior, la dosificación disminuyó con respecto a la pilocarpina y no se presentaron efecto secundarios, tales como limitaciones para conducir y leer o dolor ocular, aunque refirieron sabor amargo cinco pacientes (26,31 % e irritación conjuntival dos pacientes (10,52 % relacionados con la dorzolamida. A mediano plazo se obtiene disminución de presión intraocular con dorzolamida como con la pilocarpina combinadas con el timolol. La dorzolamida demostró menos interferencia con la calidad de vida que la pilocarpinaThe objective of this study was to evaluate the efficacy of combined therapy with timolol and dorzolamide compared to timolol and pilocarpine. Thirty eight patients with primary open-angle glaucoma were

  18. Application du carisbamate comme agent neuroprotecteur et modificateur de l’épileptogénèse dans le modèle Lithium-Pilocarpine : évaluation de l’expression protéique et des altérations neurochimiques cérébrales

    OpenAIRE

    Marques Carneiro Da Silva, Jose Eduardo

    2015-01-01

    The carisbamate is the 1st molecule showing an epileptogenesis modifying effect, that about 50% of treated animals develops absence instead limbic seizures, commonly seen in the pilocarpine model. The aim of this thesis was to study the changes that follows carisbamate treatment. Therefore, we made a brain activity cartography, by labelling c-Fos protein, and we quantified the concentrations of amino acids and monoamines in hippocampus, thalamus and piriform cortex, 4h after the status epilep...

  19. Vigabatrina aumenta atividade da superóxido dismutase no corpo estriado de ratos após crises convulsivas induzidas pela pilocarpina Vigabratine increases superoxide dismutase activity in striatum of rat after pilocarpine-induced seizures

    Directory of Open Access Journals (Sweden)

    Rivelilson Mendes de Freitas

    2010-01-01

    ES: Os resultados do presente estudo indicam que durante as crises convulsivas ocorrem alterações comportamentais, entretanto, não foram verificadas mudanças na atividade da SOD durante a fase aguda dessas crises. Esses dados sugerem que os efeitos anticonvulsivantes da vigabatrina podem ser decorrentes da neuromodulação da SOD. No entanto, serão realizados novos estudos neurofarmacológicos para o esclarecimento do mecanismo de ação da vigabatrina no modelo de epilepsia induzido pela pilocarpina.BACKGROUND: Pilocarpine-induced seizures have been suggested to be mediated by increases in oxidative stress. Current studies have suggested that antioxidant compounds may afford some level of neuroprotection against the neurotoxicity of seizures. OBJECTIVES: This study investigated the pharmacological actions of vigabatrin on behavioral changes and superoxide dismutase (SOD activity in striatum of adult rats. METHODS: Adult rats (2 months old were used in the experiments and divided into four groups. The first was treated with 0.9% saline (control group. The second group was treated with pilocarpine (400 mg/kg, i.p., P400 group. The third group received vigabatrin alone (500 mg/kg, i.p., VGB group and the fourth group was treated with vigabatrin (500 mg/kg, i.p. and 30 minutes later received pilocarpine (400 mg/kg, i.p., VGB + P400 group. The animals which had seizures and status epilepticus (SE and did not die within 24 hours of observation were sacrificed to perform the neurochemical studies. RESULTS: Behavioral studies showed that the administration of pilocarpine produces peripheral cholinergic signs, tremors and stereotyped movements in all animals. An amount of 75% of those rats developed to seizures and SE. In turn, the pre-treatment with vigabatrin produced a 50% reduction in the rate of seizures and SE. Regarding the neurochemical studies, there were no changes in the striatal SOD activity in P400 group as compared to the control group. However, in the VGB + P

  20. 莲心碱对氯化锂-匹鲁卡品致癫模型急性期皮层脑电图的影响%Effect of liensinine on electrocorticogram in lithium-pilocarpine induced acute epilepsy rats

    Institute of Scientific and Technical Information of China (English)

    吴靖; 周宏斌; 潘松青

    2016-01-01

    目的:探讨莲心碱对氯化锂—匹鲁卡品致癫大鼠模型急性期皮层脑电图的影响。方法32只 SD 大鼠随机分为生理盐水对照组(A 组,10μg)、低剂量莲心碱组(B 组,莲心碱2.5 g·L -1,25μg)、高剂量莲心碱组(C 组,莲心碱5 g·L -1,50μg)、左乙拉西坦组(D 组,100 g·L -1,1 mg),建立氯化锂—匹鲁卡品癫痫大鼠模型后侧脑室注射药物,记录各组大鼠脑电图的改变。结果莲心碱组及左乙拉西坦组与对照组相比大鼠的痫性放电频率减低,快波(β波)比率减少,慢波(δ波)比率增加;高剂量莲心碱组和左乙拉西坦组相比无统计学差异。结论莲心碱在氯化锂—匹鲁卡品癫痫大鼠模型急性期具有抗癫痫作用。%Objective To explore the effect of liensinine on electrocorticogram in lithium -pilocarpine induced acute epilepsy rats. Methods Thirty -two SD rats were randomized into four groups (8 rats in each group):epilepsy model group(group A,10 μg),low dose of liensinine group(group B,2.5 g·L -1 ,25 μg),high dose of liensinine group(group C,5 g·L -1 ,50 μg),levetiracetam group (group D,100 g·L -1 ,1 mg).Electrocorticogram of rats was recorded after lithium -pilocarpine model was induced and the drug was intra -cerebroventricular injected.Results Compared with group A,the epileptic discharge was significantly decreased in groups B,C and D (P <0.05).The proportion of δslow wave was enhanced,meanwhile βfast wave was decreased in groups B,C and D.And there was no significant difference between high dose of liensinine group and levetiracetam group.Conclusions Liensinine has the role of anti -epilepsy in acute model of epileptic rats induced by lithium -pilocarpine.

  1. Fluoro-Jade C can stain the degenerative neurons in the piriform cortex of pilocarpine-treated mice%Fluoro-Jade C揭示匹罗卡品模型中梨状皮质神经元变性

    Institute of Scientific and Technical Information of China (English)

    王莲; 魏玲; 付莉

    2012-01-01

    目的 应用Fluoro-Jade C(FJC)染色方法 在小鼠匹罗卡品癫痫模型中检测梨状皮质结构中神经元的变性情况,以了解梨状皮质结构在慢性颞叶癫痫发生中的病理变化和癫痫反复发作的神经基础.方法 雄性昆明小鼠10只(对照组5只,匹罗卡品处理组5只).处理组在癫痫持续状态后3d处死处理组小鼠.在梨状皮质水平切制冠状切片,行FJC染色,在荧光显微镜下观察FJC阳性细胞的形态和在梨状皮质中的整体分布情况.结果 在处理组,FJC染色的脑切片上可以很清楚地看到呈亮黄绿色荧光的FJC阳性细胞,呈神经元形态,胞体和突起均清晰显示.在梨状皮质和梨状内核内出现大量FJC阳性细胞,而对照组未见.结论 在小鼠匹罗卡品癫痫模型中运用FJC染色技术显示梨状皮质内发生了大量神经元变性,此研究有利于更好地理解颞叶癫痫中中枢神经系统所发生的长期病理变化和自发反复发作的癫痫机制.%Objective To detect degenerative neurons by Fluoro-Jade C ( FJC ) that my' occur in the piriform cortex of pilocarpine-treated mice and to understand the neural basis of piriform cortex in long-term brain pathological changes and recurrent seizure mechanism of chronic epilepsy. Methods Male adult mice ( Kunming stain, n = 10 ) were divided into pilocarpine-treated group ( n = 5 ) and controls ( n = 5 ). Pilocarpine-treated mice were sacrificed at 3 d after status epilepticus. Brain coronal sections in the piriform cortex levels were subject to FJC staining. Finally, FJC-stained brain sections were examined under an epifluorescence microscope to identify the shape and overall distribution of the FJC positive cells in the piriform cortex. Results The FJC-positive staining cells showed a bright green color in the somas and fine processes with neuronal profiles. Numerous degenerating cells could be seen in the piriform cortex including the endopirifonn nucleus. They were not detected

  2. 毛果芸香碱刺激与非刺激条件下家兔体液中氯胺酮含量分析%STUDY ON THE CONTENT OF KETAMINE IN RABBITS BODY FLUID UNDER THE CONDITION OF PILOCARPINE STIMULUS AND NON STIMULUS

    Institute of Scientific and Technical Information of China (English)

    钱玮; 李鹏旺; 刘俊芳; 王玉瑾

    2012-01-01

    Objective: Compared the content of ketamine and norketamine in rabbits body fluid between the pilocarpine stimulus group and non stimulus group. Methods: The acute toxic rabbits were divided into two groups, one group was stimulated by pilocarpine and another wasn' t stimulated. Collected their saliva, plasma and urine at different time points after administration. The contents of ketamine and norketamine were qualitatively analyzed by GC/MS and quantitatively analyzed by GC. Compared the contents of ketamine and norketamine between two groups by repeated measure variance analysis. Results;There was no significant difference between pilocarpine stimulus group and non stimulus group (P>0.05). Conclusion:The methods of pilocarpine stimulus and non stimulus can be applied to collecting saliva for detecting ketamine. Pilocarpine stimulus method can increase salivary secretion, collection frequency and collection quantity.%目的:比较研究毛果芸香碱刺激和非刺激条件下家兔唾液、血液和尿液中氯胺酮及代谢物去甲氯胺酮的含量.方法:实验家兔分为毛果芸香碱刺激组和非刺激组,分别于氯胺酮染毒前和染毒后不同时间点收集唾液、血液和尿液.气相色谱/质谱联用(GC/MS)全扫描定性、气相色谱(GC)定量分析血液、唾液和尿液样品中氯胺酮及去甲氯胺酮的浓度.采用两因素重复测量方差分析方法分别比较刺激组和非刺激组之间氯胺酮及去甲氯胺酮在唾液、血液和尿液中平均检出量有无统计学意义.结果:毛果芸香碱刺激组和非刺激组之间氯胺酮及去甲氯胺酮在唾液、血液和尿液中平均检出量均无显著性差异(P>0.05).结论:唾液中氯胺酮的采集可用毛果芸香碱刺激或非刺激法,但刺激法可以增加唾液分泌,易采集且增加采集次数和采集量.

  3. 不同首剂匹罗卡品制作颞叶癫痫大鼠模型的研究%Using Different Initial Doses of Pilocarpine to Establish the Temporal Lobe Epilepsy Model

    Institute of Scientific and Technical Information of China (English)

    宋延民; 杨国帅; 龙莉莉; 洪秀琴; 邓景贵

    2012-01-01

    目的:研究不同首次剂量匹罗卡品对氯化锂-匹罗卡品颞叶癫痫大鼠模型诱发成功率、死亡率的影响.方法:90只SD大鼠随机分为三组,每组首次使用匹罗卡品剂量:A组10mg/kg、B组20mg/kg、C组30mg/kg;随后采用多次注射10mg/kg匹罗卡品,比较3组癫痫持续状态诱发成功率、死亡率的差异.结果:A、B、C组癫痫持续状态诱发成功率分别为66.7%、90%、93.3%.其中A组与B组之间诱发成功率差异有统计学意义,P< 0.05;C组与B组之间相比,差异无统计学意义,P>0.05.三组死亡率分别为20%、23.3%、57.1%,其中A组与B组之间差异无统计学差异,P>0.05,;C组与其它两组相比较差异有统计学差异,P<0.05.结论:首次剂量20mg匹罗卡品,然后10mg多次反复注射,癫痫持续状态诱发成功率高、死亡率低,是一种理想的颞叶癫痫模型.%Objective:We use different initial doses of pilocatpine after Lithium injections to establish the temporal lobe epilepsy models, observe the effects of different initial dose of pilocarpine on percentage of rats being induced successfully and the mortality rate. Me

  4. Lovastatin reduces neuronal cell death in hippocampal CA1 subfield after pilocarpine-induced status epilepticus: preliminary results Lovastatina reduz a lesão celular na região CA1 do hipocampo após o status epilepticus induzido pela pilocarpina: resultados preliminares

    Directory of Open Access Journals (Sweden)

    Pauline Rangel

    2005-12-01

    Full Text Available OBJECTIVE: To further characterize the capacity of lovastatin to prevent hippocampal neuronal loss after pilocarpine-induced status epilepticus (SE METHOD: Adult male Wistar rats were divided into four groups: (A control rats, received neither pilocarpine nor lovastatin (n=5; (B control rats, received just lovastatin (n=5; (C rats that received just pilocarpine (n=5; (D rats that received pilocarpine and lovastatin (n=5. After pilocarpine injection (350mg/kg, i.p., only rats that displayed continuous, convulsive seizure activity were included in our study. Seizure activity was monitored behaviorally and terminated with an injection of diazepam (10 mg/kg, i.p. after 4 h of convulsive SE. The rats treated with lovastatin received two doses of 20mg/kg via an oesophagic probe immediately and 24 hours after SE induction. Seven days after pilocarpine-induced SE, all the animals were perfused and their brains were processed for histological analysis through Nissl method. RESULTS: The cell counts in the Nissl-stained sections performed within the hippocampal formation showed a significant cell loss in rats that received pilocarpine and presented SE (CA1= 26.8 ± 13.67; CA3= 38.1 ± 7.2; hilus= 43.8 ± 3.95 when compared with control group animals (Group A: CA1= 53.2 ± 9.63; CA3= 63.5 ± 13.35; hilus= 59.08 ± 10.24; Group B: CA1= 74.3 ± 8.16; CA3= 70.1 ± 3.83; hilus= 70.6 ± 5.10. The average neuronal cell number of CA1 subfield of rats that present SE and received lovastatin (44.4 ± 17.88 was statically significant increased when compared with animals that just presented SE. CONCLUSION: Lovastatin exert a neuroprotective role in the attenuation of brain damage after SE.OBJETIVO: Capacidade da lovastatina em prevenir a perda de neurônios hipocampais após o status epilepticus (SE induzido pela pilocarpina. MÉTODO: Ratos adultos Wistar foram divididos em 4 grupos: (A ratos controles que não receberam pilocarpina nem lovastatina (n=5; (B ratos

  5. Ações neuroprotetoras da vitamina C no corpo estriado de ratos após convulsões induzidas pela pilocarpina Neuroprotective actions of vitamin C in rat striatum after pilocarpine-induced seizures

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    Rivelilson Mendes de Freitas

    2010-01-01

    especially in limbic structures. OBJECTIVES: This study aimed to evaluate the neuroprotective effects of vitamin C in the histopathological changes observed in rat striatum after seizures. MATERIAL AND METHODS: Healthy Wistar rats were divided into four groups. The first group was treated with 0.9% saline (control group and the second one with pilocarpine (400 mg/kg, P400 group. Third and fourth groups were treated with vitamin C (250 mg/kg, 30 minutes before receiving P400 (P400 + VIT C group or 0.9% saline (VIT C group, respectively. After the treatments, all groups were observed for 24 hours, sacrificed and dissected out to remove their brains for histopathological analysis. RESULTS: The group P400 presented seizures that progressed to status epilepticus in 75% of the animals. Pretreatment with vitamin C produced a 35% reduction in this index. P400 and P400 + VIT C groups revealed 80% and 20% of animals with brain injury, respectively. In P400 group, lesion severity of the striatum was 50%. In turn, in striatal region of animals treated with P400 + VIT C group, we detected a reduction of 40% in the severity degree. DISCUSSION: Pilocarpine-induced seizures are installed by the cholinergic system and propagated by free radicals and by glutamatergic system, leading to brain damage. The antioxidant drugs may have therapeutic potential for epileptic patients to protect against brain injure through removing free radicals produced, suggesting that vitamin C may influence epileptogenesis and promote neuroprotective actions during seizures.

  6. Alterações agudas dos níveis de neurotransmissores em corpo estriado de ratos jovens após estado epiléptico induzido por pilocarpina Acute alterations of neurotransmitters levels in striatum of young rat after pilocarpine-induced status epilepticus

    Directory of Open Access Journals (Sweden)

    Rivelilson Mendes de Freitas

    2003-06-01

    Full Text Available Altas doses de agonista colinérgico muscarínico, pilocarpina, produzem alterações comportamentais, convulsões e estado epiléptico em ratos. O objetivo desse estudo foi verificar as alterações nas concentrações dos neurotransmissores em corpo estriado de ratos em desenvolvimento após estado epiléptico induzido pela pilocarpina. Ratas Wistar foram tratadas com uma única dose de pilocarpina (400mg/Kg; s.c.. Controles receberam salina. A concentração dos neurotransmissores foi determinada através do HPLC, no corpo estriado de ratos que no período de observação de 1 e 24h desencadearam estado epiléptico e não sobreviveram à fase aguda do quadro convulsivo. Foi observada redução nos níveis de dopamina, serotonina, ácido dihidroxifenilacético, ácido 5-hidroxiindolacético, e aumento no ácido 4-hidroxi-3-metoxi-fenilacético. Os resultados mostraram que a ativação do sistema colinérgico pode interagir com os sistemas dopaminérgico e serotonérgico nos mecanismos referentes à fase aguda do processo convulsivo.High doses of the muscarinic cholinergic agonist, pilocarpine, result in behavioural changes, seizures and status epilepticus in rats. The purpose of the present work is to invetigate the striatal neurotransmissors level in young rats after status epilepticus induced by pilocarpine. Wistar rats were treated with a single dose of pilocarpine (400mg/Kg; s.c.. Controls received saline. Young animals were closed observed for behavioural changes during 1 and 24h. In these periods, the animals that developed status epilepticus and didn't survive this acute phase of seizures had the brains removed and striatal neurotransmissors level determined by HPLC. The concentration of dopamine, serotonine, dihydroxyphenylacetic acid, 5-hydroxyindolacetic acid was reduced and an increase in 4-hydroxy-3-methoxy-phenylacetic acid was observed. These results suggest that cholinergic activation can interage with dopaminergic and

  7. The observation of mossy fiber sprouting in hippocampus zone of rat eoilepsy models induced by pilocarpine%匹鲁卡品癫痫模型慢性期大鼠海马苔藓纤维出芽的观察

    Institute of Scientific and Technical Information of China (English)

    陈丽丽; 黄靓妹; 詹红艳; 曹亦宾

    2011-01-01

    Objective To investigate the onset of rots epilepsy after pilocarpine injection and mossy fiber sprouting in hippocampus zone. Methods 60 rats were randomly divided into two groups: epilepsy group and control group, 30 rats in each group. The neuronal damage, mossy fiber sprouting were detected by Nissl and neo-Timm staining. The ECG was performed in the day of experiment for all the rats. Results After the rats were injected with pilocarpine,93% of them developed epilepsy,and most of them showed spontaneous onset during the chronic period. Timm stainning results showed that there was a significant difference in the neuron deletion in epilepsy group between CA3 region and dentate hilus region, and between CA3 region and CA1 region ( P <0.05 ) ,and there was a significant difference in silver staining granules scoring of mossy fiber sprouting between epilepsy group and control group ( P < 0.05 ). Conclusion The epilepsy models can be induced by pilocarpine injection in rots,with basic pathological characteristics of human epilepsy, which is a useful and simple way for studying epilepsy.%目的 观察腹腔注射匹鲁卡品后大鼠痫性发作及海马苔藓出芽.方法 Wistra大鼠60只随机分为模型组和对照组,每组30只,均进行Nissl染色及Timm银染.并于实验当天行脑电图检查.结果 腹腔注射匹鲁卡品后大鼠可见急性癫痫发作,观察慢性期自发性发作.Nissl染色示模型组各区评分两两比较,结果 示CA3、门区与CA1区神经元缺失差异有统计学意义(P<0.05).Timm银染示2组苔藓纤维出芽Timm银染颗粒评分比较,差异有统计学意义(P<0.05).结论 腹腔注射匹鲁卡品简单复制了人类癫痫的基本病理特征,是研究癫痫的一种简便方法.

  8. 匹罗卡品致痫大鼠慢性期海马神经元突触重建的实验研究%Neuronal synaptic reconstruction in hippocampus in chronic phase of pilocarpine-treated rats

    Institute of Scientific and Technical Information of China (English)

    易芳; 穆塔森; 龙莉莉; 李艺; 李蜀渝; 吴志国; 肖波

    2011-01-01

    Objective To explore the aberrant formation of excitatory and inhibitory circuit rearrangements of hippoeampus in temporal lobe epilepsy.Methods Pilocarpine-induced animal model was established.At around Day 60 post-modeling,retrograde tracer fluorogold(FG)was injected in vivo into CA1 and CA3 areas of hippocampus by stereotaxic apparatus.Immunohistochemistry of FG was used to observe the aberrant excitatory circuit rearrangements.Double immunofluorescence with NPY(neuropeptide Y)and FG was performed to observe the aberrant inhibitory circuit rearrangements.Results After an iniection of FG into CA1 area.the FG-labeled pyramidal cells could be observed distantly from the zone of dye soread in CA1 area.CA3 area and subiculm.And the FG-labeled non-principal neurons could be seen in stratum oriens of CA1 and hilus in experimental group.Double immunofluorescence revealed that the FG-labeled NPY interneurons were located distantly from the zone of dye spread in CA1 area.CA3 area and hilus in experimental rats.When injection was administered in CA3 area.the FG-labeled pyramidal cells were visible in the whole CA3 area and hilus in both groups.Some pyramidal cells were present in CA1 in experimental group.Also some FG-labeled non-principle cells were foand in hilus and distantly from the zone of dve spread in CA1 area,And the FG-labeled NPY neurons could be seen in hilus in experimental rats.Conclusion Aberrant excitatory and inhibitory synaptic reconstruction exist in hippocampus in chronic phase of temporal lobe epilepsy,including excitatory synaptic connections among pyramidal cells in CA1 area.pyramidal cells between CAl and subiculum and pyramidal cells between CA1 and CA3,inhibitory synaptie connections among dendritie intemeurons in CA1 area,CA3 to CA1,hilus to CA1 and hilus to CA3area,These circuit arrangements may play an important role in the pathogenesis of epilepsy.%目的 探讨颞叶癫痫海马的异常兴奋性与抑制性突触联系变化.方法 建立

  9. Inhibitory action of antioxidants (ascorbic acid or α-tocopherol on seizures and brain damage induced by pilocarpine in rats Ação inibitória de antioxidantes (ácido ascórbico e α-tocoferol nas convulsões e dano cerebral em ratos induzidos pela pilocarpina

    Directory of Open Access Journals (Sweden)

    Adriana da Rocha Tomé

    2010-06-01

    Full Text Available Temporal lobe epilepsy is the most common form of epilepsy in humans. Oxidative stress is a mechanism of cell death induced by seizures. Antioxidant compounds have neuroprotective effects due to their ability to inhibit free radical production. The objectives of this work were to comparatively study the inhibitory action of antioxidants (ascorbic acid or α-tocopherol on behavioral changes and brain damage induced by high doses of pilocarpine, aiming to further clarify the mechanism of action of these antioxidant compounds. In order to determinate neuroprotective effects, we studied the effects of ascorbic acid (250 or 500 mg/kg, i.p. and α-tocopherol (200 or 400 mg/kg, i.p. on the behavior and brain lesions observed after seizures induced by pilocarpine (400 mg/kg, i.p., P400 model in rats. Ascorbic acid or α-tocopherol injections prior to pilocarpine suppressed behavioral seizure episodes. These findings suggested that free radicals can be produced during brain damage induced by seizures. In the P400 model, ascorbic acid and α-tocopherol significantly decreased cerebral damage percentage. Antioxidant compounds can exert neuroprotective effects associated with inhibition of free radical production. These results highlighted the promising therapeutic potential of ascorbic acid and α-tocopherol in treatments for neurodegenerative diseases.A epilepsia de lobo temporal é a mais comum forma de epilepsia em humanos. O estresse oxidativo é um dos mecanismos de morte celular induzida pelas crises convulsivas. Os compostos antioxidantes apresentam efeitos neuroprotetores devido à sua capacidade de inibir a produção de radicais livres. Os objetivos do presente trabalho foram estudar de forma comparativa a ação inibitória de antioxidantes (ácido ascórbico e α-tocoferol sobre as alterações comportamentais e histopatológicas no hipocampo de ratos após convulsões induzidas pela pilocarpina. A fim de determinar os efeitos neuroprotetores

  10. 匹罗卡品癫(癎)模型中蛋白质netrin-1的表达与苔藓纤维出芽的动态变化%The expression of netrin-1 and the process of mossy fiber sprouting after pilocarpine-in-duced epilepsy model in rats

    Institute of Scientific and Technical Information of China (English)

    郑佳丽; 杨金升; 石向群; 马亚杰; 刘学娟

    2011-01-01

    Objective:To explore the correlation between axon guiding cues netrin-1 and the process of aberrant mossy fiber sprouting (MFS) following status epilepticus(SE)in rats. Methods: The temporal lobe epilepsy(TLE)animal model was established by lithium-pilocarpine in rats. The process of aberrant MFS and the expression of netrin-1 was observed with Timm and immunohistochemical staining. Results:In the 2nd and 4th weeks after seizure,netrin-1 immunostaining showed an increase of netrin-1 in the dentate gyrus and MFS was found after TLE. Conclusion: The netrin-1 may be involved in the process of MFS.%目的:探讨神经诱向因子蛋白质netrin-1在癫(癎)持续状态后海马苔藓纤维出芽(MFS)中的作用.方法:氯化锂-匹罗卡品建立大鼠颞叶癫(癎)(TLE)模型,采用Timm染色和免疫组化的方法分别检测MFS和netrin-1在大鼠海马组织中的表达.结果:TLE组大鼠在模型形成的第2周和第4周,海马齿状回内netrin-1的表达较正常对照组明显增加,并可见到MFS,穿越齿状回颗粒细胞层到达内分子层,并形成一条致密的层状带.结论:癫(癎)状态后在海马齿状回netrin-1的表达上调,证明其可能参与了癫(癎)后MFS过程.

  11. Effects of EES on apoptotic neurons in rat hippocampus after lithium-pilocarpine induced status epilepticus%全蝎醇提物对Li-Pilo癫痫持续状态大鼠海马神经细胞凋亡的影响

    Institute of Scientific and Technical Information of China (English)

    喻良; 孙红斌; 梁益; 谢彦; 何保明; 许飞

    2009-01-01

    目的 探讨全蝎醇提物(Ethanol Extracts of Scorpion,EES)对大鼠癫痫持续状态(SE)后海马神经元凋亡的影响.方法 建立氯化锂.匹罗卡品癫痫持续状态(Lithium-Pilocarpine induced status epilepticus,Li-Pilo SE)模型.使用TUNEL技术观察正常对照组、Li-Piio SE模型组、丙戊酸钠组(VPA)和EES低(L)、中(M)、高(H)剂量组大鼠SE后6h、24h、48h、72h和7d海马CA_1区和CA_3区TUNEL阳性细胞的动态变化,并进行组间比较.结果 正常对照组未见TUNEL阳性细胞.造模各组大鼠SE后6h海马可见部分TUNEL阳性细胞,主要分布在CA_1、CA_3区,其中模型组和EES(L)组72h达高峰,而VPA、EES(M)和EES(H)组高峰提前到SE后48h,以后各组逐渐下降.VPA、EES(M)和EES(H)组SE后各观察时间点TUNEL阳性细胞数较模型组极显著减少(P0.05),其中VPA和EES(H)组各时间点TUNEL阳性细胞数减少较EES(M)组(P0.05).结论 EES能防止Li-Pilo SE大鼠海马神经元凋亡,并呈明显的量-效关系,高剂量EES抗凋亡作用与VPA相近.

  12. 多药转运蛋白对匹罗卡品癫(癎)大鼠模型脑内拉莫三嗪浓度的影响%Impact of multi-drug transporters on regulation of concentration of lamotrigine in hippocampal extracellular fluid in rat after pilocarpine-induced seizures

    Institute of Scientific and Technical Information of China (English)

    马爱梅; 张守文; 刘玉玺; 胡风云

    2009-01-01

    Objective To investigate the impact of multi-drug transporters including P-glycoprotein (PGP) and multi-drug resistance associated protein (MRP) on concentration of lamotrigine in the extracellular fluid in hippocampus of epilepsy rat models induced by pilocarpine, and to deduce the multi-drug resistance mechanisms in refractory epilepsy. Methods The epilepsy rat models were established by repeated administration (by ip) of pilocarpine. A microdialysis probe was placed into the hippocampus of the epileptic rats and dialysate was collected at five time-points from 30--150 minutes after systemic injections of lamotrigine (10 mg/kg). The concentration of lamotrigine in the extracellular fluid in the hippocampus was determined by high-performance liquid chromatography (HPLC). Then PGP inhibitor verapamil and MRP inhibitor probenecid was added individually through microdialysis probe and the concentration of lamotrigine was detected again. Results Compared with control group (0. 41 ± 0. 10 in 60 minutes, 0. 50 ±0.04 in 90 minutes, 0. 39 ±0. 09 in 120 minutes and 0. 30±0.06 in 150 minutes), verapamil significantly increased the concentration of lamotrigine in extracellular fluid of the hippocampus 60--150 minutes (0. 65 ±0. 11, 0. 84 ± 0. 09, 0. 70± 0. 09 and 0. 58 ± 0. 08 respectively) after injection (F value were 5.01, 8.61, 10. 23 and 7.89, all P < 0. 05) and probenecid also enhanced the concentration of lamotrigine 90--150 minutes (0. 75 ± 0. 09, 0. 58±0. 10 and 0. 49±0. 07) after injection (F = 6. 58, 4. 56, 4. 75, all P < 0. 05). Conclusions Penetration of lamotrigine through blood-brain barrier in pilocarpine induced epilepsy rats is restricted by PGP and MRP, resulting in decreased concentration of lamotrigine in the extracellular fluid of the hippocampus. Therefore, increasing expression of PGP and MRP in brains of epilepsy patients might be an important mechanism involved in multi-drug resistance in refractory epilepsy.%目的 观察多药转运蛋

  13. Influence of Valproate on Surviving Neurons in Hippocampus of Rats with Epilepsy by Lithium-Pilocarpine%丙戊酸对氯化锂-毛果芸香碱致癫(癎)大鼠海马神经元存活的影响

    Institute of Scientific and Technical Information of China (English)

    崔晓普; 蒋莉; 张晓萍

    2011-01-01

    目的 探讨丙戊酸(VPA)对氯化锂-毛果芸香碱致癫(癎)大鼠神经元的保护作用.方法 出生35 d雄性Wistar大鼠分为空白对照组、癫(癎)模型组和VPA干预组,制作氯化锂-毛果芸香碱癫(癎)发作模型,VPA经口灌胃给药,每次350 mg·kg-1.VPA连续给药5 d后处死动物,取大鼠脑组织尼氏染色检测存活神经元,免疫组织化学检测脑源性神经营养因子(BDNF)蛋白和乙酰化H3组蛋白表达.结果 1.空白对照组海马形态学结构正常,细胞完整,神经元多呈三角形,核较大,胞质里可见深染尼氏小体;癫(癎)模型组大鼠海马CA1、CA3区可见神经元缺失、变性、坏死;尼氏小体数量较少,甚至出现尼氏小体溶解、消失;VPA干预组神经元坏死较少,改变较轻.2.癫(癎)模型组海马CA1区存活神经元数目显著少于空白对照组和VPA干预组;VPA干预组存活神经元数目较癫(癎)模型组增加(P<0.01),但仍少于空白对照组(P<0.01);癫(癎)模型组海马CA3区存活神经元数目显著少于空白对照组和VPA干预组(Pa<0.01);VPA干预组存活神经元数目较癫(癎)模型组增加(P<0.01),但仍少于空白对照组(P<0.01).3.癫(癎)模型组和VPA干预组海马CA1、CA3区BDNF蛋白的表达均较空白对照组高(P<0.05,0.01),但VPA干预组的表达高于癫(癎)模型组(P<0.05,0.01).4.VPA干预组CA1、CA3区乙酰化H3组蛋白表达较癫(癎)模型组和空白对照组均显著增加(Pa<0.01),癫(癎)模型组乙酰化H3组蛋白表达亦高于空白对照组(P<0.01).结论 VPA对氯化锂-毛果芸香碱致癫(癎)大鼠海马CA1、CA3区神经元具有保护作用;海马CA1 、CA3区乙酰化H3组蛋白表达增加,进而调控BDNF的表达增加,可能是VPA对癫(癎)后大鼠脑保护作用机制之一.%Objective To explore the neuroprotective effect of valproate on neurons in hippocampus of epileptic model rat induced by lithium - pilocarpine.Methods Epileptic seizure model was induced in male

  14. Pilocarpine诱导小鼠癫痫持续状态发作后海马神经元的兴奋激活、损伤和死亡%Neuron activation, degeneration and death in the hippocampus of mice after pilocarpine induced status epilepticus

    Institute of Scientific and Technical Information of China (English)

    刘建新; 唐锋儒; 刘勇

    2011-01-01

    目的:研究癫痫持续状态发作后海马神经元兴奋激活、细胞损伤和细胞死亡的发生和相互关系.方法:采用pilocarpine诱导Swiss小鼠癫痫持续状态(status epilepticus,SE)模型,分别以c-Fos,Fluoro Jade B和CFV染色分析SE后不同时间点齿状回和CA1区锥体细胞的兴奋激活、损伤和细胞存活状况.结果:在齿状回颗粒细胞层,c-Fos阳性细胞在SE后1,2和24 h增多(P<0.01或0.05),但各组齿状回颗粒细胞层均无明显Fluoro Jade B阳性细胞,CFV染色标记的阳性细胞数量在对照组和各实验组之间差异无统计学意义(P>0.05);门区神经元在SE后没有明显的c-Fos诱导表达,但SE后2和24 h,门区Fluoro Jade B阳性细胞数量较对照组增多(P<0.01),CFV染色显示SE后1d门区残存的神经元数量较对照组减少(P<0.01);CA1区锥体细胞层c-Fos阳性细胞数量在SE后30 min,1,2和24 h后增多(P<0.01或0.05),Fluoro Jade B 阳性细胞数量在SE后2和24 h也较对照组增多(P<0.01),但CFV染色CA1区锥体细胞数量在各组间差异无统计学意义(P>0.05).结论:齿状回颗粒细胞、门区中间神经元以及CA1区锥体细胞在SE后的兴奋激活、颗粒细胞的损伤和死亡之间无直接的必然关系.%Objective To examine the occurrence of neuron activation,neurodegeneraion and cell death,and the correlation among them in the hippocampus after status epilepticus.Methods CFV,Fluoro Jade B and c-Fos staining were done at multiple time points after pilocarpine induced status epilepticus.Results In the stratum granulosum of dentate gyrus,c-Fos positive neurons increased significantly at 1 h,2 h and 1 d after status epilepticus (P <0.01 or 0.05).However,almost no Fluoro Jade B staining cell was found in the stratum granulosum in the experiment and control groups,and no obvious difference was shown on the numbers of CFV staining cells in this area among all groups.In the hilus of dentate gyrus of different groups

  15. Effects of percutaneous trigeminal nerve electrical stimulation on behavior and the ability of learning and memory of epileptic rats induced by Pilocarpine%经皮三叉神经电刺激对匹罗卡品诱导的癫痫大鼠的行为及学习记忆能力的影响

    Institute of Scientific and Technical Information of China (English)

    王倩倩; 刘益民; 钟平; 张雷; 王玉

    2015-01-01

    Objective To observe the effects of percutaneous trigeminal nerve electrical stimulation on behavior and the ability of learning and memory of epileptic rats induced by Pilocarpine.Methods Thirty healthy male rats were randomly divided into normal control group, epilepsy model group and trigeminal nerve stimulation group ( TNS group) .Pilocarpine was used to inducestatus epileptics ( SE) in rats.TNS group rats were treated with percutaneous trigeminal nerve stimulation consecutive for 4 weeks.At 2 weeks after SE model established, video monitor were employed to record the frequency of spontaneous recurrence seizures ( SRS) , the intensity and duration of seizures. At 4 weeks after SE model established, rats in each group were taken Morris water maze test.The incubation period of rats finding underwater platform in the acquired training was recorded.The percentage of time and distance of rats searching target quadrant and the number of times through the platform were also recorded.Results Rats in control group didn''t show abnormal behavior throughout this experiment.During the process of animal model making, a rat died in epilepsy model group while 2 in TNS group.During-EEG monitoring, 2 rats in epilepsy model group died. Compared with epilepsy model group, the frequency of spontaneous recurrence seizure and the intensity of seizure significantly decreased, the average time of seizure significantly shortened in TNS group ( P<0.05 -0.01 ) in 2 weeks.In acquired training experiment, with the number of training increased, the average escape latent of experimental animals gradually reduced, the escape latent of rats in epilepsy model group were significantly prolonged compared with control group rats at each time-point ( all P<0.01 ) , while the average escape latency was significantly reduced in TNS group rats than epilepsy model group (P<0.05-0.01).Compared to control group, the percentage of time search and the percentage of navigation distance in the target

  16. Effect of Ghrelin on nuclear factor-κB and tumor necrosis factor-α in the cerebral cortex of immature rats with pilocarpine-induced epilepsy%Ghrelin对匹罗卡品诱导癫(癎)大鼠大脑皮层核因子-κB和肿瘤坏死因子-α表达的影响

    Institute of Scientific and Technical Information of China (English)

    张瑞云; 王清义; 李培国; 隋风轩; 王华

    2010-01-01

    目的 探讨Ghrelin治疗匹罗卡品诱导的癫(癎)大鼠大脑皮层核因子(NF)-κB和肿瘤坏死因子(TNF)-α基因和蛋白表达水平的变化.方法 建立匹罗卡品诱导的癫(癎)大鼠模型,将模型分为模型组、生理盐水组和Ghrelin治疗组,同时设正常对照组,比较各组大鼠大脑皮层NF-κB和TNF-α蛋白和基因表达水平.结果 模型组大鼠脑组织NF-κB和TNF-α蛋白和基因表达均增多,正常对照组两者表达较少;Ghrelin治疗组大鼠脑组织NF-κB和TNF-α蛋白和基因表达水平均比模型组和生理盐水组明显降低,差异有显著性(P<0.05).结论 Ghrelin可能通过降低癫(癎)大鼠大脑皮层NF-κB和TNF-α蛋白和基因表达水平,减轻皮层神经细胞的炎症反应,达到对神经细胞的保护作用.%Objective To explore the changes of gene and protein expressions of nuclear factor-κB(NF-κB) and tumor necrosis factor-α (TNF-α) in immature rats with pilocarpine-induced epilepsy treated with ghrelin. Methods The pllocarpine-induced epilepsy model in immatured rats were built, then the rats were divided into three groups: Ghrelin-treated group, saline-treated group and model group, meanwhile the normal control group was set. The NF-κB and TNF-α levels of gene and protein in the cerebral cortex of immature rats were detected. Results The expression levels of gene and protein of NF-κB and TNF-α were increased in model group,but decreased in the normal control group;NF-κB and TNF-α levels in Ghrelin treated group were obviously lower than those of saline-treated group and model group(P < 0. 05). Conclusion The protective mechanism of Ghrelin for nerve cell is cutting down the expressions of NF-κB and TNF-α in the cerebral cortex of immature rats with epilepsy and lessening inflammatory reaction in neurocytes.

  17. Effects of Low-frequency Repetitive Transcranial Magnetic Stimulation on Expressions of Hippocampus CA3 Region Annexin A7 in Rats after Pilocarpine-induced Seizures%低频重复经颅磁刺激的抗(癎)作用及其对癫(癎)大鼠海马CA3区膜连蛋白A7表达的影响

    Institute of Scientific and Technical Information of China (English)

    余琴; 王莉; 余巨明; 贾朝均

    2012-01-01

    目的:观察低频重复经颅磁刺激(rTMS)对大鼠(癎)性发作行为及海马CA3区膜连蛋白A7表达的影响.方法:取85只健康雄性SD大鼠,按预处理方式将其分成rTMS组(rTMS刺激+毛果芸香碱致(癎))、对照组(假刺激+毛果芸香碱致(癎))及生理盐水对照组(假刺激+生理盐水).各组大鼠经相应处理后,rTMS组和对照组(各n=30)大鼠制作氯化锂-毛果芸香碱癫(癎)持续状态(SE)模型;生理盐水对照组(n=25)则腹腔注射生理盐水.观察各组大鼠行为表现及SE潜伏期,应用免疫组化法观察膜连蛋白A7表达的动态变化(6h、24h、1周、3周、6周).结果:①rTMS组SE潜伏期为(41.37±5.45)min,与对照组(23.86±4.42)min比较明显延长(P<0.01);②海马CA3区膜连蛋白A7阳性细胞数在各时间点均为对照组最多,rTMS组次之,生理盐水对照组最少(均P<0.05).但是膜连蛋白A7的表达随时间变化的趋势rTMS组与对照组明显不同.结论:低频rTMS有一定抗(癎)作用;低频rTMS可影响大鼠海马CA3区膜连蛋白A7表达并呈现独特的动态变化特点.%Aim: To observe the effects of low-frequency repetitive transcranial magnetic stimulation (rTMS) on rat behavior and the expressions of hippocampus CA3 region annexin A7 in rats after pilocarpine-induced seizures, and explore the possible anti-epileptic mechanisms of rTMS. Methods: 85 healthy male SD rats were randomly divided into rTMS group (rTMS+pilocarpine), control group (sham stimulation+ pilocarpine), and NS control group (sham stimulation+normal saline), with 30 rats in each of rTMS group and control group, and 25 rats in NS control group. After consecutive corresponding stimulation for 2 weeks respectively, the status epilepticus rat models were established in rTMS group and control group, their behavior were observed and the latent time to status epilepticus was measured. The rats in all groups were killed in different time (6 h, 24 h, 1 week, 3 weeks, 6 weeks

  18. Effect of serotonin depletion on seizures learning-memory in pilocarpine-induced epileptic rats%脑内5-羟色胺减少对致痫大鼠癫痫发作及学习记忆的影响

    Institute of Scientific and Technical Information of China (English)

    吕晓钗; 黄华品; 林婉挥; 陈圣根

    2012-01-01

    目的:于中脑正中中缝核局部微量注射5,7-二羟色胺(5,7-DHT),探讨5-羟色胺(5-HT)与癫痫的关系及匹罗卡品(PILO)致痫大鼠学习记忆改变的可能机制.方法:成年SD大鼠随机分为PILO组、PILO+5,7-DHT组、空白对照组三组,然后根据是否出现癫痫持续状态(SE)再将PILO组分成:PILO+ SE组和PILO-SE组两亚组;利用视频脑电图观察大鼠癫痫发作及皮层脑电变化;运用Morris水迷宫测评大鼠空间学习记忆水平;最后运用免疫组化法观察大鼠中缝核5-HT能神经元.结果:大鼠予以5,7-DHT( PILO+ 5,7- DHT组)处理后造模成功率、死亡率及慢性期自发性发作频率均增高;与空白组比较PILO+ SE组中缝核5-HT能神经元数目有所下降(P<0.05),而PILO+5,7-DHT组下降更明显(P<0.01);与空白组比较PILO+ SE组平均逃避潜伏期延长、穿越平台次数减少、原平台象限停留时间缩短(P<0.05),而与PILO+ SE组比较PILO+ 5,7-DHT组变化不明显.结论:脑内5-H水平的降低容易诱发癫痫发作,尚不能认为癫痫大鼠合并认知功能障碍与脑内5-HT水平下降有关.%Objective: To investigate the relationship between serotonin(5-HT)and epilepsy and the mechanism of learning-memory in pi-locaipine(PILO)-induced epileptic rats after5,7-dihydroxytryplamine (5,7-DHT) microinjection in median raphe nucleus. Methods: Adult S D rats were randomly divided into 3 groups: PIL0 group, PTL0 + 5,7-DHT group, vehicle control group; PIL0 group was divided into two groups by status epilepticus(SE): PIL0+ SE group and HL0-SE group. The rats' seizures and cortex electroencephalography(EEG) were observed by vedio EEC. The rats' spatial learning-memory was evaluated by Morris water maze. Finally, serotonergic neuron in raphe nuclei was observed by immunohistochemisty. Results: After treatment of 5,7-DHT (PIL0 + 5,7-DHT group), the success rate, the mortality and the frequency of chronic spontaneous seizures in pilocarpine

  19. Downregulation of BK channel expression in the pilocarpine model of temporal lobe epilepsy

    OpenAIRE

    Pacheco Otalora, Luis F.; Hernandez, Eder F.; Arshadmansab, Massoud F.; rancisco, Sebastian F; Willis, Michael; Ermolinsky, Boris; Zarei, Masoud; Knaus, Hans-Guenther; Garrido-Sanabria, Emilio R.

    2008-01-01

    In the hippocampus, BK channels are preferentially localized in presynaptic glutamatergic terminals including mossy fibers where they are thought to play an important role regulating excessive glutamate release during hyperactive states. Large conductance calcium-activated potassium channels (BK, MaxiK, Slo) have recently been implicated in the pathogenesis of genetic epilepsy. However, the role of BK channels in acquired mesial temporal lobe epilepsy (MTLE) remains unknown. Here we used immu...

  20. Exposure to Mozart music reduces cognitive impairment in pilocarpine-induced status epilepticus rats

    National Research Council Canada - National Science Library

    Xing, Yingshou; Qin, Yi; Jing, Wei; Zhang, Yunxiang; Wang, Yanran; Guo, Daqing; Xia, Yang; Yao, Dezhong

    2016-01-01

    .... Here we evaluated spatial cognition changes at different epileptogenesis stages in rats of this model and explored the effects of long-term Mozart music exposure on the recovery of cognitive ability...

  1. Increased excitability and metabolism in pilocarpine induced epileptic rats: effect of Bacopa monnieri.

    Science.gov (United States)

    Mathew, Jobin; Paul, Jes; Nandhu, M S; Paulose, C S

    2010-09-01

    We have evaluated the acetylcholine esterase and malate dehydrogenase activity in the muscle, epinephrine, norepinephrine, insulin and T3 content in the serum of epileptic rats. Acetylcholine esterase and malate dehydrogenase activity increased in the muscle and decreased in the heart of the epileptic rats compared to control. Insulin and T3 content were increased significantly in the serum of the epileptic rats. Our results suggest that repetitive seizures resulted in increased metabolism and excitability in epileptic rats. Bacopa monnieri and Bacoside-A treatment prevents the occurrence of seizures there by reducing the impairment on peripheral nervous system.

  2. Spatial memory deficits in juvenile rats with pilocarpine induced temporal lobe epilepsy

    Directory of Open Access Journals (Sweden)

    Orbán-Kis K

    2014-10-01

    Full Text Available One of the most frequent forms of epilepsy in humans is temporal lobe epilepsy. Characteristic to this form of the disease is the frequent pharmacoresistance and the association with behavioural disorders and cognitive impairment. The objective of our study was to establish the degree of cognitive impairment in a rat model of temporal lobe epilepsy after an initial epileptogenic exposure but before of the onset of the effect of long-duration epilepsy.

  3. Pulsed direct and constant direct currents in the pilocarpine iontophoresis sweat chloride test.

    Science.gov (United States)

    Gomez, Carla Cristina Souza; Servidoni, Maria de Fatima; Marson, Fernando Augusto de Lima; Canavezi, Paulo Jose Coelho; Vinagre, Adriana Mendes; Costa, Eduardo Tavares; Ribeiro, Antonio Fernando; Ribeiro, Maria Angela Gonçalves de Oliveira; Toro, Adyleia Aparecida Dalbo Contrera; Pavan, Celia Regina; Rondon, Michelle Vivine Sá Dos Santos; Lorena, Sonia Leticia Silva; Vieria, Francisco Ubaldi; Ribeiro, Jose Dirceu

    2014-12-13

    The classic sweat test (CST) is the golden standard for cystic fibrosis (CF) diagnosis. Then, our aim was compare the production and volume of sweat, and side effects caused by pulsed direct current (PDC) and constant direct current (CDC). To determine the optimal stimulation time (ST) for the sweat collection. To verify the PDC as CF diagnosis option. Prospective study with cross-sectional experimental intervention. Experiment 1 (right arm): PDC and CDC. ST at 10 min and sweat collected at 30 min. Currents of 0.5; 0.75; 1.0 and 1.5 mA and frequencies of 0, 200, 1,000 and 5,000 Hz applied. Experiment 2 (left arm): current of 1.0 mA, ST at 5 and 10 min and sweat collected at 15 and 30 min with frequencies of 0; 200; 1,000 and 5,000 Hz applied Experiments 1 and 2 were performed with current density (CD) from 0.07 to 0.21 mA/cm2. Experiment 3: PDC was used in typical CF patients with two CFTR mutations screened and or with CF diagnosis by rectal biopsy and patients with atypical CF. 48 subjects (79.16% female) with average of 29.54 ± 8.87 years old were enrolled. There was no statistical difference between the interaction of frequency and current in the sweat weight (p = 0.7488). Individually, positive association was achieved between weight sweat and stimulation frequency (p = 0.0088); and current (p = 0.0025). The sweat production was higher for 10 min of stimulation (p = 0.0023). The sweat collection was better for 30 min (p = 0.0019). The skin impedance was not influenced by ST and sweat collection (p > 0.05). The current frequency was inversely associated with the skin impedance (p < 0.0001). The skin temperature measured before stimulation was higher than after (p < 0.0001). In Experiment 3 (29 subjects) the PDC showed better kappa index compared to CDC (0.9218 versus 0.5205, respectively). The performance of the CST with CDC and PDC with CD of 0.14 to 0.21 mA/cm2 showed efficacy in steps of stimulation and collection of sweat, without side effects. The optimal stimulation time and sweat collection were, respectively, 10 and 30 min.

  4. Physical activity and neuroprotection in adult mice after pilocarpine induced status epilepticus

    OpenAIRE

    Cesar Renato Sartori

    2005-01-01

    Resumo: O modelo de epilepsia induzida por pilocarpina em camundongos reproduz a Epilepsia do Lobo Temporal (ELT) em humanos. Animais submetidos à indução de status epilepticus apresentam alterações comportamentais, eletroencefalográficas e lesão neuronal compatíveis com esta condição. Estudos recentes relatam relevantes efeitos positivos da prática de atividade física sobre o sistema nervoso tanto em humanos como em modelos animais. Dentre estes efeitos figuram o aumento da sobrevivência neu...

  5. Drug: D08376 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available 31) Gastric acid secretion hsa04972(1131) Pancreatic secretion Enzyme: CYP2A6 [HSA:1548] map07054 Antiglaucoma...ents Pilocarpine D08376 Pilocarpine borate Ophthalmic Agents Ophthalmic Antiglaucoma Agents Pilocarpine D083

  6. Deterruination of Pilocarpine nitrate eye drops by polarimetry%旋光法测定硝酸毛果芸香硷滴眼液含量

    Institute of Scientific and Technical Information of China (English)

    母琼梅; 何继祥

    2001-01-01

    目的:测定硝酸毛果芸香硷滴眼液含量.方法:采用旋光法测定.结果:硝酸毛果芸香硷浓度在5-25mg.ml-1范围内旋光度与浓度呈良好线性关系,r=1.0000.平均回收率为99.3%,RSD为0.4%(n=5).结论:本法快速、准确可靠.

  7. Diurnal Variation Has Effect on Differential Gene Expression Analysis in the Hippocampus of the Pilocarpine-Induced Model of Mesial Temporal Lobe Epilepsy.

    Directory of Open Access Journals (Sweden)

    Evelin Antonieli da Silva Santos

    Full Text Available The molecular mechanisms underlying epileptogenesis have been widely investigated by differential gene expression approach, especially RT-qPCR methodology. However, controversial findings highlight the occurrence of unpredictable sources of variance in the experimental designs. Here, we investigated if diurnal rhythms of transcript's levels may impact on differential gene expression analysis in hippocampus of rats with experimental epilepsy. For this, we have selected six core clock genes (Per1, Per3, Bmal1, Clock, Cry1 and Cry2, whose rhythmic expression pattern in hippocampus had been previously reported. Initially, we identified Tubb2a/Rplp1 and Tubb2a/Ppia as suitable normalizers for circadian studies in hippocampus of rats maintained to 12:12 hour light:dark (LD cycle. Next, we confirmed the temporal profiling of Per1, Per3, Bmal1, Cry1 and Cry2 mRNA levels in the hippocampus of naive rats by both Acrophase and CircWave statistical tests for circadian analysis. Finally, we showed that temporal differences of sampling can change experimental results for Per1, Per3, Bmal1, Cry1 and Cry2, but not for Clock, which was consistently decreased in rats with epilepsy in all comparison to the naive group. In conclusion, our study demonstrates it is mandatory to consider diurnal oscillations, in order to avoid erroneous conclusions in gene expression analysis in hippocampus of rats with epilepsy. Investigators, therefore, should be aware that genes with circadian expression could be out of phase in different animals of experimental and control groups. Moreover, our results indicate that a sub-expression of Clock may be involved in epileptogenicity, although the functional significance of this remains to be investigated.

  8. MicroRNA profiles in hippocampal granule cells and plasma of rats with pilocarpine-induced epilepsy--comparison with human epileptic samples

    DEFF Research Database (Denmark)

    Roncon, Paolo; Soukupovà, Marie; Binaschi, Anna

    2015-01-01

    The identification of biomarkers of the transformation of normal to epileptic tissue would help to stratify patients at risk of epilepsy following brain injury, and inform new treatment strategies. MicroRNAs (miRNAs) are an attractive option in this direction. In this study, miRNA microarrays wer...

  9. Enhanced pyridoxal 5'-phosphate synthetic enzyme immunoreactivities do not contribute to GABAergic inhibition in the rat hippocampus following pilocarpine-induced status epilepticus.

    Science.gov (United States)

    Kwak, S-E; Kim, J-E; Kim, D-W; Kwon, O-S; Choi, S-Y; Kang, T-C

    2009-03-31

    To comprehend the role of pyridoxal 5'-phosphate (PLP) in epilepsy or seizure, we investigated whether the expressions of two PLP synthetic enzymes (pyridoxal kinase, PLK; pyridoxine-5'-phosphate oxidase, PNPO) are altered in the hippocampus and whether changes in paired-pulse responses in the hippocampus are associated with altered PLP synthetic enzyme expressions following status epilepticus (SE). PLK and PNPO immunoreactivities were significantly increased in the rat hippocampus accompanied by reductions in paired-pulse inhibition at 1 day and 1 week after SE. Four weeks after SE, PLK and PNPO immunoreactivities in dentate granule cells were similar to those in control animals, while their immunoreactivities were markedly reduced in Cornu Ammonis 1 (CA1) pyramidal cells due to neuronal loss. Linear regression analysis identified a direct proportional relationship between PLK/PNPO immunoreactivity and normalized population spike amplitude ratio in the dentate gyrus and the CA1 region as excluded the data obtained from 4 weeks after SE. These findings indicate that the upregulation of PLK and PNPO immunoreactivities in principal neurons may not be involved in gamma-aminobutyric acid (GABA)ergic inhibition, but rather in enhanced excitability during epileptogenic periods.

  10. Drug: D05478 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available n hsa04972(1131) Pancreatic secretion Enzyme: CYP2A6 [HSA:1548] map07054 Antiglaucoma...rpine D05478 Pilocarpine nitrate (USP) Ophthalmic Agents Ophthalmic Antiglaucoma Agents Pilocarpine D05478 P

  11. 锂-匹罗卡品癫痫模型中大鼠苔藓纤维发芽机制研究%Study on the Mechanisms of the Rat Mossy Fiber Sprouting in Lithium-Pilocarpine Epilepsy Model

    Institute of Scientific and Technical Information of China (English)

    杨银升; 韩虹

    2009-01-01

    目的:通过观察锂-匹罗卡品癫痫模型中大鼠实验性癫痫发作时的行为、脑电图及病理生理变化,探讨苔藓纤维发芽(MFS)在癫痫中的发病机制.方法:利用锂-匹罗卡品(Li-PC)制作大鼠急性癫痫持续状态(SE)的癫痫模型.造模后7 d、14 d、28 d,进行苔藓纤维发芽评分.结果:模型组大鼠2周时可见MFS穿过齿状回内分子层颗粒细胞层,进入内分子层及CA3区锥体细胞始层黑色颗粒形成,4周时形成连续密集颗粒带,MFS评分显著高于生理盐水对照组(P<0.05).结论:急性期SE神经元损伤致苔藓纤维发芽,可能是慢性颞叶癫痫的病理基础.

  12. Glucose utilization in the brain during acute seizure is a useful biomarker for the evaluation of anticonvulsants: effect of methyl ethyl ketone in lithium-pilocarpine status epilepticus rats

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, Akifumi [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan)], E-mail: yamaaki@sahs.med.osaka-u.ac.jp; Momosaki, Sotaro; Hosoi, Rie [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan); Abe, Kohji [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan); Developmental Research Laboratories, Shionogi and Co., Ltd., Toyonaka, Osaka, 561-0825 (Japan); Yamaguchi, Masatoshi [Faculty of Pharmaceutical Sciences, Fukuoka University, Johnan, Fukuoka 814-0180 (Japan); Inoue, Osamu [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan)

    2009-11-15

    Enhancement of glucose utilization in the brain has been well known during acute seizure in various kinds of animal model of epilepsy. This enhancement of glucose utilization might be related to neural damage in these animal models. Recently, we found that methyl ethyl ketone (MEK) had both anticonvulsive and neuroprotective effects in lithium-pilocapine (Li-pilo) status epilepticus (SE) rat. In this article, we measured the uptake of [{sup 14}C]2-deoxyglucose ([{sup 14}C]DG) in the Li-pilo SE and Li-pilo SE with MEK rat brain in order to assess whether the glucose utilization was a useful biomarker for the detection of efficacy of anticonvulsive compounds. Significant increase of [{sup 14}C]DG uptake (45 min after the injection) in the cerebral cortex, hippocampus, amygdala and thalamus during acute seizure induced by Li-pilo were observed. On the other hand, the initial uptake of [{sup 14}C]DG (1 min after the injection) in the Li-pilo SE rats was not different from the control rats. Therefore, the enhancement of glucose metabolism during acute seizure was due to the facilitation of the rate of phosphorylation process of [{sup 14}C]DG in the brain. Pretreatment with MEK (8 mmol/kg) completely abolished the enhancement of glucose utilization in the Li-pilo SE rats. The present results indicated that glucose utilization in the brain during acute seizure might be a useful biomarker for the evaluation of efficacy of anticonvulsive compounds.

  13. 硝酸毛果芸香碱滴眼液中苯扎氯铵含量测定%Determination of Benzalkonium Chloride in Pilocarpine Nitrate Eye Drops

    Institute of Scientific and Technical Information of China (English)

    黄志元; 朱样根

    2012-01-01

    目的:用高效液相色谱法(HPLC)测定硝酸毛果芸香碱滴眼液中苯扎氯铵的含量.方法:色谱柱为CN(Hypersil CN,150 mm × 4.6 mm,5 μm);流动相为0.03 mol/L磷酸二氢钠溶液(用1 mol/L氢氧化钠溶液调节pH至7.0)-乙腈(57∶43);流速为2 mL/min;检测波长为214 nm;柱温为35℃;苯扎氯铵C12、C14、C16之间的分离度应符合规定.结果:苯扎氯铵在40 ~ 160 μg/mL内线性关系良好,Y = 12.228 1 X + 6.881 4(r = 0.999 9,n = 6),回收率为99.7% ~ 101.9%,RSD = 0.66%.结论:该法操作简便,快速、准确、灵敏度高,重复性好,可用于硝酸毛果芸香碱滴眼液中苯扎氯铵的含量测定.

  14. The expression of Rho kinase 2 in the brain of rat after pilocarpine-induced status epilepticus%Rho激酶2在急性期癫痫大鼠脑内表达变化的研究

    Institute of Scientific and Technical Information of China (English)

    靳俊功; 张华; 刘备; 武昊; 李焕发; 杨倩丽

    2013-01-01

    目的 观察Rbo激酶2(ROCK2)在癫痫大鼠脑内的表达情况及其抑制剂法舒地尔对癫痫大鼠脑电图的影响.方法 75只SD大鼠随机分入正常对照组、6h组、1d组、3d组、5d组、7d组、空白组、癫痫组和法舒地尔组.腹腔注射氯化锂—匹鲁卡品建立癫痫模型.通过免疫组织化学和Westem blot方法,比较各组大鼠颞叶、海马区ROCK2表达的差异;对空白组、癫痫组及法舒地尔组大鼠进行脑电监测,分析脑电图变化.结果 在海马组织中,3d组、5d组ROCK2的表达水平较对照组升高,差异有统计学意义(P<0.05);在颞叶脑组织中,3d组、5d组、7d组ROCK2的表达水平较对照组均明显升高,差异有统计学意义(P<0.05).免疫荧光结果显示正常组大鼠与实验组大鼠的海马组织中ROCK2(绿色)与NeuN(红色)在神经元中共表达;颞叶脑组织中ROCK2(绿色)与GFAP(红色)在星形胶质细胞中共表达.脑电监测结果显示空白组大鼠脑电正常.癫痫组大鼠在给予匹鲁卡品约28min后出现痫样放电波形.与癫痫组大鼠相比,法舒地尔组大鼠出现痫样波形的潜伏时间明显延长,频率明显降慢、幅度明显降低(P<0.05)).结论 癫痫发作将上调大鼠颞叶和海马区ROCK2的表达,ROCK2抑制剂法舒地尔具有一定的抗癫痫作用.

  15. The dynamic study of Li-Pilocarpine epilepsy model on rats by MRI and volumetry of hippocampus%大鼠Li-Pilocarpine癫痫模型的MRI与海马体积测量动态研究

    Institute of Scientific and Technical Information of China (English)

    尹建忠; 祁吉; 倪红艳; 郭文梅

    2004-01-01

    目的:观察Li-Pilocarpine大鼠癫痫模型在慢性期出现反复自发性癫痫发作过程中MRI与海马体积的进行性变化,探索癫痫发生的病理生理机制.方法:分别在成功诱发癫痫持续状态大鼠的3小时~8周的不同时间点分别行MRI成像与海马体积测量,并与正常对照组相比较,观察急性期至慢性期过程中的动态变化.结果:在急性期最早累及杏仁核、内嗅皮层和梨状皮层,进而出现海马的水肿.两周后出现海马的进行性萎缩,海马体积在慢性期萎缩至正常的1/2左右.结论:癫痫持续状态后的脑损伤包括多个受累部位,既累及海马,也累及其它边缘系统,如杏仁核和邻近的梨状皮层、内嗅皮层、丘脑及大脑皮层感觉运动区.包括海马在内的边缘系统的改变更为复杂.

  16. Effects of UPP on PSD95/NR2B expression in rats with pilocarpine-induced epilepsy%UPP对匹罗卡品致痫大鼠PSD95/NR2B表达的影响

    Institute of Scientific and Technical Information of China (English)

    张杰; 陈阳美

    2010-01-01

    目的 研究泛素-蛋白酶体途径(UPP)对匹罗卡品致痫大鼠海马突触后致密物质-95(PSD95)/ N-甲基-D-天(门)冬氨酸2B受体(NR2B)表达的影响及其在癫痫发生和发展中的作用.方法 用免疫荧光方法检测匹罗卡品致痫大鼠及经UPP抑制剂(MG-132)预处理大鼠海马PSD95/NR2B的表达,并观察组织病理学变化.结果 MG-132能明显抑制匹罗卡品致痫大鼠海马PSD95/NR2B表达下调,并且明显加重致痫大鼠海马神经元损伤.结论 UPP能调控匹罗卡品致痫大鼠海马PSD95/NR2B的表达.

  17. Comparison of Iontophoretic Lidocaine to EMLA Cream for Pain Reduction Prior to Intravenous Fannulation in Adults

    Science.gov (United States)

    2000-10-01

    1957, used iontophoresis of local anesthetics for the treatment of trigeminal neuralgia . Gibson and Cooke, in 1959, used lidocaine and pilocarpine...al. 1994). Other clinical uses of EMLA cream include: lumbar puncture, epidural injections, drug reservoir injections, skin surgery, genitourinary...surgery, ear, nose, throat, oral surgery, lithotripsy, and there are some anecdotal reports of its use for the treatment of postherpetic neuralgia

  18. Drug: D00525 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available 9 D00525.gif Pilocarpus [TAX:77016] Antiglaucoma agent; Cholinergic [ophthalmic] [DS:H00612] Same as: C07474...tic secretion Enzyme: CYP2A6 [HSA:1548] map07054 Antiglaucoma agents map07220 Cholinergic and anticholinergi...cation [BR:br08302] Dental and Oral Agents Pilocarpine D00525 Pilocarpine (JAN/USP) Ophthalmic Agents Ophthalmic Antiglaucoma

  19. Does angiogenesis play a role in the establishment of mesial temporal lobe epilepsy?

    Science.gov (United States)

    Benini, Ruba; Roth, Raquel; Khoja, Zehra; Avoli, Massimo; Wintermark, Pia

    2016-04-01

    Mesial temporal lobe epilepsy (MTLE) is a focal epileptic disorder that is frequently associated with hippocampal sclerosis. This study investigated whether blocking angiogenesis prevents the development of seizures and hippocampal atrophy in the pilocarpine rat model of MTLE. To block angiogenesis, a subset of animals were given sunitinib orally. Continuous video recordings were performed to identify seizures. Brains were then extracted and sectioned, and hippocampal surfaces and angiogenesis were assessed. After a latent period of 6.6 ± 2.6 days, the sham-treated pilocarpine rats presented convulsive seizures, while the pilocarpine rats treated with sunitinib did not develop seizures. Sham-treated pilocarpine rats but not sunitinib-treated pilocarpine rats had significantly smaller hippocampi. Endothelial cell counts in sham-treated pilocarpine rats were significantly greater than in controls and sunitinib-treated pilocarpine rats. Blocking angiogenesis immediately following the initial insult in this animal model prevented thus angiogenesis and hippocampal atrophy and averted the development of clinical seizures.

  20. Drug: D02200 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available D02200 Drug Pilocarpine hydrochloride (JP16/USP); Isopto carpine (TN); Pilopine HS ...(TN); Sanpilo (TN) C11H16N2O2. HCl 244.0979 244.7179 D02200.gif Cholinergic [ophthalmic] [DS:H00612] Therape...n hsa04972(1131) Pancreatic secretion Enzyme: CYP2A6 [HSA:1548] map07054 Antiglaucoma agents map07220 Cholin...ents 1312 Miotic agents D02200 Pilocarpine hydrochloride (JP16/USP) 2 Agents affecting individual organs 23 ...Digestive organ agents 239 Miscellaneous 2399 Others D02200 Pilocarpine hydrochloride (JP16/USP) Anatomical

  1. Antiseizure Activity of Hydro-Ethanol Leaf Extract of Ficus Thonningh ...

    African Journals Online (AJOL)

    AFRICAN JOURNALS ONLINE (AJOL) · Journals · Advanced Search · USING ... Pentylenetetrazole, maximal electroshock and pilocarpine seizure models were ... Keywords: Seizures, antiseizure, Ficus thonningii leaf extract, albino mice ...

  2. Accidental mydriasis from blue nightshade "lipstick".

    Science.gov (United States)

    Rubinfeld, R S; Currie, J N

    1987-03-01

    A 7-year-old girl presented with bilaterally dilated pupils, nausea, and vomiting 2 days after head trauma. Pilocarpine pupil testing led to the correct diagnosis of pharmacologic pupillary dilation from an unexpected and unusual source of plant poisoning, Solanum dulcamara (blue nightshade). In patients with internal ophthalmoplegia, awareness of the possibility of pharmacologic mydriasis and correct use of topical pilocarpine testing can preclude the necessity for neuroradiologic and invasive diagnostic studies, even in cases with atypical or complex presentations.

  3. Sexual response in female rats with status epilepticus

    OpenAIRE

    2013-01-01

    Purpose Female sexual function is complex and may be disrupted by disease, in particular epilepsy. Chronic seizures in women can have adverse effects on reproductive function, but it has been difficult to dissociate the effects of epilepsy from those related to anticonvulsant medications. the purpose of this study was to evaluate sexual behavior in female rats submitted to pilocarpine-induced status epilepticus (SE). Methods Adult female Wistar rats were given saline or pilocarpine (350mg/kg,...

  4. A comparative study of amygdala Kindling and LiCl-pilocarpine model of epilepsy in rat%杏仁核电点燃癫(痫)模型与氯化锂-匹罗卡品大鼠癫(痫)模型的对比研究

    Institute of Scientific and Technical Information of China (English)

    陈姝璇; 王丽琨; 伍国锋

    2016-01-01

    目的:探讨不同类型癫(痫)大鼠在急性期和慢性期反复自发性发作时、发作前后脑电图(EEG)相应指标改变以及行为学改变.方法:选取雄性SD大鼠40只,随机分为A、B、C、D4组.A组10只为杏仁核电点燃癫(痫)模型(不作电刺激)对照组;B组:10只,为制作杏仁核电刺激点燃癫(痫)模型组;C组:10只,氯化锂-匹罗卡品癫(痫)模型(只注生理盐水)对照组;D组:10只,制作氯化锂-匹罗卡品癫(痫)模型组.结果:D组慢性期发作时EEG频率与急性期频率相比差异有统计学意义(P<0.05),与B组各时期相比差异有统计学意义(P<0.05);D组慢性期发作后3 min发作频率与B组急性期相比差异有统计学意义(P<0.05);A组与C组在频率、波幅方面比较差异无统计学意义(P>0.05).结论:B、D两种不同类型的癫(痫)大鼠不同时期EEG和行为学都会发生相应的改变.

  5. Protection of pyrrolidine dithiocarbamate on the piriform cortex of pilocarpine-induced status epileptic rats%核因子-κB抑制剂吡咯烷二硫氨基甲酸盐对毛果芸香碱诱导癫痫大鼠梨状皮层的保护性作用

    Institute of Scientific and Technical Information of China (English)

    徐海元; 徐晓云; 吕日琅

    2013-01-01

    目的 观察核因子-κB(NF-κB)抑制剂吡咯烷二硫氮基甲酸盐(PDTC)对毛果芸香碱诱导癫痫大鼠梨状皮层的保护性作用.方法 将26只正常雄性SD大鼠随机分为3组:正常对照组(NS组,n=6)、毛果芸香碱诱导癫痫组(SE组,n=10)和PDTC干预组(PDTC组,n=10).SE组一次性腹腔注射毛果芸香碱诱导大鼠癫痫发作;PDTC干预组分别于造模前24 h、20 min、SE发作后24 h腹腔注射PDTC 100 mg·kg-;NS组给予等量生理盐水.于造模成功后48 h处死各组大鼠,采用Nissl染色和Fluoro-Jade C (FJC)染色法分别检测并比较各组大鼠梨状皮层存活神经细胞情况和退行性神经元数目.结果 Nissl染色结果显示,SE组大鼠梨状皮层大量神经细胞丢失,胞体结构完整性遭到破坏.FJG染色结果显示,SE组FJC阳性细胞数目明显增多;经PDTC干预后可明显改善梨状皮层神经损伤,FJC阳性细胞数明显减少,差异有统计学意义(P<0.05).结论 PDTC对毛果芸香碱诱导癫痫大鼠梨状皮层损伤具有明显的保护性作用.

  6. Effects of Carbamazepineon on the Hippocampal BrdU/NeuN space memory in Lithium Pilocarpine-induced Rat Model of Seizure%卡马西平对成年癫癎大鼠海马齿状回BrdU/NeuN表达水平及其对空间记忆的影响

    Institute of Scientific and Technical Information of China (English)

    权青云; 杨嫣华; 李宏图; 李峰; 赵晓娟; 林芝惠

    2012-01-01

    Objective To study the effects of carbamazepinc on the expression of BrdU/NcuN in the hippocampal dentate gyrus of adult rats after lithium pilocarpinc induced seizures,and its effect on space memory. Methods The adult rats were injected with lithium and pilocarpinc to induce status cpilcpticus. Bromodc-oxyuridinc was injected to label the newborn inherent neural precursor cells in hippocampus dentate gyrus. By using immunofluorcsccncc, the expression of BrdU/NcuN was observed on the 28th days after injection of Br-dU. Space memory function performance was assessed with behavioral analysis. Results (1) Carbamazepinc increased the number of matured neurons in the dentate gyrus in the rat model of status cpilcpticus(P?). 05). (2)Carbamazepinc improved space memory of adult rats after lithium pilocarpinc induced seizures (P<0. 01). Conclusions The effects of carbamazepinc on the expression of BrdU/NcuN in the hippocampal dentate gyrus of adult rats after lithium pilocarpinc induced seizures arc possibly one of the mechanisms of improving epilepsy space memory function.%目的 研究卡马西平对成年癫癎大鼠海马齿状回新生神经元的影响及其与空间记忆之间的关系.方法 采用氯化锂和匹罗卡品联合诱导大鼠癫痫 间模型,利用5-溴脱氧尿苷嘧啶与神经元核性蛋白双标记观察海马齿状回内源性神经前体细胞分化为成熟神经元的情况;利用行为学分析评价大鼠的空间记忆.结果 (1)卡马西平可增加癫癎大鼠海马齿状回新生成熟神经元的数量(P<0.05);(2)卡马西平对癫癎大鼠的空间记忆有明显改善作用(P<0.01).结论 卡马西平增加癫癎大鼠海马齿状回新生成熟神经元形成,是其改善癫癎大鼠空间记忆的可能机制之一.

  7. The Increase of Endogenous Digitalis-like Factor Release and Inhabitation of Renal Cortical Na+-K+ ATPase with Intracerebroventricular Administration of Pilocarpin%侧脑室注射匹罗卡品引起内源性洋地黄素释放抑制肾皮质Na+-K+ ATP酶

    Institute of Scientific and Technical Information of China (English)

    王勇; 张成标

    2007-01-01

    目的:探讨激活SD大鼠脑内胆碱能M受体对内源性洋地黄素(EDLF)释放及肾皮质Na+-K+ ATP酶活性的影响.方法:侧脑室注射M受体激动剂匹罗卡品,放射免疫法测定血清EDLF水平,化学比色法测定肾皮质Na+-K+ ATP酶活性.结果:①侧脑室注射匹罗卡品引起大鼠血清EDLF水平显著升高(P<0.01),肾皮质Na+-K+ ATP酶活性抑制(P<0.01).②侧脑室注射阿托品可阻断匹罗卡品引起的血清EDLF水平升高和肾皮质Na+-K+ ATP酶活性抑制效应.结论:脑内胆碱能系统可通过M受体介导参与EDLF释放的调节.

  8. Quantitative changes and axonal sprouting of PV positive interneurons in the hippocampus of epileptic rats induced by pilocarpine%匹罗卡品致痫大鼠的海马PV中间神经元数目变化及其轴突出芽

    Institute of Scientific and Technical Information of China (English)

    龙莉莉; 肖波; 宋延民; 易芳; 李蜀渝; 龙小艳; 陈锶

    2011-01-01

    目的 探讨PV(parvalbumin,微清蛋白)中间神经元在颞叶癫痫的发生和痫性损伤后脑组织的自我修复中的作用.方法 建立匹罗卡品致痫模型.免疫组化方法检测各时间点海马不同区域PV中间神经元的数目变化及其轴突出芽情况;结合FJB(Fluoro-Jade B)行免疫荧光双标特异性检测SE(status epilepticus,癫痫持续状态)后60 d海马不同区域PV中间神经元及其轴树突的变性情况.结果 实验组海马门区及CA区域PV阳性神经元数目在SE后早期均无明显变化(P>0.05).门区PV神经元在SE后15 d时数目下降(对照组17.1±2.67,实验组15.0±3.06,P<0.05),在SE后60 d明显下降(对照组16.6±2.75,实验组9.7±2.88,P<0.01).海马CA1区PV中间神经元数目在SE后60 d明显增多(对照组4.1±0.86,实验组6.0±1.18,P<0.01),可见相应增多的PV阳性纤维.海马CA3区PV阳性神经元数目在SE后60d明显增多(对照组4.6±0.89,实验组6.1±1.64,P<0.01),并可见相应增多的PV阳性纤维.各亚区均未见明显亦被FJB标记的PV中间神经元.结论 海马PV中间神经元的数目变化及其轴突出芽可能在颞叶癫痫的发生和痫性损伤后脑组织的自我修复中起重要作用,PV中间神经元的缺失可能与细胞变性死亡无关.%Objective To investigate the roles of PV positive interneurons in the generation of temporal lobe epilepsy and the compensation of the brain tissue after seisures. Methods Pitocarpine-induced animal model was founded.Immunohistochemistry method was used to detect number changes and axonal sprouting of PV positive interneurons in different domains of the hippocampus at different time points. Degeneration of PV positive interneurons and their neurophils on 60d after epileptic status( SE ) was detected by the technique of double immunofluorescence combined with FJB. Results No significant changes of PV-IR neurons was present in the hilus and CA regions in the acute and resting phase( P > 0. 05 ).The number of hilar PV-IR neurons decreased at 15days post-SE ( control 17. 1 ± 2. 67, post-SE 15.0 ± 3.06, P < 0. 05 ),and decreased significantly at 60days post-SE( control 16.6 ±2. 75 ,post-SE 9. 7 ±2. 88 ,P <0. 01 ). PV-IR neurons in CA1 region increased significantly at 60days post-SE( control 4. 1 ±0. 86 ,post-SE 6. 0 ± 1. 18 ,P <0. 01 ) , and increased PV-IR fibers were also observed. PV-IR neurons in CA3 region increased significantly at 60days post-SE( control 4. 6 ±0. 89 ,postSE 6. 1 ± 1.64,P <0. 01 ) , with correspondingly increased PV-IR fibers also seen. We examined PV-IR and FJB doublestaining in serial sections at 60d post-SE. Double-immunofluorescenee revealed there were no neurons double-labeled for Fluoro-Jade B and PV in each subfield. Conclusions Quantitative changes of PV interneurons and their axonal sprouting may play an important role in the generation of temporal lobe epilepsy and the compensation of the brain tissue after seizures, loss of PV interneurons may be not due to the cell's degeneration and death.

  9. 匹罗卡品致疒间大鼠海马GAP-43与TrkB基因表达及其意义%Expression and its significance of GAP-43 and TrkB mRNA in the rat hippocampus after epilepsy induced by pilocarpine

    Institute of Scientific and Technical Information of China (English)

    李国良; 肖波; 谢光洁; 章蓓; 李昌奇; 伍校琼

    2003-01-01

    目的探讨生长相关蛋白(GAP-43)和脑源性神经营养因子(BDNF)受体TrkB基因在匹罗卡品致疒间大鼠海马的表达及其意义.方法应用原位杂交组织化学方法研究匹罗卡品(PILO)致疒间大鼠海马GAP-43及TrkB mRNA表达的变化.结果匹罗卡品致癫疒间持续状态后3~6小时,海马齿状回颗粒细胞、CA3区及CA1区锥体细胞层TrkB mRNA表达显著高于对照组(P<0.01或0.05);在慢性期第7~30天,呈现第2次表达增强.致疒间后6~12小时,正常状态下并不表达GAP-43的大鼠海马颗粒细胞其GAP-43 mRNA表达较对照组显著增高(P<0.01),24小时~7天表达减少,在癫疒间慢性期表达再次高于对照组.结论 GAP-43及TrkB是颞叶癫疒间病理基础--海马苔藓纤维出芽的重要分子机制;BDNF对苔藓纤维的作用部分是通过GAP-43实现的.

  10. Number changes and axonal sprouting of somatostatin positive interneurons in the hippocampus of pilocarpine-induced epileptic rats%匹罗卡品致(癎)大鼠海马中表达生长抑素的中间神经元数目变化及其轴突出芽

    Institute of Scientific and Technical Information of China (English)

    冯莉; 龙莉莉; 肖波; 龙小艳; 李蜀渝; 易芳; 陈锶; 吴小妹

    2009-01-01

    Objective To investigate the roles of somatostatin(SS)positive intemeurons in the development and compensation of temporal lobe epilepsy.Methods Piloearpine-induced epilepsy rat model was established.Immunohistochemistry method was used to detect number changes and axonal sprouting of SS positive intemeurons in different domains of the hippocampus at difierent time points.Degeneration of SS positive interneurons and their neurophils were detected by the double immunofluorescence staining with SS and Fluoro-Jade B(FJB)at 7 and 60 days after status epilepticus (SE).Results In the exoerimental rat group,the number of SS positive neurons decreased in each hippocampal domain,and it reached the lowest at 7 days post-SE(There were 11.1±3.3 in hilus,2.8±0.9 in CA1region and 1.8±0.7 in CA1region,t=13.519,9.644 and 8.808,all P<0.01).In chronic phase,the number of SS neurons gradually recovered,and exceeded the control group in CA1 area at 60 days post-SE(12.8±1.5 vs 8.8±1.3,t=-4.506,P<0.01),however,the number of SS neurons in the hilus(25.5±4.6)and CA1 area(4.8±0.8)remained significantly less than normal levels(t value were 4.691 and 3.953.both P<0.01).Increased SS positive fibers were found in the lacunosum-molecular (1m)layer and outer molecular layer of dentate gyrus after 30 days post-SE,and numerous SS positive fibers were seen threnghout the layers of area CA1 at 60 days post-SE.Double immunofluuorescence revealed that a few SS positive interneurons and fibers were also labeled by FJB in area CA1 at 7 days post-SE and in CA domain/hilus at 60 days post-SE.Conclusions SS intemeurons loss plays an important role in the development of temporal lobe epilepsy.The loss is partially caIlsed by the degeneration and death of neurons;SS positive neurophils increase within area CA1 in chronic phase may play a significant role in the generation and compensation of temporal lobe epilepsy.%目的 探讨表达生长抑素(SS)的中间神经元在颞叶癫(癎)的发生和自我修复中的作用.方法 建立匹罗卡品致(癎)大鼠模型.免疫组织化学方法检测各时间点海马不同区域SS中间神经元的数目变化及其轴突出芽情况;结合Fluoro-Jade B(FJB)行免疫荧光双标方法特异性检测大鼠癫(癎)持续状态(status epilepticus,SE)后7 d和60 d海马不同区域SS中间神经元及其轴、树突的变性情况.结果 实验组大鼠海马各区Ss阳性神经元数目均在SE后7 d减至最低(门区11.1±3.3,CA1区2.8±0.9,CA1区1.8±0.7,t=13.519、9.644、8.808,均P<0.01),慢性期开始部分恢复,SE后60 d时CA1区SS神经元数目(12.8±1.5)超过对照组(8.8±1.3,t=-4.506,P<0.01),但门区和CA3区SS神经元数目(分别为25.5±4.6和4.8±0.8)仍明显低于正常水平(t=4.691、3.953,P<0.01);sE后30 d大鼠海马回腔隙.分子层(lacunosum-moleculare,lm)和齿状回外分子层出现大量SS染色阳性纤维,60 d时海马CA1区全层均可见大量增多的SS阳性纤维.实验组中SE后7 d的海马CA1区、60 d的CA1、CA1区和门区均可观察到少部分被FJB染色的SS中间神经元胞体及其轴、树突.结论 SS中间神经元的缺失在颞叶癫(癎)的发生中起重要作用,其缺失部分是由于神经元的变性死亡所致;慢性期CA,区SS阳性纤维大量增多,可能在颞叶癫(癎)的发生和自我修复中起重要作用.

  11. 匹罗卡品致痫大鼠海马神经肽Y中间神经元数目变化及其轴突出芽%Number changes and axonal sprouting of neuropeptide Y interneurons in the hippocampus of pilocarpine-induced rats

    Institute of Scientific and Technical Information of China (English)

    吴志国; 龙莉莉; 肖波; 陈锶; 易芳

    2009-01-01

    目的:探讨神经肽Y(neuropeptide Y,NPY)中间神经元在颞叶癫痫的发生和自我修复中的作用.方法:建立匹罗卡品致痫模型,应用免疫组织化学技术动态观察大鼠海马NPY中间神经元的数目变化及其轴突出芽.结果:实验组大鼠腹腔注射氯化锂-匹罗卡品后,癫痫持续状态(status epilepticus, SE)诱发成功率为92.9%,死亡率19.2%.免疫组织化学结果显示,实验组大鼠海马门区NPY中间神经元数目在SE后下降,至7 d时降至最低(P0.05);SE后30 d齿状回分子层可见增多的NPY阳性纤维.结论:NPY中间神经元在不同部位不同时段对颞叶癫痫所致损伤的敏感性不同,NPY中间神经元的缺失在颞叶癫痫发生中起重要作用,NPY中间神经元的轴突出芽在颞叶癫痫的自我修复中起作用.

  12. Tonic Pupil Following Traumatic Hyphema: Case Report

    Directory of Open Access Journals (Sweden)

    Şaban Gönül

    2013-04-01

    Full Text Available A 7-year-old girl was admitted to our clinic after crash injury with air gun pellet in her right eye. There was an intense anterior chamber reaction and hyphema on the biomicroscopic examination. During the control examination after treatment of hyphema including cyclopentolate 1%, prednisolone acetate 1% and lomefloxacin 0.3%, anisocoria and mydriasis in the right eye were observed and the difference between both pupils was less in darkness. The case was diagnosed as tonic pupil following trauma, and diluted pilocarpine 0.125% test was performed. On diluted pilocarpine test, right pupil responded excessively to pilocarpine compared with the other pupil. As in our case, in cases with anisocoria following blunt trauma to orbit, tonic pupil should be keep in mind. (Turk J Ophthalmol 2013; 43: 132-4

  13. Knockdown of Carboxypeptidase A6 in Zebrafish Larvae Reduces Response to Seizure-Inducing Drugs and Causes Changes in the Level of mRNAs Encoding Signaling Molecules.

    Directory of Open Access Journals (Sweden)

    Mark William Lopes

    Full Text Available Carboxypeptidase A6 (CPA6 is an extracellular matrix metallocarboxypeptidase that modulates peptide and protein function by removal of hydrophobic C-terminal amino acids. Mutations in the human CPA6 gene that reduce enzymatic activity in the extracellular matrix are associated with febrile seizures, temporal lobe epilepsy, and juvenile myoclonic epilepsy. The characterization of these human mutations suggests a dominant mode of inheritance by haploinsufficiency through loss of function mutations, however the total number of humans with pathologic mutations in CPA6 identified to date remains small. To better understand the relationship between CPA6 and seizures we investigated the effects of morpholino knockdown of cpa6 mRNA in zebrafish (Danio rerio larvae. Knockdown of cpa6 mRNA resulted in resistance to the effect of seizure-inducing drugs pentylenetetrazole and pilocarpine on swimming behaviors. Knockdown of cpa6 mRNA also reduced the levels of mRNAs encoding neuropeptide precursors (bdnf, npy, chga, pcsk1nl, tac1, nts, edn1, a neuropeptide processing enzyme (cpe, transcription factor (c-fos, and molecules implicated in glutamatergic signaling (grin1a and slc1a2b. Treatment of zebrafish embryos with 60 mM pilocarpine for 1 hour led to reductions in levels of many of the same mRNAs when measured 1 day after pilocarpine exposure, except for c-fos which was elevated 1 day after pilocarpine treatment. Pilocarpine treatment, like cpa6 knockdown, led to a reduced sensitivity to pentylenetetrazole when tested 1 day after pilocarpine treatment. Taken together, these results add to mounting evidence that peptidergic systems participate in the biological effects of seizure-inducing drugs, and are the first in vivo demonstration of the molecular and behavioral consequences of cpa6 insufficiency.

  14. Growth hormone deficiency and hyperthermia during exercise

    DEFF Research Database (Denmark)

    Juul, A; Hjortskov, N; Jepsen, Leif

    1995-01-01

    Sweat secretion is often disturbed in patients with GH secretory disorders. Hyperhidrosis is a classic feature of acromegaly, and it has recently been shown that GH-deficient patients exhibit decreased sweating capacity after pilocarpine stimulation of the skin. Thus, patients with GH-deficiency ......Sweat secretion is often disturbed in patients with GH secretory disorders. Hyperhidrosis is a classic feature of acromegaly, and it has recently been shown that GH-deficient patients exhibit decreased sweating capacity after pilocarpine stimulation of the skin. Thus, patients with GH...

  15. Twenty-four-hour intraocular pressure patterns in a symptomatic patient after ab interno trabeculotomy surgery

    Directory of Open Access Journals (Sweden)

    Mansouri K

    2014-11-01

    Full Text Available Kaweh Mansouri,1 Felipe A Medeiros,2 Robert N Weinreb2 1Glaucoma Sector, Department of Ophthalmology, Geneva University Hospitals, Geneva, Switzerland; 2Hamilton Glaucoma Center and Department of Ophthalmology, University of California, San Diego, La Jolla, CA, USA Abstract: We report the results of repeated ambulatory continuous 24-hour intraocular pressure (IOP monitoring with a contact lens sensor (CLS in a glaucoma patient with ocular pain after ab interno trabeculotomy (Trabectome™ surgery. Our findings show that a combined prostaglandin–pilocarpine treatment reduced nighttime IOP peaks and relieved the patient’s symptoms. Keywords: 24-hour, Trabectome contact lens sensor, prostaglandin–pilocarpine treatment

  16. Ocular hypotensive effects of a Rho-associated protein kinase inhibitor in rabbits

    Directory of Open Access Journals (Sweden)

    Kamaruddin MI

    2017-03-01

    Full Text Available Muhammad Irfan Kamaruddin,1,2 Momoko Nakamura-Shibasaki,1 Yu Mizuno,1 Yoshiaki Kiuchi1 1Department of Ophthalmology and Visual Sciences, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan; 2Department of Ophthalmology, Hasanuddin University, Makassar, South Sulawesi, Indonesia Purpose: Ripasudil is a novel Rho-associated protein kinase inhibitor that is used to treat ocular hypertension. However, the comparison of the intraocular pressure (IOP-lowering effects between ripasudil alone and other ocular hypotensive drugs has not been studied thoroughly. The purpose of this study is to examine the ocular hypotensive effects of 0.4% ripasudil, 2% pilocarpine, 0.5% timolol and 0.1% dorzolamide in rabbits. We also studied the IOP changes when 0.4% ripasudil was combined with 2% pilocarpine, 0.5% timolol or 0.1% dorzolamide. Methods: One drop of saline solution, 0.4% ripasudil, 0.5% timolol, 2% pilocarpine or 1% dorzolamide or a combination of these agents was applied topically to the left eyes of eight healthy albino rabbits. Posttreatment changes in the IOP of albino rabbits were monitored using a rebound tonometer over a 5-h time course. Changes in IOP after application of saline served as the control. One-way analysis of variance and Dunnett’s post hoc tests were used for statistical analyses. Results: After topical instillation, 0.4% ripasudil resulted in significant decreases in IOP at 0.5 and 1 h compared with the control group. Treatment with timolol, pilocarpine or dorzolamide had no significant effect on IOP. Treatment with timolol, pilocarpine or dorzolamide in combination with ripasudil resulted in significant reductions in IOP at 1 h. However, none of these agents enhanced the IOP-lowering effects of ripasudil. Conclusion: Ripasudil has stronger IOP-lowering effects than timolol, pilocarpine or dorzolamide hypotensive agents in our rabbit model. Addition of timolol, pilocarpine or dorzolamide did not enhance

  17. Growth hormone deficiency and hyperthermia during exercise

    DEFF Research Database (Denmark)

    Juul, A; Hjortskov, N; Jepsen, Leif

    1995-01-01

    Sweat secretion is often disturbed in patients with GH secretory disorders. Hyperhidrosis is a classic feature of acromegaly, and it has recently been shown that GH-deficient patients exhibit decreased sweating capacity after pilocarpine stimulation of the skin. Thus, patients with GH-deficiency ......Sweat secretion is often disturbed in patients with GH secretory disorders. Hyperhidrosis is a classic feature of acromegaly, and it has recently been shown that GH-deficient patients exhibit decreased sweating capacity after pilocarpine stimulation of the skin. Thus, patients with GH...

  18. Plasma cysteine/cystine redox couple disruption in animal models of temporal lobe epilepsy

    Directory of Open Access Journals (Sweden)

    Li-Ping Liang

    2016-10-01

    Full Text Available Currently the field of epilepsy lacks peripheral blood-based biomarkers that could predict the onset or progression of chronic seizures following an epileptogenic injury. Thiol/disulfide ratios have been shown to provide a sensitive means of assessing the systemic redox potential in tissue and plasma. In this study, we utilized a rapid, simple and reliable method for simultaneous determination of several thiol-containing amino acids in plasma using HPLC with electrochemical detection in kainic acid (KA and pilocarpine rat models of epilepsy. In contrast to GSH and GSSG levels, the levels of cysteine (Cys were decreased by 42% and 62% and cystine (Cyss were increased by 46% and 23% in the plasma of KA- and pilocarpine-injected rats, respectively after 48 h. In chronically epileptic rats, plasma cysteine was decreased by 40.4% and 37.7%, and plasma GSSG increased by 33.8% and 35.0% following KA and pilocarpine, respectively. Treatment of rats with a catalytic antioxidant, 60 min after KA or pilocarpine significant attenuated the decrease of plasma Cys/Cyss ratios at the 48 h time point in both models. These observations suggest that the decreased cysteine and ratio of Cys/Cyss in plasma could potentially serve as redox biomarkers in temporal lobe epilepsy.

  19. Pupillary capture of the optic in secondary piggyback implantation.

    Science.gov (United States)

    Gayton, J L; Sanders, V; Van Der Karr, M

    2001-09-01

    Two patients who received a minus-power intraocular lens implanted as a secondary piggyback to correct pseudophakic myopia experienced pupillary optic capture following dilation in the early postoperative period. Both cases were successfully managed by pressing the optic back into the ciliary sulcus and constricting the pupil with pilocarpine.

  20. Linking crenarchaeal and bacterial nitrification to anammox in the Black Sea

    DEFF Research Database (Denmark)

    Lam, Phyllis; Jensen, Marlene Mark; Lavik, Gaute

    2007-01-01

    Generation of dentate granule cells and its modulation by glutamate receptor antagonists, growth factors and pilocarpine-induced seizure-like activity was investigated in rat hippocampal slice cultures derived from 1-week-old rats and grown for 2 weeks. Focussing on the dentate granule cell layer...

  1. Muscarinic receptor stimulation increases tolerance of rat salivary gland function to radiation damage

    NARCIS (Netherlands)

    Coppes, RP; Vissink, A; Zeilstra, LJW; Konings, AWT

    1997-01-01

    Purpose: To investigate if muscarinic receptor-stimulated activation of the PLC/PIP2 second messenger pathway prior to irradiation increases the radiotolerance of rat salivary gland. Materials and methods: Rats were treated with pilocarpine, methacholine, reserpine, methacholine plus reserpine, or

  2. DIFFERENT EFFECTS OF MUSCARINIC AGONISTS IN RAT SUPERIOR CERVICAL-GANGLION AND HIPPOCAMPAL SLICES

    NARCIS (Netherlands)

    BODDEKE, HWGM

    1991-01-01

    In this study the effects of muscarinic antagonists and agonists on M1 muscarinic receptors in the isolated rat superior cervical ganglion and the rat hippocampal slice were investigated. Oxotremorine and APE but not pilocarpine, McN-A-343 or 4-Cl-McN-A-343 induced small M2 muscarinic

  3. Circadian clock-dependent increase in salivary IgA secretion modulated by sympathetic receptor activation in mice.

    Science.gov (United States)

    Wada, Misaki; Orihara, Kanami; Kamagata, Mayo; Hama, Koki; Sasaki, Hiroyuki; Haraguchi, Atsushi; Miyakawa, Hiroki; Nakao, Atsuhito; Shibata, Shigenobu

    2017-08-18

    The salivary gland is rhythmically controlled by sympathetic nerve activation from the suprachiasmatic nucleus (SCN), which functions as the main oscillator of circadian rhythms. In humans, salivary IgA concentrations reflect circadian rhythmicity, which peak during sleep. However, the mechanisms controlling this rhythmicity are not well understood. Therefore, we examined whether the timing of parasympathetic (pilocarpine) or sympathetic (norepinephrine; NE) activation affects IgA secretion in the saliva. The concentrations of saliva IgA modulated by pilocarpine activation or by a combination of pilocarpine and NE activation were the highest in the middle of the light period, independent of saliva flow rate. The circadian rhythm of IgA secretion was weakened by an SCN lesion and Clock gene mutation, suggesting the importance of the SCN and Clock gene on this rhythm. Adrenoceptor antagonists blocked both NE- and pilocarpine-induced basal secretion of IgA. Dimeric IgA binds to the polymeric immunoglobulin receptor (pIgR) on the basolateral surface of epithelial cells and forms the IgA-pIgR complex. The circadian rhythm of Pigr abundance peaked during the light period, suggesting pIgR expression upon rhythmic secretion of IgA. We speculate that activation of sympathetic nerves during sleep may protect from bacterial access to the epithelial surface through enhanced secretion of IgA.

  4. Neuroprotection and Anti-Epileptogenesis with a Mitochondria-Targeted Antioxidant

    Science.gov (United States)

    2013-12-01

    Epileptogenesis with a Mitochondria-Targeted Antioxidant PRINCIPAL INVESTIGATOR: Jeffrey H. Goodman, Ph.D. CONTRACTING ORGANIZATION: Research...antiepiletogenic properties of a mitochondrial-targeted antioxidant , SS-31 using the pilocarpine (Pilo) model of status epilepticus (SE), the kindling seizure...SUBJECT TERMS Neuroprotection, antiepileptogenesis, antioxidant 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES

  5. Environ: E00402 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available 4] Pilocarpus jaborandi, Pilocarpus pennatifolius [TAX:77017], Pilocarpus [TAX:77016] Rutaceae (rue family) ...Pilocarpus small leaf Major component: Pilocarpine [CPD:C07474] Crude drugs [BR:br08305] Dicot plants: rosids Rutaceae (rue family) E00402 Jaborandi leaf ...

  6. ACUTE IRRADIATION EFFECTS ON MORPHOLOGY AND FUNCTION OF RAT SUBMANDIBULAR GLANDS

    NARCIS (Netherlands)

    VISSINK, A; KALICHARAN, D; GRAVENMADE, EJ; JONGEBLOED, WL; LIGEON, EE; NIEUWENHUIS, P; KONINGS, AWT

    1991-01-01

    In this study the morphologic and functional changes were compared after irradiation (single dose, 15 Gy) of rat submandibular salivary glands. Before and 1-10 days after local irradiation of the gland region, samples of submandibular saliva were collected after stimulation by pilocarpine. At the sa

  7. Muscarinic receptor stimulation increases tolerance of rat salivary gland function to radiation damage

    NARCIS (Netherlands)

    Coppes, RP; Vissink, A; Zeilstra, LJW; Konings, AWT

    1997-01-01

    Purpose: To investigate if muscarinic receptor-stimulated activation of the PLC/PIP2 second messenger pathway prior to irradiation increases the radiotolerance of rat salivary gland. Materials and methods: Rats were treated with pilocarpine, methacholine, reserpine, methacholine plus reserpine, or a

  8. Neo-Timm staining in the thalamus of chronically epileptic rats

    Directory of Open Access Journals (Sweden)

    Hamani C.

    2005-01-01

    Full Text Available The thalamus is an important modulator of seizures and is severely affected in cholinergic models of epilepsy. In the present study, chronically epileptic rats had their brains processed for neo-Timm and acetylcholinesterase two months after the induction of status epilepticus with pilocarpine. Both controls and pilocarpine-treated animals presented neo-Timm staining in the anterodorsal nucleus, laterodorsal nucleus, reticular nucleus, most intralaminar nuclei, nucleus reuniens, and rhomboid nucleus of the thalamus, as well as in the zona incerta. The intensity of neo-Timm staining was similar in control and pilocarpine-treated rats, except for the nucleus reuniens and the rhomboid nucleus, which had a lower intensity of staining in the epileptic group. In animal models of temporal lobe epilepsy, zinc seems to modulate glutamate release and to decrease seizure activity. In this context, a reduction of neo-Timm-stained terminals in the midline thalamus could ultimately result in an increased excitatory activity, not only within its related nuclei, but also in anatomical structures that receive their efferent connections. This might contribute to the pathological substrate observed in chronic pilocarpine-treated epileptic animals.

  9. Effect of P2X(7) receptor knockout on exocrine secretion of pancreas, salivary glands and lacrimal glands

    DEFF Research Database (Denmark)

    Novak, Ivana; Jans, Ida M; Wohlfahrt, Louise

    2010-01-01

    the P2X(7) receptors affect fluid secretion in pancreas, salivary glands and tear glands. We monitored gland secretions in in vivo preparations of wild-type and P2X(7)(-/-) (Pfizer) mice stimulated with pilocarpine. In cell preparations from pancreas, parotid and lacrimal glands we measured ATP release...

  10. Status Epilepticus Induced Spontaneous Dentate Gyrus Spikes: In Vivo Current Source Density Analysis.

    Science.gov (United States)

    Flynn, Sean P; Barriere, Sylvain; Barrier, Sylvain; Scott, Rod C; Lenck-Santini, Pierre-Pascal; Holmes, Gregory L

    2015-01-01

    The dentate gyrus is considered to function as an inhibitory gate limiting excitatory input to the hippocampus. Following status epilepticus (SE), this gating function is reduced and granule cells become hyper-excitable. Dentate spikes (DS) are large amplitude potentials observed in the dentate gyrus (DG) of normal animals. DS are associated with membrane depolarization of granule cells, increased activity of hilar interneurons and suppression of CA3 and CA1 pyramidal cell firing. Therefore, DS could act as an anti-excitatory mechanism. Because of the altered gating function of the dentate gyrus following SE, we sought to investigate how DS are affected following pilocarpine-induced SE. Two weeks following lithium-pilocarpine SE induction, hippocampal EEG was recorded in male Sprague-Dawley rats with 16-channel silicon probes under urethane anesthesia. Probes were placed dorso-ventrally to encompass either CA1-CA3 or CA1-DG layers. Large amplitude spikes were detected from EEG recordings and subject to current source density analysis. Probe placement was verified histologically to evaluate the anatomical localization of current sinks and the origin of DS. In 9 of 11 pilocarpine-treated animals and two controls, DS were confirmed with large current sinks in the molecular layer of the dentate gyrus. DS frequency was significantly increased in pilocarpine-treated animals compared to controls. Additionally, in pilocarpine-treated animals, DS displayed current sinks in the outer, middle and/or inner molecular layers. However, there was no difference in the frequency of events when comparing between layers. This suggests that following SE, DS can be generated by input from medial and lateral entorhinal cortex, or within the dentate gyrus. DS were associated with an increase in multiunit activity in the granule cell layer, but no change in CA1. These results suggest that following SE there is an increase in DS activity, potentially arising from hyperexcitability along the

  11. Expressional analysis of the astrocytic Kir4.1 channel in a pilocarpine–induced temporal lobe epilepsy model

    Science.gov (United States)

    Nagao, Yuki; Harada, Yuya; Mukai, Takahiro; Shimizu, Saki; Okuda, Aoi; Fujimoto, Megumi; Ono, Asuka; Sakagami, Yoshihisa; Ohno, Yukihiro

    2013-01-01

    The inwardly rectifying potassium (Kir) channel Kir4.1 in brain astrocytes mediates spatial K+ buffering and regulates neural activities. Recent studies have shown that loss-of-function mutations in the human gene KCNJ10 encoding Kir4.1 cause epileptic seizures, suggesting a close relationship between the Kir4.1 channel function and epileptogenesis. Here, we performed expressional analysis of Kir4.1 in a pilocarpine-induced rat model of temporal lobe epilepsy (TLE) to explore the role of Kir4.1 channels in modifying TLE epileptogenesis. Treatment of rats with pilocarpine (350 mg/kg, i.p.) induced acute status epilepticus, which subsequently caused spontaneous seizures 7–8 weeks after the pilocarpine treatment. Western blot analysis revealed that TLE rats (interictal condition) showed significantly higher levels of Kir4.1 than the control animals in the cerebral cortex, striatum, and hypothalamus. However, the expression of other Kir subunits, Kir5.1 and Kir2.1, remained unaltered. Immunohistochemical analysis illustrated that Kir4.1-immunoreactivity-positive astrocytes in the pilocarpine-induced TLE model were markedly increased in most of the brain regions examined, concomitant with an increase in the number of glial fibrillary acidic protein (GFAP)-positive astrocytes. In addition, Kir4.1 expression ratios relative to the number of astrocytes (Kir4.1-positive cells/GFAP-positive cells) were region-specifically elevated in the amygdala (i.e., medial and cortical amygdaloid nuclei) and sensory cortex. The present study demonstrated for the first time that the expression of astrocytic Kir4.1 channels was elevated in a pilocarpine-induced TLE model, especially in the amygdala, suggesting that astrocytic Kir4.1 channels play a role in modifying TLE epileptogenesis, possibly by acting as an inhibitory compensatory mechanism. PMID:23922547

  12. Intervention of laser periphery iridectomy to posterior iris bowing in high myopic eyes

    Institute of Scientific and Technical Information of China (English)

    WANG Hong-tao; WANG Ning-li; LI Shu-ning

    2012-01-01

    Background For some high myopic patients with posterior iris bowing,laser periphery iridectomy should be performed pre-operation to prevent pupil block glaucoma if these patients would have phakic intraocular lens implantation to correct high myopia.So we had the opportunity to analysis the influence of laser iridectomy on posterior iris bowing.Methods Eighteen high myopic patients with posterior iris bowing (11 males and 7 females) were involved in the study in Beijing Tongren Eye Center from March 2008 to July 2008.Phakic intraocular lens were implanted to correct their ametropia.The mean age was (32±6) years (range,25-40 years).The center anterior chamber depth,the pupil diameter,the posterior iris bowing depth and the anterior chamber angle were measured with anterior segment coherence tomography (AS-OCT) under the normal condition,myosis condition induced by 2% pilocarpine,laser periphery iridectomy after myosis,and 2% pilocarpine eluting condition respectively.Results There was no significant difference of center anterior chamber depth under the four conditions (P=0.512).The pupil constricted after pilocarpine (P=0.001).After laser iridectomy performed and pilocarpine eluted,posterior iris bowing depth reduced more than that in normal condition (P=0.003).The anterior chamber angle reduced significantly after laser periphery iridectomy and pilocarpine eluted (P=0.012).Conclusion Laser periphery iridectomy can reduce the posterior iris bowing,which might be due to the change in aqueous circulate pathway.

  13. Status Epilepticus Induced Spontaneous Dentate Gyrus Spikes: In Vivo Current Source Density Analysis.

    Directory of Open Access Journals (Sweden)

    Sean P Flynn

    Full Text Available The dentate gyrus is considered to function as an inhibitory gate limiting excitatory input to the hippocampus. Following status epilepticus (SE, this gating function is reduced and granule cells become hyper-excitable. Dentate spikes (DS are large amplitude potentials observed in the dentate gyrus (DG of normal animals. DS are associated with membrane depolarization of granule cells, increased activity of hilar interneurons and suppression of CA3 and CA1 pyramidal cell firing. Therefore, DS could act as an anti-excitatory mechanism. Because of the altered gating function of the dentate gyrus following SE, we sought to investigate how DS are affected following pilocarpine-induced SE. Two weeks following lithium-pilocarpine SE induction, hippocampal EEG was recorded in male Sprague-Dawley rats with 16-channel silicon probes under urethane anesthesia. Probes were placed dorso-ventrally to encompass either CA1-CA3 or CA1-DG layers. Large amplitude spikes were detected from EEG recordings and subject to current source density analysis. Probe placement was verified histologically to evaluate the anatomical localization of current sinks and the origin of DS. In 9 of 11 pilocarpine-treated animals and two controls, DS were confirmed with large current sinks in the molecular layer of the dentate gyrus. DS frequency was significantly increased in pilocarpine-treated animals compared to controls. Additionally, in pilocarpine-treated animals, DS displayed current sinks in the outer, middle and/or inner molecular layers. However, there was no difference in the frequency of events when comparing between layers. This suggests that following SE, DS can be generated by input from medial and lateral entorhinal cortex, or within the dentate gyrus. DS were associated with an increase in multiunit activity in the granule cell layer, but no change in CA1. These results suggest that following SE there is an increase in DS activity, potentially arising from

  14. Effects of Modulating M3 Muscarinic Receptor Activity on Azoxymethane-Induced Liver Injury in Mice

    Science.gov (United States)

    Khurana, Sandeep; Jadeja, Ravirajsinh; Twadell, William; Cheng, Kunrong; Rachakonda, Vikrant; Saxena, Neeraj; Raufman, Jean-Pierre

    2013-01-01

    Previously, we reported that azoxymethane (AOM)-induced liver injury is robustly exacerbated in M3 muscarinic receptor (M3R)-deficient mice. We used the same mouse model to test the hypothesis that selective pharmacological modulation of M3R activity regulates the liver injury response. Initial experiments confirmed that giving a selective M3R antagonist, darifenacin, to AOM-treated mice mimicked M3R gene ablation. Compared to vehicle controls, mice treated with the M3R antagonist had reduced survival and increased liver nodularity and fibrosis. We next assessed AOM-induced liver injury in mice treated with a selective M3R agonist, pilocarpine. After pilocarpine treatment, stimulation of post-M3R signaling in the liver was evidenced by ERK and AKT activation. In contrast to the damaging effects of the M3R antagonist, administering pilocarpine to AOM-treated mice significantly attenuated hepatic stellate cell activation, collagen deposition, bile ductule proliferation, and liver fibrosis and nodularity. As anticipated from these findings, livers from pilocarpine-treated mice exhibited reduced expression of key players in fibrosis (α1 collagen, α-smooth muscle actin, TGF-β1, PGDF, TGF-β1R, PGDFR) and decreased mRNA levels for molecules that regulate extracellular matrix formation (TIMP-1, TIMP-2, MMP-2, MMP-13). Cleaved caspase-3, nitrotyrosine and BrdU immunostaining provided evidence that pilocarpine treatment reduced hepatocyte apoptosis and oxidative stress, while increasing hepatocyte proliferation. Collectively, these findings identify several downstream mechanisms whereby M3R activation ameliorates toxic liver injury. These novel observations provide a proof-of-principle that selectively stimulating M3R activation to prevent or diminish liver injury is a therapeutic strategy worthy of further investigation. PMID:23707755

  15. Status epilepticus induction has prolonged effects on the efficacy of antiepileptic drugs in the 6-Hz seizure model.

    Science.gov (United States)

    Leclercq, Karine; Kaminski, Rafal M

    2015-08-01

    Several factors may influence the efficacy of antiepileptic drugs (AEDs) in patients with epilepsy, and treatment resistance could be related to genetics, neuronal network alterations, and modification of drug transporters or targets. Consequently, preclinical models used for the identification of potential new, more efficacious AEDs should reflect at least a few of these factors. Previous studies indicate that induction of status epilepticus (SE) may alter drug efficacy and that this effect could be long-lasting. In this context, we wanted to assess the protective effects of mechanistically diverse AEDs in mice subjected to pilocarpine-induced SE in another seizure model. We first determined seizure thresholds in mice subjected to pilocarpine-induced SE in the 6-Hz model, 2 weeks and 8 weeks following SE. We then evaluated the protective effects of mechanistically diverse AEDs in post-SE and control animals. No major differences in 6-Hz seizure susceptibility were observed between control groups, while the seizure threshold of pilocarpine mice at 8 weeks after SE was higher than at 2 weeks and higher than in control groups. Treatment with AEDs revealed major differences in drug response depending on their mechanism of action. Diazepam produced a dose-dependent protection against 6-Hz seizures in control and pilocarpine mice, both at 2 weeks and 8 weeks after SE, but with a more pronounced increase in potency in post-SE animals at 2 weeks. Levetiracetam induced a potent and dose-dependent protection in pilocarpine mice, 2 weeks after SE, while its protective effects were observed only at much higher doses in control mice. Its potency decreased in post-SE mice at 8 weeks and was very limited (30% protection at the highest tested dose) in the control group. Carbamazepine induced a dose-dependent protection at 2 weeks in control mice but only limited effect (50% at the highest tested dose) in pilocarpine mice. Its efficacy deeply decreased in post-SE mice at 8 weeks

  16. Anticonvulsant Activity of Extracts of Plectranthus barbatus Leaves in Mice

    Directory of Open Access Journals (Sweden)

    Luciana Cristina Borges Fernandes

    2012-01-01

    Full Text Available Plectranthus barbatus is a medicinal plant used to treat a wide range of disorders including seizure. However, the anticonvulsant activity of this plant has not been studied in depth. We therefore sought to evaluate the anticonvulsant activity of a hydroalcoholic extract of P. barbatus leaves on seizures induced by strychnine sulphate (2.0 mg/kg and pilocarpine (600 mg/kg in mice. The extract was administered orally at 1, 10, 30, and 100 mg/kg. We report that the P. barbatus extract had marked anticonvulsant activity against strychnine-induced convulsions, but was quite ineffective against pilocarpine-induced convulsions. Further experiments will be required to identify the active molecules(s and their mechanism(s of action.

  17. Behavioral impairments in rats with chronic epilepsy suggest comorbidity between epilepsy and attention deficit/hyperactivity disorder.

    Science.gov (United States)

    Pineda, Eduardo; Jentsch, J David; Shin, Don; Griesbach, Grace; Sankar, Raman; Mazarati, Andrey

    2014-02-01

    Attention deficit/hyperactivity disorder (ADHD) is encountered among patients with epilepsy at a significantly higher rate than in the general population. Mechanisms of epilepsy-ADHD comorbidity remain largely unknown. We investigated whether a model of chronic epilepsy in rats produces signs of ADHD, and thus, whether it can be used for studying mechanisms of this comorbidity. Epilepsy was induced in male Wistar rats via pilocarpine status epilepticus. Half of the animals exhibited chronic ADHD-like abnormalities, particularly increased impulsivity and diminished attention in the lateralized reaction-time task. These impairments correlated with the suppressed noradrenergic transmission in locus coeruleus outputs. The other half of animals exhibited depressive behavior in the forced swimming test congruently with the diminished serotonergic transmission in raphe nucleus outputs. Attention deficit/hyperactivity disorder and depressive behavior appeared mutually exclusive. Therefore, the pilocarpine model of epilepsy affords a system for reproducing and studying mechanisms of comorbidity between epilepsy and both ADHD and/or depression.

  18. Subsensitivity to cholinoceptor stimulation of the human iris sphincter in situ following acute and chronic administration of cholinomimetic miotic drugs.

    Science.gov (United States)

    Smith, S. A.; Smith, S. E.

    1980-01-01

    1 Maximal pupillary miosis was obtained with single topical applications of 4 cholinomimetic drugs in therapeutic concentrations to normal human subjects. 2 When the pupil had recovered from the miosis, there remained a reduced light reflex response of 22.7% at 24 h after aceclidine, 18.0% at 31 h after pilocarpine, 10.3% at 48 h after physostigmine and 4.9% at 7 h after arecoline. 3 This reduced sensitivity to light was accompanied by an overshoot of the resting pupil diameter and, after aceclidine miosis, a reduced response to a second application of miotic. 4 Similar findings were observed in glaucoma patients following withdrawal of chronic pilocarpine therapy. 5 It is suggested that the slowly reversible after-effects of acute and chronic administration of cholinomimetic miotics can be explained by desensitization of iris sphincter cholinoceptors. PMID:6105002

  19. Controlled release gelatin hydrogels and lyophilisates with potential application as ocular inserts

    Energy Technology Data Exchange (ETDEWEB)

    Natu, Madalina V [Department of Chemical Engineering, University of Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra (Portugal); Sardinha, Jose P [Department of Chemical Engineering, University of Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra (Portugal); Correia, IlIdio J [Centro de Investigacao em Ciencias da Saude, Faculdade de Ciencias da Saude, Universidade da Beira Interior, Covilha (Portugal); Gil, M H [Department of Chemical Engineering, University of Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra (Portugal)

    2007-12-15

    Hydrogels and lyophilisates were obtained by chemical crosslinking of gelatin using N-hydroxysuccinimide and N, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride. The systems were characterized with respect to the degree of crosslinking, morphology, water uptake, in vitro drug release and biocompatibility studies. Pilocarpine hydrochloride, a drug for the treatment of glaucoma, was loaded by soaking in an aqueous solution containing the drug. In vitro, the released drug percentage varied between 29.2% and 99.2% in 8 h of study. The release data were fitted to the Korsmeyer-Peppas equation to calculate the release exponent, which indicated anomalous transport for the release of pilocarpine. The corneal endothelial cell culture tests indicated that the prepared biomaterials are not cytotoxic.

  20. Diet Composition Exacerbrates or Attenuates Soman Toxicity in Rats: Implied Metabolic Control of Nerve Agent Toxicity

    Science.gov (United States)

    2011-01-01

    approximated 50 % in the standard and choline-enriched diet groups, but equaled 87% in the ketogenic diet group. Body weight loss was significantly...epileptic disorders (6 Hz audiogenic seizures, pentylenetetrazole [PTZ], kainic acid [KA], pilocarpine, etc.), the KD has been shown to elevate seizure...elevated sugar intake (glucose or high fructose corn syrup in drinking water) exacerbates the toxicity of parathion poisoning, an organophos- phorus

  1. Efficacy of anti-inflammatory therapy in a model of acute seizures and in a population of pediatric drug resistant epileptics.

    Directory of Open Access Journals (Sweden)

    Nicola Marchi

    Full Text Available Targeting pro-inflammatory events to reduce seizures is gaining momentum. Experimentally, antagonism of inflammatory processes and of blood-brain barrier (BBB damage has been demonstrated to be beneficial in reducing status epilepticus (SE. Clinically, a role of inflammation in the pathophysiology of drug resistant epilepsies is suspected. However, the use anti-inflammatory drug such as glucocorticosteroids (GCs is limited to selected pediatric epileptic syndromes and spasms. Lack of animal data may be one of the reasons for the limited use of GCs in epilepsy. We evaluated the effect of the CG dexamethasone in reducing the onset and the severity of pilocarpine SE in rats. We assessed BBB integrity by measuring serum S100β and Evans Blue brain extravasation. Electrophysiological monitoring and hematologic measurements (WBCs and IL-1β were performed. We reviewed the effect of add on dexamethasone treatment on a population of pediatric patients affected by drug resistant epilepsy. We excluded subjects affected by West, Landau-Kleffner or Lennox-Gastaut syndromes and Rasmussen encephalitis, known to respond to GCs or adrenocorticotropic hormone (ACTH. The effect of two additional GCs, methylprednisolone and hydrocortisone, was also reviewed in this population. When dexamethasone treatment preceded exposure to the convulsive agent pilocarpine, the number of rats developing status epilepticus (SE was reduced. When SE developed, the time-to-onset was significantly delayed compared to pilocarpine alone and mortality associated with pilocarpine-SE was abolished. Dexamethasone significantly protected the BBB from damage. The clinical study included pediatric drug resistant epileptic subjects receiving add on GC treatments. Decreased seizure frequency (≥ 50% or interruption of status epilepticus was observed in the majority of the subjects, regardless of the underlying pathology. Our experimental results point to a seizure-reducing effect of

  2. Proceedings of the International Symposium on Subtypes of Muscarinic Receptors 2 (2nd) Held in Boston, Massachusetts on August 22-24 1985

    Science.gov (United States)

    1986-04-30

    pilocarpine CCh (1.0 mm,) stimulated incorporation of [2P] into and 4-(lo-chlorophenylcarbamyloxy)-2-butynyltrimethyl- synaptosomal phosphatidic acid in...evoked acid secretion, receptor subtype on gastric smooth muscle using selective antagonists, and have examined its regulation by guanine nucleotides...submucous plexus and their-roles in m-ucosal ion transport) 4 A. -~eli - 23 Amechanism of muscarinic excitation in dissociated smooth muscle cells

  3. Layer-specific modulation of entorhinal cortical excitability by presubiculum in a rat model of temporal lobe epilepsy

    OpenAIRE

    Abbasi, Saad; Sanjay S Kumar

    2015-01-01

    Temporal lobe epilepsy (TLE) is the most common form of epilepsy in adults and is often refractory to antiepileptic medications. The medial entorhinal area (MEA) is affected in TLE but mechanisms underlying hyperexcitability of MEA neurons require further elucidation. Previous studies suggest that inputs from the presubiculum (PrS) contribute to MEA pathophysiology. We assessed electrophysiologically how PrS influences MEA excitability using the rat pilocarpine model of TLE. PrS-MEA connectiv...

  4. Peripheral Sweat Gland Function Improves With Humid Heat Acclimation

    Science.gov (United States)

    2009-04-01

    via pilocarpine iontophoresis on the 128 MJ. Buono et al. / Journal ofThennal Biology 34 (2009) 127-130 proximal half of the flexor surface of both...forearms. The reported value is the mean of both forearms. The iontophoresis current was fixed at 1.5 rnA for 10 min, and was applied using a Wescor...discs (Pilogel, Wescor). Sweat was collected for 15 min immediately after iontophoresis using a Macroduct sweat collec- tor (Wescor) according to the

  5. Countermeasures to Heat Stress in Women

    Science.gov (United States)

    1996-10-01

    the Ta. Hence, both the low-moisture permeability and the high-insulating properties of CP clothing prevented heat loss through normal avenues, and...and in response to pilocarpine iontophoresis (27). Since a greater SR confers a larger capacity for heat loss via evaporation, it was uncertain if...M., DeCristofano, B., Speckman, K., & Sawka, M. (1990). Evaluation of three commercial microclimate cooling systems. Aviation, Space, and

  6. The prophylaxis and Therapy of Adhesions of the Addominal Cavity With Homotransplantion of Preserved Peritoneum - USSR

    Science.gov (United States)

    2007-11-02

    for incarcerated right inguinal hernia . Patients condition of average severity. Temperature 37.-6°, No particular symptoms for heart or lungs...Pilocarpine test before the operation was positive. Emergency operation on the day of admission. Under local in- filtration anesthesia , the abdominal...carried out prior to the operation proved to be sharply positive. Diagnosis — chronic appendicitis. Operation on 22 June. Under local anesthesia

  7. Studies of Micellar Electrokinetic Chromatography as an Analytical Technique in Pharmaceutical Analysis - an Industrial Perspective

    OpenAIRE

    Stubberud, Karin

    2002-01-01

    Studies have been performed to evaluate the use of micellar electrokinetic chromatography (MEKC), one mode of capillary electrophoresis (CE), as an analytical technique in industrial pharmaceutical analysis. The potential for using chemometrics for the optimisation of MEKC methods has also been studied as well as the possibilities of coupling MEKC with mass spectrometry (MS). Two methods were developed, one for the determination of ibuprofen and codeine and another for pilocarpine, together ...

  8. Management of xerostomia and salivary gland hypofunction.

    Science.gov (United States)

    Ram, Saravanan; Kumar, Satish; Navazesh, Mahvash

    2011-09-01

    Xerostomia and salivary gland hypofunction are conditions that have been associated with increased prevalence of caries, periodontitis, and candidiasis. Oral health care providers must be aware of the etiologies and clinical manifestations of salivary gland hypofunction in order to identify patients with this condition and to prevent its potential complications. The various modalities available to manage this condition range from frequent sips of water to the intake of systemic medications like pilocarpine or cevimeline.

  9. Anticonvulsant and proconvulsant roles of nitric oxide in experimental epilepsy models

    Directory of Open Access Journals (Sweden)

    Del-Bel E.A.

    1997-01-01

    Full Text Available The effect of acute (120 mg/kg and chronic (25 mg/kg, twice a day, for 4 days intraperitonial injection of the nitric oxide (NO synthase (NOS inhibitor NG-nitro-L-arginine (L-NOARG was evaluated on seizure induction by drugs such as pilocarpine and pentylenetetrazole (PTZ and by sound stimulation of audiogenic seizure-resistant (R and audiogenic seizure-susceptible (S rats. Seizures were elicited by a subconvulsant dose of pilocarpine (100 mg/kg only after NOS inhibition. NOS inhibition also simultaneously potentiated the severity of PTZ-induced limbic seizures (60 mg/kg and protected against PTZ-induced tonic seizures (80 mg/kg. The audiogenic seizure susceptibility of S or R rats did not change after similar treatments. In conclusion, proconvulsant effects of NOS inhibition are suggested to occur in the pilocarpine model and in the limbic components of PTZ-induced seizures, while an anticonvulsant role is suggested for the tonic seizures induced by higher doses of PTZ, revealing inhibitor-specific interactions with convulsant dose and also confirming the hypothesis that the effects of NOS inhibitors vary with the model of seizure

  10. Administration of copper reduced the hyper-excitability of neurons in CA1 hippocampal slices from epileptic rats.

    Science.gov (United States)

    Leiva, Juan; Infante, Claudio

    2016-04-01

    Copper as a trace metal is involved in several neurodegenerative illnesses, such as Menkes, Wilson's, Alzheimer's, amyotrophic lateral sclerosis (ALS), and Creutzfeldt-Jakob. Electrophysiological evidence indicates that acute perfusion of copper can inhibit long-term synaptic potentiation in hippocampal slices. The objective of this work is to determine whether Cu perfusion can perturb synaptic transmission in hippocampal slices derived from pilocarpine treated epileptic rats. Field potential (FP) recordings of the CA1 neurons of rats with chronic epilepsy showed voltage and response duration decrease following copper sulfate perfusion. However, voltage and response duration were higher after removing copper by washing. The discharge frequency of the CA1 neurons of hippocampal slices from non-epileptic control rats was increased after acute perfusion of 10 μM of pilocarpine. This increase was blocked by administering copper sulphate 10 μM. Krebs-Ringer solution washing re-established the discharges, with a higher frequency than that provoked by pilocarpine perfusion. We discuss the blocking effect of copper and the synaptic hyper-excitability generated by its removal.

  11. Early life status epilepticus and stress have distinct and sex-specific effects on learning, subsequent seizure outcomes, including anticonvulsant response to phenobarbital.

    Science.gov (United States)

    Akman, Ozlem; Moshé, Solomon L; Galanopoulou, Aristea S

    2015-02-01

    Neonatal status epilepticus (SE) is often associated with adverse cognitive and epilepsy outcomes. We investigate the effects of three episodes of kainic acid-induced SE (3KA-SE) and maternal separation in immature rats on subsequent learning, seizure susceptibility, and consequences, and the anticonvulsant effects of phenobarbital, according to sex, type, and age at early life (EL) event. 3KA-SE or maternal separation was induced on postnatal days (PN) 4-6 or 14-16. Rats were tested on Barnes maze (PN16-19), or lithium-pilocarpine SE (PN19) or flurothyl seizures (PN32). The anticonvulsant effects of phenobarbital (20 or 40 mg/kg/rat, intraperitoneally) pretreatment were tested on flurothyl seizures. FluoroJadeB staining assessed hippocampal injury. 3KA-SE or separation on PN4-6 caused more transient learning delays in males and did not alter lithium-pilocarpine SE latencies, but aggravated its outcomes in females. Anticonvulsant effects of phenobarbital were preserved and potentiated in specific groups depending on sex, type, and age at EL event. Early life 3KA-SE and maternal separation cause more but transient cognitive deficits in males but aggravate the consequences of subsequent lithium-pilocarpine SE in females. In contrast, on flurothyl seizures, EL events showed either beneficial or no effect, depending on gender, type, and age at EL events. © 2014 John Wiley & Sons Ltd.

  12. Network excitability in a model of chronic temporal lobe epilepsy critically depends on SK channel-mediated AHP currents.

    Science.gov (United States)

    Schulz, Robert; Kirschstein, Timo; Brehme, Hannes; Porath, Katrin; Mikkat, Ulrike; Köhling, Rüdiger

    2012-01-01

    Hippocampal CA1 pyramidal neurons generate an after-hyperpolarization (AHP) whose medium component is thought to be generated by small-conductance Ca(2+)-activated K(+) channels (SK channels). Neuronal excitability is increased in epilepsy, and the AHP in turn is fundamentally involved in regulation of cellular excitability. We therefore investigated the involvement of the SK channel-mediated AHP in controlling cell and network excitability in the pilocarpine model epilepsy. Both acutely isolated CA1 pyramidal cells and isolated hippocampal slices were investigated in terms of the impact of SK channel-mediated AHP on hyperexcitability. Our findings show that pilocarpine-treated chronically epileptic rats exhibit significantly reduced SK channel-mediated hyperpolarizing outward current which was accompanied by a significant decrease in the somatic AHP. Paradoxically, inhibiting SK channels strongly exacerbated 0-Mg(2+)-induced epileptiform activity in slices from pilocarpine-treated animals, while having a significantly smaller effect in control tissue. This suggests that in chronically epileptic tissue, network excitability very critically depends on the remaining SK-channel mediated AHP. Additional real-time RT-PCR and semiquantitative Western blot experiments revealed that both the SK2 channel transcript and protein were significantly downregulated in the epileptic CA1 region. We conclude that SK2 channels are down-regulated in chronic epilepsy underlying the impaired SK channel function in CA1 pyramidal cells, and a further reduction of the remaining critical mass of SK channels results in an acute network decompensation.

  13. Progressive brain damage, synaptic reorganization and NMDA activation in a model of epileptogenic cortical dysplasia.

    Directory of Open Access Journals (Sweden)

    Francesca Colciaghi

    Full Text Available Whether severe epilepsy could be a progressive disorder remains as yet unresolved. We previously demonstrated in a rat model of acquired focal cortical dysplasia, the methylazoxymethanol/pilocarpine - MAM/pilocarpine - rats, that the occurrence of status epilepticus (SE and subsequent seizures fostered a pathologic process capable of modifying the morphology of cortical pyramidal neurons and NMDA receptor expression/localization. We have here extended our analysis by evaluating neocortical and hippocampal changes in MAM/pilocarpine rats at different epilepsy stages, from few days after onset up to six months of chronic epilepsy. Our findings indicate that the process triggered by SE and subsequent seizures in the malformed brain i is steadily progressive, deeply altering neocortical and hippocampal morphology, with atrophy of neocortex and CA regions and progressive increase of granule cell layer dispersion; ii changes dramatically the fine morphology of neurons in neocortex and hippocampus, by increasing cell size and decreasing both dendrite arborization and spine density; iii induces reorganization of glutamatergic and GABAergic networks in both neocortex and hippocampus, favoring excitatory vs inhibitory input; iv activates NMDA regulatory subunits. Taken together, our data indicate that, at least in experimental models of brain malformations, severe seizure activity, i.e., SE plus recurrent seizures, may lead to a widespread, steadily progressive architectural, neuronal and synaptic reorganization in the brain. They also suggest the mechanistic relevance of glutamate/NMDA hyper-activation in the seizure-related brain pathologic plasticity.

  14. Inositol-deficient food augments a behavioral effect of long-term lithium treatment mediated by inositol monophosphatase inhibition: an animal model with relevance for bipolar disorder.

    Science.gov (United States)

    Shtein, Liza; Agam, Galila; Belmaker, R H; Bersudsky, Yuly

    2015-04-01

    Lithium treatment in rodents markedly enhances cholinergic agonists such as pilocarpine. This effect can be reversed in a stereospecific manner by administration of inositol, suggesting that the effect of lithium is caused by inositol monophosphatase inhibition and consequent inositol depletion. If so, inositol-deficient food would be expected to enhance lithium effects. Inositol-deficient food was prepared from inositol-free ingredients. Mice with a homozygote knockout of the inositol monophosphatase 1 gene unable to synthesize inositol endogenously and mimicking lithium-treated animals were fed this diet or a control diet. Lithium-treated wild-type animals were also treated with the inositol-deficient diet or control diet. Pilocarpine was administered after 1 week of treatment, and behavior including seizures was assessed using rating scale. Inositol-deficient food-treated animals, both lithium treated and with inositol monophosphatase 1 knockout, had significantly elevated cholinergic behavior rating and significantly increased or earlier seizures compared with the controls. The effect of inositol-deficient food supports the role of inositol depletion in the effects of lithium on pilocarpine-induced behavior. However, the relevance of this behavior to other more mood-related effects of lithium is not clear.

  15. X-ray microanalysis of exocrine glands in animal models for cystic fibrosis

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, R.M.R.; Roomans, G.M.

    1985-01-01

    Elemental distribution and ultrastructure of the submandibular gland, the parotid gland and the pancreas were investigated in three suggested animal models of the disease cystic fibrosis: the chronically reserpinized rat, the chronically isoproterenol-treated rat, and the chronically pilocarpine-treated rat. To elucidate the cellular mechanism underlying the effects of these treatments, chronic effects of specific alpha - and beta -adrenergic agonists, as well as acute effects of reserpine and various agonists were also investigated. Reserpine, isoproterenol, and pilocarpine cause an increase in the calcium concentration in submandibular gland acinar cells, due to an increased calcium content of the intracellular mucus. In the parotid gland, reserpine and isoproterenol cause a decrease of the calcium concentration in acinar cells, due to a lower calcium content of the zymogen granules. In the submandibular gland, a decreased cellular Na concentration was noted after chronic treatment with isoproterenol or pilocarpine, and after a single dose of reserpine or isoproterenol. Ultrastructural changes in the exocrine glands investigated included excessive accumulation of intracellular secretory material and formation of abnormal uncondensed secretion granules. A common pattern in the animal models appears to be (1) inhibition of secretion resulting in intracellular accumulation of secretory material, (2) synthesis of secretory macromolecules with altered cation-binding properties.

  16. Effects of thyroxine and dexamethasone on rat submandibular glands

    Energy Technology Data Exchange (ETDEWEB)

    Sagulin, G.B.; Roomans, G.M. (Karolinska Institutet, Huddinge (Sweden))

    1989-08-01

    Glucocorticoids and thyroxine are known to have a marked effect on the flow rate and protein composition of rat parotid saliva in hormonally intact animals. In the present study, the effects of a one-week treatment of male rats with dexamethasone and thyroxine were studied by electron microscopy and x-ray micro-analysis, and by measurement of the flow rate and determination of the chemical composition of pilocarpine-induced submandibular saliva. Thyroxine had the most extensive effects on the submandibular gland. The acinar cells were enlarged and filled with mucus; the cellular calcium concentration was significantly increased. The flow rate of the submandibular saliva was significantly reduced compared with that in saline-injected control animals. Thyroxine caused an increase in the concentrations of protein, total calcium, and potassium in the saliva. Dexamethasone had no significant effects on gland ultrastructure or on the elemental composition of the acinar cells; flow rate was not affected, but the concentrations of protein, calcium, and potassium were significantly increased. The effects of dexamethasone and thyroxine on the flow rate and protein composition of pilocarpine-induced rat submandibular saliva differ from those reported earlier for rat parotid saliva after simultaneous stimulation with pilocarpine and isoproterenol.

  17. Effects of Benzodiazepines on Acinar and Myoepithelial Cells

    Science.gov (United States)

    Mattioli, Tatiana M. F.; Alanis, Luciana R. A.; Sapelli, Silvana da Silva; de Lima, Antonio A. S.; de Noronha, Lucia; Rosa, Edvaldo A. R.; Althobaiti, Yusuf S.; Almalki, Atiah H.; Sari, Youssef; Ignacio, Sergio A.; Johann, Aline C. B. R.; Gregio, Ana M. T.

    2016-01-01

    Background: Benzodiazepines (BZDs), the most commonly prescribed psychotropic drugs with anxiolytic action, may cause hyposalivation. It has been previously shown that BZDs can cause hypertrophy and decrease the acini cell number. In this study, we investigated the effects of BZDs and pilocarpine on rat parotid glands, specifically on acinar, ductal, and myoepithelial cells. Methods: Ninety male Wistar rats were divided into nine groups. Control groups received a saline solution for 30 days (C30) and 60 days (C60), and pilocarpine (PILO) for 60 days. Experimental groups received lorazepam (L30) and midazolam (M30) for 30 days. Another group (LS60 or MS60) received lorazepam or midazolam for 30 days, respectively, and saline for additional 30 days. Finally, other groups (LP60 or MP60) received either lorazepam or midazolam for 30 days, respectively, and pilocarpine for additional 30 days. The expression of calponin in myoepithelial cells and the proliferating cell nuclear antigen (PCNA) in acinar and ductal cells were evaluated. Results: Animals treated with lorazepam showed an increase in the number of positive staining cells for calponin as compared to control animals (p acinar and ductal cells and a decrease in the positive staining cells for calponin as compared to midazolam administered with saline (MS60). Conclusion: We found that myoepithelial cells might be more sensitive to the effects of BZD than acinar and ductal cells in rat parotid glands. PMID:27445812

  18. Protective Effects of Cannabidiol against Seizures and Neuronal Death in a Rat Model of Mesial Temporal Lobe Epilepsy.

    Science.gov (United States)

    Do Val-da Silva, Raquel A; Peixoto-Santos, Jose E; Kandratavicius, Ludmyla; De Ross, Jana B; Esteves, Ingrid; De Martinis, Bruno S; Alves, Marcela N R; Scandiuzzi, Renata C; Hallak, Jaime E C; Zuardi, Antonio W; Crippa, Jose A; Leite, Joao P

    2017-01-01

    The present study reports the behavioral, electrophysiological, and neuropathological effects of cannabidiol (CBD), a major non-psychotropic constituent of Cannabis sativa, in the intrahippocampal pilocarpine-induced status epilepticus (SE) rat model. CBD was administered before pilocarpine-induced SE (group SE+CBDp) or before and after SE (group SE+CBDt), and compared to rats submitted only to SE (SE group), CBD, or vehicle (VH group). Groups were evaluated during SE (behavioral and electrophysiological analysis), as well as at days one and three post-SE (exploratory activity, electrophysiological analysis, neuron density, and neuron degeneration). Compared to SE group, SE+CBD groups (SE+CBDp and SE+CBDt) had increased SE latency, diminished SE severity, increased contralateral afterdischarge latency and decreased relative powers in delta (0.5-4 Hz) and theta (4-10 Hz) bands. Only SE+CBDp had increased vertical exploratory activity 1-day post SE and decreased contralateral relative power in delta 3 days after SE, when compared to SE group. SE+CBD groups also showed decreased neurodegeneration in the hilus and CA3, and higher neuron density in granule cell layer, hilus, CA3, and CA1, when compared to SE group. Our findings demonstrate anticonvulsant and neuroprotective effects of CBD preventive treatment in the intrahippocampal pilocarpine epilepsy model, either as single or multiple administrations, reinforcing the potential role of CBD in the treatment of epileptic disorders.

  19. Morphological alterations in newly born dentate gyrus granule cells that emerge after status epilepticus contribute to make them less excitable.

    Directory of Open Access Journals (Sweden)

    Julián Tejada

    Full Text Available Computer simulations of external current stimulations of dentate gyrus granule cells of rats with Status Epilepticus induced by pilocarpine and control rats were used to evaluate whether morphological differences alone between these cells have an impact on their electrophysiological behavior. The cell models were constructed using morphological information from tridimensional reconstructions with Neurolucida software. To evaluate the effect of morphology differences alone, ion channel conductances, densities and distributions over the dendritic trees of dentate gyrus granule cells were the same for all models. External simulated currents were injected in randomly chosen dendrites belonging to one of three different areas of dentate gyrus granule cell molecular layer: inner molecular layer, medial molecular layer and outer molecular layer. Somatic membrane potentials were recorded to determine firing frequencies and inter-spike intervals. The results show that morphologically altered granule cells from pilocarpine-induced epileptic rats are less excitable than control cells, especially when they are stimulated in the inner molecular layer, which is the target area for mossy fibers that sprout after pilocarpine-induced cell degeneration. This suggests that morphological alterations may act as a protective mechanism to allow dentate gyrus granule cells to cope with the increase of stimulation caused by mossy fiber sprouting.

  20. Chronic Trigeminal Nerve Stimulation Protects Against Seizures, Cognitive Impairments, Hippocampal Apoptosis, and Inflammatory Responses in Epileptic Rats.

    Science.gov (United States)

    Wang, Qian-Qian; Zhu, Li-Jun; Wang, Xian-Hong; Zuo, Jian; He, Hui-Yan; Tian, Miao-Miao; Wang, Lei; Liang, Gui-Ling; Wang, Yu

    2016-05-01

    Trigeminal nerve stimulation (TNS) has recently been demonstrated effective in the treatment of epilepsy and mood disorders. Here, we aim to determine the effects of TNS on epileptogenesis, cognitive function, and the associated hippocampal apoptosis and inflammatory responses. Rats were injected with pilocarpine to produce status epilepticus (SE) and the following chronic epilepsy. After SE induction, TNS treatment was conducted for 4 consecutive weeks. A pilocarpine re-injection was then used to induce a seizure in the epileptic rats. The hippocampal neuronal apoptosis induced by seizure was assessed by TUNEL staining and inflammatory responses by immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). The spontaneous recurrent seizure (SRS) number was counted through video monitoring, and the cognitive function assessed through Morris Water Maze (MWM) test. TNS treatment attenuated the SRS attacks and improved the cognitive impairment in epileptic rats. A pilocarpine re-injection resulted in less hippocampal neuronal apoptosis and reduced level of interleukin-1 beta (IL-1β), tumor necrosis factor-α (TNF-α), and microglial activation in epileptic rats with TNS treatment in comparison to the epileptic rats without TNS treatment. It is concluded that TNS treatment shortly after SE not only protected against the chronic spontaneous seizures but also improved cognitive impairments. These antiepileptic properties of TNS may be related to its attenuating effects on hippocampal apoptosis and pro-inflammatory responses.

  1. Muscarinic regulation of Kenyon cell dendritic arborizations in adult worker honey bees.

    Science.gov (United States)

    Dobrin, Scott E; Herlihy, J Daniel; Robinson, Gene E; Fahrbach, Susan E

    2011-09-01

    The experience of foraging under natural conditions increases the volume of mushroom body neuropil in worker honey bees. A comparable increase in neuropil volume results from treatment of worker honey bees with pilocarpine, an agonist for muscarinic-type cholinergic receptors. A component of the neuropil growth induced by foraging experience is growth of dendrites in the collar region of the calyces. We show here, via analysis of Golgi-impregnated collar Kenyon cells with wedge arborizations, that significant increases in standard measures of dendritic complexity were also found in worker honey bees treated with pilocarpine. This result suggests that signaling via muscarinic-type receptors promotes the increase in Kenyon cell dendritic complexity associated with foraging. Treatment of worker honey bees with scopolamine, a muscarinic inhibitor, inhibited some aspects of dendritic growth. Spine density on the Kenyon cell dendrites varied with sampling location, with the distal portion of the dendritic field having greater total spine density than either the proximal or medial section. This observation may be functionally significant because of the stratified organization of projections from visual centers to the dendritic arborizations of the collar Kenyon cells. Pilocarpine treatment had no effect on the distribution of spines on dendrites of the collar Kenyon cells.

  2. Syntheses and evaluation of anticonvulsant activity of novel branched alkyl carbamates.

    Science.gov (United States)

    Hen, Naama; Bialer, Meir; Yagen, Boris

    2012-03-22

    A novel class of 19 carbamates was synthesized, and their anticonvulsant activity was comparatively evaluated in the rat maximal electroshock (MES) and subcutaneous metrazol (scMet) seizure tests and pilocarpine-induced status epilepticus (SE) model. In spite of the alkyl-carbamates' close structural features, only compounds 34, 38, and 40 were active at the MES test. The analogues 2-ethyl-3-methyl-butyl-carbamate (34) and 2-ethyl-3-methyl-pentyl-carbamate (38) also exhibited potent activity in the pilocarpine-SE model 30 min postseizure onset. Extending the aliphatic side chains of homologous carbamates from 7 to 8 (34 to 35) and from 8 to 9 carbons in the homologues 38 and 43 decreased the activity in the pilocarpine-SE model from ED(50) = 81 mg/kg (34) to 94 mg/kg (35) and from 96 mg/kg (38) to 114 mg/kg (43), respectively. The most potent carbamate, phenyl-ethyl-carbamate (47) (MES ED(50) = 16 mg/kg) contains an aromatic moiety in its structure. Compounds 34, 38, 40, and 47 offer the optimal efficacy-safety profile and, consequently, are promising candidates for development as new antiepileptics.

  3. Neuronal damage and memory deficits after seizures are reversed by ascorbic acid? O dano neuronal e o déficit de memória após convulsões são revertidos pelo ácido ascórbico?

    Directory of Open Access Journals (Sweden)

    Adriana da Rocha Tomé

    2010-08-01

    Full Text Available The objective of the present study was to evaluate the neuroprotective effects of ascorbic acid (AA in rats, against the neuronal damage and memory deficit caused by seizures. Wistar rats were treated with 0.9% saline (i.p., control group, ascorbic acid (500 mg/kg, i.p., AA group, pilocarpine (400 mg/kg, i.p., pilocarpine group, and the association of ascorbic acid (500 mg/kg, i.p. plus pilocarpine (400 mg/kg, i.p., 30 min before of administration of ascorbic acid (AA plus pilocarpine group. After the treatments all groups were observed for 24 h. Pilocarpine group presented seizures which progressed to status epilepticus in 75% of the animals. Pretreatment with AA led to a reduction of 50% of this rate. Results showed that pretreatment with AA did not alter reference memory when compared to a control group. In the working memory task, we observed a significant day's effect with important differences between control, pilocarpine and AA plus pilocarpine groups. Pilocarpine and AA plus pilocarpine groups had 81 and 16% of animals with brain injury, respectively. In the hippocampus of pilocarpine animals, it was detected an injury of 60%. As for the animals tested with AA plus pilocarpine, the hippocampal region of the group had a reduction of 43% in hippocampal lesion. Our findings suggest that seizures caused cognitive dysfunction and neuronal damage that might be related, at least in part, to the neurological problems presented by epileptic patients. AA can reverse cognitive dysfunction observed in rats with seizures as well as decrease neuronal injury in rat hippocampus.O objetivo do presente estudo foi avaliar o efeito neuroprotetor do ácido ascórbico (AA, contra o dano neuronal e o déficit de memória em ratos causados pelas convulsões. Ratos Wistar foram tratados com solução salina a 0,9% (i.p., grupo controle, ácido ascórbico (500 mg/kg, i.p., grupo AA, pilocarpina (400 mg/kg, i.p., grupo pilocarpina, e a associação de ácido asc

  4. Effect of acetylcholine receptors on the pain-related electrical activities in the hippocampal CA3 region of morphine-addicted rats

    Directory of Open Access Journals (Sweden)

    Guan Zeng Li

    2015-07-01

    Full Text Available Objective(s:To determine the effect of acetylcholine (ACh, pilocarpine, and atropine on pain evoked responses of pain excited neurons (PEN and pain inhibited neurons (PIN in hippocampal CA3 region of morphine addicted rats. Materials and Methods:Female Wistar rats, weighing between 230-260 g were used in this study. Morphine addicted rats were generated by subcutaneous injection of increasing concentrations of morphine hydrochloride for six days. Trains of electrical impulses applied to the sciatic nerve were used as noxious stimulation and the evoked electrical activities of PEN or PIN in hippocampal CA3 area were recorded using extracellular electrophysiological recording techniques in hippocampal slices. The effect of acetylcholine receptor stimulation byACh, the muscarinic agonist pilocarpine, and the muscarinic antagonist atropine on the pain evoked responses of pain related electrical activities was analyzed in hippocampal CA3 area of morphine addicted rats. Results:Intra-CA3 microinjection of ACh (2 μg/1 μl or pilocarpine (2 μg/1 μl decreased the discharge frequency and prolonged the firing latency of PEN, but increased the discharge frequency and shortened the firing inhibitory duration (ID of PIN. The intra-CA3 administration of atropine (0.5 μg/1 μl produced opposite effect. The peak activity of cholinergic modulators was 2 to 4 min later in morphine addicted rats compared to peak activity previously observed in normal rats. Conclusion: ACh dependent modulation of noxious stimulation exists in hippocampal CA3 area of morphine addicted rats. Morphine treatment may shift the sensitivity of pain related neurons towards a delayed response to muscarinergic neurotransmission in hippocampal CA3 region.

  5. Akt pathway activation and increased neuropeptide Y mRNA expression in the rat hippocampus: implications for seizure blockade.

    Science.gov (United States)

    Goto, Eduardo M; Silva, Marcelo de Paula; Perosa, Sandra R; Argañaraz, Gustavo A; Pesquero, João B; Cavalheiro, Esper A; Naffah-Mazzacoratti, Maria G; Teixeira, Vicente P C; Silva, José A

    2010-04-01

    The aim of this study was to analyze the expression of survival-related molecules such Akt and integrin-linked kinase (ILK) to evaluate Akt pathway activation in epileptogenesis process. Furthermore, was also investigated the mRNA expression of neuropeptide Y, a considered antiepileptic neuropeptide, in the pilocarpine-induced epilepsy. Male Wistar rats were submitted to the pilocarpine model of epilepsy. Hippocampi were removed 6h (acute phase), 12h (late acute), 5d (silent) and 60d (chronic) after status epilepticus (SE) onset, and from animals that received pilocarpine but did not develop SE (partial group). Hippocampi collected were used to specify mRNA expression using Real-Time PCR. Immunohistochemistry assay was employed to place ILK distribution in the hippocampus and Western blot technique was used to determine Akt activation level. A decrease in ILK mRNA content was found during acute (0.39+/-0.03) and chronic (0.48+/-0.06) periods when compared to control group (0.87+/-0.10). Protein levels of ILK were also diminished during both periods. Partial group showed increased ILK mRNA expression (0.80+/-0.06) when compared with animals in the acute stage. Silent group had ILK mRNA and immunoreactivity similar to control group. Western blot assay showed an augmentation in Akt activation in silent period (0.52+/-0.03) in comparison with control group (0.44+/-0.01). Neuropeptide Y mRNA expression increased in the partial group (1.67+/-0.22) and in the silent phase (1.45+/-0.29) when compared to control group (0.36+/-0.12). Results suggest that neuropeptide Y (as anticonvulsant) might act in protective mechanisms occurred during epileptic phenomena. Together with ILK expression and Akt activation, these molecules could be involved in hippocampal neuroprotection in epilepsy. Copyright 2009 Elsevier Ltd. All rights reserved.

  6. Pilocarpus spp.: a survey of its chemical constituents and biological activities Pilocarpus spp.: revisão sobre sua constituição química e atividades biológicas

    Directory of Open Access Journals (Sweden)

    Ana Paula Santos

    2004-06-01

    Full Text Available Pilocarpus species have been exploited as the only source of the imidazole alkaloid pilocarpine (used in glaucoma treatment, since its isolation up to the present. Almost all Pilocarpusspecies are native from Brazil. Because of the medicinal importance of pilocarpine, several of them are in the path of extinction. Other secondary metabolites, such as coumarins, flavonoids and terpenes, were described for Pilocarpusspecies. In this review the secondary metabolites, other than pilocarpine, isolated from Pilocarpusspecies and their biological activities were compiled. Although the variety of structures and the importance of the biological activities described in literature for Pilocarpusspecies this is an unexploited field of research in Natural Products and Pharmacology.Espécies de Pilocarpus têm sido exploradas como única fonte do alcalóide imidazólico pilocarpina (utilizado no tratamento do glaucoma desde o isolamento dessa substância até os dias atuais. A maioria das espécies de Pilocarpus conhecida é nativa do Brasil e, devido à importância medicinal que a pilocarpina possui e ao desmatamento, várias se encontram em risco de extinção. Outros metabólitos secundários entre os quais cumarinas, flavonóides e terpenos foram descritos em espécies desse gênero. Nesta revisão foram relacionados os metabólitos secundários isolados em diversas espécies de Pilocarpus bem como suas atividades biológicas. Apesar da variedade de estruturas e as importantes atividades biológicas já descritas na literatura para as outras classes de metabólitos secundários, ainda há um vasto campo de estudo para as espécies de Pilocarpus.

  7. Delayed damage after radiation therapy for head and neck cancer

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Yoshiyuki [Osaka Dental Univ., Hirakata (Japan)

    2000-03-01

    I investigated radiation damage, including osteoradionecrosis, arising from tooth extraction in fields that had received radiation therapy for head and neck cancer, and evaluated the effectiveness of pilocarpine for xerostomia. Between January 1990 and April 1996, I examined 30 patients for bone changes after tooth extraction in fields irradiated at the Department of Oral Radiology, Osaka Dental University Hospital. Nineteen of the patients had been treated for nasopharyngeal cancer and 11 for oropharyngeal cancer. Between January and April 1996, 4 additional patients were given pilocarpine hydrochloride (3-mg, 6-mg and 9-mg of KSS-694 orally three times a day) for 12 weeks and evaluated every 4 weeks as a base line. One had been treated for nasopharyngeal carcinoma, two for cancer of the cheek and one for an unknown carcinoma. Eighteen of the patients (11 with nasopharyngeal carcinoma and 7 with oropharyngeal carcinoma) had extractions. Use of preoperative and postoperative radiographs indicated that damage to the bone following tooth extraction after radiation exposure was related to whether antibiotics were administered the day before the extraction, whether forceps or elevators were used, and whether the tooth was in the field of radiation. Xerostomia improved in all 4 of the patients who received 6-mg or 9-mg of pilocarpine. It improved saliva production and relieved the symptoms of xerostomia after radiation therapy for head and neck cancer, although there were minor side effects such as fever. This information can be used to improve the oral environment of patients who have received radiation therapy for head and neck cancer, and to better understand their oral environment. (author)

  8. Acetylcholinesterase loosens the brain's cholinergic anti-inflammatory response and promotes epileptogenesis

    Directory of Open Access Journals (Sweden)

    Yehudit eGnatek

    2012-05-01

    Full Text Available Recent studies show a key role of brain inflammation in epilepsy. However, the mechanisms controlling brain immune response are only partly understood. In the periphery, acetylcholine (ACh release by the vagus nerve restrains inflammation by inhibiting the activation of leukocytes. Recent reports suggested a similar anti-inflammatory effect for ACh in the brain. Since brain cholinergic dysfunction are documented in epileptic animals, we explored changes in brain cholinergic gene expression and associated immune response during pilocarpine-induced epileptogenesis. Levels of acetylcholinesterase (AChE and inflammatory markers were measured using real-time RT-PCR, in-situ hybridization and immunostaining in wild type (WT and transgenic mice over-expressing the "synaptic" splice variant AChE-S (TgS. One month following pilocarpine, mice were video-monitored for spontaneous seizures. To test directly the effect of ACh on the brain's innate immune response, cytokines expression levels were measured in acute brain slices treated with cholinergic agents. We report a robust upregulation of AChE as early as 48 hrs following pilocarpine-induced status epilepticus (SE. AChE was expressed in hippocampal neurons, microglia and endothelial cells but rarely in astrocytes. TgS mice overexpressing AChE showed constitutive increased microglial activation, elevated levels of pro-inflammatory cytokines 48 hrs after SE and accelerated epileptogenesis compared to their WT counterparts. Finally we show a direct, muscarine-receptor dependant, nicotine-receptor independent anti-inflammatory effect of ACh in brain slices maintained ex vivo. Our work demonstrates for the first time, that ACh directly suppresses brain innate immune response and that AChE up-regulation after SE is associated with enhanced immune response, facilitating the epileptogenic process. Our results highlight the cholinergic system as a potential new target for the prevention of seizures and epilepsy.

  9. A reduced susceptibility to chemoconvulsant stimulation in adenylyl cyclase 8 knockout mice

    Science.gov (United States)

    Chen, Xia; Dong, Guoying; Zheng, Changhong; Wang, Hongbing; Yun, Wenwei; Zhou, Xianju

    2015-01-01

    Objective Adenylyl cyclases (ACs) catalyze the synthesis of cAMP from ATP, and cAMP signaling affects a large number of neuronal processes. Ca2+-stimualted adenylyl cyclase 8 (AC8) expressed in the CNS plays a role in synaptic plasticity, drug addiction and ethanol sensitivity, and chronic pain. This study was to aim at examining the contributions of AC8 to epileptogenesis. Methods In this study, we observed the seizure behavior induced by kainic acid (20mg/kg or 30mg/kg) or pilocarpine (350mg/kg) in AC8 KO and wild-type mice. Next we injected kainic acid or pilocarpine to induce status epilepticus (SE), and examined neuronal degeneration (by Fluoro-Jade B staining) and mossy fiber sprouting (by Timm staining) 24 hr and 2 weeks after SE termination in the hippocampus, respectively. Finally, 15min after intraperitoneal injection of kainic acid (30mg/kg), we examined phosphor-ERK1/2 in the hippocampus by western blot and immunochemistry staining. Results We first observed that AC8 KO mutants display reduced susceptibility (including seizure latency and episodes) to two chemoconvulsants, kainic acid and pilocarpine. Moreover, we found that degenerative neurons and mossy fiber sprouting induced by chemoconvulsants were significant decreased in the hippocampus. Further, western blot and immunochemistry analysis revealed that the MAPK signaling in the hippocampus was attenuated in kainic acid-injected AC8 KO mice. Conclusion AC8 is involved in epileptogenesis, and may serve as a potential target for the treatment of epilepsy. PMID:26656781

  10. Carbachol as miotic agent in intra-ocular lens implant surgery

    Directory of Open Access Journals (Sweden)

    Agarwal Jitendra

    1988-01-01

    Full Text Available The intra cameral use of 0.025% carbachol as a miotic agent in anterior chamber intraocular lens im-plant surgery us reported in 15 cases. Carbachol produced prompt and effective moisis and was found to be harmless and non-irritating to the anterior chamber structures. A rebound dialatation of the pipit was noticed in the post operative period in 3 of our first 5 cases where only carbachol was used. For prolonged miosis instillation of a more powerful miotic like pilocarpine is recommended at the completion of surgery:

  11. Glutamate carboxypeptidase II (GCPII) inhibitor displays anti-glutamate and anti-cocaine effects in an invertebrate assay.

    Science.gov (United States)

    Tallarida, Chris; Song, Kevin; Raffa, Robert B; Rawls, Scott M

    2012-06-01

    Glutamate carboxypeptidase II (GCPII) inhibitors are promising anti-glutamatergic and anti-addictive agents. We hypothesized that a GCPII inhibitor 2 (phosphonomethyl) pentanedioic acid (2-PMPA) would display anti-stereotypical activity in planarians. Experiments revealed that 2-PMPA displayed no overt behavioral activity by itself but attenuated stereotypical counts (C-shape hyperkinesias) elicited by four compounds (2-PMPA rank order potency: glutamate>NMDA>pilocarpine>cocaine). These data suggest GCPII inhibitors display broad-spectrum efficacy against behavioral activity produced by glutamatergic and non-glutamatergic compounds in an invertebrate assay.

  12. Disruption of TrkB-mediated PLCγ signaling inhibits limbic epileptogenesis

    OpenAIRE

    He, XP; Pan, E; C. Sciarretta; Minichiello, L.; McNamara, JO

    2010-01-01

    The BDNF receptor, TrkB, is critical to limbic epileptogenesis, but the responsible downstream signaling pathways are unknown. We hypothesized that TrkB-dependent activation of PLCγ1 signaling is the key pathway and tested this in trkBPLC/PLC mice carrying a mutation (Y816F) that uncouples TrkB from PLCγ1. Biochemical measures revealed activation of both TrkB and PLCγ1 in hippocampi in the pilocarpine and kindling models in WT mice. PLCγ1 activation was decreased in hippocampi isolated from t...

  13. Essential oils from Brazilian rutaceae. Part I. Genus Pilocardus

    Energy Technology Data Exchange (ETDEWEB)

    Craveiro, A.A.; Andrade, C.H.S.; Matos, F.J.A.; Alencar, J.W.

    1979-11-01

    The leaves of jaborandi are commercially exploited in the Brazilian Northeast for industrial extraction of pilocarpine, an alkaloid with potent cholinergic activity. The leaves also contain an essential oil whose composition is registered in the literature in a confusing and incomplete way. Chemical re-examination of the essential oil from five distinct Pilocarpus species was conducted together with an analysis of the leaves used by local industry. It was found to contain terpenes, sesquiterpenes and ketones. Some of them are reported for the first time in the genus.

  14. Systematic review of miscellaneous agents for the management of oral mucositis in cancer patients

    DEFF Research Database (Denmark)

    Jensen, Siri Beier; Jarvis, Virginia; Zadik, Yehuda

    2013-01-01

    : A total of 32 papers across 10 interventions were examined. New suggestions were developed against the use of systemic pilocarpine administered orally for prevention of OM during RT in head and neck cancer patients and in patients receiving high-dose chemotherapy, with or without total body irradiation......, prior to hematopoietic stem cell transplantation. A suggestion was also made against the use of systemic pentoxifylline administered orally for the prevention of OM in patients undergoing bone marrow transplantation. No guideline was possible for any other agent reviewed due to inadequate and...

  15. Tonic pupil in leprosy

    Directory of Open Access Journals (Sweden)

    Marco Aurélio Lana-Peixoto

    2014-12-01

    Full Text Available Pupil abnormalities in leprosy usually result from chronic iritis with loss of stroma, iris miosis, a sluggish reaction to light, and poor dilation in response to anticholinergic mydriatics. We report two patients with long-standing lepromatous leprosy who developed tonic pupils characterized by mydriasis, absence of reaction to light and hypersensitivity to weak cholinergic solution. Examination revealed iritis and iris atrophy. In both cases, instillation of dilute 0.1% pilocarpine caused miosis in the affected eyes. Tonic pupil occurs in many conditions, but its association with leprosy had not been previously reported.

  16. Histology of the iris in geese and ducks photosensitized by ingestion of Ammi majus seeds.

    Science.gov (United States)

    Barishak, Y R; Beemer, A M; Egyed, M N; Shlosberg, A; Eilat, A

    1975-09-01

    Geese and ducks were photosensitized by the ingestion of Ammi majus seeds, and exposure to sunlight. Mydriasis was a characteristic clinical feature of this syndrome in both species. Histologically the iris of the affected birds showed vacuolisation and varying degrees of atrophy of the muscle of the sphincter pupillae. The effect of pilocarpine and physostigmine on the normal and mydriatic eyes was studied. The possible mode of action of photosensitization and the significance of these findings in the light of the use of psoralens in human medicine is discussed.

  17. Degeneration and regeneration of GABAergic interneurons in the dentate gyrus of adult mice in experimental models of epilepsy.

    Science.gov (United States)

    Wei, Dong; Yang, Fang; Wang, Ying; Yang, Feng; Wu, Chen; Wu, Sheng-Xi; Jiang, Wen

    2015-01-01

    Mounting evidence showed that GABAergic interneurons play an important role in the generation of seizures by regulating excitatory/inhibitory balance in the hippocampus; however, there is a continuous debate regarding the alteration in the number of hippocampal GABAergic interneurons during epileptogenesis. Here, we investigated the degeneration and regeneration of GABAergic interneurons in the dentate gyrus during epileptogenesis using glutamic acid decarboxylase-green fluorescence protein (GAD67-GFP) knock-in mice. Pentylenetetrazol (PTZ)-induced chronic kindling model and lithium-pilocarpine-induced status epilepticus (SE) model were used in this study. We found a progressive loss of GABAergic interneurons in the dentate gyrus during post-SE epileptogenesis rather than PTZ kindling. Both types of epileptogenic insults significantly promoted the proliferation of neural progenitor cells in the dentate gyrus; however, compared to 80% neuronal differentiation ratio in the control group, there was a remarkable decrease in PTZ kindling and pilocarpine models, that is 58% and 29%, respectively. Double/triple immunofluorescence labeling revealed no newborn neurons colabeled with GFP in both intact and epileptic dentate gyrus. In addition, valproate (a first-line antiepileptic drug) treatment prevented the loss of GABAergic interneurons but still failed to induce the regeneration of GAD67-positive interneurons in the dentate gyrus during post-SE epileptogenesis. These results indicate that degeneration of GABAergic interneurons may depend on the type of epileptogenic insult and that no newborn GABAergic interneurons occur in the adult dentate gyrus during epileptogenesis. © 2014 John Wiley & Sons Ltd.

  18. Seizure susceptibility of neuropeptide-Y null mutant mice in amygdala kindling and chemical-induced seizure models.

    Science.gov (United States)

    Shannon, Harlan E; Yang, Lijuan

    2004-01-01

    Neuropeptide Y (NPY) administered exogenously is anticonvulsant, and, NPY null mutant mice are more susceptible to kainate-induced seizures. In order to better understand the potential role of NPY in epileptogenesis, the present studies investigated the development of amygdala kindling, post-kindling seizure thresholds, and anticonvulsant effects of carbamazepine and levetiracetam in 129S6/SvEv NPY(+/+) and NPY(-/-) mice. In addition, susceptibility to pilocarpine- and kainate-induced seizures was compared in NPY(+/+) and (-/-) mice. The rate of amygdala kindling development did not differ in the NPY(-/-) and NPY(+/+) mice either when kindling stimuli were presented once daily for at least 20 days, or, 12 times daily for 2 days. However, during kindling development, the NPY(-/-) mice had higher seizure severity scores and longer afterdischarge durations than the NPY(+/+) mice. Post-kindling, the NPY(-/-) mice had markedly lower afterdischarge thresholds and longer afterdischarge durations than NPY (+/+) mice. Carbamazepine and levetiracetam increased the seizure thresholds of both NPY (-/-) and (+/+) mice. In addition, NPY (-/-) mice had lower thresholds for both kainate- and pilocarpine-induced seizures. The present results in amygdala kindling and chemical seizure models suggest that NPY may play a more prominent role in determining seizure thresholds and severity of seizures than in events leading to epileptogenesis. In addition, a lack of NPY does not appear to confer drug-resistance in that carbamazepine and levetiracetam were anticonvulsant in both wild type (WT) and NPY null mutant mice.

  19. A functional and chemical study of radiation effects on rat parotid and submandibular/sublingual glands

    Energy Technology Data Exchange (ETDEWEB)

    Vissink, A.; S-Gravenmade, E.J.; Ligeon, E.E.; Konings, W.T. (Univ. of Groningen (Netherlands))

    1990-12-01

    The aim of this study was to monitor composition and rate of secretion of rat parotid and submandibular/sublingual saliva following local single doses of X-rays ranging from 5 to 20 Gy. Pilocarpine-stimulated samples of parotid and submandibular/sublingual saliva were simultaneously collected with miniaturized Lashley cups before and 1-30 days after irradiation. The lag phase (period between injection of pilocarpine and start of the secretion) and flow rate were recorded and the concentrations of sodium, potassium, calcium, phosphate, and amylase were measured. With increasing dose and time, the salivary flow rate as well as sodium concentration decreased, while potassium concentrations increased throughout the follow-up period. The lag phase and the concentration of amylase reached their maximum at 3 and 10 days after irradiation, respectively. The changes in lag phase and flow rate were most obvious after doses of 15 or 20 Gy and showed a great similarity for parotid and submandibular/sublingual saliva. No dose-response relationship was observed for the changes in concentrations of calcium and phosphate. It is concluded that for radiation doses of 10 Gy and above, irreversible changes (lag phase, flow rate, potassium, sodium) were observed. A saturation of the irradiation effects (lag phase, flow rate) seems to exist at doses larger than 15 Gy. No significant differences were observed between the radiation-induced functional changes in parotid and submandibular/sublingual salivary gland tissue.

  20. Flight and walking in locusts-cholinergic co-activation, temporal coupling and its modulation by biogenic amines.

    Science.gov (United States)

    Rillich, Jan; Stevenson, Paul A; Pflueger, Hans-Joachim

    2013-01-01

    Walking and flying in locusts are exemplary rhythmical behaviors generated by central pattern generators (CPG) that are tuned in intact animals by phasic sensory inputs. Although these two behaviors are mutually exclusive and controlled by independent CPGs, leg movements during flight can be coupled to the flight rhythm. To investigate potential central coupling between the underlying CPGs, we used the muscarinic agonist pilocarpine and the amines octopamine and tyramine to initiate fictive flight and walking in deafferented locust preparations. Our data illustrate that fictive walking is readily evoked by comparatively lower concentrations of pilocarpine, whereas higher concentrations are required to elicit fictive flight. Interestingly, fictive flight did not suppress fictive walking so that the two patterns were produced simultaneously. Frequently, leg motor units were temporally coupled to the flight rhythm, so that each spike in a step cycle volley occurred synchronously with wing motor units firing at flight rhythm frequency. Similarly, tyramine also induced fictive walking and flight, but mostly without any coupling between the two rhythms. Octopamine in contrast readily evoked fictive flight but generally failed to elicit fictive walking. Despite this, numerous leg motor units were recruited, whereby each was temporarily coupled to the flight rhythm. Our results support the notion that the CPGs for walking and flight are largely independent, but that coupling can be entrained by aminergic modulation. We speculate that octopamine biases the whole motor machinery of a locust to flight whereas tyramine primarily promotes walking.

  1. Acetylcholine plays an antinociceptive role by modulating pain-induced discharges of pain-related neurons in the caudate putamen of rats.

    Science.gov (United States)

    Li, Chun-Mei; Zhang, Da-Ming; Yang, Chun-Xiao; Ma, Xu; Gao, He-Ren; Zhang, Duo; Xu, Man-Ying

    2014-02-12

    The caudate putamen (CPu) has been suggested to be involved in nociceptive modulation. Some neurotransmitters, including acetylcholine (ACh), participate in pain modulation in the central nervous system. However, the active mechanism of ACh on the pain-related neurons in the CPu remains unclear. This study aimed to investigate the effects of the cholinergic agonists ACh and pilocarpine and the muscarinic ACh receptor antagonist atropine on the pain-induced response of pain-related neurons in the CPu of Wistar rats. Trains of electrical impulses applied to the sciatic nerve of rat were used as the noxious stimulus. The electrical activities of pain-excited neurons (PENs) or pain-inhibited neurons (PINs) in the CPu were recorded by a glass microelectrode. Our results showed that an intra-CPu injection of 4 μg/2 μl ACh or pilocarpine decreased and increased the pain-induced discharge frequency in the PENs and PINs, respectively. Intra-CPu administration of 1 μg/2 μl atropine produced the opposite effect on these neurons. These findings indicate that ACh may play an analgesic role by affecting the electric activities of PENs and PINs, and the muscarinic pathway may be involved in the modulation of pain perception in the CPu.

  2. Stereoselectivity of satropane, a novel tropane analog, on iris muscarinic receptor activation and intraocular hypotension

    Institute of Scientific and Technical Information of China (English)

    Liang ZHU; Hong-zhuan CHEN; Li-min YANG; Yong-yao CUI; Pei-li ZHENG; Yin-yao NIU; Hao WANG; Yang LU; Qiu-shi REN; Pi-jing WEI

    2008-01-01

    Aim: To study the stereoselectivity of satropane (3-paramethylbenzene sulfonyloxy-6-acetoxy tropane), a novel tropane analog, on iris muscarinic receptor activation and intraocular hypotension. Methods: The assays for radioligand-receptor binding, the contractile responses of isolated iris muscle, the miosis response, and the intraocular hypotension of the enantiomers of satropane were investigated. Results: In the binding analysis, S(-)satropane (lesatropane) completely com-peted against the [3H]quinuclydinyl benzilate-labeled ligand at muscarinic recep-tors in the iris muscle, whereas R(+)satropane failed to completely compete. In an isolated iris contractile assay, R,S(±)satropane and S(-)satropane produced a concentration-dependent contractile response with similar efficacy and potency to that of carbachol. R(+)satropane did not induce any contractile response. In the pupil diameter measurement assay in vivo, S(-)satropane induced miosis much more effectively than pilocarpine, while R(+)satropane failed to produce any miosis. In the water loading-induced and methylcellulose-induced ocular hypertensive models, S(-)satropane, but not R(+)satropane, significantly suppressed intraocu-lar pressure at a much lower concentration than pilocarpine. Conclusion: The ago-nistic and hypotensive properties of satropane on rabbit eyes are stereoselective, with the S(-)isomer being its active form.

  3. Anti-epileptic effects of focal micro-injection of excitatory amino acid antagonists.

    Science.gov (United States)

    Meldrum, B; Millan, M; Patel, S; de Sarro, G

    1988-01-01

    The role of excitatory synaptic activity at various brain regions in the development and spread of seizure activity has been investigated by the focal microinjection of 2-amino-7-phosphono-heptanoate (2-APH), a selective antagonist at the N-methyl-D-aspartate preferring receptor, or gamma-D-glutamyl-aminomethyl sulphonate (GAMS), a partially selective antagonist at the kainate receptor. In genetically epilepsy prone rats the seizure response to a loud sound in most effectively suppressed by focal injections of 2-APH, 0.1-1.0 nmol, in the inferior colliculus. Protection is also seen after injections of 2-APH, 25 nmoles, in the substantia nigra (pars reticulata) or the midbrain reticular formation. Motor limbic seizures induced by pilocarpine, 380 mg/kg intraperitoneally, are prevented by prior injection into the substantia nigra, pars reticulata, or the entopeduncular nucleus, of 2-APH, 10 nmol or 10 pmol, respectively. Similar protection follows the injection of 2-APH, 1-5 pmol in the piriform cortex. The convulsant effects of pilocarpine are also blocked by the focal injection of GAMS, 10 nmol in the entopeduncular nucleus. This experimental approach can indicate critical sites at which seizure activity is initiated in particular models (e.g., inferior colliculus in sound-induced seizures, and piriform cortex in limbic seizures) and the pathways controlling seizure expression, such as the basal ganglia outputs. It also identifies specific receptors at which anticonvulsant drugs may operate.

  4. Reciprocal regulation of epileptiform neuronal oscillations and electrical synapses in the rat hippocampus.

    Science.gov (United States)

    Kinjo, Erika R; Higa, Guilherme S V; Morya, Edgard; Valle, Angela C; Kihara, Alexandre H; Britto, Luiz R G

    2014-01-01

    Gap junction (GJ) channels have been recognized as an important mechanism for synchronizing neuronal networks. Herein, we investigated the participation of GJ channels in the pilocarpine-induced status epilepticus (SE) by analyzing electrophysiological activity following the blockade of connexins (Cx)-mediated communication. In addition, we examined the regulation of gene expression, protein levels, phosphorylation profile and distribution of neuronal Cx36, Cx45 and glial Cx43 in the rat hippocampus during the acute and latent periods. Electrophysiological recordings revealed that the GJ blockade anticipates the occurrence of low voltage oscillations and promotes a marked reduction of power in all analyzed frequencies.Cx36 gene expression and protein levels remained stable in acute and latent periods, whereas upregulation of Cx45 gene expression and protein redistribution were detected in the latent period. We also observed upregulation of Cx43 mRNA levels followed by changes in the phosphorylation profile and protein accumulation. Taken together, our results indisputably revealed that GJ communication participates in the epileptiform activity induced by pilocarpine. Moreover, considering that specific Cxs undergo alterations through acute and latent periods, this study indicates that the control of GJ communication may represent a focus in reliable anti-epileptogenic strategies.

  5. Effect of pregabalin on apoptotic regulatory genes in hippocampus of rats with chronic temporal lobe epilepsy

    Directory of Open Access Journals (Sweden)

    ZHANG Yi-dan

    2012-04-01

    Full Text Available Objective To observe the effect of pregabalin on the expression of Bcl-2 and Bax in hippocampus of chronic epileptic rats induced by pilocarpine, to explore the anti-epileptic pharmacology mechanism of pregabalin, and its anti-apoptotic effect on hippocampal neurons of rats. Methods The model of chronic temporal lobe epileptic rats induced by lithium-pilocarpine was established, then the rats in pregabalin treatment group received intraperitoneal injection of pregabalin (40 mg/kg once daily for three weeks. The expression of Bcl-2 and Bax in hippocampus of all rats was detected by immunohistochemical technique and Western blotting. Results Compared with normal saline group rats, the expression of Bcl-2 and Bax in hippocampus of rats with chronic temporal lobe epilepsy was significantly increased (P = 0.000, for all. Pregabalin can down-regulate the expression of Bax and up-regulate the expression of Bcl-2 in hippocampus of rats compared to model group rats (P = 0.000, for all. Conclusion Pregabalin may have the effects of inhibiting cell apoptosis and protecting neurons through lowing Bax level and increasing Bcl-2 level in hippocampus of chronic temporal lobe epileptic rats.

  6. Time Course of Pupil Center Location after Ocular Drug Application.

    Science.gov (United States)

    Hoang, Tony A; Macdonnell, Jacqueline E; Mangan, Michelle C; Monsour, Cindy S; Polwattage, Buddhika L; Wilson, Sarah F; Suheimat, Marwan; Atchison, David A

    2016-06-01

    To investigate the time course of pupil centration after application of common topical ocular drugs. Single drops of 2.5% phenylephrine hydrochloride, 1% tropicamide, and 2% pilocarpine hydrochloride were applied on different days to the right eyes of 12 participants. Anterior eye images were captured, at 5-min intervals for an hour, using an infrared-sensitive camera. The images were analyzed to determine pupil diameter and pupil center, the latter with respect to the limbal center. As a control, natural pupil size and pupil center were determined under different illuminances. Pupil centers of natural pupils shifted temporally as pupils dilated. At common pupil sizes, drug-induced pupil centers were different from natural pupil centers. Phenylephrine produced a center shift in the nasal and inferior directions that peaked after a mean of 30 min, whereas dilation was continuing up to 60 min. Tropicamide produced transient center shifts in the nasal and inferior directions that peaked at about 10 min before reducing toward baseline values, whereas dilation reached a peak at about 25 min. Pilocarpine produced a small sustained superior shift that, like constriction, reached a peak after about 25 min. Application of topical ophthalmic drugs cause shifts in pupil center that do not match those produced by natural changes in pupil size and that, in the cases of phenylephrine and tropicamide, follow a different time course than the pupil size changes.

  7. Cannabidiol exerts anti-convulsant effects in animal models of temporal lobe and partial seizures.

    Science.gov (United States)

    Jones, Nicholas A; Glyn, Sarah E; Akiyama, Satoshi; Hill, Thomas D M; Hill, Andrew J; Weston, Samantha E; Burnett, Matthew D A; Yamasaki, Yuki; Stephens, Gary J; Whalley, Benjamin J; Williams, Claire M

    2012-06-01

    Cannabis sativa has been associated with contradictory effects upon seizure states despite its medicinal use by numerous people with epilepsy. We have recently shown that the phytocannabinoid cannabidiol (CBD) reduces seizure severity and lethality in the well-established in vivo model of pentylenetetrazole-induced generalised seizures, suggesting that earlier, small-scale clinical trials examining CBD effects in people with epilepsy warrant renewed attention. Here, we report the effects of pure CBD (1, 10 and 100mg/kg) in two other established rodent seizure models, the acute pilocarpine model of temporal lobe seizure and the penicillin model of partial seizure. Seizure activity was video recorded and scored offline using model-specific seizure severity scales. In the pilocarpine model CBD (all doses) significantly reduced the percentage of animals experiencing the most severe seizures. In the penicillin model, CBD (≥ 10 mg/kg) significantly decreased the percentage mortality as a result of seizures; CBD (all doses) also decreased the percentage of animals experiencing the most severe tonic-clonic seizures. These results extend the anti-convulsant profile of CBD; when combined with a reported absence of psychoactive effects, this evidence strongly supports CBD as a therapeutic candidate for a diverse range of human epilepsies. Copyright © 2012 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.

  8. State and parameter estimation of a neural mass model from electrophysiological signals during the status epilepticus.

    Science.gov (United States)

    López-Cuevas, Armando; Castillo-Toledo, Bernardino; Medina-Ceja, Laura; Ventura-Mejía, Consuelo

    2015-06-01

    Status epilepticus is an emergency condition in patients with prolonged seizure or recurrent seizures without full recovery between them. The pathophysiological mechanisms of status epilepticus are not well established. With this argument, we use a computational modeling approach combined with in vivo electrophysiological data obtained from an experimental model of status epilepticus to infer about changes that may lead to a seizure. Special emphasis is done to analyze parameter changes during or after pilocarpine administration. A cubature Kalman filter is utilized to estimate parameters and states of the model in real time from the observed electrophysiological signals. It was observed that during basal activity (before pilocarpine administration) the parameters presented a standard deviation below 30% of the mean value, while during SE activity, the parameters presented variations larger than 200% of the mean value with respect to basal state. The ratio of excitation-inhibition, increased during SE activity by 80% with respect to the transition state, and reaches the lowest value during cessation. In addition, a progression between low and fast inhibitions before or during this condition was found. This method can be implemented in real time, which is particularly important for the design of stimulation devices that attempt to stop seizures. These changes in the parameters analyzed during seizure activity can lead to better understanding of the mechanisms of epilepsy and to improve its treatments.

  9. Mucoadhesive ophthalmic vehicles: evaluation of polymeric low-viscosity formulations.

    Science.gov (United States)

    Saettone, M F; Monti, D; Torracca, M T; Chetoni, P

    1994-01-01

    A series of polyanionic natural or semi-synthetic polymers (polygalacturonic acid, hyaluronic acid, carboxymethylamylose, carboxymethylchitin, chondroitin sulfate, heparan sulfate and mesoglycan) were evaluated as potential mucoadhesive carriers for ophthalmic drugs. Solutions containing cyclopentolate (CY) or pilocarpine (PI) as salts (or polyanionic complexes) with the acidic polymers, all showing a low viscosity, were tested for miotic (resp. mydriatic) activity in albino rabbits. In the case of some polymeric complexes, small but significant increases of the areas under the activity vs. time curves (AUC) over reference cyclopentolate hydrochloride (CYHC1) or pilocarpine nitrate (PINO3) vehicles, and significant AUC decreases after removal of precorneal mucin by treatment with N-acetylcysteine were observed. A correlation was found between these data, considered indicative of the occurrence of a mucoadhesive interaction "in vivo", and "in vitro" viscometric data expressing the polymers-mucin force of interaction. The advantages and limitations of the mucoadhesive non-viscous approach in the formulation of ophthalmic vehicles are presented and discussed.

  10. Etiology matters - Genomic DNA Methylation Patterns in Three Rat Models of Acquired Epilepsy.

    Science.gov (United States)

    Dębski, Konrad J; Pitkanen, Asla; Puhakka, Noora; Bot, Anna M; Khurana, Ishant; Harikrishnan, K N; Ziemann, Mark; Kaspi, Antony; El-Osta, Assam; Lukasiuk, Katarzyna; Kobow, Katja

    2016-05-09

    This study tested the hypothesis that acquired epileptogenesis is accompanied by DNA methylation changes independent of etiology. We investigated DNA methylation and gene expression in the hippocampal CA3/dentate gyrus fields at 3 months following epileptogenic injury in three experimental models of epilepsy: focal amygdala stimulation, systemic pilocarpine injection, or lateral fluid-percussion induced traumatic brain injury (TBI) in rats. In the models studies, DNA methylation and gene expression profiles distinguished controls from injured animals. We observed consistent increased methylation in gene bodies and hypomethylation at non-genic regions. We did not find a common methylation signature in all three different models and few regions common to any two models. Our data provide evidence that genome-wide alteration of DNA methylation signatures is a general pathomechanism associated with epileptogenesis and epilepsy in experimental animal models, but the broad pathophysiological differences between models (i.e. pilocarpine, amygdala stimulation, and post-TBI) are reflected in distinct etiology-dependent DNA methylation patterns.

  11. Etiology matters – Genomic DNA Methylation Patterns in Three Rat Models of Acquired Epilepsy

    Science.gov (United States)

    Dębski, Konrad J.; Pitkanen, Asla; Puhakka, Noora; Bot, Anna M.; Khurana, Ishant; Harikrishnan, KN; Ziemann, Mark; Kaspi, Antony; El-Osta, Assam; Lukasiuk, Katarzyna; Kobow, Katja

    2016-01-01

    This study tested the hypothesis that acquired epileptogenesis is accompanied by DNA methylation changes independent of etiology. We investigated DNA methylation and gene expression in the hippocampal CA3/dentate gyrus fields at 3 months following epileptogenic injury in three experimental models of epilepsy: focal amygdala stimulation, systemic pilocarpine injection, or lateral fluid-percussion induced traumatic brain injury (TBI) in rats. In the models studies, DNA methylation and gene expression profiles distinguished controls from injured animals. We observed consistent increased methylation in gene bodies and hypomethylation at non-genic regions. We did not find a common methylation signature in all three different models and few regions common to any two models. Our data provide evidence that genome-wide alteration of DNA methylation signatures is a general pathomechanism associated with epileptogenesis and epilepsy in experimental animal models, but the broad pathophysiological differences between models (i.e. pilocarpine, amygdala stimulation, and post-TBI) are reflected in distinct etiology-dependent DNA methylation patterns. PMID:27157830

  12. Flight and walking in locusts-cholinergic co-activation, temporal coupling and its modulation by biogenic amines.

    Directory of Open Access Journals (Sweden)

    Jan Rillich

    Full Text Available Walking and flying in locusts are exemplary rhythmical behaviors generated by central pattern generators (CPG that are tuned in intact animals by phasic sensory inputs. Although these two behaviors are mutually exclusive and controlled by independent CPGs, leg movements during flight can be coupled to the flight rhythm. To investigate potential central coupling between the underlying CPGs, we used the muscarinic agonist pilocarpine and the amines octopamine and tyramine to initiate fictive flight and walking in deafferented locust preparations. Our data illustrate that fictive walking is readily evoked by comparatively lower concentrations of pilocarpine, whereas higher concentrations are required to elicit fictive flight. Interestingly, fictive flight did not suppress fictive walking so that the two patterns were produced simultaneously. Frequently, leg motor units were temporally coupled to the flight rhythm, so that each spike in a step cycle volley occurred synchronously with wing motor units firing at flight rhythm frequency. Similarly, tyramine also induced fictive walking and flight, but mostly without any coupling between the two rhythms. Octopamine in contrast readily evoked fictive flight but generally failed to elicit fictive walking. Despite this, numerous leg motor units were recruited, whereby each was temporarily coupled to the flight rhythm. Our results support the notion that the CPGs for walking and flight are largely independent, but that coupling can be entrained by aminergic modulation. We speculate that octopamine biases the whole motor machinery of a locust to flight whereas tyramine primarily promotes walking.

  13. Fine structure of bat deep posterior lingual glands (von Ebner's)

    Science.gov (United States)

    Azzali, G; Gatti, R; Bucci, G; Orlandini, G

    1989-10-01

    We studied the morphology and ultrastructure of the bat (Pipistrellus k.k. and Rhinolophus f.e.) deep posterior lingual glands (Ebner's glands) during hibernation, summer and after stimulation with pilocarpine. Ebner's glands are formed by serous tubulo-alveolar adenomeres and by an excretory system organized in intercalated ducts, long excretory ducts and a main excretory duct. The latter opens in the vallum which surrounds the circumvallate papillae and in the groove of the foliate papillae. The secretory cells, which lack basal folds, show abundant and dense granules (PAS+, Alcian blue -), microvilli (scarce during hibernation), a Golgi apparatus (well developed during summer and after stimulation with pilocarpine), a large nucleus and RER cisternae stacked at the basal pole. Centrioles, lipid droplets, heterogeneous bodies (in content and density, probably lipofuscin bodies), lysosomal multivesicular bodies and large, dense granules with a microcrystalline structure were also encountered. The lateral membranes of adjacent cells are joined by desmosomes; their interdigitations are neither numerous nor prominent during summer. Microfilaments, often gathered in small bundles, lie in the lateral, peripheral cytoplasm without any relation with desmosomes. In summer and particularly after stimulation with pilocarpine, the apical pole of the secretory cells is characterized by many long microvilli, pedunculated hyaloplasmic protrusions and secretory granules. During hibernation the lumen is filled with secretory material. Myoepithelial cells are arranged among secretory cells or between them and the basal lamina. The short intercalated ducts show similarities with the analogous ducts of the parotid gland. Striated ducts are absent. Excretory ducts are endowed with: a) an inner layer of cuboidal cells characterized by poorly developed cytoplasmic organelles, rare dense granules and a few small microvilli; b) an outer layer of basal cells lying on the basal lamina

  14. Human fetal brain-derived neural stem/progenitor cells grafted into the adult epileptic brain restrain seizures in rat models of temporal lobe epilepsy.

    Science.gov (United States)

    Lee, Haejin; Yun, Seokhwan; Kim, Il-Sun; Lee, Il-Shin; Shin, Jeong Eun; Park, Soo Chul; Kim, Won-Joo; Park, Kook In

    2014-01-01

    Cell transplantation has been suggested as an alternative therapy for temporal lobe epilepsy (TLE) because this can suppress spontaneous recurrent seizures in animal models. To evaluate the therapeutic potential of human neural stem/progenitor cells (huNSPCs) for treating TLE, we transplanted huNSPCs, derived from an aborted fetal telencephalon at 13 weeks of gestation and expanded in culture as neurospheres over a long time period, into the epileptic hippocampus of fully kindled and pilocarpine-treated adult rats exhibiting TLE. In vitro, huNSPCs not only produced all three central nervous system neural cell types, but also differentiated into ganglionic eminences-derived γ-aminobutyric acid (GABA)-ergic interneurons and released GABA in response to the depolarization induced by a high K+ medium. NSPC grafting reduced behavioral seizure duration, afterdischarge duration on electroencephalograms, and seizure stage in the kindling model, as well as the frequency and the duration of spontaneous recurrent motor seizures in pilocarpine-induced animals. However, NSPC grafting neither improved spatial learning or memory function in pilocarpine-treated animals. Following transplantation, grafted cells showed extensive migration around the injection site, robust engraftment, and long-term survival, along with differentiation into β-tubulin III+ neurons (∼34%), APC-CC1+ oligodendrocytes (∼28%), and GFAP+ astrocytes (∼8%). Furthermore, among donor-derived cells, ∼24% produced GABA. Additionally, to explain the effect of seizure suppression after NSPC grafting, we examined the anticonvulsant glial cell-derived neurotrophic factor (GDNF) levels in host hippocampal astrocytes and mossy fiber sprouting into the supragranular layer of the dentate gyrus in the epileptic brain. Grafted cells restored the expression of GDNF in host astrocytes but did not reverse the mossy fiber sprouting, eliminating the latter as potential mechanism. These results suggest that human fetal

  15. Neurogenesis induced by seizures in the dentate gyrus is not related to mossy fiber sprouting but is age dependent in developing rats A neurogênese induzida por crises no giro denteado não está relacionada ao brotamento de fibras musgosas, mas é dependente da idade, em ratos durante o desenvolvimento

    Directory of Open Access Journals (Sweden)

    Yaima del Carmen Garrido Sanabria

    2008-12-01

    Full Text Available Neurogenesis in the dentate gyrus (DG has attracted attention since abnormal supragranular mossy fiber sprouting occurs in the same region, in temporal lobe epilepsy. Thus, we submitted developing rats to pilocarpine-induced status epilepticus (SE to study the relationship between neurogenesis and mossy fiber sprouting. Groups were submitted to SE at: I-P9, II-P7, P8 and P9, III-P17 e IV-P21. Neurogenesis was quantified using BrdU protocol and confirmed through double staining, using neuronal pentraxin. Other animals were monitored by video system until P120 and their brain was studied (Timm and Nissl staining. The neurogenesis at P17 (p=0.007 and P21 (p=0.006 were increased. However, only P21 group showed recurrent seizures and the mossy fiber sprouting in the same region, during adult life, while P17 did not. Thus, our results suggest that neurogenesis is not related to mossy fiber sprouting neither to recurrent spontaneous seizures in pilocarpine model.A neurogênese no giro dentado tem atraído atenção já que ela ocorre na mesma região do hipocampo que o brotamento das fibras musgosas, na epilepsia do lobo temporal. Assim, submetemos ratos em desenvolvimento ao status epilepticus induzido (SE por pilocarpine. Grupos foram submetidos em I-P9, II-P7, P8, P9; III-P17 e IV-P21. A neurogênese foi observada usando o protocolo do BrdU e confirmada por dupla marcação com pentraxina neuronal. Outros animais foram monitorados até P120 e seus cérebros analisados (Nissl e Timm. A neurogênese nos grupos P17 (p=0,007 e P21 (p=0,006 aumentaram. Entretanto, o P21 apresentou crises espontâneas e brotamento de fibras musgosas, na mesma região onde ocorreu a neurogênese, enquanto o grupo P17 apresentou somente aumento na neurogênese. Assim, nossos resultados sugerem que o fenômeno da neurogênese não está relacionado com o brotamento de fibras musgosas nem com o aparecimento de crises espontâneas e recorrentes no modelo da pilocarpina.

  16. The protective effect of curcumin on hippocampus of epileptic mouse%姜黄素对癫痫小鼠海马脑损伤的保护作用

    Institute of Scientific and Technical Information of China (English)

    孟伟; 杨锦青; 鞠培新; 贾本智; 李洪秀

    2012-01-01

    Objective To investigate the effects of curcumin on newborn neurons and apoptosis in epileptic mouse hippocampus. Method Experimental animals (pre-treatment with curcumin) were injected (i.p.) with a dose of 300 mg/Kg of pilocarpine to induce seizure, 72 h after pilocarpine treatment, doublecortin (DCX) iminunotiistoehemistry and TUNEL staining were used to detect newborn neurons and apoptosis in epileptic hippocampus. Results Compared with controls, there were less DCX positive cells in granular cell layer of the model group and curcumin treated model group, whereas more DCX positive cells in curcumin treated model group than in model group. TUNEL staining indicated that less TUNEL positive cells in the dentate gyrus (DG) were found in curcunrin treated model group than in model group. Conclusion Curcumin has a protective effect on hippocampus neurons of the pilocarpine-induced epileptic mouse.%目的 研究姜黄素对匹罗卡品诱导的癫痫小鼠海马新生神经元和细胞凋亡的影响.方法 姜黄素预处理后,小鼠腹腔注射匹罗卡品建立小鼠癫痫模型,应用新生神经元标记物双皮层蛋白( doublecortin,DCX)免疫组织化学染色及TUNEL染色对造模后72h的模型小鼠海马进行检测.结果 DCX免疫组织化学染色结果表明,与对照组相比,模型组及姜黄素处理模型组小鼠海马齿状回DCX阳性细胞明显减少;与模型组相比,姜黄素处理模型组小鼠海马齿状回DCX阳性细胞明显增多.TUNEL染色结果表明,与对照组相比,模型组及姜黄素处理模型组小鼠海马齿状回TUNEL阳性细胞明显增多;与模型组相比,姜黄素处理模型组小鼠海马齿状回TUNEL阳性细胞明显减少.结论 姜黄素可能对匹罗卡品诱导的癫痫小鼠海马神经元有保护作用.

  17. Surviving mossy cells enlarge and receive more excitatory synaptic input in a mouse model of temporal lobe epilepsy.

    Science.gov (United States)

    Zhang, Wei; Thamattoor, Ajoy K; LeRoy, Christopher; Buckmaster, Paul S

    2015-05-01

    Numerous hypotheses of temporal lobe epileptogenesis have been proposed, and several involve hippocampal mossy cells. Building on previous hypotheses we sought to test the possibility that after epileptogenic injuries surviving mossy cells develop into super-connected seizure-generating hub cells. If so, they might require more cellular machinery and consequently have larger somata, elongate their dendrites to receive more synaptic input, and display higher frequencies of miniature excitatory synaptic currents (mEPSCs). To test these possibilities pilocarpine-treated mice were evaluated using GluR2-immunocytochemistry, whole-cell recording, and biocytin-labeling. Epileptic pilocarpine-treated mice displayed substantial loss of GluR2-positive hilar neurons. Somata of surviving neurons were 1.4-times larger than in controls. Biocytin-labeled mossy cells also were larger in epileptic mice, but dendritic length per cell was not significantly different. The average frequency of mEPSCs of mossy cells recorded in the presence of tetrodotoxin and bicuculline was 3.2-times higher in epileptic pilocarpine-treated mice as compared to controls. Other parameters of mEPSCs were similar in both groups. Average input resistance of mossy cells in epileptic mice was reduced to 63% of controls, which is consistent with larger somata and would tend to make surviving mossy cells less excitable. Other intrinsic physiological characteristics examined were similar in both groups. Increased excitatory synaptic input is consistent with the hypothesis that surviving mossy cells develop into aberrantly super-connected seizure-generating hub cells, and soma hypertrophy is indirectly consistent with the possibility of axon sprouting. However, no obvious evidence of hyperexcitable intrinsic physiology was found. Furthermore, similar hypertrophy and hyper-connectivity has been reported for other neuron types in the dentate gyrus, suggesting mossy cells are not unique in this regard. Thus

  18. Decreased sweating in seven patients with Laron syndrome

    DEFF Research Database (Denmark)

    Main, K M; Price, D A; Savage, M O;

    1993-01-01

    Previous studies have shown that sweat secretion was reduced in patients with GH deficiency and increased during GH treatment, indicating an influence of GH on sweat gland function. Thus, patients with GH deficiency have impaired thermoregulation. We report on sweat secretion rates (SSRs) in seven...... patients with Laron syndrome, measured by pilocarpine iontophoresis. The patients had significantly lower SSRs than healthy children matched for sex and pubertal stage (P sweat electrolyte concentrations (P ...). These observations further supported the hypothesis that sweat gland function in humans is under the influence of the GH-insulin-like growth factor-I axis. It remains to be seen whether the decrease in SSR also leads to altered thermoregulation in patients with Laron syndrome....

  19. Hilar somatostatin interneuron loss reduces dentate gyrus inhibition in a mouse model of temporal lobe epilepsy.

    Science.gov (United States)

    Hofmann, Gabrielle; Balgooyen, Laura; Mattis, Joanna; Deisseroth, Karl; Buckmaster, Paul S

    2016-06-01

    In patients with temporal lobe epilepsy, seizures usually start in the hippocampus, and dentate granule cells are hyperexcitable. Somatostatin interneurons are a major subpopulation of inhibitory neurons in the dentate gyrus, and many are lost in patients and animal models. However, surviving somatostatin interneurons sprout axon collaterals and form new synapses, so the net effect on granule cell inhibition remains unclear. The present study uses optogenetics to activate hilar somatostatin interneurons and measure the inhibitory effect on dentate gyrus perforant path-evoked local field potential responses in a mouse model of temporal lobe epilepsy. In controls, light activation of hilar somatostatin interneurons inhibited evoked responses up to 40%. Epileptic pilocarpine-treated mice exhibited loss of hilar somatostatin interneurons and less light-induced inhibition of evoked responses. These findings suggest that severe epilepsy-related loss of hilar somatostatin interneurons can overwhelm the surviving interneurons' capacity to compensate by sprouting axon collaterals. Wiley Periodicals, Inc. © 2016 International League Against Epilepsy.

  20. Temporal lobe epilepsy and social behavior: an animal model for autism?

    Science.gov (United States)

    Marin, Joao Carlos M; Moura, Paula J; Cysneiros, Roberta M; Colugnati, Diego B; Cavalheiro, Esper A; Scorza, Fulvio A; Xavier, Gilberto F; Zilbovicius, Monica; Mercadante, Marcos T

    2008-07-01

    Social behavior depends on the integrity of social brain circuitry. The temporal lobe is an important part of the social brain, and manifests morphological and functional alterations in autism spectrum disorders (ASD). Rats with temporal lobe epilepsy (TLE), induced with pilocarpine, were subjected to a social discrimination test that has been used to investigate potential animal models of ASD, and the results were compared with those for the control group. Rats with TLE exhibited fewer social behaviors than controls. No differences were observed in nonsocial behavior between groups. The results suggest an important role for the temporal lobe in regulating social behaviors. This animal model might be used to explore some questions about ASD pathophysiology.

  1. Distinct rhythmic locomotor patterns can be generated by a simple adaptive neural circuit: biology, simulation, and VLSI implementation.

    Science.gov (United States)

    Ryckebusch, S; Wehr, M; Laurent, G

    1994-12-01

    Rhythmic motor patterns can be induced in leg motor neurons of isolated locust thoracic ganglia by bath application of pilocarpine. We observed that the relative phases of levators and depressors differed in the three thoracic ganglia. Assuming that the central pattern generating circuits underlying these three segmental rhythms are probably very similar, we developed a simple model circuit that can produce any one of the three activity patterns and characteristic phase relationships by modifying a single synaptic weight. We show results of a computer simulation of this circuit using the neuronal simulator NeuraLOG/Spike. We built and tested an analog VLSI circuit implementation of this model circuit that exhibits the same range of "behaviors" as the computer simulation. This multidisciplinary strategy will be useful to explore the dynamics of central pattern generating networks coupled to physical actuators, and ultimately should allow the design of biologically realistic walking robots.

  2. Aberrant hippocampal neurogenesis contributes to epilepsy and associated cognitive decline.

    Science.gov (United States)

    Cho, Kyung-Ok; Lybrand, Zane R; Ito, Naoki; Brulet, Rebecca; Tafacory, Farrah; Zhang, Ling; Good, Levi; Ure, Kerstin; Kernie, Steven G; Birnbaum, Shari G; Scharfman, Helen E; Eisch, Amelia J; Hsieh, Jenny

    2015-03-26

    Acute seizures after a severe brain insult can often lead to epilepsy and cognitive impairment. Aberrant hippocampal neurogenesis follows the insult but the role of adult-generated neurons in the development of chronic seizures or associated cognitive deficits remains to be determined. Here we show that the ablation of adult neurogenesis before pilocarpine-induced acute seizures in mice leads to a reduction in chronic seizure frequency. We also show that ablation of neurogenesis normalizes epilepsy-associated cognitive deficits. Remarkably, the effect of ablating adult neurogenesis before acute seizures is long lasting as it suppresses chronic seizure frequency for nearly 1 year. These findings establish a key role of neurogenesis in chronic seizure development and associated memory impairment and suggest that targeting aberrant hippocampal neurogenesis may reduce recurrent seizures and restore cognitive function following a pro-epileptic brain insult.

  3. Behavioral deficit and decreased GABA receptor functional regulation in the hippocampus of epileptic rats: effect of Bacopa monnieri.

    Science.gov (United States)

    Mathew, Jobin; Gangadharan, Gireesh; Kuruvilla, Korah P; Paulose, C S

    2011-01-01

    In the present study, alterations of the General GABA and GABA(A) receptors in the hippocampus of pilocarpine-induced temporal lobe epileptic rats and the therapeutic application of Bacopa monnieri and its active component Bacoside-A were investigated. Bacopa monnieri (Linn.) is a herbaceous plant belonging to the family Scrophulariaceae. Hippocampus is the major region of the brain belonging to the limbic system and plays an important role in epileptogenesis, memory and learning. Scatchard analysis of [³H]GABA and [³H]bicuculline in the hippocampus of the epileptic rat showed significant decrease in B(max) (P Bacoside-A treatment reverses all these changes near to control. Our results suggest that decreased GABA receptors in the hippocampus have an important role in epilepsy associated behavioral deficit, Bacopa monnieri and Bacoside-A have clinical significance in the management of epilepsy.

  4. 本刊参考文献格式

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    专著主要作者.文献题名[M].出版地:出版者,出版年:起止页码.例:凌沛学.透明质酸[M].北京:中国轻工业出版社,2000:3-12.专著(或论文集)中析出文献析出文献主要作者.析出文献题名[M或C]//专著主要责任者.专著题名.出版地:出版者,出版年:析出文献的页码.例:Ling P X,Guo X P,Zhang T M,et al.Efficacy promoting effect ofhyaluronan on pilocarpine nitrate eye drops[C]//Kennedy J F.Hyaluronan(vol 2).Cambridge:Woodhead,2002:257-260.

  5. Neuroaminidase reduces interictal spikes in a rat temporal lobe epilepsy model.

    Science.gov (United States)

    Isaev, Dmytro; Zhao, Qian; Kleen, Jonathan K; Lenck-Santini, Pierre Pascal; Adstamongkonkul, Dusit; Isaeva, Elena; Holmes, Gregory L

    2011-03-01

    Interictal spikes have been implicated in epileptogenesis and cognitive dysfunction in epilepsy. Unfortunately, antiepileptic drugs have shown poor efficacy in suppressing interictal discharges; novel therapies are needed. Surface charge on neuronal membranes provides a novel target for abolishing interictal spikes. This property can be modulated through the use of neuraminidase, an enzyme that decreases the amount of negatively charged sialic acid. In the present report we determined whether applying neuraminidase to brains of rats with a history of status epilepticus would reduce number of interictal discharges. Following pilocarpine-induced status epilepticus, rats received intrahippocampal injections of neuraminidase, which significantly decreased the number of interictal spikes recorded in the CA1 region. This study provides evidence that sialic acid degradation can reduce the number of interictal spikes. Furthermore, the results suggest that modifying surface charge created by negatively charged sialic acid may provide new opportunities for reducing aberrant epileptiform events in epilepsy.

  6. PENGARUH EKSTRAK BEBERAPA TANAMAN OBAT TERHADAP USUS TERISOLASI

    Directory of Open Access Journals (Sweden)

    B. Dzulkarnain

    2012-09-01

    Full Text Available The extraction of Anacardium occidentale L.leaves, Aegle marmelos Corr leaves and wood bark, Acorus calamus L. tuber and Desmodium triquetrum D.C. leaves has been tested on the isolated rabbit and guinea pig intestine. The extraction of A. occidentale L. leaves stimulated the isolated rabbit and guinea pig intestine which may due to the anacardic acid content. No consistent influence was seen by the extraction of A.marmelos Corr. leaves and wood bark. The A. calamus L. tuber extraction decreases the isolated intestine activities which is of the atropine-like type not antihistamin one. This may explain the use as antidysentri agent from the motility point of view. The D. triquetrum D.C. leaves extraction stimulated the isolated intestine which has a pilocarpine and histamine-like activity but does not exclude a seretonine-like action.

  7. Amentoflavone protects hippocampal neurons: anti-inflammatory, antioxidative, and antiapoptotic effects

    Directory of Open Access Journals (Sweden)

    Zhen Zhang

    2015-01-01

    Full Text Available Amentoflavone is a natural biflavone compound with many biological properties, including anti-inflammatory, antioxidative, and neuroprotective effects. We presumed that amentoflavone exerts a neuroprotective effect in epilepsy models. Prior to model establishment, mice were intragastrically administered 25 mg/kg amentoflavone for 3 consecutive days. Amentoflavone effectively prevented pilocarpine-induced epilepsy in a mouse kindling model, suppressed nuclear factor-κB activation and expression, inhibited excessive discharge of hippocampal neurons resulting in a reduction in epileptic seizures, shortened attack time, and diminished loss and apoptosis of hippocampal neurons. Results suggested that amentoflavone protected hippocampal neurons in epilepsy mice via anti-inflammation, antioxidation, and antiapoptosis, and then effectively prevented the occurrence of seizures.

  8. Spontaneous recurrent seizures in rats: an experimental model of partial epilepsy.

    Science.gov (United States)

    Leite, J P; Bortolotto, Z A; Cavalheiro, E A

    1990-01-01

    Seizures induced by pilocarpine (PILO) have proven to be a useful procedure for investigating the basic mechanisms essential for generation, spread and motor expression of seizures in rodents. Here we report the long-term effects of PILO in rats. Following PILO (380 mg/kg, IP), 3 distinct phases were observed: 1) an acute period which lasted 1-2 days which corresponds to the pattern of repetitive seizures and status epilepticus; 2) a silent period (4-44 days) characterized by a progressive return to normal EEG and behavior; and 3) a period of recurrent seizures which started 5-45 days after PILO and lasted up to 120 days. These seizures lasted up to 50-60 sec, recurred 2-3 times per week and were more frequent during the light period of the light-dark cycle. These serial events offer a new method to induce spontaneous recurrent seizures in rats.

  9. Detection of visual signals by rats: effects of chlordiazepoxide and cholinergic and adrenergic drugs on sustained attention.

    Science.gov (United States)

    Bushnell, P J; Oshiro, W M; Padnos, B K

    1997-12-01

    Central cholinergic and adrenergic pathways support the attentional processes necessary for detecting and reporting temporally unpredictable stimuli. To assess the functional effects of pharmacological manipulations of these pathways, male Long-Evans rats performed a two-choice, discrete-trial signal-detection task in which food was provided for pressing one lever after presentation of a signal (a 300-ms light flash), and for pressing a second lever at the end of a trial lacking a signal. Seven signal intensities were presented during each 1-h session in a pseudo-random order across three 100-trial blocks. After acquisition of a stable performance baseline, the acute effects of chlordiazepoxide (0, 3, 5, 8 mg/kg i.p.), pilocarpine (0, 1.0, 1.8, 3.0 mg/kg s.c.), scopolamine 0, 0.030, 0.056, 0.100 mg/kg s.c.), nicotine (0, 0.08, 0.25, 0.75 mg/kg s.c.), mecamylamine (0, 1.8, 3.0, 5.6 mg/kg i.p.), clonidine (0, 0.003, 0.010, 0.030 mg/kg s.c.), and idazoxan (0, 1, 3, 10 mg/kg s.c.) were assessed. Five measures of performance were analyzed: response failures; the proportion of "hits" [P(hit): the proportion of correct responses on signal trials]; the proportion of "false alarms" [P(fa): the proportion of incorrect responses on non-signal trials]; and response times (RT) for hits and for correct rejections. All drugs which slowed responding affected RT for hits and correct rejections equivalently, suggesting little or no influence of motor slowing on choice accuracy. Chlordiazepoxide reduced P(hit) at low signal intensities only, without affecting P(fa) or RT, consistent with sensory impairment (reduced visual sensitivity). All other drugs except nicotine reduced P(hit) at high signal intensities preferentially, suggesting a non-visual source of the impairment. Scopolamine, mecamylamine and clonidine affected both P(hit) and P(fa); pilocarpine and idazoxan reduced P(hit) without affecting P(fa). Nicotine at 0.75 mg/kg decreased P(hit) in the first block of trials; at 0

  10. Nematode cholinergic pharmacology

    Energy Technology Data Exchange (ETDEWEB)

    Segerberg, M.A.

    1989-01-01

    Nematode acetylcholine (ACh) receptors were characterized using both biochemical and electrophysiological techniques, including: (1) receptor binding studies in crude homogenates of the free-living nematode Caenorhabditis elegans and the parasitic nematode Ascaris lumbricoides with the high-affinity probe ({sup 3}H)N-methylscopolamine (({sup 3}H)NMS) which binds to muscarinic receptors in many vertebrate and invertebrate tissues (2) measurement of depolarization and contraction induced by a variety of cholinergic agents, including N-methylscopolamine (NMS), in an innervated dorsal muscle strip preparation of Ascaris; (3) examination of the antagonistic actions of d-tubocurarine (dTC) and NMS at dorsal neuromuscular junction; (4) measurement of input resistance changes in Ascaris commissural motorneurons induced by ACh, dTC, NMS, pilocarpine and other cholinergic drugs.

  11. SINUSITIS, BRONCHIECTASIS, AND FLATUS IN A SUMATRAN ORANGUTAN (PONGO ABELII): COULD THIS BE CYSTIC FIBROSIS?

    Science.gov (United States)

    Stringer, Elizabeth; Cossaboon, Cindy; Han, Sushan; Taylor-Cousar, Jennifer L

    2016-03-01

    A 31-yr-old male Sumatran orangutan (Pongo abelii) presented with 14 yr of chronic purulent nasal drainage and cough with intermittent exacerbation of symptoms requiring systemic antibiotic treatment. He was diagnosed with a cystic fibrosis (CF)-like condition. Evaluation consisted of bronchoscopy with bronchoalveolar lavage, culture, and computed tomography scanning of the sinuses and chest. Although the presence of low fecal elastase activity increased the suspicion for a diagnosis of CF, pilocarpine iontophoresis with sweat collection and analysis ("sweat testing") was inconclusive. Medical management included twice-daily nebulization with bronchodilators and alternating month inhaled antibiotics, pancreatic enzyme replacement therapy, and simethicone as needed. Sinopulmonary and gastrointestinal symptoms improved substantially with treatment. Several years later, the animal died acutely of colonic volvulus. Necropsy and histopathology confirmed CF-like lung disease with chronic air sacculitis.

  12. Attenuation of cocaine's reinforcing and discriminative stimulus effects via muscarinic M1 acetylcholine receptor stimulation

    DEFF Research Database (Denmark)

    Thomsen, Morgane; Conn, P Jeffrey; Lindsley, Craig

    2010-01-01

    Muscarinic cholinergic receptors modulate dopaminergic function in brain pathways thought to mediate cocaine's abuse-related effects. Here, we sought to confirm and extend in the mouse species findings that nonselective muscarinic receptor antagonists can enhance cocaine's discriminative stimulus....... More importantly, we tested the hypothesis that muscarinic receptor agonists with varied receptor subtype selectivity can blunt cocaine's discriminative stimulus and reinforcing effects; we hypothesized a critical role for the M(1) and/or M(4) receptor subtypes in this modulation. Mice were trained...... to discriminate cocaine from saline, or to self-administer intravenous cocaine chronically. The nonselective muscarinic antagonists scopolamine and methylscopolamine, the nonselective muscarinic agonists oxotremorine and pilocarpine, the M(1)/M(4)-preferring agonist xanomeline, the putative M(1)-selective agonist...

  13. Chlordiazepoxide and diazepam induced mouse killing by rats.

    Science.gov (United States)

    Leaf, R C; Wnek, D J; Gay, P E; Corcia, R M; Lamon, S

    1975-10-14

    Chlordiazepoxide HCl, at dose levels from 2.5 mg/kg to 80 mg/kg, significantly increased the low base rates of mouse killing (3-9%) observed in large samples (N = 100/dose) of Holtzman strain albino male rats. Maximal killing rates were obtained at doses from 7.5 mg/kg to 20 mg/kg. Diazepam was equally effective, and several times more potent than chlordiazepoxide. Pentobarbital did not increase killing. Killing induced by chlordiazepoxide was blocked by d-amphetamine SO4, but not by l-amphetamine, at dose levels similar to those that block undrugged killing in this strain (ED50 = 1.5 mg/kg). Unlike pilocarpine-induced killing, the effects of chlordiazepoxide were not increased or decreased significantly by either peripherally or centrally active anticholinergic drugs, over wide dose ranges of these agents; nor were the effects of chlordiazepoxide increased by repeated daily administration.

  14. Antimycobacterial activity of chemically defined natural substances from the Caribbean flora in Guadeloupe.

    Science.gov (United States)

    Rastogi, N; Abaul, J; Goh, K S; Devallois, A; Philogène, E; Bourgeois, P

    1998-04-01

    Eight chemically defined, naturally occurring compounds were extracted from the tropical flora of the Caribbean island of Guadeloupe: pilocarpine, an alkaloid from Pilocarpus racemosus; heraclenol and isomeranzin, coumarins from Triphasia trifolia; lochnerin, an indole alkaloid from Rauwolfia biauriculata; ibogaine and voacangine, indole alkaloids from Tabernaemontana citrifolia; texalin, an oxazole from Amyris elemifera; and canellal, a sesquiterpene dialdehyde from Canella winterana. An essential oil fraction from Canella winterana was also tested. The antimycobacterial activity of these substances was tested against Mycobacterium tuberculosis, M. avium and M. kansasii using the Middlebrook 7H11 agar medium, the Bactec 460-TB radiometric methodology, and determination of bacterial viable counts. Three compounds, namely ibogaine, voacangine and texalin, showed antimycobacterial activity. Investigations on the structure-modification and structure-activity relationships of these compounds may help determine new targets for future drug development.

  15. Reconstruction of the adenosine system by bone marrow-derived mesenchymal stem cell transplantation

    Institute of Scientific and Technical Information of China (English)

    Huicong Kang; Qi Hu; Xiaoyan Liu; Yinhe Liu; Feng Xu; Xiang Li; Suiqiang Zhu

    2012-01-01

    In the present study, we transplanted bone marrow-derived mesenchymal stem cells into the CA3 area of the hippocampus of chronic epilepsy rats kindled by lithium chloride-pilocarpine. Immunofluorescence and western blotting revealed an increase in adenosine A1 receptor expression and a decrease in adenosine A2a receptor expression in the brain tissues of epileptic rats 3 months after transplantation. Moreover, the imbalance in the A1 adenosine receptor/A2a adenosine receptor ratio was improved. Electroencephalograms showed that frequency and amplitude of spikes in the hippocampus and frontal lobe were reduced. These results suggested that mesenchymal stem cell transplantation can reconstruct the normal function of the adenosine system in the brain and greatly improve epileptiform discharges.

  16. Role of adenosine in the antiepileptic effects of deep brain stimulation

    Science.gov (United States)

    Miranda, Maisa F.; Hamani, Clement; de Almeida, Antônio-Carlos G.; Amorim, Beatriz O.; Macedo, Carlos E.; Fernandes, Maria José S.; Nobrega, José N.; Aarão, Mayra C.; Madureira, Ana Paula; Rodrigues, Antônio M.; Andersen, Monica L.; Tufik, Sergio; Mello, Luiz E.; Covolan, Luciene

    2014-01-01

    Despite the effectiveness of anterior thalamic nucleus (AN) deep brain stimulation (DBS) for the treatment of epilepsy, mechanisms responsible for the antiepileptic effects of this therapy remain elusive. As adenosine modulates neuronal excitability and seizure activity in animal models, we hypothesized that this nucleoside could be one of the substrates involved in the effects of AN DBS. We applied 5 days of stimulation to rats rendered chronically epileptic by pilocarpine injections and recorded epileptiform activity in hippocampal slices. We found that slices from animals given DBS had reduced hippocampal excitability and were less susceptible to develop ictal activity. In live animals, AN DBS significantly increased adenosine levels in the hippocampus as measured by microdialysis. The reduced excitability of DBS in vitro was completely abolished in animals pre-treated with A1 receptor antagonists and was strongly potentiated by A1 receptor agonists. We conclude that some of the antiepileptic effects of DBS may be mediated by adenosine. PMID:25324724

  17. The function study of entorhinal cortex neuron GABAA receptor in an epilepsia rat model%癫痫大鼠内嗅皮层神经元GABAA受体功能的研究

    Institute of Scientific and Technical Information of China (English)

    李桀; 孙杨; 刘备; 王超; 井晓荣; 梁秦川; 李焕发; 张华; 高国栋

    2011-01-01

    Objective Established the lithium chloride-pilocarpine inducing epilepsia rat model, and initially study about the entorhinal cortex neuron GABAA receptor by patch clamp of whole-cell mode. Methods The SD rats were divided into control group and experiment group randomly. Intraperitoneal injection of lithium chloride-pilocarpine have been used in experiment group, and physiologic saline in control group. Observed the ethology characteristic and recorded the attenuation tendency of GABAa receptor electric current in whole cell mode. Results The e-lectric current attenuation tendency of GABAA Receptor of Epilepsia Rat greatly intensify in the experiment group. The group diveded have significant difference by variance test and interclass analyse (P<0. 05), and the datas of every point of time in experiment group and control group also have significant difference by t-text analyse(P<0. 05). Conclusion The electric current attenuation tendency of Entorhinal Cortex Neuron GABAA Receptor of lithium chloride-pilocarpine inducing epilepsia rat have intensified. This phenomenon could be the possible mechanism of lithium chloride-pilocarpine inducing epilepsia rat and hint that GABAA receptor may contribute to the epileptic attack and brain injured after epileptic attack.%目的 介绍建立氯化锂-匹鲁卡品致痫大鼠模型的方法,并且通过全细胞膜片钳记录,初步研究其内嗅皮层神经元GABAA受体功能.方法 将所有SD大鼠随机分为对照组和实验组.实验组大鼠腹腔注射氯化锂以及匹鲁卡品,对照组注射生理盐水,观察其行为学特征,并用全细胞膜片钳记录GABAA受体电流的衰减趋势.结果 与对照组相比,实验组癫痫大鼠内嗅皮层神经元的GABAA受体电流的衰减加剧.方差检验进行组间分析,分组的作用是有差异的(P<0.001),;固定时间,对每个时间点上的处理组和对照组进行t检验,分组都有统计学意义(P<0.001).结论 锂-匹罗卡品致病大鼠

  18. Influence of sex and growth hormone deficiency on sweating

    DEFF Research Database (Denmark)

    Main, K; Nilsson, K O; Skakkebaek, N E

    1991-01-01

    than 0.001, respectively). There was a significant increase in SSR from prepuberty to puberty (p less than 0.001) for both sexes. The children with GH deficiency, all pre-pubertal, showed significantly reduced SSR (p less than 0.001) compared with the healthy children (median values: 32.8 vs 80.0 mg 30...... min-1). We conclude that (a) sweat secretion pattern in children shows a significant sex difference and (b) sweating in children is dependent on growth hormone.......Sweat secretion rate (SSR) was measured by the pilocarpine iontophoresis test in (a) 254 healthy children and adolescents (aged 6.0 to 19.2 years, mean age 11.2 years); in (b) 58 healthy adults (aged 20.4 to 75.2 years, mean age 37.6 years); and in (c) eight prepubertal patients with growth hormone...

  19. Impaired executive functions in experimental model of temporal lobe epilepsy

    Directory of Open Access Journals (Sweden)

    Fabiane Ochai Ramos

    2016-06-01

    Full Text Available ABSTRACT Objective The present study aimed to investigate cognitive and behavioural changes consistent with attention deficit hyperactivity disorder (ADHD -like behavior in male Wistar rats with temporal lobe epilepsy (TLE. Method Male Wistar rats at 25 day of age were submitted to animal model of TLE by pilocarpine injection (350 mg/kg, ip and a control group received saline 0.9%. The animals were continuously video monitored up to the end of experiments. The behavioural tests (open field, elevated plus maze and operant conditioning box started from 60 days postnatal. Results Animals with TLE exhibited elevated locomotor activity, reduced level of anxiety-related behavior, impulsivity and impaired visuospatial working memory. Conclusion Taken as a whole, we concluded that animals with TLE exhibited some cognitive and behavioural changes consistent with ADHD-like behavior.

  20. Unique behavioral characteristics and microRNA signatures in a drug resistant epilepsy model.

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    Jangsup Moon

    Full Text Available BACKGROUND: Pharmacoresistance is a major issue in the treatment of epilepsy. However, the mechanism underlying pharmacoresistance to antiepileptic drugs (AEDs is still unclear, and few animal models have been established for studying drug resistant epilepsy (DRE. In our study, spontaneous recurrent seizures (SRSs were investigated by video-EEG monitoring during the entire procedure. METHODS/PRINCIPAL FINDINGS: In the mouse pilocarpine-induced epilepsy model, we administered levetiracetam (LEV and valproate (VPA in sequence. AED-responsive and AED-resistant mice were naturally selected after 7-day treatment of LEV and VPA. Behavioral tests (open field, object exploration, elevated plus maze, and light-dark transition test and a microRNA microarray test were performed. Among the 37 epileptic mice with SRS, 23 showed significantly fewer SRSs during administration of LEV (n = 16, LEV sensitive (LS group or VPA (n = 7, LEV resistant/VPA sensitive (LRVS group, while 7 epileptic mice did not show any amelioration with either of the AEDs (n = 7, multidrug resistant (MDR group. On the behavioral assessment, MDR mice displayed distinctive behaviors in the object exploration and elevated plus maze tests, which were not observed in the LS group. Expression of miRNA was altered in LS and MDR groups, and we identified 4 miRNAs (miR-206, miR-374, miR-468, and miR-142-5p, which were differently modulated in the MDR group versus both control and LS groups. CONCLUSION: This is the first study to identify a pharmacoresistant subgroup, resistant to 2 AEDs, in the pilocarpine-induced epilepsy model. We hypothesize that modulation of the identified miRNAs may play a key role in developing pharmacoresistance and behavioral alterations in the MDR group.

  1. Pregabalin attenuates excitotoxicity in diabetes.

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    Chin-Wei Huang

    Full Text Available Diabetes can exacerbate seizures and worsen seizure-related brain damage. In the present study, we aimed to determine whether the standard antiepileptic drug pregabalin (PGB protects against pilocarpine-induced seizures and excitotoxicity in diabetes. Adult male Sprague-Dawley rats were divided into either a streptozotocin (STZ-induced diabetes group or a normal saline (NS group. Both groups were further divided into subgroups that were treated intravenously with either PGB (15 mg/kg or a vehicle; all groups were treated with subcutaneous pilocarpine (60 mg/kg to induce seizures. To evaluate spontaneous recurrent seizures (SRS, PGB-pretreated rats were fed rat chow containing oral PGB (450 mg for 28 consecutive days; vehicle-pretreated rats were fed regular chow. SRS frequency was monitored for 2 weeks from post-status epilepticus day 15. We evaluated both acute neuronal loss and chronic mossy fiber sprouting in the CA3 area. In addition, we performed patch clamp recordings to study evoked excitatory postsynaptic currents (eEPSCs in hippocampal CA1 neurons for both vehicle-treated rats with SRS. Finally, we used an RNA interference knockdown method for Kir6.2 in a hippocampal cell line to evaluate PGB's effects in the presence of high-dose ATP. We found that compared to vehicle-treated rats, PGB-treated rats showed less severe acute seizure activity, reduced acute neuronal loss, and chronic mossy fiber sprouting. In the vehicle-treated STZ rats, eEPSC amplitude was significantly lower after PGB administration, but glibenclamide reversed this effect. The RNA interference study confirmed that PGB could counteract the ATP-sensitive potassium channel (KATP-closing effect of high-dose ATP. By opening KATP, PGB protects against neuronal excitotoxicity, and is therefore a potential antiepileptogenic in diabetes. These findings might help develop a clinical algorithm for treating patients with epilepsy and comorbid metabolic disorders.

  2. Salivary Alterations in Rats with Experimental Chronic Kidney Disease.

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    Ana Carolina Romero

    Full Text Available This study aimed to analyze changes in saliva composition and salivary secretion process of rats with chronic kidney disease induced by 5/6 nephrectomy to set the foundation for salivary studies related to CKD.CKD was induced in Wistar rats via 5/6 nephrectomy. Blood and saliva samples were collected from Control, Sham and CKD groups at 8 and 12 weeks after the surgery. Salivation was stimulated via intraperitoneal injections of pilocarpine (1.0 mg/Kg body weight or isoproterenol (5.0 mg/Kg body weight. Saliva was collected and immediately stored at -80°C until analysis. The salivary flow rate, total protein, amylase and peroxidase activities, and urea concentrations were measured. The blood urea nitrogen (BUN and serum creatinine concentrations were also evaluated.Increases in BUN and serum creatinine concentrations were observed in the CKD groups. Amylase activity was significantly reduced in response to both stimuli in the CKD groups at 8 weeks and increased in the CKD groups at 12 weeks in response to isoproterenol stimulus. The peroxidase activities of the CKD groups were significantly reduced in response to isoproterenol stimulation and were increased at 12 weeks in response to pilocarpine stimulation. Salivary urea was significantly increased in the CKD groups at 8 weeks in response to the isoproterenol stimuli and at 12 weeks in response to both salivary agonists.The pattern of alterations observed in this experimental model is similar to those observed in patients and clearly demonstrates the viability of 5/6 nephrectomy as an experimental model in future studies to understand the alterations in salivary compositions and in salivary glands that are elicited by CKD.

  3. Perirhinal cortex and temporal lobe epilepsy

    Directory of Open Access Journals (Sweden)

    Giuseppe eBiagini

    2013-08-01

    Full Text Available The perirhinal cortex – which is interconnected with several limbic structures and is intimately involved in learning and memory - plays major roles in pathological processes such as the kindling phenomenon of epileptogenesis and the spread of limbic seizures. Both features may be relevant to the pathophysiology of mesial temporal lobe epilepsy that represents the most refractory adult form of epilepsy with up to 30% of patients not achieving adequate seizure control. Compared to other limbic structures such as the hippocampus or the entorhinal cortex, the perirhinal area remains understudied and, in particular, detailed information on its dysfunctional characteristics remains scarce; this lack of information may be due to the fact that the perirhinal cortex is not grossly damaged in mesial temporal lobe epilepsy and in models mimicking this epileptic disorder. However, we have recently identified in pilocarpine-treated epileptic rats the presence of selective losses of interneuron subtypes along with increased synaptic excitability. In this review we: (i highlight the fundamental electrophysiological properties of perirhinal cortex neurons; (ii briefly stress the mechanisms underlying epileptiform synchronization in perirhinal cortex networks following epileptogenic pharmacological manipulations; and (iii focus on the changes in neuronal excitability and cytoarchitecture of the perirhinal cortex occurring in the pilocarpine model of mesial temporal lobe epilepsy. Overall, these data indicate that perirhinal cortex networks are hyperexcitable in an animal model of temporal lobe epilepsy, and that this condition is associated with a selective cellular damage that is characterized by an age-dependent sensitivity of interneurons to precipitating injuries, such as status epilepticus.

  4. Experimental Models of Status Epilepticus and Neuronal Injury for Evaluation of Therapeutic Interventions

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    Ramkumar Kuruba

    2013-09-01

    Full Text Available This article describes current experimental models of status epilepticus (SE and neuronal injury for use in the screening of new therapeutic agents. Epilepsy is a common neurological disorder characterized by recurrent unprovoked seizures. SE is an emergency condition associated with continuous seizures lasting more than 30 min. It causes significant mortality and morbidity. SE can cause devastating damage to the brain leading to cognitive impairment and increased risk of epilepsy. Benzodiazepines are the first-line drugs for the treatment of SE, however, many people exhibit partial or complete resistance due to a breakdown of GABA inhibition. Therefore, new drugs with neuroprotective effects against the SE-induced neuronal injury and degeneration are desirable. Animal models are used to study the pathophysiology of SE and for the discovery of newer anticonvulsants. In SE paradigms, seizures are induced in rodents by chemical agents or by electrical stimulation of brain structures. Electrical stimulation includes perforant path and self-sustaining stimulation models. Pharmacological models include kainic acid, pilocarpine, flurothyl, organophosphates and other convulsants that induce SE in rodents. Neuronal injury occurs within the initial SE episode, and animals exhibit cognitive dysfunction and spontaneous seizures several weeks after this precipitating event. Current SE models have potential applications but have some limitations. In general, the experimental SE model should be analogous to the human seizure state and it should share very similar neuropathological mechanisms. The pilocarpine and diisopropylfluorophosphate models are associated with prolonged, diazepam-insensitive seizures and neurodegeneration and therefore represent paradigms of refractory SE. Novel mechanism-based or clinically relevant models are essential to identify new therapies for SE and neuroprotective interventions.

  5. Urokinase-type plasminogen activator deficiency has little effect on seizure susceptibility and acquired epilepsy phenotype but reduces spontaneous exploration in mice.

    Science.gov (United States)

    Rantala, J; Kemppainen, S; Ndode-Ekane, X E; Lahtinen, L; Bolkvadze, Tamuna; Gurevicius, K; Tanila, H; Pitkänen, A

    2015-01-01

    Urokinase-type plasminogen activator (uPA), a serine protease, converts plasminogen to plasmin. Activation of plasmin leads to degradation of the extracellular matrix, which is critical for tissue recovery, angiogenesis, cell migration, and axonal and synaptic plasticity. We hypothesized that uPA deficiency would cause an abnormal neurophenotype and would lead to exacerbated epileptogenesis after brain injury. Wild-type (Wt) and uPA-/- mice underwent a battery of neurologic behavioral tests evaluating general reactivity, spontaneous exploratory activity, motor coordination, pain threshold, fear and anxiety, and memory. We placed particular emphasis on the effect of uPA deficiency on seizure susceptibility, including the response to convulsants (pentylenetetrazol, kainate, or pilocarpine) and kainate-induced epileptogenesis and epilepsy. The uPA-/- mice showed no motor or sensory impairment compared with the Wt mice. Hippocampus-dependent spatial memory also remained intact. The uPA-/- mice, however, exhibited reduced exploratory activity and an enhanced response to a tone stimulus (p<0.05 compared with the Wt mice). The urokinase-type plasminogen activator deficient mice showed no increase in spontaneous or evoked epileptiform electrographic activity. Rather, the response to pilocarpine administration was reduced compared with the Wt mice (p<0.05). Also, the epileptogenesis and the epilepsy phenotype after intrahippocampal kainate injection were similar to those in the Wt mice. Taken together, uPA deficiency led to diminished interest in the environmental surroundings and enhanced emotional reactivity to unexpected aversive stimuli. Urokinase-type plasminogen activator deficiency was not associated with enhanced seizure susceptibility or worsened poststatus epilepticus epilepsy phenotype.

  6. Salivary Alterations in Rats with Experimental Chronic Kidney Disease

    Science.gov (United States)

    Romero, Ana Carolina; Bergamaschi, Cassia Toledo; de Souza, Douglas Nesadal; Nogueira, Fernando Neves

    2016-01-01

    Objective This study aimed to analyze changes in saliva composition and salivary secretion process of rats with chronic kidney disease induced by 5/6 nephrectomy to set the foundation for salivary studies related to CKD. Methods CKD was induced in Wistar rats via 5/6 nephrectomy. Blood and saliva samples were collected from Control, Sham and CKD groups at 8 and 12 weeks after the surgery. Salivation was stimulated via intraperitoneal injections of pilocarpine (1.0 mg/Kg body weight) or isoproterenol (5.0 mg/Kg body weight). Saliva was collected and immediately stored at -80°C until analysis. The salivary flow rate, total protein, amylase and peroxidase activities, and urea concentrations were measured. The blood urea nitrogen (BUN) and serum creatinine concentrations were also evaluated. Results Increases in BUN and serum creatinine concentrations were observed in the CKD groups. Amylase activity was significantly reduced in response to both stimuli in the CKD groups at 8 weeks and increased in the CKD groups at 12 weeks in response to isoproterenol stimulus. The peroxidase activities of the CKD groups were significantly reduced in response to isoproterenol stimulation and were increased at 12 weeks in response to pilocarpine stimulation. Salivary urea was significantly increased in the CKD groups at 8 weeks in response to the isoproterenol stimuli and at 12 weeks in response to both salivary agonists. Conclusions The pattern of alterations observed in this experimental model is similar to those observed in patients and clearly demonstrates the viability of 5/6 nephrectomy as an experimental model in future studies to understand the alterations in salivary compositions and in salivary glands that are elicited by CKD. PMID:26859883

  7. Late treatment with choline alfoscerate (l-alpha glycerylphosphorylcholine, α-GPC) increases hippocampal neurogenesis and provides protection against seizure-induced neuronal death and cognitive impairment.

    Science.gov (United States)

    Lee, Song Hee; Choi, Bo Young; Kim, Jin Hee; Kho, A Ra; Sohn, Min; Song, Hong Ki; Choi, Hui Chul; Suh, Sang Won

    2017-01-01

    Choline alfoscerate (α-GPC) is a common choline compound and acetylcholine precursor in the brain, which has been shown to be effective in the treatment of Alzheimer's disease and dementia. α-GPC has been shown to enhance memory and cognitive function in stroke and Alzheimer's patients but currently remains untested in patients suffering from epilepsy. This study aimed to evaluate whether α-GPC treatment after seizure can ameliorate seizure-induced cognitive impairment and neuronal injury. The potential therapeutic effects of α-GPC on seizure-induced cognitive impairment were tested in an animal model of pilocarpine-induced seizure. Seizures were induced by intraperitoneal injection of pilocarpine (25mg/kg) in male rats. α-GPC (250mg/kg) was injected into the intramuscular space once daily for one or three weeks from immediately after seizure, or from 3 weeks after the seizure onset for 3 weeks. Here we found that immediate 1-week treatment of α-GPC showed no neuroprotective effects and neurogenesis. Immediate 3-week treatment of α-GPC showed neuroprotective effect but no effect on neurogenesis. To evaluate the effect of late treatment of α-GPC on cognitive impairment following seizure, rats were injected α-GPC from 3 weeks after seizure for 3 weeks and subjected to a water maze test. In the present study, we found that administration of α-GPC starting at 3 weeks after seizure improved cognitive function through reduced neuronal death and BBB disruption, and increased neurogenesis. Therefore, α-GPC injection may serve as a beneficial treatment for improvement of cognitive function in epilepsy patients.

  8. Unilateral Adie's tonic pupil and viral hepatitis: Report of two cases

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    Karadžić Jelena

    2015-01-01

    Full Text Available Introduction. Adie’s (tonic pupil is a neuro-ophthalmological disorder characterized by a tonically dilated pupil, which is unresponsive to light. It is caused by damage to postganglionic fibers of the parasympathetic innervation of the eye, usually by a viral or bacterial infection. Adie’s syndrome includes diminished deep tendon reflexes. Outline of Cases. We report data of a 59-year-old female with unequal pupil sizes. She complained of blurred vision and headache mainly while reading. She had a 35-year history of hepatitis B and liver cirrhosis. On exam, left pupil was mydriatic and there was no response to light and at slit lamp we saw segments of the sphincter constrict. We performed 0.125% pilocarpine test and there was a remarkable reduction of size in the left pupil. The second case is a 55-year-old female who was referred to the University Eye Clinic because of a headache and mydriatic left pupil. She had diabetes mellitus type 2, as well as hepatitis A virus 20 years earlier. On exam, the left pupil was mydriatic, with no response to light. Test with diluted pilocarpine was positive. Neurological examinations revealed no abnormality in either case so we excluded Adie’s syndrome. Conclusion. Adie’s tonic pupil is benign neuro-ophthalmological disorder of unknown etiology. Most patients commonly present no symptoms and anisocoria is noticed accidentally. Although the etiology is unknown, there are some conditions that cause tonic pupil. It may be a part of a syndrome in which tonic pupil is associated with absent deep tendon reflexes.

  9. Cannabinoid and nitric oxide signaling interplay in the modulation of hippocampal hyperexcitability: Study on electrophysiological and behavioral models of temporal lobe epilepsy in the rat.

    Science.gov (United States)

    Carletti, F; Gambino, G; Rizzo, V; Ferraro, G; Sardo, P

    2015-09-10

    A growing bulk of evidence suggests that cannabinoid system plays a pivotal role in the control of hyperexcitability phenomena. Notwithstanding, the anticonvulsant action of cannabinoids has not been fully addressed, in particular the involvement of potential cellular neuromodulators, for instance nitric oxide. In the current study, we focused on two distinct rat models of temporal lobe epilepsy, the Maximal Dentate Activation and the pilocarpine-induced acute seizures, providing both electrophysiological and behavioral data on cannabinoid and nitrergic system interplay. We evaluated the antiepileptic effects of WIN 55,212-2, (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl) pyrrolo[1,2,3-de]-1,4-benzoxazin-6-Yl]-1-naphthalenylmethanone (WIN), a CB agonist, and of 7-Nitroindazole (7NI), a preferential neuronal nitric oxide synthase (nNOS) inhibitor, at different doses, alone and in combination. MDA study showed that these drugs protected animals in a dose-dependent manner from electrically induced epileptiform discharges. In pilocarpine model, a dose-related activity of 7NI and WIN: a) decreased the behavioral scoring, used to describe the severity of chemically induced acute seizures; b) affected latency of the onset of acute convulsions; c) dampened mortality rate. Interestingly, the combination of the treatments brought to light that individually ineffective doses of WIN turn into effective when nNOS activity is pharmacologically inhibited in both experimental conditions. This effect is mediated by CB1 receptor since the co-administration of N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251), a CB1 receptor specific antagonist, thwarted the 7NI-WIN convergent action. In the light of this, our findings suggest a putative antagonism between CBr-activated pathway and NO signaling in the context of neuronal hyperexcitability and contribute to elucidate possible synaptic processes underlying neuroprotective

  10. Improving access to intestinal stem cells as a step toward intestinal gene transfer.

    Science.gov (United States)

    Sandberg, J W; Lau, C; Jacomino, M; Finegold, M; Henning, S J

    1994-03-01

    In previous studies exploring the intestinal epithelium as a potential site for somatic gene therapy, we concluded that the mucus lining the intestine constitutes a significant barrier to any attempts at gene transfer via the lumenal route. The mucus problem is aggravated by the fact that the epithelial stem cells, which are the logical target for gene transfer, are located deep in the intestinal crypts. The goals of the current study were to develop procedures that would improve accessibility to the intestinal stem cells and which would effect in vivo mucus removal without damaging the underlying epithelium. Initial experiments involved evaluation of the use of distension to improve accessibility to the intestinal crypts and the use of the mucolytic agents dithiothreitol (DTT) and N-acetyl-cysteine (NAC) versus a control solution of phosphate-buffered saline (PBS) for mucus removal. Catheters were inserted in each end of 3-cm terminal ileal segments in anesthetized rats. Two milliliters of agent was instilled into the clamped segment for 2 min, removed, and repeated. Lumenal distension resulted in shortened villi with wider intervillus spacing, thereby improving crypt access. Both NAC and DTT washes removed significant mucus between the villi but failed to reach the crypt lumen. To enhance mucus release from the crypt lumen, pilocarpine was selected due to its cholinergic properties and preferential binding to muscarinic receptors on crypt goblet cells. Pilocarpine given intraperitoneally 30 min prior to the mucolytic or PBS wash resulted in significant eradication of mucus down into the crypt lumen. This effect was still evident 3-4 hr later provided the intestine remained undisturbed.

  11. Beneficial influence of physical exercise following status epilepticus in the immature brain of rats.

    Science.gov (United States)

    Gomes, F G Novaes; Gomes Da Silva, S; Cavalheiro, E A; Arida, R M

    2014-08-22

    Studies in adult animals have demonstrated a beneficial effect of physical exercise on epileptic insults. Although the effects of physical exercise on the mature nervous system are well documented, its influence on the developing nervous system subjected to injuries in childhood has been little explored. The purpose of our study was to investigate whether a physical exercise program applied during brain development could influence the hippocampal plasticity of rats submitted to status epilepticus (SE) induced by pilocarpine model at two different ages of the postnatal period. Male Wistar rats aged 18 (P18) and 28 (P28) days were randomly divided into four groups: Control (CTRL), Exercise (EX), SE (SE) and SE Exercise (SE/EX) (n=17 per group). After the aerobic exercise program, histological and behavioral (water maze) analyses were performed. Our results showed that only animals subjected to pilocarpine-induced SE at P28 presented spontaneous seizures during the observational period. A significant reduction in seizure frequency was observed in the SE/EX group compared to the SE group. In adulthood, animals submitted to early-life SE displayed impairment in long-term memory in the water maze task, while the exercise program reversed this deficit. Reduced mossy fiber sprouting in the dentate gyrus was noted in animals that presented spontaneous seizures (SE/EX vs SE). Exercise increased cell proliferation (Ki-67 staining) and anti-apoptotic response (bcl-2 staining) and reduced pro-apoptotic response (Bax staining) in animals of both ages of SE induction (P18/28). Exercise also modified the brain-derived neurotrophic factor (BDNF) levels in EX and SE/EX animals. Our findings indicate that in animals subjected to SE in the postnatal period a physical exercise program brings about beneficial effects on seizure frequency and hippocampal plasticity in later stages of life.

  12. Nucleus accumbens neurotransmission and effort-related choice behavior in food motivation: effects of drugs acting on dopamine, adenosine, and muscarinic acetylcholine receptors.

    Science.gov (United States)

    Nunes, Eric J; Randall, Patrick A; Podurgiel, Samantha; Correa, Mercè; Salamone, John D

    2013-11-01

    Mesolimbic dopamine (DA) is a critical component of the brain circuitry regulating behavioral activation and effort-related processes. Although nucleus accumbens (NAc) DA depletions or antagonism leave aspects of appetite and primary food motivation intact, rats with impaired DA transmission reallocate their instrumental behavior away from food-reinforced tasks with high response requirements, and instead select less effortful food-seeking behaviors. Previous work showed that adenosine A2A antagonists can reverse the effects of DA D2 antagonists on effort-related choice, and that stimulation of adenosine A2A receptors produces behavioral effects that are similar to those induced by DA antagonism. The present review summarizes the literature on the role of NAc DA and adenosine in effort-related processes, and also presents original data on the effects of local stimulation of muscarinic acetylcholine receptors in NAc core. Local injections of the muscarinic agonist pilocarpine directly into NAc core produces shifts in effort-related choice behavior similar to those induced by DA antagonism or A2A receptor stimulation, decreasing lever pressing but increasing chow intake in rats responding on a concurrent fixed ratio/chow feeding choice task. In contrast, injections into a neostriatal control site dorsal to the NAc were ineffective. The actions of pilocarpine on this task were attenuated by co-administration of the muscarinic antagonist scopolamine. Thus, drugs that act on DA, adenosine A2A, and muscarinic receptors regulate effort-related choice behavior, which may have implications for the treatment of psychiatric symptoms such as psychomotor slowing, fatigue or anergia that can be observed in depression and other disorders. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Involvement of upregulation of miR-210 in a rat epilepsy model

    Science.gov (United States)

    Chen, Licheng; Zheng, Hao; Zhang, Shimeng

    2016-01-01

    Epilepsy is a common type of neurological disorder with complex etiology. The mechanisms are still not clear. MicroRNAs are endogenous noncoding RNAs with many physiological activities. Multiple microRNAs were abnormally expressed in status epilepticus, including miR-210. In this study, we applied lithium chloride and pilocarpine to induce epileptic activity and aimed to disclose the potential mechanisms. Our data showed that miR-210 was significantly upregulated in hippocampus one day after modeling (P<0.05 vs control) and the high expression of miR-210 lasted for at least 30 days. By contrast, γ-aminobutyric acid (GABA) level significantly decreased concurrently after modeling (P<0.05 vs control). To question whether miR-210 could be a potential therapeutic target for epilepsy, miR-210 inhibitor was administrated through intrahippocampal injection after epilepsy modeling. Our data showed that morphological changes of hippocampal neurons and apoptosis triggered by epilepsy were mitigated by miR-210 inhibition. More importantly, the expressions of GABA-related proteins, including GABAA receptor α1, glutamate decarboxylase, and GABA transporter 1, were significantly elevated after epilepsy modeling in both mRNA and protein levels 3 days postmodeling (P<0.05 vs control), which were mitigated by miR-210 inhibitor treatment (P<0.05 vs model). In addition, epilepsy-induced upregulation of GABA transaminase was alleviated by miR-210 inhibitor. Taken together, these data implicated potential roles of miR-210 in lithium chloride–pilocarpine-induced epilepsy model and miR-210 could serve as a potential therapeutic target in status epilepticus. PMID:27471387

  14. Tumor necrosis factor-α-mediated threonine 435 phosphorylation of p65 nuclear factor-κB subunit in endothelial cells induces vasogenic edema and neutrophil infiltration in the rat piriform cortex following status epilepticus

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    Kim Ji-Eun

    2012-01-01

    Full Text Available Abstract Background Status epilepticus (SE induces severe vasogenic edema in the piriform cortex (PC accompanied by neuronal and astroglial damages. To elucidate the mechanism of SE-induced vasogenic edema, we investigated the roles of tumor necrosis factor (TNF-α in blood-brain barrier (BBB disruption during vasogenic edema and its related events in rat epilepsy models provoked by pilocarpine-induced SE. Methods SE was induced by pilocarpine in rats that were intracerebroventricularly infused with saline-, and soluble TNF p55 receptor (sTNFp55R prior to SE induction. Thereafter, we performed Fluoro-Jade B staining and immunohistochemical studies for TNF-α and NF-κB subunits. Results Following SE, most activated microglia showed strong TNF-α immunoreactivity. In addition, TNF p75 receptor expression was detected in endothelial cells as well as astrocytes. In addition, only p65-Thr435 phosphorylation was increased in endothelial cells accompanied by SMI-71 expression (an endothelial barrier antigen. Neutralization of TNF-α by soluble TNF p55 receptor (sTNFp55R infusion attenuated SE-induced vasogenic edema and neuronal damages via inhibition of p65-Thr435 phosphorylation in endothelial cells. Furthermore, sTNFp55R infusion reduced SE-induced neutrophil infiltration in the PC. Conclusion These findings suggest that impairments of endothelial cell functions via TNF-α-mediated p65-Thr 485 NF-κB phosphorylation may be involved in SE-induced vasogenic edema. Subsequently, vasogenic edema results in extensive neutrophil infiltration and neuronal-astroglial loss.

  15. Altered Expression Pattern of Acid-Sensing Ion Channel Isoforms in Piriform Cortex After Seizures.

    Science.gov (United States)

    Wu, Hao; Wang, Chao; Liu, Bei; Li, Huanfa; Zhang, Yu; Dong, Shan; Gao, Guodong; Zhang, Hua

    2016-04-01

    The piriform cortex (PC) is highly susceptible to chemical and electrical seizure induction. Epileptiform activity is associated with an acid shift in extracellular pH, suggesting that acid-sensing ion channels (ASICs) expressed by PC neurons may contribute to this enhanced epileptogenic potential. In epileptic rats and surgical samples from patients with medial temporal lobe epilepsy (TLE), PC layer II ASIC1a-immunopositive neurons appeared swollen with dendritic elongation, and there was loss of ASIC1a-positive neurons in layer III, consistent with enhanced vulnerability to TLE-induced plasticity and cell death. In rats, pilocarpine-induced seizures led to transient downregulation of ASIC1a and concomitant upregulation of ASIC2a in the first few days post-seizure. These changes in expression may be due to seizure-induced oxidative stress as a similar reciprocal change in ASIC1a, and ASIC2a expression was observed in PC12 cells following H2O2 application. The proportion of ASIC1a/ASIC2a heteromers was reduced in the acute phase following status epilepticus (SE) but increased during the latent phase when rats developed spontaneous seizures. Knockdown of ASIC2a by RNAi reduced dendritic length and spine density in primary neurons, suggesting that seizure-induced upregulation of ASIC2a contributes to dendritic lengthening in PC layer II in rats. Administration of the ASIC inhibitor amiloride before pilocarpine reduced the proportion of rats reaching Racine level IV seizures, protected layer II and III neurons, and prolonged survival in the acute phase following SE. Our findings suggest that ASICs may enhance susceptibility to epileptogenesis in the PC. Inhibition of ASICs, particularly ASIC2a, may suppress seizures originating in the PC.

  16. Spontaneous recurrent seizures in rats: amino acid and monoamine determination in the hippocampus.

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    Cavalheiro, E A; Fernandes, M J; Turski, L; Naffah-Mazzacoratti, M G

    1994-01-01

    Rats subjected to structural brain damage induced by sustained convulsions triggered by systemic administration of pilocarpine (PILO) are a useful model for investigation of the mechanisms essential for seizure generation and spread in rodents. After PILO administration, three distinct phases are observed: (a) an acute period of 1-2 days' duration corresponding to a pattern of repetitive limbic seizures and status epilepticus; (b) a seizure-free (silent) period characterized by a progressive return to normal EEG and behavior of 4-44 days' duration; and (c) a period of spontaneous recurrent seizures (SRS) starting 5-45 days after PILO administration and lasting throughout the animal's life. PILO (320-350 mg/kg intraperitoneally, i.p.) was administered to rats, and the content of hippocampal monoamines and amino acids was measured in the acute, silent, and SRS periods by liquid chromatography. Norepinephrine (NE) level was decreased during all periods whereas dopamine (DA) content was increased. Serotonin (5-hydroxytryptamine, 5-HT) was increased only in the acute period. Utilization rate measurement of monoamines showed increased NE consumption and decreased DA consumption during all phases. 5-HT utilization rate was increased only in the acute period. Amino acid content showed a decrease in aspartate (ASP) and glutamate (GLU) concentrations associated with increased gamma-aminobutyric acid (GABA) level during the acute period. The silent phase was characterized by a decrease in glycine (GLY) and GABA levels and an increase in GLU concentration. The SRS period showed an increase in all amino acid concentrations. These findings show important neurochemical changes in the course of establishment of an epileptic focus after brain damage induced by status epilepticus triggered by pilocarpine.

  17. Enhancement of inhibitory neurotransmission and inhibition of excitatory mechanisms underlie the anticonvulsant effects of Mallotus oppositifolius

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    Kennedy Kwami Edem Kukuia

    2016-01-01

    Full Text Available Context: Mallotus oppositifolius is a shrub that is used traditionally to treat epilepsy, but its potential has not been scientifically validated. Aims: This study investigated the anticonvulsant properties and possible mechanism of action of the 70% v/v hydroalcoholic extract of the leaves of M. oppositifolius.Materials and Methods: Inprinting control region (ICR mice (25–30 g were pretreated with the M. oppositifolius leaf extract (10–100 mg/kg before administering the respective convulsants (pentylenetetrazole [PTZ], picrotoxin [PTX], strychnine [STR], 4-aminopyridine [4-AP], and pilocarpine. The effect of the extract in maximal electroshock seizure (MES model was investigated also. Statistical Analysis: Data were presented as mean ± standard error of the mean and were analyzed with one-way analysis of variance (ANOVA or two-way ANOVA where appropriate with Newman–Keuls or Bonferroni post hoc test respectively. P< 0.05 was considered significant. Results: In both PTX and PTZ test, extract delayed the onset of seizures and reduced the frequency and duration of seizures. In the STR-induced seizure test, the extract significantly delayed the onset of seizures and reduced the duration of seizures. The extract also delayed the onset of clonic and tonic seizures as well as increasing the survival of mice in the 4-AP-induced seizure test. It further reduced the duration of tonic limb extensions in the MES test. In the pilocarpine-induced status epilepticus, the extract significantly delayed the onset of clonic convulsions and reduced the frequency and duration of seizures. Moreover, the anticonvulsant effect of the extract was attenuated by flumazenil, a benzodiazepine/gamma-aminobutyric acid (GABA receptor antagonist. Conclusion: These findings show that the extract has anticonvulsant effect possible mediated by GABAergic, glycinergic neurotransmission, and potassium channel conductions. It may also be acting by antagonizing muscarinic

  18. Interictal spike frequency varies with ovarian cycle stage in a rat model of epilepsy.

    Science.gov (United States)

    D'Amour, James; Magagna-Poveda, Alejandra; Moretto, Jillian; Friedman, Daniel; LaFrancois, John J; Pearce, Patrice; Fenton, Andre A; MacLusky, Neil J; Scharfman, Helen E

    2015-07-01

    In catamenial epilepsy, seizures exhibit a cyclic pattern that parallels the menstrual cycle. Many studies suggest that catamenial seizures are caused by fluctuations in gonadal hormones during the menstrual cycle, but this has been difficult to study in rodent models of epilepsy because the ovarian cycle in rodents, called the estrous cycle, is disrupted by severe seizures. Thus, when epilepsy is severe, estrous cycles become irregular or stop. Therefore, we modified kainic acid (KA)- and pilocarpine-induced status epilepticus (SE) models of epilepsy so that seizures were rare for the first months after SE, and conducted video-EEG during this time. The results showed that interictal spikes (IIS) occurred intermittently. All rats with regular 4-day estrous cycles had IIS that waxed and waned with the estrous cycle. The association between the estrous cycle and IIS was strong: if the estrous cycles became irregular transiently, IIS frequency also became irregular, and when the estrous cycle resumed its 4-day pattern, IIS frequency did also. Furthermore, when rats were ovariectomized, or males were recorded, IIS frequency did not show a 4-day pattern. Systemic administration of an estrogen receptor antagonist stopped the estrous cycle transiently, accompanied by transient irregularity of the IIS pattern. Eventually all animals developed severe, frequent seizures and at that time both the estrous cycle and the IIS became irregular. We conclude that the estrous cycle entrains IIS in the modified KA and pilocarpine SE models of epilepsy. The data suggest that the ovarian cycle influences more aspects of epilepsy than seizure susceptibility.

  19. Various ketogenic diets can differently support brain resistance against experimentally evoked seizures and seizure-induced elemental anomalies of hippocampal formation.

    Science.gov (United States)

    Chwiej, J; Patulska, A; Skoczen, A; Matusiak, K; Janeczko, K; Ciarach, M; Simon, R; Setkowicz, Z

    2017-07-01

    In this paper the influence of two different ketogenic diets (KDs) on the seizure-evoked elemental anomalies of hippocampal formation was examined. To achieve this purpose normal and pilocarpine treated rats previously fed with one of the two high fat and carbohydrate restricted diets were compared with animals on standard laboratory diet. The ketogenic ratios of the examined KDs were equal to 5:1 (KD1) and 9:1 (KD2). KD1 and standard diet fed animals presented similar patterns of seizure-evoked elemental changes in hippocampal formation. Also the analysis of behavioral data recorded after pilocarpine injection did not show any significant differences in intensity and duration of seizures between KD1 and standard diet fed animals. Higher ketogenic ratio KD2 introduced in the normal hippocampal formation prolonged changes in the accumulation of P, K, Zn and Ca. Despite this, both the intensity and duration of seizures were significantly reduced in rats fed with KD2 which suggests that its saving action on the nerve tissue may protect brain from seizure propagation. Also seizure-evoked elemental anomalies in KD2 animals were different than those observed for rats both on KD1 and standard diets. The comparison of seizure experiencing and normal rats on KD2, did not show any statistically significant differences in elemental composition of CA1 and H hippocampal areas whilst in CA3 area only Zn level changed as a result of seizures. DG was the area mostly affected by seizures in KD2 fed rats but areal densities of all examined elements increased in this hippocampal region. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. A Phase II Study of Submandibular Gland Transfer Prior to Radiation for Prevention of Radiation-induced Xerostomia in Head-and-Neck Cancer (RTOG 0244)

    Energy Technology Data Exchange (ETDEWEB)

    Jha, Naresh, E-mail: naresh.jha@albertahealthservices.ca [University of Alberta, Cross Cancer Institute, Edmonton, Alberta (Canada); Harris, Jonathan [Radiation Therapy Oncology Group Statistical Center, Philadelphia, Pennsylvania (United States); Seikaly, Hadi [University of Alberta, Edmonton, Alberta (Canada); Jacobs, John R. [Wayne State University School of Medicine, Detroit, Michigan (United States); McEwan, A.J.B. [University of Alberta, Cross Cancer Institute, Edmonton, Alberta (Canada); Robbins, K. Thomas [St. John' s Hospital Cancer Institute, Springfield, Illinois (United States); Grecula, John [Ohio State University Medical Center, Columbus, Ohio (United States); Sharma, Anand K. [Medical University of South Carolina, Charleston, South Carolina (United States); Ang, K. Kian [University of Texas MD Anderson Cancer Center, Houston, Texas (United States)

    2012-10-01

    Purpose: We report the results of a phase II study to determine the reproducibility of a submandibular salivary gland transfer (SGT) surgical technique for prevention of radiation (XRT)-induced xerostomia in a multi-institutional setting and to assess severity of xerostomia. Methods and Materials: Eligible patients had surgery for primary, neck dissection, and SGT, followed by XRT, during which the transferred salivary gland was shielded. Intensity modulated radiation therapy, amifostine, and pilocarpine were not allowed, but postoperative chemotherapy was allowed. Each operation was reviewed by 2 reviewers and radiation by 1 reviewer. If 13 or more (of 43) were 'not per protocol,' then the technique would be considered not reproducible as per study design. The secondary endpoint was the rate of acute xerostomia, grade 2 or higher, and a rate of {<=}51% was acceptable. Results: Forty-four of the total 49 patients were analyzable: male (81.8%), oropharynx (63.6%), stage IV (61.4%), median age 56.5 years. SGT was 'per protocol' or within acceptable variation in 34 patients (77.3%) and XRT in 79.5%. Nine patients (20.9%) developed grade 2 acute xerostomia; 2 had grade 0-1 xerostomia (4.7%) but started on amifostine/pilocarpine. Treatment for these 11 patients (25.6%) was considered a failure for the xerostomia endpoint. Thirteen patients died; median follow-up for 31 surviving patients was 2.9 years. Two-year overall and disease-free survival rates were 76.4% and 71.7%, respectively. Conclusions: The technique of submandibular SGT is reproducible in a multicenter setting. Seventy-four percent of patients were prevented from XRT-induced acute xerostomia.

  1. Skin Tattoos Alter Sweat Rate and Na+ Concentration.

    Science.gov (United States)

    Luetkemeier, Maurie Joe; Hanisko, Joseph Michael; Aho, Kyle Mathiew

    2017-07-01

    The popularity of tattoos has increased tremendously in the last 10 yr particularly among athletes and military personnel. The tattooing process involves permanently depositing ink under the skin at a similar depth as eccrine sweat glands (3-5 mm). The purpose of this study was to compare the sweat rate and sweat Na concentration of tattooed versus nontattooed skin. The participants were 10 healthy men (age = 21 ± 1 yr), all with a unilateral tattoo covering a circular area at least 5.2 cm. Sweat was stimulated by iontophoresis using agar gel disks impregnated with 0.5% pilocarpine nitrate. The nontattooed skin was located contralateral to the position of the tattooed skin. The disks used to collect sweat were composed of Tygon® tubing wound into a spiral so that the sweat was pulled into the tubing by capillary action. The sweat rate was determined by weighing the disk before and after sweat collection. The sweat Na concentration was determined by flame photometry. The mean sweat rate from tattooed skin was significantly less than nontattooed skin (0.18 ± 0.15 vs 0.35 ± 0.25 mg·cm·min; P = 0.001). All 10 participants generated less sweat from tattooed skin than nontattooed skin and the effect size was -0.79. The mean sweat Na concentration from tattooed skin was significantly higher than nontattooed skin (69.1 ± 28.9 vs 42.6 ± 15.2 mmol·L; P = 0.02). Nine of 10 participants had higher sweat Na concentration from tattooed skin than nontattooed skin, and the effect size was 1.01. Tattooed skin generated less sweat and a higher Na concentration than nontattooed skin when stimulated by pilocarpine iontophoresis.

  2. Changes in the sensitivity of GABAA current rundown to drug treatments in a model of temporal lobe epilepsy

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    Pierangelo eCifelli

    2013-07-01

    Full Text Available The pharmacological treatment of mesial temporal lobe epilepsy (mTLE, the most common epileptic syndrome in adults, is still unsatisfactory, as one third of the patients are or become refractory to antiepileptic agents. Refractoriness may depend upon drug-induced alterations, but the disease per se may also undergo a progressive evolution that affects the sensitivity to drugs. mTLE has been shown to be associated with a dysfunction of the inhibitory signaling mediated by GABAA receptors. In particular, the repetitive activation of GABAA receptors produces a use-dependent decrease (rundown of the evoked currents (IGABA, which is markedly enhanced in the hippocampus and cortex of drug-resistant mTLE patients. This phenomenon has been also observed in the pilocarpine model, where the increased IGABA rundown is observed in the hippocampus at the time of the first spontaneous seizure, then extends to the cortex and remains constant in the chronic phase of the disease. Here, we examined the sensitivity of IGABA to pharmacological modulation. We focused on the antiepileptic agent levetiracetam and on the neurotrophin BDNF, which were previously reported to attenuate mTLE-induced increased rundown in the chronic human tissue. In the pilocarpine model, BDNF displayed a paramount effect, decreasing rundown in the hippocampus at the time of the first seizure, as well as in the hippocampus and cortex in the chronic period. In contrast, levetiracetam did not affect rundown in the hippocampus, but attenuated it in the cortex. Interestingly, this effect of levetiracetam was also observed on the still unaltered rundown observed in the cortex at the time of the first spontaneous seizure. These data suggest that the sensitivity of GABAA receptors to pharmacological interventions undergoes changes during the natural history of mTLE, implicating that the site of seizure initiation and the timing of treatment may highly affect the therapeutic outcome.

  3. Sulforaphane is anticonvulsant and improves mitochondrial function.

    Science.gov (United States)

    Carrasco-Pozo, Catalina; Tan, Kah Ni; Borges, Karin

    2015-12-01

    The nuclear factor erythroid 2-related factor 2 pathway (Nrf2) has been previously identified to protect the brain against various impacts. Here, we investigated the effect of the Nrf2 activator sulforaphane in various seizure models and hippocampal mitochondrial bioenergetics. We found that daily injections of sulforaphane for 5 days elevated the seizure thresholds to 6 Hz stimulation and fluorothyl-, but not pentylenetetrazole-induced tonic seizures and protected mice against pilocarpine-induced status epilepticus (SE). Also, sulforaphane increased the antioxidant defences within hippocampal formations and blood plasma. In addition, sulforaphane treatment reduced the extent of hippocampal lipid peroxidation 24 h post-SE and protected hippocampal mitochondria against SE-induced reduction in state 2 and uncoupler-stimulated state 3 respiration. SE-mediated partial loss of rotenone-sensitive and complex II-driven respiration was reduced, consistent with the enhanced activities of complexes I and II in sulforaphane-treated SE mice. In mitochondria isolated from both no SE and SE mice, sulforaphane increased state 3 respiration and respiration linked to ATP synthesis, which may contribute to its anticonvulsant and antioxidant effects by providing more ATP for cellular vital and protective functions. However, sulforaphane did not prevent SE-induced hippocampal cell death. In conclusion, sulforaphane and/or Nrf2 activation are viable anticonvulsant strategies, which are antioxidant and enhance mitochondrial function, especially the ability to produce ATP. Sulforaphane was anticonvulsant in two acute mouse models of epilepsy and protected mice against pilocarpine-induced status epilepticus (SE). We also found antioxidant effects of sulforaphane in mouse plasma and hippocampal formations, exhibited by increased catalase and superoxide dismutase (SOD) activity, as well as increased abilities of hippocampal mitochondria to produce ATP. These effects likely underlie

  4. Persistent reduction of hippocampal glutamine synthetase expression after status epilepticus in immature rats.

    Science.gov (United States)

    van der Hel, W Saskia; Hessel, Ellen V S; Bos, Ineke W M; Mulder, Sandra D; Verlinde, Suzanne A M W; van Eijsden, Pieter; de Graan, Pierre N E

    2014-12-01

    Mesiotemporal sclerosis (MTS), the most frequent form of drug-resistant temporal lobe epilepsy, often develops after an initial precipitating injury affecting the immature brain. To analyse early processes in epileptogenesis we used the juvenile pilocarpine model to study status epilepticus (SE)-induced changes in expression of key components in the glutamate-glutamine cycle, known to be affected in MTS patients. SE was induced by Li(+) /pilocarpine injection in 21-day-old rats. At 2-19 weeks after SE hippocampal protein expression was analysed by immunohistochemistry and neuron damage by FluoroJade staining. Spontaneous seizures occurred in at least 44% of animals 15-18 weeks after SE. As expected in this model, we did not observe loss of principal hippocampal neurons. Neuron damage was most pronounced in the hilus, where we also detected progressive loss of parvalbumin-positive GABAergic interneurons. Hilar neuron loss (or end-folium sclerosis), a common feature in patients with MTS, was accompanied by a progressively decreased glutamine synthetase (GS)-immunoreactivity from 2 (-15%) to 19 weeks (-33.5%) after SE. Immunoreactivity for excitatory amino-acid transporters, vesicular glutamate transporter 1 and glial fibrillary acidic protein was unaffected. Our data show that SE elicited in 21-day-old rats induces a progressive reduction in hilar GS expression without affecting other key components of the glutamate-glutamine cycle. Reduced expression of glial enzyme GS was first detected 2 weeks after SE, and thus clearly before spontaneous recurrent seizures occurred. These results support the hypothesis that reduced GS expression is an early event in the development of hippocampal sclerosis in MTS patients and emphasize the importance of astrocytes in early epileptogenesis.

  5. 柯尔莫哥罗夫熵对大鼠痫性发作的预报价值%Value of Kolmogorov entropy on the prediction of seizure in epileptic rats

    Institute of Scientific and Technical Information of China (English)

    王顺先; 马英; 王寅旭

    2011-01-01

    Objective To investigate the application of Kolmogorov entropy (KE) in epileptic rats induced by lithium-pilocarpine,and to explore its value in predicting epileptic seizures. Methods Twenty-four Sprague-Dawley(SD) rats were randomly divided into 3 groups; a normal group, a control group and epileptic group. Acute chemical models were made by lithium-pilocarpine. The rats were received continuous electroencephalographic(EEG) monitoring by scalp surface electrode. Changes of KE of EEG signal in the epileptic group were analysis in the whole seizure process and compared with those in the normal group and control group. Results In the whole seizure process of the rats induced by lithium -pilocarpine, KE began to decline during the pre-ictal period and dropped sharply during the ictal period . KE in the pre-ictal and ictal period declined apparently comparing with that in the non -ictal period. KE in the epileptic group had significant differences during the ictal and pre -ictal period compared with that in the normal group and the control group (P < 0. 05 ). Conclusions Changes of KE reveal the changes of the complex ictal EEG signals,and may be useful to predict epileptic seizure.%目的 研究柯尔莫哥罗夫熵(KE)在氯化锂-匹鲁卡品致痫大鼠脑电监测中的应用,探讨运用KE预报痫性发作的可行性.方法 将24只健康雄性SD大鼠随机分为3组,分别为正常组、对照组和致痫组,致痫组采用氯化锂-匹鲁卡品制作急性痫性发作模型,利用头皮电极连续记录大鼠脑电信号,运用KE对瘸性发作大鼠全过程的脑电信号进行分析,并与正常组及注射生理盐水的对照组对比分析.结果 在整个痫性发作过程中,KE在发作前期开始下降,而在痫性发作期显著下降,与发作间期比较,发作前期与发作期的KE明显降低.致痫组痫性发作期和发作前期的KE值与正常组对比差异具有统计学意义(P<0.05).结论 KE的变化提示痫性发作过程脑电

  6. Study of video-electroencephalogram on epilepsy model of rats%癫痫大鼠模型的视频脑电图研究

    Institute of Scientific and Technical Information of China (English)

    张宁; 李文强; 杜好瑞; 王夏红

    2011-01-01

    目的 探讨癫痫大鼠模型的视频脑电图检测方法,为评价癫痫大鼠模型寻找实验室依据.方法 构建氯化锂和匹罗卡品急性癫痫大鼠模型,利用视频脑电监测仪观察脑电图动态变化过程.结果 大鼠腹腔未注射氯化锂和匹罗卡品药物前脑电图各个导联均表现为9 ~12 Hz的α波或17~19 Hz的β波,波幅20~ 80 μV,即基础波.未观察到棘波、尖波等癫痫样波.注入氯化锂和匹罗卡品后平均5~ 80 min中央区出现散在的尖波、棘波为先兆期,然后逐渐波及到各个导联,可见棘波、尖波及短程放电,强直-阵挛期表现为棘波、尖波、多棘-慢波及棘慢波长程放电.结论 视频脑电图可作为评价癫痫大鼠模型有效的检测手段.%Objective To explore the methods of detecting video-electroencephalogram (V-EEG) in the epilepsy rat model,in order to find the laboratory index to appraise the epilepsy rat model. Methods The rat model of acute epilepsy which was induced by lithium chloride and pilocarpine was established, the dynamic changes of electroencephalogram ( EEG) were investigated by using the V-EEG monitoring. Results The rats' EEG of all standard leads showed a wave of 9 -12 Hz or β wave of 17 - 19 Hz before injecting lithium chloride and pilocarpine into abdominal cavity,amplitude was 20-80 μ,V,namely basic wave. The spikes wave and sharp epilepsy wave were not been observed. When the rats were injected with lithium and pilocarpine,sporadic spike wave and sharp epilepsy wave appeared average 5 -80 minutes on central region,it was premonitory period,gradually spread to all standard leads,spike wave,sharp wave and short-range discharge were visible. The EEG showed spike wave,sharp wave,multitude spike-slow wave,and long-range discharge of spike-slow wave in rigidity-clonus period. Conclusion V-EEG can be used as an effective test facility for the epilepsy rat model.

  7. Níveis dos neurotransmissores estriatais durante o estado epiléptico Striatal monoamines levels during status epilepticus

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    Rivelilson Mendes de Freitas

    2003-01-01

    Full Text Available O objetivo desse estudo foi verificar os níveis dos neurotransmissores estriatais de ratas adultas durante o estado epiléptico induzido pela pilocarpina. Ratas wistar foram tratadas com uma única dose de pilocarpina (400 mg/kg por via subcutânea (S.C.; P400 e os controles receberam salina. A concentração dos neurotransmissores foi determinada através do HPLC eletroquímico, no corpo estriado de ratas que no período de observação de 1 hora desencadearam estado epiléptico e que sobreviveram à fase aguda do quadro convulsivo. Foi observada redução nos níveis de dopamina, serotonina, ácido diidroxifenilacético e aumento na concentração do ácido 5-hidroxiindolacético. Nenhuma alteração foi observada no 4-hidroxi-3-metoxi-fenilacético. Os resultados sugerem que a ativação do sistema colinérgico pode interagir com os sistemas dopaminérgico e serotonérgico nos mecanismos referentes à fase aguda do processo convulsivo no corpo estriado de ratos desenvolvidos.The purpose of the present work to investigate the striatal neurotransmissors level in adult rats after status epilepticus induced by pilocarpine. Wistar rats were treated with a single dose of pilocarpine (400 mg/kg; s.c.; P400 and the controls received saline. Adult animals were closed observed for behavioural changes during 1h. In this period, the animals that developed status epilepticus and survive this acute phase of seizures had the brains removed and striatal neurotransmissors level determiden by HPLC. The concentration of dopamine, serotonine, dihydroxyphenylacetic acid was reduced and an concentration increase in 5-hydroxyindolacetic acid. Didn't observed alteration in 4-hydroxy-3-methoxy-phenylacetic acid. These results suggest that cholinergic activation can interage with dopaminergic and serotonergic systems in acute phase of the convulsive process in rat mature striatum.

  8. Aspirin attenuates spontaneous recurrent seizures in the chronically epileptic mice.

    Science.gov (United States)

    Zhu, Kun; Hu, Ming; Yuan, Bo; Liu, Jian-Xin; Liu, Yong

    2017-08-01

    Neuroinflammatory processes are pathologic hallmarks of both experimental and human epilepsy, and could be implicated in the neuronal hyperexcitability. Aspirin represents one of the non-selective nonsteroidal anti-inflammatory drugs with fewer side effects in long-term application. This study was carried out to assess the anti-epileptic effects of aspirin when administered during the chronic stage of temporal lobe epilepsy [TLE] in mice. The alteration of hippocampal neurogenesis was also examined for raising a possible mechanism underlying the protective effect of anti-inflammatory treatment in the TLE. Two months after pilocarpine-induced status epilepticus, the chronically epileptic mice were treated with aspirin (20 mg, 60 mg or 80 mg/kg) once a day for 10 weeks. Spontaneous recurrent seizures were monitored by video camera for 2 weeks. To evaluate the profile of hippocampal neurogenesis, the newly generated cells in the dentate gyrus were labeled by the proliferation marker BrdU. The newborn neurons that extended axons to CA3 area were visualized by cholera toxin B subunit retrograde tracing. Administration of aspirin with a dosage of 60 mg or 80 mg/kg initiated at 2 months after pilocarpine-induced status epilepticus significantly reduced the frequency and duration of spontaneous recurrent seizures. Aspirin treatment also increased the number of newborn neurons with anatomic integration through improving the survival of the newly generated cells. Aspirin treatment during the chronic stage of TLE could attenuate the spontaneous recurrent seizures in mice. Promotion of hippocampal neurogenesis and inhibition of COX-PGE2 pathway might partly contribute to this anti-epileptic effect. Highlights • Aspirin attenuates spontaneous recurrent seizures of chronically epileptic mice • Aspirin increases neurogenesis of chronically epileptic hippocampus by improving the survival of newly generated cells • Promotion of hippocampal neurogenesis and inhibition

  9. Cannabidivarin is anticonvulsant in mouse and rat

    Science.gov (United States)

    Hill, AJ; Mercier, MS; Hill, TDM; Glyn, SE; Jones, NA; Yamasaki, Y; Futamura, T; Duncan, M; Stott, CG; Stephens, GJ; Williams, CM; Whalley, BJ

    2012-01-01

    Background and Purpose Phytocannabinoids in Cannabis sativa have diverse pharmacological targets extending beyond cannabinoid receptors and several exert notable anticonvulsant effects. For the first time, we investigated the anticonvulsant profile of the phytocannabinoid cannabidivarin (CBDV) in vitro and in in vivo seizure models. Experimental Approach The effect of CBDV (1–100 μM) on epileptiform local field potentials (LFPs) induced in rat hippocampal brain slices by 4-aminopyridine (4-AP) application or Mg2+-free conditions was assessed by in vitro multi-electrode array recordings. Additionally, the anticonvulsant profile of CBDV (50–200 mg·kg−1) in vivo was investigated in four rodent seizure models: maximal electroshock (mES) and audiogenic seizures in mice, and pentylenetetrazole (PTZ) and pilocarpine-induced seizures in rats. The effects of CBDV in combination with commonly used antiepileptic drugs on rat seizures were investigated. Finally, the motor side effect profile of CBDV was investigated using static beam and grip strength assays. Key Results CBDV significantly attenuated status epilepticus-like epileptiform LFPs induced by 4-AP and Mg2+-free conditions. CBDV had significant anticonvulsant effects on the mES (≥100 mg·kg−1), audiogenic (≥50 mg·kg−1) and PTZ-induced seizures (≥100 mg·kg−1). CBDV (200 mg·kg−1) alone had no effect against pilocarpine-induced seizures, but significantly attenuated these seizures when administered with valproate or phenobarbital at this dose. CBDV had no effect on motor function. Conclusions and Implications These results indicate that CBDV is an effective anticonvulsant in a broad range of seizure models. Also it did not significantly affect normal motor function and, therefore, merits further investigation as a novel anti-epileptic in chronic epilepsy models. Linked Articles This article is part of a themed section on Cannabinoids. To view the other articles in this section visit http

  10. 吗啡成瘾及癫痫造模对ICR小鼠学习记忆能力的相关性研究%Correlation between Morphine addiction, epilepsy model and learning and memorizing of ICR mice

    Institute of Scientific and Technical Information of China (English)

    林梅; 黄昶荃; 李永红; 彭小东; 林红; 杨波

    2012-01-01

    目的:观察吗啡成瘾及癫痫对ICR小鼠的空间学习记忆能力的影响是否具有相关性.方法:将40只小鼠随机分成吗啡成瘾继发癫痫组、吗啡成瘾组、癫痫组和空白对照组,每组10只.用连续注射吗啡建立吗啡成瘾模型,用匹罗卡品(Pilocarpine,PLO)建立癫痫模型.用水迷宫实验检测各组小鼠的记忆能力.结果:吗啡成瘾继发癫痫小鼠与癫痫小鼠水迷宫逃避潜伏期时间相对比明显延长(P<0.05).结论:吗啡成瘾与癫痫对ICR小鼠的学习记忆能力损害具有协同作用.%0bjeclive-.To create Morphine addiction model hy conditioned place preference firstly and then to create epilepsy model on the basis of Moq>hine addiction,finally to study whether Morphine addiction and epilepsy exert effects on spatial learning and memorizing of institute of cancer research(ICR) mice from behavioral hy water maze. Methods-,Totally 40 ICR mice having excellent performance by early platform of natural preference and training of water maze were selected. All these mice were equally divided into Morphine addiction complicated with epilepsy group. Morphine addiction group, epilepsy group and control group(n=10). Seven days continuous injection of Morphine was used to create Morphine addiction model while Pilocarpine was injected to create epilepsy model. Water maze test was performed to observe the learning and memorizing of mice. Results:Water maze escape latency was longer in Morphine addiction complicated epilepsy mice than in epilepsy mice, Conclusion; Morphine addition and epilepsy exert synergistic effect on learning and memory of ICR mice.

  11. A pupila na fase crônica da doença de Chagas e reação à pilocarpina e à fenilefrina

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    João Antonio Prata

    1996-12-01

    Full Text Available Com objetivo de desenvolver metodologia para avaliar alterações pupilares na fase crônica da doença de Chagas em área endêmica, foram examinados dez pacientes chagãsicos e dez controles, pareados quanto ao sexo, idade e cor. O diâmetro e área pupilar, determinados com recursos de projeção e topografia, foram comparados nos dois grupos. Ambas as pupilas foram fotografadas, simultaneamente, com iluminação padronizada. De cada indivíduo foram feitas três fotos sucessivas: inicial, apôs 30 minutos da instilação de colírio depilocaipina a 0,1% e apôs 30 minutos da instilação de colírio de fenilefrina a 3%. As pupilas dos chagãsicos diferiram das dos controles, de forma estatisticamente significante: maiores diâmetros e áreas iniciais; irregularidades nos contornos; maiores reduções percentuais em diâmetro e área apôs pilocarpina; maiores aumentos percentuais em diâmetro e área após fenilefrina. A metodogia foi considerada satisfatória e os resultados sugerem alterações no sistema nervoso autônomo ocular nos chagásicos.To develop a method to analyse pupillary disturbances in patients with chronic Chagas disease in an endemic area, ten chagasic and ten normal subjects were matched according to sex, age and race. Pupillary diameter and area ivere determined using projection and topography techniques and compared between groups. Both pupils were visualised simultaneously. In each case three photographs were taken under standardised illumination. The first photo was obtained without medication, the second, 30 minutes after instillation of -0,1% pilocarpine and the last 30 minutes after instillation of 3% phenylephrine (60 minutes after pilocarpine. Pupils of chagasic patients had a statistically significant greater initial diameter and area, irregularity of the pupil border, greater percentual reduction in diameter and area after pilocaipine 0.1% and greater percentual increase in diameter and area after 3

  12. Enantioselective pharmacodynamic and pharmacokinetic analysis of two chiral CNS-active carbamate derivatives of valproic acid.

    Science.gov (United States)

    Shekh-Ahmad, Tawfeeq; Mawasi, Hafiz; McDonough, John H; Finnell, Richard H; Wlodarczyk, Bogdan J; Yavin, Eylon; Bialer, Meir

    2014-12-01

    2-Ethyl-3-methylbutyl-carbamate (EMC) and 2-isopropylpentyl-carbamate (IPC) are among the most potent anticonvulsant carbamate derivatives of valproic acid. EMC and IPC are chiral compounds. Consequently, the aim of the current study was to comparatively evaluate the pharmacokinetic (PK) and pharmacodynamic (PD anticonvulsant activity) profile of EMC and IPC individual enantiomers. The anticonvulsant activity of EMC and IPC individual enantiomers was evaluated in several anticonvulsant rodent models including maximal electroshock (MES), 6 Hz psychomotor, subcutaneous (pentylenetetrazole) (scMet), and the pilocarpine-induced and soman-induced status epilepticus (SE). The PK-PD relationship of EMC and IPC individual enantiomers was evaluated following intraperitoneal administration (50 mg/kg) to rats. Induction of neural tube defects (NTDs) was evaluated in a mouse strain that was highly susceptible to teratogen-induced NTDs. In mice and rats, (2S)-EMC exhibited anticonvulsant activity similar to that of racemic EMC in the MES and scMet tests, whereas in the 6 Hz test, racemic EMC was more potent than its two individual enantiomers. Racemic EMC exhibited a potent activity in the soman-induced SE model when administered 5 and 20 min after seizure onset with median effective dose (ED50 ) values of 33 and 48 mg/kg, respectively. (2R)-IPC and (2S)-IPC exhibited ED50 values similar to those of racemic IPC in the mouse and rat MES and scMet models. (2R)-IPC had similar ED50 values on the 6 Hz tests. Racemic IPC had an ED50 value of 107 mg/kg in the pilocarpine-induced SE model when given 30 min after seizure onset. Racemic EMC and IPC and their enantiomers had similar clearance (3.8-5.5 L/h/kg) and short half-life (<1 h). EMC and its enantiomers did not cause NTDs at doses 3-10 times higher than their anticonvulsant ED50 values. EMC and IPC did not exhibit enantioselective PK, a fact that may contribute to their nonenantioselective activity in any of the anticonvulsant

  13. Inhibition of the prostaglandin receptor EP2 following status epilepticus reduces delayed mortality and brain inflammation.

    Science.gov (United States)

    Jiang, Jianxiong; Quan, Yi; Ganesh, Thota; Pouliot, Wendy A; Dudek, F Edward; Dingledine, Raymond

    2013-02-26

    Prostaglandin E2 is now widely recognized to play critical roles in brain inflammation and injury, although the responsible prostaglandin receptors have not been fully identified. We developed a potent and selective antagonist for the prostaglandin E2 receptor subtype EP2, TG6-10-1, with a sufficient pharmacokinetic profile to be used in vivo. We found that in the mouse pilocarpine model of status epilepticus (SE), systemic administration of TG6-10-1 completely recapitulates the effects of conditional ablation of cyclooxygenase-2 from principal forebrain neurons, namely reduced delayed mortality, accelerated recovery from weight loss, reduced brain inflammation, prevention of blood-brain barrier opening, and neuroprotection in the hippocampus, without modifying seizures acutely. Prolonged SE in humans causes high mortality and morbidity that are associated with brain inflammation and injury, but currently the only effective treatment is to stop the seizures quickly enough with anticonvulsants to prevent brain damage. Our results suggest that the prostaglandin receptor EP2 is critically involved in neuroinflammation and neurodegeneration, and point to EP2 receptor antagonism as an adjunctive therapeutic strategy to treat SE.

  14. Relationship between seizure frequency and number of neuronal and non-neuronal cells in the hippocampus throughout the life of rats with epilepsy.

    Science.gov (United States)

    Lopim, Glauber Menezes; Vannucci Campos, Diego; Gomes da Silva, Sérgio; de Almeida, Alexandre Aparecido; Lent, Roberto; Cavalheiro, Esper Abrão; Arida, Ricardo Mario

    2016-03-01

    The relationship between seizure frequency and cell death has been a subject of controversy. To tackle this issue, we determined the frequency of seizures and the total number of hippocampal cells throughout the life of rats with epilepsy using the pilocarpine model. Seizure frequency varied in animals with epilepsy according to which period of life they were in, with a progressive increase in the number of seizures until 180 days (sixth months) of epileptic life followed by a decrease (330 days-eleventh month) and subsequently stabilization of seizures. Cell counts by means of isotropic fractionation showed a reduction in the number of hippocampal neuronal cells following 30, 90, 180 and 360 days of spontaneous recurrent seizures (SRS) in rats compared to their controls (about 25%-30% of neuronal cell reduction). In addition, animals with 360 days of SRS showed a reduction in the number of neuronal cells when compared with animals with 90 and 180 days of seizures. The total number of hippocampal non-neuronal cells was reduced in rats with epilepsy after 30 days of SRS, but no significant alteration was observed on the 90th, 180th and 360th days. The total number of neuronal cells was negatively correlated with seizure frequency, indicating an association between occurrence of epileptic seizures throughout life and neuronal loss. In sum, our results add novel data to the literature concerning the time-course of SRS and hippocampal cell number throughout epileptic life.

  15. Optogenetic delay of status epilepticus onset in an in vivo rodent epilepsy model.

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    Inna Sukhotinsky

    Full Text Available Epilepsy is a devastating disease, currently treated with medications, surgery or electrical stimulation. None of these approaches is totally effective and our ability to control seizures remains limited and complicated by frequent side effects. The emerging revolutionary technique of optogenetics enables manipulation of the activity of specific neuronal populations in vivo with exquisite spatiotemporal resolution using light. We used optogenetic approaches to test the role of hippocampal excitatory neurons in the lithium-pilocarpine model of acute elicited seizures in awake behaving rats. Hippocampal pyramidal neurons were transduced in vivo with a virus carrying an enhanced halorhodopsin (eNpHR, a yellow light activated chloride pump, and acute seizure progression was then monitored behaviorally and electrophysiologically in the presence and absence of illumination delivered via an optical fiber. Inhibition of those neurons with illumination prior to seizure onset significantly delayed electrographic and behavioral initiation of status epilepticus, and altered the dynamics of ictal activity development. These results reveal an essential role of hippocampal excitatory neurons in this model of ictogenesis and illustrate the power of optogenetic approaches for elucidation of seizure mechanisms. This early success in controlling seizures also suggests future therapeutic avenues.

  16. Elevated Expression of Acid-Sensing Ion Channel 3 Inhibits Epilepsy via Activation of Interneurons.

    Science.gov (United States)

    Cao, Qingqing; Wang, Wei; Gu, Juan; Jiang, Guohui; Bian, Xiling; Wang, Kewei; Xu, Zucai; Li, Jie; Chen, Guojun; Wang, Xuefeng

    2016-01-01

    Recent studies have indicated that acid-sensing ion channels may play a significant role in the termination of epilepsy. In particular, acid-sensing ion channel 3 (ASIC3) is expressed in the central nervous system and is most sensitive to extracellular pH. However, whether ASIC3 plays a role in epilepsy is unknown. In this study, qRT-PCR, Western blot, immunohistochemistry, double immunofluorescence labeling, and slice recordings were used. We first detected elevated ASIC3 expression patterns in the brains of temporal lobe epilepsy patients and epileptic rats. ASIC3 was expressed in neurons and glia in both humans and in an experimental model of epilepsy, and ASIC3 was colocalized with inhibitory GABAergic interneurons. By blocking ASIC3 with its antagonist APETx2, we observed that injected APETx2 shortened the latency to seizure and increased the incidence of generalized tonic clonic seizure compared to the control group in models of both pilocarpine- and pentylenetetrazole (PTZ)-induced seizures. Additionally, blocking ASIC3 significantly decreased the frequency of action potential (AP) firing in interneurons. Moreover, APETx2 significantly reduced the amplitudes and frequencies of miniature inhibitory postsynaptic currents (mIPSCs) while showed no differences with the APETx2 + bicuculline group and the bicuculline group. These findings suggest that elevated levels of ASIC3 may serve as an anti-epileptic mechanism via postsynaptic mechanisms in interneurons. It could represent a novel therapeutic strategy for epilepsy treatment.

  17. Effects of 5-fluorouracil on the secretory process of the rat parotid gland

    Energy Technology Data Exchange (ETDEWEB)

    Sandborg, R.R.

    1986-01-01

    Experimental animals were injected intraperitoneally with 100 mg/kg 5-fluorouracil for three days. The total volume, amylase and protein content of cannulated parotid saliva were determined following stimulation with either 5 mg/kg pilocarpine or 5 mg/kg isoproterenol in experimental, pair-fed , and control animals. Saliva from experimental animals was significantly lower in volume, amylase and protein content than both control groups. 5-fluorouracil treatment reduced the total glandular amylase per unit DNA in both unstimulated and isoproterenol-stimulated parotid glands. Decreased protein synthesis may be the mechanism underlying depleted secretory protein stores since the contents of isolated secretory granules from experimental parotid glands contained less radiolabelled protein than either control group and whole gland homogenates showed marked reductions in the activities of three lysosomal enzymes and total RNA content. Experimental animals contained less labelled protein in their secretory granules than controls, but secreted a greater proportion of their total glandular radiolabelled secretory protein into saliva relative to amylase suggesting that newly synthesized secretory proteins are preferentially secreted.

  18. Behavioral and electroencephalographic analysis of seizures induced by intrahippocampal injection of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera

    Directory of Open Access Journals (Sweden)

    A.N.C. Santana

    2001-06-01

    Full Text Available In this study, the behavioral and electroencephalographic (EEG analysis of seizures induced by the intrahippocampal injection in rats of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera, was determined. The first alterations occurred during microinjection of granulitoxin (8 µg into the dorsal hippocampus and consisted of seizure activity that began in the hippocampus and spread rapidly to the occipital cortex. This activity lasted 20-30 s, and during this period the rats presented immobility. During the first 40-50 min after its administration, three to four other similar short EEG seizure periods occurred and the rats presented the following behavioral alterations: akinesia, facial automatisms, head tremor, salivation, rearing, jumping, barrel-rolling, wet dog shakes and forelimb clonic movements. Within 40-50 min, the status epilepticus was established and lasted 8-12 h. These results are similar to those observed in the acute phase of the pilocarpine model of temporal lobe epilepsy and suggest that granulitoxin may be a useful tool not only to study the sodium channels, but also to develop a new experimental model of status epilepticus.

  19. Central generation of grooming motor patterns and interlimb coordination in locusts.

    Science.gov (United States)

    Berkowitz, A; Laurent, G

    1996-12-15

    Coordinated bursts of leg motoneuron activity were evoked in locusts with deefferented legs by tactile stimulation of sites that evoke grooming behavior. This suggests that insect thoracic ganglia contain central pattern generators for directed leg movements. Motoneuron recordings were made from metathoracic and mesothoracic nerves, after eliminating all leg motor innervation, as well as all input from the brain, subesophageal ganglion, and prothoracic ganglion. Strong, brief trochanteral levator motoneuron bursts occurred, together with silence of the slow and fast trochanteral depressor motoneurons and activation of the common inhibitor motoneuron. The metathoracic slow tibial extensor motoneuron was active in a pattern distinct from its activity during walking or during rhythms evoked by the muscarinic agonist pilocarpine. Preparations in which the metathoracic ganglion was isolated from all other ganglia could still produce fictive motor patterns in response to tactile stimulation of metathoracic locations. Bursts of trochanteral levator and depressor motoneurons were clearly coordinated between the left and right metathoracic hemiganglia and also between the mesothoracic and the ipsilateral metathoracic ganglia. These data provide clear evidence for centrally generated interlimb coordination in an insect.

  20. Anticonvulsant and related neuropharmacological effects of the whole plant extract of Synedrella nodiflora (L. Gaertn (Asteraceae

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    Patrick Amoateng

    2012-01-01

    Full Text Available Purpose: The plant Synedrella nodiflora (L Gaertn is traditionally used by some Ghanaian communities to treat epilepsy. To determine if this use has merit, we studied the anticonvulsant and other neuropharmacological effects of a hydro-ethanolic extract of the whole plant using murine models. Materials and Methods: The anticonvulsant effect of the extract (10-1000 mg/kg was tested on the pentylenetetrazole-, picrotoxin-, and pilocarpine-induced seizure models and PTZ-kindling in mice/rats. The effect of the extract was also tested on motor coordination using the rota-rod. Results: The results obtained revealed that the extract possesses anticonvulsant effects in all the experimental models of seizures tested as it significantly reduced the latencies to myoclonic jerks and seizures as well as seizure duration and the percentage severity. The extract was also found to cause motor incoordination at the higher dose of 1000 mg/kg. Conclusions: In summary, the hydro-ethanolic extract of the whole plant of S. nodiflora possesses anticonvulsant effects, possibly through an interaction with GABAergic transmission and antioxidant mechanisms and muscle relaxant effects. These findings thus provide scientific evidence in support of the traditional use of the plant in the management of epilepsy.

  1. The effect of pupil size on stimulation of the melanopsin containing retinal ganglion cells, as evaluated by monochromatic pupillometry

    DEFF Research Database (Denmark)

    Nissen, Claus Jeppe; Sander, Birgit; Lund-Andersen, Henrik

    2012-01-01

    Purpose: To evaluate the influence of the size of the light exposed pupil in one eye on the pupillary light reflex of the other eye. Method: Using a monochromatic pupillometer, the left eye in each of 10 healthy subjects was exposed to 20¿s of monochromatic light of luminance 300¿cd/m(2), first red...... (660¿nm) and in a following session, blue (470¿nm) light. The consensual pupillary diameter in the right eye was continuously measured before, during, and after light exposure. Subsequently, Tropicamide 1% or Pilocarpine 2% was instilled into the left eye and when the pupil was either maximally dilated...... or contracted, the entire sequence of red and blue light exposure repeated. After at least 3¿days, when the effect of the eye drop had subsided, the entire experiment was repeated, this time employing the other substance. Results: Prior dilatation of the left pupil augmented the post light contraction to blue...

  2. Screening of intraocular pressure before routine pupil dilation for retinal photography: Clinical case report

    Directory of Open Access Journals (Sweden)

    Lap-kin Chiang

    2016-12-01

    Full Text Available Introduction: Pharmacologic dilation of the pupil results in twice the sensitivity of detection of diabetic retinopathy compared with undilated retinal examination. The potential risk of acute angle-closure glaucoma after pupil dilation has been hypothesized to be higher in Asian patients with diabetes mellitus. Clinical case: A 61-year-old man with diabetes mellitus and hypertension was incidentally found to have elevated intraocular pressure (IOP before routine retinal photography. He was asymptomatic and the visual acuity was 0.67 for both eyes. An ophthalmologist later found he had anatomical narrow-angle borderline glaucoma. Topical administration of pilocarpine and oral administration of acetazolamide were initiated, and laser iridotomy was later performed. IOP screening: Among 1736 diabetic and/or hypertensive patients who underwent IOP screening, 31 patients (1.8% had IOP of any eye persistently higher than 21 mm Hg on at least two occasions. The mean (standard deviation IOP of the right eye was 24.1 (2.1 mm Hg, while that of the left eye was 24.6 (2.5 mm Hg. Four patients (12.9% were found to have glaucoma, and treatment was initiated by an ophthalmologist. Therefore further study should be conducted to evaluate the cost-effectiveness of IOP screening among this group of patients.

  3. Chemical functionalization of hyaluronic acid for drug delivery applications

    Energy Technology Data Exchange (ETDEWEB)

    Vasi, Ana-Maria [“Gheorghe Asachi” Technical University of Iasi, Faculty of Chemical Engineering and Environmental Protection, 73 Prof. dr. docent Dimitrie Mangeron Street, 700050 Iasi (Romania); Popa, Marcel Ionel, E-mail: mipopa@ch.tuiasi.ro [“Gheorghe Asachi” Technical University of Iasi, Faculty of Chemical Engineering and Environmental Protection, 73 Prof. dr. docent Dimitrie Mangeron Street, 700050 Iasi (Romania); Butnaru, Maria [“Grigore T. Popa” University of Medicine Pharmacy, Faculty of Medical Bioengineering, 9-13 Kogalniceanu Street, 700454 Iasi (Romania); Dodi, Gianina [“Gheorghe Asachi” Technical University of Iasi, Faculty of Chemical Engineering and Environmental Protection, 73 Prof. dr. docent Dimitrie Mangeron Street, 700050 Iasi (Romania); SCIENT — Research Center for Instrumental Analysis, S.C. CROMATEC PLUS, 18 Sos. Cotroceni, 060114 Bucharest (Romania); Verestiuc, Liliana [“Grigore T. Popa” University of Medicine Pharmacy, Faculty of Medical Bioengineering, 9-13 Kogalniceanu Street, 700454 Iasi (Romania)

    2014-05-01

    Functionalized hyaluronic acid (HA) derivatives were obtained by ring opening mechanism of maleic anhydride (MA). FTIR and H{sup 1} NMR spectroscopy were used to confirm the chemical linkage of MA on the hyaluronic acid chains. Thermal analysis (TG-DTG and DSC) and GPC data for the new products revealed the formation of new functional groups, without significant changes in molecular weight and thermal stability. New gels based on hyaluronic acid modified derivatives were obtained by acrylic acid copolymerization in the presence of a redox initiation system. The resulted circular and interconnected pores of the gels were visualized by SEM. The release profiles of an ophthalmic model drug, pilocarpine from tested gels were studied in simulated media. Evaluation of the cytotoxicity and cell proliferation properties indicates the potential of the new systems to be used in contact with biological media in drug delivery applications. - Highlights: • New functionalized hyaluronic acid was prepared by ring opening of maleic anhydride. • Gels with circular pores based on acrylic acid copolymerization were formulated. • In vitro drug loading/release profile was evaluated in simulated ophthalmic media. • The cytotoxicity indicates the potential of derivatives to be used in vivo.

  4. VEGF regulates hippocampal neurogenesis and reverses cognitive deficits in immature rats after status epilepticus through the VEGF R2 signaling pathway.

    Science.gov (United States)

    Han, Wei; Song, Xiaojie; He, Rong; Li, Tianyi; Cheng, Li; Xie, Lingling; Chen, Hengsheng; Jiang, Li

    2017-02-10

    Epilepsy is the most common chronic disease in children, who exhibit a higher risk for status epilepticus (SE) than adults. Hippocampal neurogenesis is altered by epilepsy, particularly in the immature brain, which may influence cognitive development. Vascular endothelial growth factor (VEGF) represents an attractive target to modulate brain function at the neurovascular interface and is a double-edged sword in seizures. We used the lithium-pilocarpine-induced epilepsy model in immature Sprague-Dawley rats to study the effects of VEGF on hippocampal neurogenesis in the acute phase and on long-term cognitive behaviors in immature rats following status epilepticus (SE). VEGF correlates with cell proliferation in the immature brain after SE. By preprocessing VEGF in the lateral ventricles prior to the induction of the SE model, we found that VEGF increased the proliferation of neural stem cells (NSCs) and promoted the migration of newly generated cells via the VEGF receptor 2 (VEGFR2) signaling pathway. VEGF also inhibited cell loss and reversed the cognitive deficits that accompany SE. Based on our results, VEGF positively contributes to the initial stages of neurogenesis and alleviates cognitive deficits following seizures; moreover, the VEGF/VEGFR2 signaling pathway may provide a novel treatment strategy for epilepsy.

  5. The mast cell stabilizer sodium cromoglycate reduces histamine release and status epilepticus-induced neuronal damage in the rat hippocampus.

    Science.gov (United States)

    Valle-Dorado, María Guadalupe; Santana-Gómez, César Emmanuel; Orozco-Suárez, Sandra Adela; Rocha, Luisa

    2015-05-01

    Experiments were designed to evaluate changes in the histamine release, mast cell number and neuronal damage in hippocampus induced by status epilepticus. We also evaluated if sodium cromoglycate, a stabilizer of mast cells with a possible stabilizing effect on the membrane of neurons, was able to prevent the release of histamine, γ-aminobutyric acid (GABA) and glutamate during the status epilepticus. During microdialysis experiments, rats were treated with saline (SS-SE) or sodium cromoglycate (CG-SE) and 30 min later received the administration of pilocarpine to induce status epilepticus. Twenty-four hours after the status epilepticus, the brains were used to determine the neuronal damage and the number of mast cells in hippocampus. During the status epilepticus, SS-SE group showed an enhanced release of histamine (138.5%, p = 0.005), GABA (331 ± 91%, p ≤ 0.001) and glutamate (467%, p ≤ 0.001), even after diazepam administration. One day after the status epilepticus, SS-SE group demonstrated increased number of mast cells in Stratum pyramidale of CA1 (88%, p histamine (but not GABA and glutamate) release, lower number of mast cells (p = 0.008) and reduced neuronal damage in hippocampus. Our data revealed that histamine, possibly from mast cells, is released in hippocampus during the status epilepticus. This effect may be involved in the subsequent neuronal damage and is diminished with sodium cromoglycate pretreatment.

  6. Status epilepticus during early development disrupts sexual behavior in adult female rats: recovery with sexual experience.

    Science.gov (United States)

    Coria-Avila, Genaro Alfonso; Paredes-Ramos, Pedro; Galán, Ricardo; Herrera-Covarrubias, Deissy; López-Meraz, Maria-Leonor

    2014-05-01

    Female sexual behavior is sensitive to stress and diseases. Some studies have shown that status epilepticus (SE) can affect sexual proceptivity and receptivity in female rats and also increases reject responses towards males. However, epidemiologic studies indicate that SE is more frequent in young individuals. Herein, we assessed the effects of SE in infant females on their sexual behavior during adulthood. Thirteen-day-old (P13) rat pups received intraperitoneal injections of lithium chloride (3 mEq/kg). Twenty hours later, at P14, SE was induced by subcutaneous injection of pilocarpine hydrochloride (100 mg/kg s.c.). Control animals were given an equal volume of saline subcutaneously. The animals were weaned at P21 and, later in adulthood, were ovariectomized and hormone-primed with estradiol+progesterone, and their sexual behavior assessed during 4 separate trials of 30 min each with a stud male. Our results indicate that proceptive behaviors (solicitations and hops and darts) were impaired during the first trial, but no alterations were observed for receptivity and attractivity. By trial 3, all SE females displayed normal proceptivity. These results indicate that SE in infancy readily affects proceptivity in a reversible manner. We discuss the role of sexual experience in recovery.

  7. Abnormal expression of stathmin 1 in brain tissue of patients with intractable temporal lobe epilepsy and a rat model.

    Science.gov (United States)

    Zhao, Fenghua; Hu, Yida; Zhang, Ying; Zhu, Qiong; Zhang, Xiaogang; Luo, Jing; Xu, Yali; Wang, Xuefeng

    2012-09-01

    Microtubule dynamics have been shown to contribute to neurite outgrowth, branching, and guidance. Stathmin 1 is a potent microtubule-destabilizing factor that is involved in the regulation of microtubule dynamics and plays an essential role in neurite elongation and synaptic plasticity. Here, we investigate the expression of stathmin 1 in the brain tissues of patients with intractable temporal lobe epilepsy (TLE) and experimental animals using immunohistochemistry, immunofluorescence and western blotting. We obtained 32 temporal neocortex tissue samples from patients with intractable TLE and 12 histologically normal temporal lobe tissues as controls. In addition, 48 Sprague Dawley rats were randomly divided into six groups, including one control group and five groups with epilepsy induced by lithium chloride-pilocarpine. Hippocampal and temporal lobe tissues were obtained from control and epileptic rats on Days 1, 7, 14, 30, and 60 after kindling. Stathmin 1 was mainly expressed in the neuronal membrane and cytoplasm in the human controls, and its expression levels were significantly higher in patients with intractable TLE. Moreover, stathmin 1 was also expressed in the neurons of both the control and the experimental rats. Stathmin 1 expression was decreased in the experimental animals from 1 to 14 days postseizure and then significantly increased at Days 30 and 60 compared with the control group. Many protruding neuronal processes were observed in the TLE patients and in the chronic stage epileptic rats. These data suggest that stathmin 1 may participate in the abnormal network reorganization of synapses and contribute to the pathogenesis of TLE.

  8. Endogenous rhythm and pattern-generating circuit interactions in cockroach motor centres

    Directory of Open Access Journals (Sweden)

    Izhak David

    2016-09-01

    Full Text Available Cockroaches are rapid and stable runners whose gaits emerge from the intricate, and not fully resolved, interplay between endogenous oscillatory pattern-generating networks and sensory feedback that shapes their rhythmic output. Here we studied the endogenous motor output of a brainless, deafferented preparation. We monitored the pilocarpine-induced rhythmic activity of levator and depressor motor neurons in the mesothoracic and metathoracic segments in order to reveal the oscillatory networks’ architecture and interactions. Data analyses included phase relations, latencies between and overlaps of rhythmic bursts, spike frequencies, and the dependence of these parameters on cycle frequency. We found that, overall, ipsilateral connections are stronger than contralateral ones. Our findings revealed asymmetries in connectivity among the different ganglia, in which meta-to-mesothoracic ascending coupling is stronger than meso-to-metathoracic descending coupling. Within-ganglion coupling between the metathoracic hemiganglia is stronger than that in the mesothoracic ganglion. We also report differences in the role and mode of operation of homologue network units (manifested by levator and depressor nerve activity. Many observed characteristics are similar to those exhibited by intact animals, suggesting a dominant role for feedforward control in cockroach locomotion. Based on these data we posit a connectivity scheme among components of the locomotion pattern generating system.

  9. Disruption of TrkB-mediated phospholipase Cgamma signaling inhibits limbic epileptogenesis.

    Science.gov (United States)

    He, Xiao Ping; Pan, Enhui; Sciarretta, Carla; Minichiello, Liliana; McNamara, James O

    2010-05-05

    The BDNF receptor, TrkB, is critical to limbic epileptogenesis, but the responsible downstream signaling pathways are unknown. We hypothesized that TrkB-dependent activation of phospholipase Cgamma1 (PLCgamma1) signaling is the key pathway and tested this in trkB(PLC/PLC) mice carrying a mutation (Y816F) that uncouples TrkB from PLCgamma1. Biochemical measures revealed activation of both TrkB and PLCgamma1 in hippocampi in the pilocarpine and kindling models in wild-type mice. PLCgamma1 activation was decreased in hippocampi isolated from trkB(PLC/PLC) compared with control mice. Epileptogenesis assessed by development of kindling was inhibited in trkB(PLC/PLC) compared with control mice. Long-term potentiation of the mossy fiber-CA3 pyramid synapse was impaired in slices of trkB(PLC/PLC) mice. We conclude that TrkB-dependent activation of PLCgamma1 signaling is an important molecular mechanism of limbic epileptogenesis. Elucidating signaling pathways activated by a cell membrane receptor in animal models of CNS disorders promises to reveal novel targets for specific and effective therapeutic intervention.

  10. Disease-modifying effect of intravenous immunoglobulin in an experimental model of epilepsy

    Science.gov (United States)

    Chen, Min; Arumugam, Thiruma V.; Leanage, Gayeshika; Tieng, Quang M.; Yadav, Ashwin; Ullmann, Jeremy F. P.; She, David T.; Truong, Vy; Ruitenberg, Marc J.; Reutens, David C.

    2017-01-01

    Novel therapies that prevent or modify the development of epilepsy following an initiating brain insult could significantly reduce the burden of this disease. In light of evidence that immune mechanisms play an important role in generating and maintaining the epileptic condition, we evaluated the effect of a well-established immunomodulatory treatment, intravenous immunoglobulin (IVIg), on the development of epilepsy in an experimental model of epileptogenesis. In separate experiments, IVIg was administered either before (pre-treatment) or after (post-treatment) the onset of pilocarpine status epilepticus (SE). Our results show that both pre- and post-treatment with IVIg attenuated acute inflammation in the SE model. Specifically, IVIg reduced local activation of glial cells, complement system activation, and blood-brain barrier damage (BBB), which are all thought to play important roles in the development of epilepsy. Importantly, post-treatment with IVIg was also found to reduce the frequency and duration of subsequent spontaneous recurrent seizures as detected by chronic video-electroencephalographic (video-EEG) recordings. This finding supports a novel application for IVIg, specifically its repurposing as a disease-modifying therapy in epilepsy. PMID:28074934

  11. Stimulation of acid secretion and phosphoinositol production by rat parietal cell muscarinic M sub 2 receptors

    Energy Technology Data Exchange (ETDEWEB)

    Pfeiffer, A.; Rochlitz, H.; Herz, A.; Paumgartner, G. (Univ. of Munich (West Germany))

    1988-04-01

    The muscarinic receptor system involved in hydrogen production by enriched rat gastric parietal cells was investigated. Muscarinic receptor density determined by (N-methyl-{sup 3}H)scopolamine binding was 8,100/cell. The receptor appeared to be of the M{sub 2} muscarinic receptor subtype, since it had a low affinity (K{sub d} 189 nM) for the M{sub 1} receptor antagonist pirenzepine compared with atropine. Receptor activation by carbachol rapidly augmented levels of polyphosphoinositides, indicating an activation of phospholipase C. The dose-response relations for the increase in inositol phosphates closely paralleled the binding of carbachol to muscarinic receptors. The inositol phosphate response was antagonized by pirenzepine with a K{sub i} of 177 nM. the stimulation of inositol phosphate levels by carbachol correlated well with the stimulation of ({sup 14}C)aminopyrine uptake, determine as an index of acid secretion. The muscarinic agonists oxotremorine, pilocarpine, and bethanechol elicited partial increases in inositol phosphates at maximal drug concentrations, and these partial increases correlated with their ability to stimulate ({sup 14}C)aminopyrine uptake. These data indicate that inositolpolyphosphates may be a second messenger of M{sub 2} receptors stimulating acid secretion.

  12. Pharmacological and ionic characterizations of the muscarinic receptors modulating (/sup 3/H)acetylcholine release from rat cortical synaptosomes

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, E.M.; Otero, D.H.

    1985-05-01

    The muscarinic receptors that modulate acetylcholine release from rat cortical synaptosomes were characterized with respect to sensitivity to drugs that act selectively at M1 or M2 receptor subtypes, as well as to changes in ionic strength and membrane potential. The modulatory receptors appear to be of the M2 type, since they are activated by carbachol, acetylcholine, methacholine, oxotremorine, and bethanechol, but not by pilocarpine, and are blocked by atropine, scopolamine, and gallamine (at high concentrations), but not by pirenzepine or dicyclomine. The ED50S for carbachol, acetylcholine, and oxotremorine are less than 10 microM, suggesting that the high affinity state of the receptor is functional. High ionic strength induced by raising the NaCl concentration has no effect on agonist (oxotremorine) potency, but increases the efficacy of this compound, which disagrees with receptor-binding studies. On the other hand, depolarization with either KCl or with veratridine (20 microM) reduces agonist potencies by approximately an order of magnitude, suggesting a potential mechanism for receptor regulation.

  13. M1/M2 muscarinic receptor selectivity using potassium (K/sup +/)-stimulated release of (/sup 3/H)-dopamine (DA) and (/sup 14/C)-acetyl-choline (ACH) in striatum

    Energy Technology Data Exchange (ETDEWEB)

    DeHaven, D.L.; Steranka, L.R.

    1986-03-05

    Raiteri et al have suggested that muscarinic receptor subtypes can be differentiated in striatal synaptosomes by the release of DA (M1) or ACh (M2). The authors attempted to replicate this finding and to characterize responses of selective and non-selective cholinergic agonists and antagonists using K+-stimulated release of transmitters from rat striatal slices. The non-selective agonists ACh, carbachol and oxotremorine stimulated release of (/sup 3/H)-DA and inhibited release of (/sup 14/C)-ACh with EC50 values of 10.6, 9.2 and 4.2 ..mu..M (DA) and 1.2, 0.77 and 0.43 ..mu..M (ACh), respectively. The M1 agonist McN-A-343-11 selectively inhibited release of DA with an EC50 value of 4.8 ..mu..M. Pilocarpine was ineffective in this system. The M1 antagonist pirenzepine reversed the effects of 10/sup -4/ M carbachol on release with an eight-fold selectivity for release of (/sup 3/H)-DA (IC50 = 0.77 ..mu..M) vs (/sup 14/C)-ACh (IC50 = 6.3 ..mu..M). These results suggest that although this system can determine relative subtype selectivities, the results obtained in this assay do not always correlate with those obtained from phosphatidyl inositol turnover or adenylate cyclase activity.

  14. Identification of four areas each enriched in a unique muscarinic receptor subtype

    Energy Technology Data Exchange (ETDEWEB)

    Hoss, W.; Ellerbrock, B.R.; Goldman, P.S.; Collins, D.A.; Messer, W.S. Jr. (Univ. of Toledo College of Pharmacy, OH (USA))

    1990-01-01

    The affinities of muscarinic agonists and antagonists were determined by autoradiography and image analysis in selected areas of the rat brain. IC{sub 50} values and Hill coefficients for the inhibition of the binding of 0.2 nM ({sup 3}H)-QNB to dentate gyrus, superior colliculus, rhomboid thalamus and substantia nigra were measured in coronal sections. Pirenzepine displayed a high affinity for receptors in the dentate gyrus and AF-DX 116, the superior colliculus. Both pirenzepine and AF-DX 116 had high affinities for the substantia nigra and low affinities for the rhomboid thalamus. Gallamine displayed a 50-fold preference for superior colliculus over dentate gyrus receptors. Amitriptyline was less selective, showing a modest preference for substantia nigra receptors and 4-DAMP was essentially nonselective. Carbachol was the most selective agonist with a 4000-fold preference for superior colliculus over dentate gyrus receptors. Other agonists except RS 86 were also selective for superior colliculus receptors in the order carbachol >> arecoline > bethanechol > McN A343 = oxotremorine = pilocarpine.

  15. [Tonic pupil caused by ischemia].

    Science.gov (United States)

    Wilhelm, H

    1989-01-01

    Tonic pupil is usually an idiopathic condition. In some cases, the cause of the ciliary ganglion lesion leading to tonic pupils is obvious. Rarely ischemia causes a lesion of the ciliary ganglion or the short ciliary nerves due to the good blood supply of the ciliary ganglion. Only two cases of tonic pupils in the course of giant cell arteritis are mentioned in the literature, but tonic pupils are probably much more common with this disease. Five cases are demonstrated here. All had associated ischemic optic neuropathy, and stagnation of the blood flow in the supratrochlear artery could be demonstrated in two cases by Doppler sonography. Tonic pupils may also occur when an oclusion of the internal carotid artery resolves, probably because of transient stasis of the orbital blood flow. In another case, tonic pupils were associated with choroidal ischemia (proved by video fluorescent angiography) of unknown origin. The diagnosis of tonic pupils was made by pharmacological testing for cholinergic hypersensitivity with 0.1% pilocarpine.

  16. Low-frequency repetitive transcranial magnetic simulation prevents chronic epileptic seizure*

    Institute of Scientific and Technical Information of China (English)

    Yinxu Wang; Xiaoming Wang; Sha Ke; Juan Tan; Litian Hu; Yaodan Zhang; Wenjuan Cui

    2013-01-01

    Although low-frequency repetitive transcranial magnetic simulation can potentially treat epilepsy, its underlying mechanism remains unclear. This study investigated the influence of low-frequency re-petitive transcranial magnetic simulation on changes in several nonlinear dynamic electroenceph-alographic parameters in rats with chronic epilepsy and explored the mechanism underlying repeti-tive transcranial magnetic simulation-induced antiepileptic effects. An epilepsy model was estab-lished using lithium-pilocarpine intraperitoneal injection into adult Sprague-Dawley rats, which were then treated with repetitive transcranial magnetic simulation for 7 consecutive days. Nonlinear elec-electroencephalographic parameters were obtained from the rats at 7, 14, and 28 days post-stimulation. Results showed significantly lower mean correlation-dimension and Kolmogo-rov-entropy values for stimulated rats than for non-stimulated rats. At 28 days, the complexity and point-wise correlation dimensional values were lower in stimulated rats. Low-frequency repetitive transcranial magnetic simulation has suppressive effects on electrical activity in epileptic rats, thus explaining its effectiveness in treating epilepsy.

  17. Optogenetic delay of status epilepticus onset in an in vivo rodent epilepsy model.

    Science.gov (United States)

    Sukhotinsky, Inna; Chan, Alexander M; Ahmed, Omar J; Rao, Vikram R; Gradinaru, Viviana; Ramakrishnan, Charu; Deisseroth, Karl; Majewska, Ania K; Cash, Sydney S

    2013-01-01

    Epilepsy is a devastating disease, currently treated with medications, surgery or electrical stimulation. None of these approaches is totally effective and our ability to control seizures remains limited and complicated by frequent side effects. The emerging revolutionary technique of optogenetics enables manipulation of the activity of specific neuronal populations in vivo with exquisite spatiotemporal resolution using light. We used optogenetic approaches to test the role of hippocampal excitatory neurons in the lithium-pilocarpine model of acute elicited seizures in awake behaving rats. Hippocampal pyramidal neurons were transduced in vivo with a virus carrying an enhanced halorhodopsin (eNpHR), a yellow light activated chloride pump, and acute seizure progression was then monitored behaviorally and electrophysiologically in the presence and absence of illumination delivered via an optical fiber. Inhibition of those neurons with illumination prior to seizure onset significantly delayed electrographic and behavioral initiation of status epilepticus, and altered the dynamics of ictal activity development. These results reveal an essential role of hippocampal excitatory neurons in this model of ictogenesis and illustrate the power of optogenetic approaches for elucidation of seizure mechanisms. This early success in controlling seizures also suggests future therapeutic avenues.

  18. BDNF modifies hippocampal KCC2 and NKCC1 expression in a temporal lobe epilepsy model.

    Science.gov (United States)

    Eftekhari, Sanaz; Mehrabi, Soraya; Soleimani, Mansooreh; Hassanzadeh, Gholamreza; Shahrokhi, Amene; Mostafavi, Hossein; Hayat, Parisa; Barati, Mahmood; Mehdizadeh, Hajar; Rahmanzadeh, Reza; Hadjighassem, Mahmoud Reza; Joghataei, Mohammad Taghi

    2014-01-01

    Excitatory GABA actions, induced by altered expression of chloride transporters (KCC2/NKCC1), can contribute to seizure generation in temporal lobe epilepsy. In the present study, we evaluated whether BDNF administration can affect KCC2/NKCC1 expression, ictogenesis and behavioral alterations in this paradigm. Status epilepticus was induced in male rats with pilocarpine, followed by a treatment of either a single high dose or multiple injections of BDNF during the latent phase of temporal lobe epilepsy. Chloride transporters expression, spontaneous recurrent seizures, and hyperexcitability post-seizural behaviors were evaluated after treatment. NKCC1 protein expression was markedly upregulated, whereas that of KCC2 was significantly downregulated in epileptic hippocampi compared to intact controls. Application of BDNF (both single high dose and multiple injections) increased KCC2 expression in epileptic hippocampi, while NKCC1 expression was downregulated exclusively by the single high dose injection of BDNF. Development of spontaneous recurrent seizures was delayed but not prevented by the treatment, and hyperexcitability behaviors were ameliorated for a short period of time. To prevent GABA-A mediated depolarization and design appropriate treatment strategies for temporal lobe epilepsy, chloride transporters can be considered as a target. Future studies are warranted to investigate any possible therapeutic effects of BDNF via altering chloride transporters expression.

  19. Epilepsy-induced electrocardiographic alterations following cardiac ischemia and reperfusion in rats

    Directory of Open Access Journals (Sweden)

    J.G.P. Tavares

    2015-02-01

    Full Text Available The present study evaluated electrocardiographic alterations in rats with epilepsy submitted to an acute myocardial infarction (AMI model induced by cardiac ischemia and reperfusion. Rats were randomly divided into two groups: control (n=12 and epilepsy (n=14. It was found that rats with epilepsy presented a significant reduction in atrioventricular block incidence following the ischemia and reperfusion procedure. In addition, significant alterations were observed in electrocardiogram intervals during the stabilization, ischemia, and reperfusion periods of rats with epilepsy compared to control rats. It was noted that rats with epilepsy presented a significant increase in the QRS interval during the stabilization period in relation to control rats (P<0.01. During the ischemia period, there was an increase in the QRS interval (P<0.05 and a reduction in the P wave and QT intervals (P<0.05 for both in rats with epilepsy compared to control rats. During the reperfusion period, a significant reduction in the QT interval (P<0.01 was verified in the epilepsy group in relation to the control group. Our results indicate that rats submitted to an epilepsy model induced by pilocarpine presented electrical conductivity alterations of cardiac tissue, mainly during an AMI episode.

  20. ALA-induced PpIX fluorescence in epileptogenic tissue

    Science.gov (United States)

    Kleen, Jonathan K.; Valdes, Pablo A.; Harris, Brent T.; Holmes, Gregory L.; Paulsen, Keith D.; Roberts, David W.

    2011-03-01

    Astrogliotic tissue displays markedly increased levels of ALA-induced PpIX fluorescence, making it useful for fluorescence-guided resection in glioma surgery. In patients with temporal lobe epilepsy (TLE) and corresponding animal models, there are areas of astrogliosis that often co-localize with the epileptic focus, which can be resected to eliminate seizures in the majority of treated patients. If this epileptogenic tissue can exhibit PpIX fluorescence that is sufficiently localized, it could potentially help identify margins in epilepsy surgery. We tested the hypothesis that ALA-induced PpIX fluorescence could visually accentuate epileptogenic tissue, using an established animal model of chronic TLE. An acute dose of pilocarpine was used to induce chronic seizure activity in a rat. This rat and a normal control were given ALA, euthanized, and brains examined post-mortem for PpIX fluorescence and neuropathology. Preliminary evidence indicates increased PpIX fluorescence in areas associated with chronic epileptic changes and seizure generation in TLE, including the hippocampus and parahippocampal areas. In addition, strong PpIX fluorescence was clearly observed in layer II of the piriform cortex, a region known for epileptic reorganization and involvement in the generation of seizures in animal studies. We are further investigating whether ALA-induced PpIX fluorescence can consistently identify epileptogenic zones, which could warrant the extension of this technique to clinical studies for use as an adjuvant guidance technology in the resection of epileptic tissue.

  1. Changes in synaptic and extrasynaptic N-methyl-D-aspartate receptor-mediated currents at early-stage epileptogenesis in adult mice

    Institute of Scientific and Technical Information of China (English)

    Juegang Ju; Sheng-tian Li

    2011-01-01

    Previous reports have shown that N-methyl-D-aspartate (NMDA) receptors are extensively involved in epilepsy genesis and recurrence.Recent studies have shown that synaptic and extrasynaptic NMDA receptors play different, or even opposing, roles in various signaling pathways, including synaptic plasticity and neuronal death.The present study analyzed changes in synaptic and extrasynaptic NMDA receptor-mediated currents during epilepsy onset.Mouse models of lithium chloride pilocarpine-induced epilepsy were established, and hippocampal slices were prepared at 24 hours after the onset of status epilepticus.Synaptic and extrasynaptic NMDA receptor-mediated excitatory post-synaptic currents (NMDA-EPSCs) were recorded in CA1 pyramidal neurons by whole-cell patch clamp technique.Results demonstrated no significant difference in rise and delay time of synaptic NMDA-EPSCs compared with normal neurons.Peak amplitude, area-to-peak ratio,and rising time of extrasynaptic NMDA-EPSCs remained unchanged, but decay of extrasynaptic NMDA-EPSCs was faster than that of normal neurons.These results suggest that extrasynaptic NMDA receptors play a role in epileptogenesis.

  2. A Low Mortality, High Morbidity Reduced Intensity Status Epilepticus (RISE) Model of Epilepsy and Epileptogenesis in the Rat

    Science.gov (United States)

    Pérès, Isabelle A. A.; Hadid, Rebecca D.; Amada, Naoki; Hill, Charlotte; Williams, Claire; Stanford, Ian M.; Morris, Christopher M.; Jones, Roland S. G.; Whalley, Benjamin J.; Woodhall, Gavin L.

    2016-01-01

    Animal models of acquired epilepsies aim to provide researchers with tools for use in understanding the processes underlying the acquisition, development and establishment of the disorder. Typically, following a systemic or local insult, vulnerable brain regions undergo a process leading to the development, over time, of spontaneous recurrent seizures. Many such models make use of a period of intense seizure activity or status epilepticus, and this may be associated with high mortality and/or global damage to large areas of the brain. These undesirable elements have driven improvements in the design of chronic epilepsy models, for example the lithium-pilocarpine epileptogenesis model. Here, we present an optimised model of chronic epilepsy that reduces mortality to 1% whilst retaining features of high epileptogenicity and development of spontaneous seizures. Using local field potential recordings from hippocampus in vitro as a probe, we show that the model does not result in significant loss of neuronal network function in area CA3 and, instead, subtle alterations in network dynamics appear during a process of epileptogenesis, which eventually leads to a chronic seizure state. The model’s features of very low mortality and high morbidity in the absence of global neuronal damage offer the chance to explore the processes underlying epileptogenesis in detail, in a population of animals not defined by their resistance to seizures, whilst acknowledging and being driven by the 3Rs (Replacement, Refinement and Reduction of animal use in scientific procedures) principles. PMID:26909803

  3. Prevention and management of radiation-induced dermatitis, mucositis, and xerostomia.

    Science.gov (United States)

    Radvansky, Lauren J; Pace, Makala B; Siddiqui, Asif

    2013-06-15

    Current strategies for preventing and managing radiation-induced dermatitis, mucositis, and xerostomia are reviewed, with an emphasis on pharmacologic interventions. Nearly two thirds of all patients with cancer receive radiation therapy during the course of treatment, frequently resulting in acute skin and mucosal toxicities. The severity of radiotherapy-associated toxicities varies according to multiple treatment- and patient-related factors (e.g., total radiation dose and dose fractionation schedule, volume of organ or tissue irradiated, use of concurrent versus sequential chemotherapy, comorbid conditions, functional performance status). Three major radiation toxicities encountered in clinical practice are (1) radiation dermatitis, typically managed with a variety of topical agents such as water-based moisturizing creams or lotions, topical steroids, antiinflammatory emulsions, and wound dressings, (2) radiation-induced oral mucositis, which can be managed through proper basic oral care practices, appropriate pain management, and the use of medicated mouthwashes and oral rinses and gels, and (3) radiation-induced xerostomia, which can be alleviated with saliva substitutes, moistening agents, and sialagogues. Pharmacists involved in the care of patients receiving radiotherapy can play an important role in optimizing symptom control, educating patients on self-care strategies, and adverse effect monitoring and reporting. Radiation-induced dermatitis, mucositis, and xerostomia can cause significant morbidity and diminished quality of life. Pharmacologic interventions for the prevention and treatment of these toxicities include topical agents for dermatitis; oral products, analgesics, and palifermin for mucositis; and amifostine, saliva substitutes, and pilocarpine for xerostomia.

  4. Effect of central muscarinic receptors on passive-avoidance learning deficits induced by prenatal pentylenetetrazol kindling in male offspring.

    Science.gov (United States)

    Pourmotabbed, A; Mahmoodi, G; Mahmoodi, S; Mohammadi-Farani, A; Nedaei, S E; Pourmotabbed, T; Pourmotabbed, T

    2014-10-24

    Occurrence of the epileptic seizures during gestation might affect the neurodevelopment of the fetus resulting in cognitive problems for the child later in life. We have previously reported that prenatal pentylenetetrazol (PTZ)-kindling induces learning and memory deficits in the children born to kindled mothers, later in life but the mechanisms involved in this processes are unknown. The cholinergic system plays a major role in learning and memory. The present study was performed to investigate the possible involvement of central muscarinic cholinergic receptors on learning and memory deficits induced by prenatal PTZ-kindling in male offspring. Pregnant Wistar rats were kindled by repetitive i.p. injection of 25mg/kg of PTZ on day 13 of their pregnancy. The effect of intracerebroventricular (ICV) microinjection of scopolamine and pilocarpine, muscarinic cholinergic receptors antagonist and agonist, respectively on passive-avoidance learning of pups were tested at 12weeks of age using shuttle-box apparatus. Our data showed that the retention latencies of pups that received scopolamine (2 or 3μg) were significantly reduced compared to those received normal saline (pkindled dams and suggest a central mechanism for the cognitive and memory dysfunction, associated with seizures during pregnancy.

  5. Age-dependent consequences of seizures and the development of temporal lobe epilepsy in the rat.

    Science.gov (United States)

    Dubé, C; da Silva Fernandes, M J; Nehlig, A

    2001-01-01

    The age-related functional changes underlying epileptogenesis remain to be clarified. In the present study, we explored the correlation between metabolic changes, neuronal damage and epileptogenesis during the acute, silent and chronic phases following status epilepticus (SE) induced by lithium-pilocarpine (Li-Pilo) in 10- (P10), 21-day-old (P21) and adult rats. Local cerebral metabolic rates for glucose (LCMRglcs) were measured by the [14C]2-deoxyglucose method during SE, the silent period and the interictal phase of the chronic period. Neurodegeneration was assessed by cresyl violet staining. During SE, LCMRglcs dramatically increased at all ages mainly in forebrain vulnerable regions. During the silent phase, in P21 and adult rats, metabolic decreases were recorded in damaged forebrain regions involved in the genesis and propagation of seizures 14 days after SE. At the end of the silent phase, P21 and adult rats exhibited metabolic increases in intact brainstem areas involved in the remote control of epilepsy. During the interictal phase of the chronic period, LCMRglcs decreased in damaged forebrain areas of adult and P21 rats that were not spontaneously epileptic, while LCMRglcs were similar to control levels in epileptic P21 rats. In P10 rats, there was no damage and no metabolic consequences at any time after SE. In conclusion, the process of epileptogenesis and its functional consequences differ in P21 and adult rats. The factors underlying these age-related differences remain to be explored. Copyright 2001 S. Karger AG, Basel

  6. Pentylenetetrazole-induced seizures in developing rats prenatally exposed to valproic acid

    Directory of Open Access Journals (Sweden)

    Angel A. Puig-Lagunes

    2016-11-01

    Full Text Available Background Epidemiological evidence indicates epilepsy is more common in patients with autism spectrum disorders (ASD (20–25% than in the general population. The aim of this project was to analyze seizure susceptibility in developing rats prenatally exposed to valproic acid (VPA as autism model. Methods Pregnant females were injected with VPA during the twelfth embryonic day. Seizures were induced in fourteen-days-old rat pups using two models of convulsions: pentylenetetrazole (PTZ and lithium-pilocarpine (Li-Pilo. Results Two subgroups with different PTZ-induced seizure susceptibility in rats exposed to VPA were found: a high susceptibility (VPA+ (28/42, seizure severity 5 and a low susceptibility (VPA− (14/42, seizure severity 2. The VPA+ subgroup exhibited an increased duration of the generalized tonic-clonic seizure (GTCS; 45 ± 2.7 min, a higher number of rats showed several GTCS (14/28 and developed status epilepticus (SE after PTZ injection (19/27 compared with control animals (36.6 ± 1.9 min; 10/39; 15/39, respectively. No differences in seizure severity, latency or duration of SE induced by Li-Pilo were detected between VPA and control animals. Discussion Prenatal VPA modifies the susceptibility to PTZ-induced seizures in developing rats, which may be linked to an alteration in the GABAergic transmission. These findings contribute to a better understanding of the comorbidity between autism and epilepsy.

  7. Autocrine/paracrine dopamine in the salivary glands of the blacklegged tick Ixodes scapularis.

    Science.gov (United States)

    Koči, Juraj; Simo, Ladislav; Park, Yoonseong

    2014-03-01

    Dopamine (DA) is known to be the most potent activator of tick salivary secretion, which is an essential component of successful tick feeding. We examined the quantitative changes of catecholamines using a method coupling high-pressure liquid chromatography with electrochemical detection (HPLC-ECD). We also investigated the levels of catecholamines conjugated to other molecules utilising appropriate methods to hydrolyse the conjugates. Three different biological samples, salivary glands, synganglia, ovaries and haemolymph were compared, and the largest quantity of DA was detected in salivary gland extracts (up to ∼100pg/tick), supporting the hypothesis that autocrine/paracrine dopamine activates salivary secretion. Quantitative changes of catecholamines in the salivary glands over the entire blood feeding duration were examined. The amount of dopamine in the salivary glands increased until the day 5 of feeding, at which the rapid engorgement phase began. We also detected a small but significant amount of norepinephrine in the salivary glands. Interestingly, saliva collected after induction of salivary secretion by the cholinergic agonist pilocarpine contained a large amount of DA sulphate with a trace amount of DA, suggesting a potential biological role of DA sulphate in tick saliva.

  8. Comparative Anticonvulsant Study of Epoxycarvone Stereoisomers

    Directory of Open Access Journals (Sweden)

    Paula Regina Rodrigues Salgado

    2015-10-01

    Full Text Available Stereoisomers of the monoterpene epoxycarvone (EC, namely (+-cis-EC, (−-cis-EC, (+-trans-EC, and (−-trans-EC, were comparatively evaluated for anticonvulsant activity in specific methodologies. In the pentylenetetrazole (PTZ-induced anticonvulsant test, all of the stereoisomers (at 300 mg/kg increased the latency to seizure onset, and afforded 100% protection against the death of the animals. In the maximal electroshock-induced seizures (MES test, prevention of tonic seizures was also verified for all of the isomers tested. However, the isomeric forms (+ and (−-trans-EC showed 25% and 12.5% inhibition of convulsions, respectively. In the pilocarpine-induced seizures test, all stereoisomers demonstrated an anticonvulsant profile, yet the stereoisomers (+ and (−-trans-EC (at 300 mg/kg showed a more pronounced effect. A strychnine-induced anticonvulsant test was performed, and none of the stereoisomers significantly increased the latency to onset of convulsions; the stereoisomers probably do not act in this pathway. However, the stereoisomers (+-cis-EC and (+-trans-EC greatly increased the latency to death of the animals, thus presenting some protection. The four EC stereoisomers show promise for anticonvulsant activity, an effect emphasized in the isomers (+-cis-EC, (+-trans-EC, and (−-trans-EC for certain parameters of the tested methodologies. These results serve as support for further research and development of antiepileptic drugs from monoterpenes.

  9. Ketogenic Diet Prevents Epileptogenesis and Disease Progression in Adult Mice and Rats

    Science.gov (United States)

    Lusardi, Theresa A.; Akula, Kiran K.; Coffman, Shayla Q.; Ruskin, David; Masino, Susan A.; Boison, Detlev

    2015-01-01

    Epilepsy is a highly prevalent seizure disorder which tends to progress in severity and become refractory to treatment. Yet no therapy is proven to halt disease progression or to prevent the development of epilepsy. Because a high fat low carbohydrate ketogenic diet (KD) augments adenosine signaling in the brain and because adenosine not only suppresses seizures but also affects epileptogenesis, we hypothesized that a ketogenic diet might prevent epileptogenesis through similar mechanisms. Here, we tested this hypothesis in two independent rodent models of epileptogenesis. Using a pentylenetetrazole kindling paradigm in mice, we first show that a KD, but not a conventional antiepileptic drug (valproic acid), suppressed kindling-epileptogenesis. Importantly, after treatment reversal, increased seizure thresholds were maintained in those animals kindled in the presence of a KD, but not in those kindled in the presence of valproic acid. Next, we tested whether a KD can halt disease progression in a clinically relevant model of progressive epilepsy. Epileptic rats that developed spontaneous recurrent seizures after a pilocarpine-induced status epilepticus were treated with a KD or control diet (CD). Whereas seizures progressed in severity and frequency in the CD-fed animals, KD-fed animals showed a prolonged reduction of seizures, which persisted after diet reversal. KD-treatment was associated with increased adenosine and decreased DNA methylation, the latter being maintained after diet discontinuation. Our findings demonstrate that a KD prevented disease progression in two mechanistically different models of epilepsy, and suggest an epigenetic mechanism underlying the therapeutic effects. PMID:26256422

  10. Anticonvulsant and behavioral effects observed in mice following treatment with an ester derivative of ferulic acid: Isopentyl ferulate.

    Science.gov (United States)

    Machado, Keylla C; Oliveira, George Laylson S; Machado, Kátia C; Islam, Md Torequl; Junior, Antonio Luiz G; De Sousa, Damião P; Freitas, Rivelilson M

    2015-12-01

    The objective of this study was to evaluate the potential anticonvulsant effect of isopentyl ferulate, a new ester derived from ferulic acid in mice (Mus musculus) subjected to two models of induced seizures. According to the results obtained, the IF at doses of 25, 50 and 75 mg/kg (i.p.) showed protective effect against induced seizures by pilocarpine (400 mg/kg, i.p.) and pentylenetetrazole (70 mg/kg, i.p.). In the two animal models of seizures, the pretreatment of the IF (25, 50 and 75 mg/kg) with flumazenil blocked the anticonvulsant effect, suggesting that the mechanism of action of this ester derived of ferulic acid may be related to activity in the benzodiazepine-binding site of the GABAA receptor (γ-aminobutyric acid, type A). In addition to the anticonvulsant effect, behavioral changes as neurotoxicity indication were assessed by using the rota rod and open field tests. The results obtained showed that the IF (25, 50 and 75 mg/kg) does not induce significant changes in locomotor activity and motor coordination when compared with the control group, unlike the results presented by diazepam. Thus, these results demonstrate a new pharmacological knowledge of IF with potential application against epileptic seizures. However, further studies are needed to elucidate other neurobiological mechanisms underlying epilepsy.

  11. Neuronal activity rapidly induces transcription of the CREB-regulated microRNA-132, in vivo.

    Science.gov (United States)

    Nudelman, Aaron S; DiRocco, Derek P; Lambert, Talley J; Garelick, Michael G; Le, Josh; Nathanson, Neil M; Storm, Daniel R

    2010-04-01

    Activity-dependent changes in gene-expression are believed to underlie the molecular representation of memory. In this study, we report that in vivo activation of neurons rapidly induces the CREB-regulated microRNA miR-132. To determine if production of miR-132 is regulated by neuronal activity its expression in mouse brain was monitored by quantitative RT-PCR (RT-qPCR). Pilocarpine-induced seizures led to a robust, rapid, and transient increase in the primary transcript of miR-132 (pri-miR-132) followed by a subsequent rise in mature microRNA (miR-132). Activation of neurons in the hippocampus, olfactory bulb, and striatum by contextual fear conditioning, odor-exposure, and cocaine-injection, respectively, also increased pri-miR-132. Induction kinetics of pri-miR-132 were monitored and found to parallel those of immediate early genes, peaking at 45 min and returning to basal levels within 2 h of stimulation. Expression levels of primary and mature-miR-132 increased significantly between postnatal Days 10 and 24. We conclude that miR-132 is an activity-dependent microRNA in vivo, and may contribute to the long-lasting proteomic changes required for experience-dependent neuronal plasticity.

  12. Epilepsy-induced electrocardiographic alterations following cardiac ischemia and reperfusion in rats

    Energy Technology Data Exchange (ETDEWEB)

    Tavares, J.G.P. [Departamento de Farmacologia, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Universidade Iguaçu, Campos V, Itaperuna, RJ (Brazil); Faculdade de Minas, Muriaé, MG (Brazil); Vasques, E.R. [Departamento de Gastroenterologia, LIM 37, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP (Brazil); Arida, R.M. [Departamento de Fisiologia, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Cavalheiro, E.A. [Departamento de Neurologia e Neurocirurgia, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Cabral, F.R.; Torres, L.B. [Hospital Israelita Albert Einstein, Instituto do Cérebro, São Paulo, SP (Brazil); Menezes-Rodrigues, F.S.; Jurkiewicz, A.; Caricati-Neto, A. [Departamento de Farmacologia, Universidade Federal de São Paulo, São Paulo, SP (Brazil); Godoy, C.M.G. [Departamento de Ciência e Tecnologia, Universidade Federal de São Paulo, São José dos Campos, SP (Brazil); Gomes da Silva, S. [Hospital Israelita Albert Einstein, Instituto do Cérebro, São Paulo, SP (Brazil); Núcleo de Pesquisas Tecnológicas, Programa Integrado em Engenharia Biomédica, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP (Brazil)

    2015-01-13

    The present study evaluated electrocardiographic alterations in rats with epilepsy submitted to an acute myocardial infarction (AMI) model induced by cardiac ischemia and reperfusion. Rats were randomly divided into two groups: control (n=12) and epilepsy (n=14). It was found that rats with epilepsy presented a significant reduction in atrioventricular block incidence following the ischemia and reperfusion procedure. In addition, significant alterations were observed in electrocardiogram intervals during the stabilization, ischemia, and reperfusion periods of rats with epilepsy compared to control rats. It was noted that rats with epilepsy presented a significant increase in the QRS interval during the stabilization period in relation to control rats (P<0.01). During the ischemia period, there was an increase in the QRS interval (P<0.05) and a reduction in the P wave and QT intervals (P<0.05 for both) in rats with epilepsy compared to control rats. During the reperfusion period, a significant reduction in the QT interval (P<0.01) was verified in the epilepsy group in relation to the control group. Our results indicate that rats submitted to an epilepsy model induced by pilocarpine presented electrical conductivity alterations of cardiac tissue, mainly during an AMI episode.

  13. PDI regulates seizure activity via NMDA receptor redox in rats.

    Science.gov (United States)

    Kim, Ji Yang; Ko, Ah-Rhem; Hyun, Hye-Won; Min, Su-Ji; Kim, Ji-Eun

    2017-02-15

    Redox modulation of cysteine residues is one of the post-translational modifications of N-methyl-D-aspartate receptor (NMDAR). Protein disulfide isomerases (PDI), an endoplasmic reticulum (ER) chaperone, plays a crucial role in catalyzing disulfide bond formation, reduction, and isomerization. In the present study, we found that PDI bound to NMDAR in the normal hippocampus, and that this binding was increased in chronic epileptic rats. In vitro thiol reductase assay revealed that PDI increased the amount of thiols on full-length recombinant NR1 protein. PDI siRNA, 5-5'-dithio-bis(2-nitrobenzoic acid) (DTNB), bacitracin and PDI antibody reduced seizure susceptibility in response to pilocarpine. In addition, PDI knockdown effectively ameliorated spontaneous seizure activity in chronic epileptic rats. Anticonvulsive effects of PDI siRNA were correlated to the reduction of the amount of free- and nitrosothiols on NMDAR, accompanied by the inhibition of PDI activity. However, PDI knockdown did not lead to alteration in basal neurotransmission or ER stress under physiological condition. These findings provide mechanistic insight into sulfhydration of disulfide bonds on NMDAR by PDI, and suggest that PDI may represent a target of potential therapeutics for epilepsy, which avoids a possible side effect on physiological receptor functionality.

  14. P2X7 receptor-mediated PARP1 activity regulates astroglial death in the rat hippocampus following status epilepticus

    Directory of Open Access Journals (Sweden)

    Ji Yang eKim

    2015-09-01

    Full Text Available Poly(ADP-ribose polymerase-1 (PARP1 plays a regulatory role in apoptosis, necrosis, and other cellular processes after injury. Recently, we revealed that PARP1 regulates the differential neuronal/astroglial responses to pilocarpine-induced status epilepticus (SE in the distinct brain regions. In addition, P2X7 receptor (P2X7R, an ATP-gated ion channel, activation accelerates astroglial apoptosis, while it attenuates clasmatodendrosis (lysosome-derived autophagic astroglial death. Therefore, we investigated whether P2X7R regulates regional specific astroglial PARP1 expression/activation in response to SE. In the present study, P2X7R activation exacerbates SE-induced astroglial apoptosis, while P2X7R inhibition attenuates it accompanied by increasing PARP1 activity in the molecular layer of the dentate gyrus following SE. In the CA1 region, however, P2X7R inhibition deteriorates SE-induced clasmatodendrosis via PARP1 activation following SE. Taken together, our findings suggest that P2X7R function may affect SE-induced astroglial death by regulating PARP1 activation/expression in regional-specific manner. Therefore, the selective modulation of P2X7R-mediated PARP1 functions may be a considerable strategy for controls in various types of cell deaths.

  15. Effect of pupil size on multifocal pattern visual evoked potentials.

    Science.gov (United States)

    Martins, Alessandra; Balachandran, Chandra; Klistorner, Alexander I; Graham, Stuart L; Billson, Francis A

    2003-08-01

    The purpose of this study was to investigate the influence of pupil diameter on the amplitude and latency of multifocal visual evoked potentials (mfVEP). The multifocal objective perimeter (Accumap; Objectivision) was used to stimulate the visual field at 56 sites extending to 32 degrees using a pseudo-random pattern stimulus. The mfVEP were recorded using bipolar occipital electrodes, 7 min/eye. Ten normal subjects were recruited from the community and one eye was randomly selected for testing. The mfVEP were recorded at four different pupil diameters (2 mm, 4 mm, 6 mm, 8 mm), obtained by applying tropicamide (0.5%) or pilocarpine (2%) in different dilutions. Appropriate refractive correction was provided to overcome cycloplegia and achieve a visual acuity of 6/7.5 or better. Analysis revealed that at most pupil diameters the normalized full field amplitude did not show significant variation, except at the most miotic pupil diameter (2 mm), where the amplitude became reduced, based on 2-way anova and Tukey's T method. There was, however, significant correlation between latency and pupil area (correlation coefficient: upper field -0.63, lower field -0.76). The results suggest that even in the presence of mydriatics or miotics, the mfVEP test can be used to assess diseases that affect amplitude, provided near correction is used. The interpretation of latency, however, must be made with caution, as a borderline conduction defect with a dilated pupil may appear normal.

  16. MRI changes and complement activation correlate with epileptogenicity in a mouse model of temporal lobe epilepsy.

    Science.gov (United States)

    Kharatishvili, Irina; Shan, Zuyao Y; She, David T; Foong, Samuel; Kurniawan, Nyoman D; Reutens, David C

    2014-03-01

    The complex pathogenesis of temporal lobe epilepsy includes neuronal and glial pathology, synaptic reorganization, and an immune response. However, the spatio-temporal pattern of structural changes in the brain that provide a substrate for seizure generation and modulate the seizure phenotype is yet to be completely elucidated. We used quantitative magnetic resonance imaging (MRI) to study structural changes triggered by status epilepticus (SE) and their association with epileptogenesis and with activation of complement component 3 (C3). SE was induced by injection of pilocarpine in CD1 mice. Quantitative diffusion-weighted imaging and T2 relaxometry was performed using a 16.4-Tesla MRI scanner at 3 h and 1, 2, 7, 14, 28, 35, and 49 days post-SE. Following longitudinal MRI examinations, spontaneous recurrent seizures and interictal spikes were quantified using continuous video-EEG monitoring. Immunohistochemical analysis of C3 expression was performed at 48 h, 7 days, and 4 months post-SE. MRI changes were dynamic, reflecting different outcomes in relation to the development of epilepsy. Apparent diffusion coefficient changes in the hippocampus at 7 days post-SE correlated with the severity of the evolving epilepsy. C3 activation was found in all stages of epileptogenesis within the areas with significant MRI changes and correlated with the severity of epileptic condition.

  17. Commentary: a skeptical view of experimental gene therapy to block epileptogenesis.

    Science.gov (United States)

    Dudek, F Edward

    2009-04-01

    Gene therapy offers exciting new options for treating epileptic seizures, and even for blocking the development of epilepsy (i.e., epileptogenesis) after a brain insult. Although the available studies provide interesting new data, the experiments discussed in this issue also have limitations and raise concerns. The criticisms offered in this commentary center around the nature of the experimental testing (e.g., changes in seizure threshold), the animal models (e.g., kindling), and the measures of epileptogenesis in those animal models with spontaneous seizures (e.g., the latent period after pilocarpine-induced status epilepticus). Another set of criticisms relate to the relative lack of positive controls showing that the actual mechanism purported to be activated via the gene-therapeutic approach has in fact been upregulated in the specific animals that show the hypothetical antiepileptic result. This commentary takes the con side in the debate, to generate constructive criticism to help direct future studies to provide increasingly stronger data to support the view that gene therapy approaches may be useful in the treatment of epilepsy.

  18. Rhythm and blues: animal models of epilepsy and depression comorbidity.

    Science.gov (United States)

    Epps, S Alisha; Weinshenker, David

    2013-01-15

    Clinical evidence shows a strong, bidirectional comorbidity between depression and epilepsy that is associated with decreased quality of life and responsivity to pharmacotherapies. At present, the neurobiological underpinnings of this comorbidity remain hazy. To complicate matters, anticonvulsant drugs can cause mood disturbances, while antidepressant drugs can lower seizure threshold, making it difficult to treat patients suffering from both depression and epilepsy. Animal models have been created to untangle the mechanisms behind the relationship between these disorders and to serve as screening tools for new therapies targeted to treat both simultaneously. These animal models are based on chemical interventions (e.g. pentylenetetrazol, kainic acid, pilocarpine), electrical stimulations (e.g. kindling, electroshock), and genetic/selective breeding paradigms (e.g. genetically epilepsy-prone rats (GEPRs), genetic absence epilepsy rat from Strasbourg (GAERS), WAG/Rij rats, swim lo-active rats (SwLo)). Studies on these animal models point to some potential mechanisms that could explain epilepsy and depression comorbidity, such as various components of the dopaminergic, noradrenergic, serotonergic, and GABAergic systems, as well as key brain regions, like the amygdala and hippocampus. These models have also been used to screen possible therapies. The purpose of the present review is to highlight the importance of animal models in research on comorbid epilepsy and depression and to explore the contributions of these models to our understanding of the mechanisms and potential treatments for these disorders.

  19. Neuronal activity rapidly induces transcription of the CREB-regulated microRNA-132, in vivo

    DEFF Research Database (Denmark)

    Nudelman, Aaron Samuel; DiRocco, Derek P; Lambert, Talley J

    2010-01-01

    of stimulation. Expression levels of primary and mature-miR-132 increased significantly between postnatal Days 10 and 24. We conclude that miR-132 is an activity-dependent microRNA in vivo, and may contribute to the long-lasting proteomic changes required for experience-dependent neuronal plasticity.......Activity-dependent changes in gene-expression are believed to underlie the molecular representation of memory. In this study, we report that in vivo activation of neurons rapidly induces the CREB-regulated microRNA miR-132. To determine if production of miR-132 is regulated by neuronal activity its...... expression in mouse brain was monitored by quantitative RT-PCR (RT-qPCR). Pilocarpine-induced seizures led to a robust, rapid, and transient increase in the primary transcript of miR-132 (pri-miR-132) followed by a subsequent rise in mature microRNA (miR-132). Activation of neurons in the hippocampus...

  20. Strontium and bromide as tracers in X-ray microanalysis of biological tissue

    Energy Technology Data Exchange (ETDEWEB)

    Wroblewski, J.; Sagstroem, S.M.; Mulders, H.; Roomans, G.M. (Karolinska Institutet, Stockholm (Sweden))

    1989-09-01

    Since energy dispersive X-ray microanalysis cannot distinguish between isotopes of the same element, alternative methods have to be used to get information similar to that obtained in experiments with radioactive tracers. In the present study, strontium was used as a tracer for calcium, and bromide as a tracer for chloride. Rats were injected with strontium chloride in vivo, and the uptake of strontium in the acinar cells of the submandibular gland was studied. Eventually a Sr/Ca ratio of 0.3 was reached. In some animals, secretion of mucus had been elicited by stimulation with isoproterenol 4 h prior to injection of strontium chloride. Exchange of calcium for strontium was enhanced by prior injection with isoproterenol. In a second experiment, rats were injected with sodium bromide, and the uptake of bromide by the submandibular acinar cells was followed in time, both in pilocarpine-stimulated and unstimulated glands. Under the experimental conditions, bromide was rapidly taken up by the cells, and the cellular Br/Cl ratio was close to that found in serum. Submandibular glands take up Br somewhat faster than other tissues (liver, heart muscle, skeletal muscle). The uptake of Br in pancreatic acinar cells was studied in vitro. These experiments showed a 1:1 ratio (molar) exchange of Cl for Br.

  1. Effect of duct obstruction on structure, elemental composition, and function of rat submandibular glands

    Energy Technology Data Exchange (ETDEWEB)

    Sagstroem, S.S.; Sagulin, G.B.; Roomans, G.M. (Univ. of Uppsala (Sweden))

    1989-06-01

    Obstruction of salivary glands occurs in association with a number of pathological conditions. It has been suggested that the major changes found in the salivary glands of patients with cystic fibrosis are due to obstruction of the excretory duct by viscous mucus. In the present study, the effect of excretory duct obstruction on structure, elemental composition and function of rat submandibular gland was investigated. Obstruction was effected by infusion of a fast-hardening protein emulsion in the main excretory duct. After 1 week, and more pronounced after 2 weeks of obstruction the number of granular duct cells had decreased in the obstructed gland. X-ray microanalysis showed an increase in Mg, Ca and K, and a decrease in Na levels in the acinar cells, compared to normal glands. The contralateral glands apparently underwent compensatory hypertrophy and showed a similar pattern of changes in elemental composition. The composition of pilocarpine-induced submandibular saliva was neither in the obstructed nor in the contralateral gland significantly different from that in control glands. However, the flow rate was somewhat lower. Hence, increase in cellular Ca levels in submandibular gland acinar cells in cystic fibrosis could be secondary to duct obstruction, but the present study does not support the hypothesis that duct obstruction would result in changes in the composition of saliva.

  2. Seasonal change in main alkaloids of jaborandi (Pilocarpus microphyllus Stapf ex Wardleworth), an economically important species from the Brazilian flora

    Science.gov (United States)

    Véras, Leiz Maria Costa; Azevedo, Iábita Fabiana Sousa; Biase, Adriele Giaretta; Costa, Joana; Oliveira, Maria Beatriz P. P.; Mafra, Isabel

    2017-01-01

    Pilocarpus microphyllus Stapf ex Wardleworth (jaborandi, Rutaceae) is one of the most important Brazilian medicinal species owing to its content of pilocarpine (PIL), an alkaloid used for treating glaucoma and xerostomia. This species contains another alkaloid, epiisopiloturine (EPI), which has demonstrated effectiveness against schistosomiasis. The aim of this work was to assess seasonal changes of PIL and EPI in three populations of cultivated P. microphyllus from northeastern Brazil over one year, including the dry and rainy seasons. Alkaloid profiles were correlated to phenotypic and genetic patterns in the morphological and molecular characterizations. PIL was the primary alkaloid and its levels differed among populations in all months except September. The S01 population (green line) showed an especially high PIL content compared to populations S02 and S03 (traditional line), which had similar alkaloid contents. PIL content gradually decreased in the three populations in the rainy season.EPI content was significantly different between the green line (S01) and the traditional line (S02 and S03).S01 had a significantly lower EPI content in all months, demonstrating that it was not the best source for EPI extraction. Inter simple sequence repeat (ISSR) markers and morphological analyses clearly separated S01 from S02 and S03, in agreement with the alkaloid results. This study shows the first correlation between the chemical, morphological, and molecular markers of P. microphyllus and highlights the potential benefits of a multidisciplinary research approach aimed at supporting both industry and conservation of natural resources. PMID:28151972

  3. Hippocampal FGF-2 and BDNF overexpression attenuates epileptogenesis-associated neuroinflammation and reduces spontaneous recurrent seizures

    Directory of Open Access Journals (Sweden)

    Osculati Francesco

    2010-11-01

    Full Text Available Abstract Under certain experimental conditions, neurotrophic factors may reduce epileptogenesis. We have previously reported that local, intrahippocampal supplementation of fibroblast growth factor-2 (FGF-2 and brain-derived neurotrophic factor (BDNF increases neurogenesis, reduces neuronal loss, and reduces the occurrence of spontaneous seizures in a model of damage-associated epilepsy. Here, we asked if these possibly anti-epileptogenic effects might involve anti-inflammatory mechanisms. Thus, we used a Herpes-based vector to supplement FGF-2 and BDNF in rat hippocampus after pilocarpine-induced status epilepticus that established an epileptogenic lesion. This model causes intense neuroinflammation, especially in the phase that precedes the occurrence of spontaneous seizures. The supplementation of FGF-2 and BDNF attenuated various parameters of inflammation, including astrocytosis, microcytosis and IL-1β expression. The effect appeared to be most prominent on IL-1β, whose expression was almost completely prevented. Further studies will be needed to elucidate the molecular mechanism(s for these effects, and for that on IL-1β in particular. Nonetheless, the concept that neurotrophic factors affect neuroinflammation in vivo may be highly relevant for the understanding of the epileptogenic process.

  4. Wilson protein expression, copper excretion and sweat production in sweat glands of Wilson disease patients and controls

    Directory of Open Access Journals (Sweden)

    Weiss Karl

    2008-07-01

    Full Text Available Abstract Background In Wilson disease, copper is not sufficiently excreted into bile due to the absence or malfunction of the Wilson protein copper ATPase in the excretory pathway of hepatocytes. Copper is found in sweat. It is unknown if the Wilson protein plays a role in copper excretion into sweat. It is the aim of this study to investigate Wilson protein expression in sweat glands and analysing its effects on copper excretion into sweat in controls and patients with Wilson disease. Methods Immunofluorescent analysis of the Wilson protein in skin samples from normal rat, LEC rat and human skin biopsies were performed. Pilocarpin-induced sweat gland stimulation by iontophoretic transfer adapted from the methods used for cystic fibrosis sweat test was used for sweat induction. Sweat volume, sweat copper concentration, serum ceruloplasmin and serum copper were analysed in 28 Wilson patients and 21 controls. Results The Wilson protein is expressed in human and rat sweat gland epithelia. Copper concentration in sweat is not significantly different between controls and Wilson patients. Wilson patients produce significantly smaller volumes of sweat compared to controls. Sweat production is partially reversible in Wilson patients under medical treatment for Wilson disease or after liver transplantation Conclusion Wilson patients show a reduced sweat production with unaltered sweat copper concentration. The Wilson protein might play an important role in physiological sweat production.

  5. Topiramate-associated acute, bilateral, angle-closure glaucoma: case report

    Directory of Open Access Journals (Sweden)

    Lucas Barasnevicius Quagliato

    2013-02-01

    Full Text Available This paper describes a topiramate induced acute bilateral angle-closure glaucoma. This rare adverse effect is an idiosyncratic reaction characterized by uveal effusion and lens forward displacement, leading to increased intraocular pressure and vision loss. We describe a 55 year-old white woman with migraine, spasmodic torticollis and essential tremor, who developed bilateral acute angle-closure glaucoma, one week after starting topiramate 25 mg/day. She was seen at the Ophthalmology Emergency Department of the Fundação João Penido Burnier (Campinas, SP, Brazil with a 4 hours history of blurry vision, ocular pain and bright flashes vision. Slit lamp examination revealed moderate conjunctival injection and corneal edema, and shallow anterior chambers. Intraocular pressure was 48 mmHg in both eyes. Fundoscopic examination findings were normal. She was treated with timolol, brimonidine, dorzolamide, pilocarpine, prednisone acetate eye drops and acetazolamide. One hour after those measures, as the intraocular pressure was 30 mmHg, she received a manitol intravenous injection and the intraocular pressure normalized. After 24 hours an iridotomy with Yag laser was performed. Topiramate was discontinued and she was totally recovered after one week.

  6. Ketogenic diet prevents epileptogenesis and disease progression in adult mice and rats.

    Science.gov (United States)

    Lusardi, Theresa A; Akula, Kiran K; Coffman, Shayla Q; Ruskin, David N; Masino, Susan A; Boison, Detlev

    2015-12-01

    Epilepsy is a highly prevalent seizure disorder which tends to progress in severity and become refractory to treatment. Yet no therapy is proven to halt disease progression or to prevent the development of epilepsy. Because a high fat low carbohydrate ketogenic diet (KD) augments adenosine signaling in the brain and because adenosine not only suppresses seizures but also affects epileptogenesis, we hypothesized that a ketogenic diet might prevent epileptogenesis through similar mechanisms. Here, we tested this hypothesis in two independent rodent models of epileptogenesis. Using a pentylenetetrazole kindling paradigm in mice, we first show that a KD, but not a conventional antiepileptic drug (valproic acid), suppressed kindling-epileptogenesis. Importantly, after treatment reversal, increased seizure thresholds were maintained in those animals kindled in the presence of a KD, but not in those kindled in the presence of valproic acid. Next, we tested whether a KD can halt disease progression in a clinically relevant model of progressive epilepsy. Epileptic rats that developed spontaneous recurrent seizures after a pilocarpine-induced status epilepticus were treated with a KD or control diet (CD). Whereas seizures progressed in severity and frequency in the CD-fed animals, KD-fed animals showed a prolonged reduction of seizures, which persisted after diet reversal. KD-treatment was associated with increased adenosine and decreased DNA methylation, the latter being maintained after diet discontinuation. Our findings demonstrate that a KD prevented disease progression in two mechanistically different models of epilepsy, and suggest an epigenetic mechanism underlying the therapeutic effects.

  7. Development of epileptiform excitability in the deep entorhinal cortex after status epilepticus

    Science.gov (United States)

    Bragin, Denis E.; Sanderson, Jennifer L.; Peterson, Steven; Connor, John A.; Müller, Wolfgang S.

    2009-01-01

    Epileptiform neuronal activity during seizures is observed in many brain areas, but its origins following status epilepticus (SE) are unclear. We have used the Li-low dose pilocarpine rat model of temporal lobe epilepsy (TLE) to examine early development of epileptiform activity in the deep entorhinal cortex (EC). We show that during the 3 week latent period that follows SE, an increasing percentage of neurons in EC layer 5 respond to a single synaptic stimulus with polysynaptic burst depolarizations. This change is paralleled by a progressive depolarizing shift of the IPSP reversal potential in layer 5 neurons, apparently caused by upregulation of the Cl- inward transporter NKCC1 and concurrent downregulation of the Cl- outward transporter KCC2, both changes favoring intracellular Cl- accumulation. Inhibiting Cl- uptake in the latent period restored more negative GABAergic reversal potentials and eliminated polysynaptic bursts. The changes in the Cl- transporters were highly specific to the deep entorhinal cortex. They did not occur in layers 1-3, perirhinal cortex, subiculum or dentate gyrus during this period. We propose that the changes in Cl- homeostasis facilitate hyperexcitability in the deep entorhinal cortex leading to epileptiform discharge there, which subsequently affects downstream cortical regions. PMID:19674083

  8. Expression of Toll-like receptor 4 in hippocampus of rat model with temporal lobe epilepsy

    Directory of Open Access Journals (Sweden)

    PAN Li-ping

    2013-12-01

    Full Text Available Objective To investigate the expression of Toll-like receptor 4 (TLR4 protein in hippocampus of rat model with temporal lobe epilepsy after status epilepticus (SE and explore its function in the pathogenesis of temporal lobe epilepsy. Methods Rat model with temporal lobe epilepsy was induced by lithium chloride (LiCl-pilocarpine. Total protein was extracted from hippocampus and rat brain slices were obtained at different time points (0, 1, 6, 12, 24, 48 h and 7, 10, 30, 50 d after SE. Western blotting and immunohistochemical staining were used for detection of the expression of TLR4 in the hippocampus. Results The results of Western blotting showed the TLR4 protein expression at 0, 1, 6, 12, 24, 48 h and 7, 10, 30 d after SE was higher than that in the control group (P 0.05. Conclusion TLR4 protein was mainly expressed in cytoplasm of pyramidal cells in CA3 area of hippocampus. TLR4 protein expression in the hippocampus was increased in varying degrees at different observation time points after SE, indicating that TLR4 may play an important role in the development of epilepsy.

  9. Status epilepticus increases mature granule cells in the molecular layer of the dentate gyrus in rats

    Institute of Scientific and Technical Information of China (English)

    Zhaoliang Liang; Fei Gao; Fajun Wang; Xiaochen Wang; Xinyu Song; Kejing Liu; Ren-Zhi Zhan

    2013-01-01

    Enhanced neurogenesis in the dentate gyrus of the hippocampus following seizure activity, especially status epilepticus, is associated with ectopic residence and aberrant integration of newborn granule cells. Hilar ectopic granule cells may be detrimental to the stability of dentate circuitry by means of their electrophysiological properties and synaptic connectivity. We hypothesized that status epilepticus also increases ectopic granule cells in the molecular layer. Status epilepticus was induced in male Sprague-Dawley rats by intraperitoneal injection of pilocarpine. Immunostaining showed that many doublecortin-positive cells were present in the molecular layer and the hilus 7 days after the induction of status epilepticus. At least 10 weeks after status epilepticus, the estimated number of cells positive for both prospero homeobox protein 1 and neuron-specific nuclear protein in the hilus was significantly increased. A similar trend was also found in the molecular layer. These findings indicate that status epilepticus can increase the numbers of mature and ectopic newborn granule cells in the molecular layer.

  10. Seizure-induced disinhibition of the HPA axis increases seizure susceptibility.

    Science.gov (United States)

    O'Toole, Kate K; Hooper, Andrew; Wakefield, Seth; Maguire, Jamie

    2014-01-01

    Stress is the most commonly reported precipitating factor for seizures. The proconvulsant actions of stress hormones are thought to mediate the effects of stress on seizure susceptibility. Interestingly, epileptic patients have increased basal levels of stress hormones, including corticotropin-releasing hormone (CRH) and corticosterone, which are further increased following seizures. Given the proconvulsant actions of stress hormones, we proposed that seizure-induced activation of the hypothalamic-pituitary-adrenal (HPA) axis may contribute to future seizure susceptibility. Consistent with this hypothesis, our data demonstrate that pharmacological induction of seizures in mice with kainic acid or pilocarpine increases circulating levels of the stress hormone, corticosterone, and exogenous corticosterone administration is sufficient to increase seizure susceptibility. However, the mechanism(s) whereby seizures activate the HPA axis remain unknown. Here we demonstrate that seizure-induced activation of the HPA axis involves compromised GABAergic control of CRH neurons, which govern HPA axis function. Following seizure activity, there is a collapse of the chloride gradient due to changes in NKCC1 and KCC2 expression, resulting in reduced amplitude of sIPSPs and even depolarizing effects of GABA on CRH neurons. Seizure-induced activation of the HPA axis results in future seizure susceptibility which can be blocked by treatment with an NKCC1 inhibitor, bumetanide, or blocking the CRH signaling with Antalarmin. These data suggest that compromised GABAergic control of CRH neurons following an initial seizure event may cause hyperexcitability of the HPA axis and increase future seizure susceptibility.

  11. Neuropeptide FF receptors as novel targets for limbic seizure attenuation.

    Science.gov (United States)

    Portelli, Jeanelle; Meurs, Alfred; Bihel, Frederic; Hammoud, Hassan; Schmitt, Martine; De Kock, Joery; Utard, Valerie; Humbert, Jean-Paul; Bertin, Isabelle; Buffel, Ine; Coppens, Jessica; Tourwe, Dirk; Maes, Veronique; De Prins, An; Vanhaecke, Tamara; Massie, Ann; Balasubramaniam, Ambikaipakan; Boon, Paul; Bourguignon, Jean-Jacques; Simonin, Frederic; Smolders, Ilse

    2015-08-01

    Neuropeptide Y (NPY) is a well established anticonvulsant and first-in-class antiepileptic neuropeptide. In this study, the controversial role of NPY1 receptors in epilepsy was reassessed by testing two highly selective NPY1 receptor ligands and a mixed NPY1/NPFF receptor antagonist BIBP3226 in a rat model for limbic seizures. While BIBP3226 significantly attenuated the pilocarpine-induced seizures, neither of the highly selective NPY1 receptor ligands altered the seizure severity. Administration of the NPFF1/NPFF2 receptor antagonist RF9 also significantly attenuated limbic seizure activity. To further prove the involvement of NPFF receptors in these seizure-modulating effects, low and high affinity antagonists for the NPFF receptors were tested. We observed that the low affinity ligand failed to exhibit anticonvulsant properties while the two high affinity ligands significantly attenuated the seizures. Continuous NPFF1 receptor agonist administration also inhibited limbic seizures whereas bolus administration of the NPFF1 receptor agonist was without effect. This suggests that continuous agonist perfusion could result in NPFF1 receptor desensitization and mimic NPFF1 receptor antagonist administration. Our data unveil for the first time the involvement of the NPFF system in the management of limbic seizures.

  12. Insidious weight gain in prepubertal seized rats treated with an atypical neuroleptic: the role of food consumption, fluid consumption, and spontaneous ambulatory activity.

    Science.gov (United States)

    St-Pierre, L S; Bubenik, G A; Parker, G H; Persinger, M A

    2009-02-01

    Extreme obesity slowly develops in female rats over the months following seizures induced by a single systemic injection of lithium and pilocarpine if the resulting limbic seizures are treated with the atypical neuroleptic acepromazine (but not with ketamine). To discern the contributions from food consumption, water consumption, and (daytime and nighttime) activity to this weight gain, these behaviors were monitored for 4 months, about 2 months after seizure induction. The results indicated that the rats that underwent the obesity procedure exhibited 50% heavier body weights and consumed 42% more food than the reference group, which included rats that had been induced to seize but treated with ketamine. There were no statistically significant differences between groups with respect to either water consumption or (daytime or nighttime) activity. Factor analyses of data for individual rats verified the dissociation between activity and weight gain for the obese rats. The results suggest that the progressive weight gains are centrally mediated and are not secondary to diminished activity or altered fluid consumption.

  13. The Role of Canonical Transient Receptor Potential Channels in Seizure and Excitotoxicity

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    Fang Zheng

    2014-04-01

    Full Text Available Canonical transient receptor potential (TRPC channels are a family of polymodal cation channels with some degree of Ca2+ permeability. Although initially thought to be channels mediating store-operated Ca2+ influx, TRPC channels can be activated by stimulation of Gq-coupled G-protein coupled receptors, or by an increase in intracellular free Ca2+ concentration. Thus, activation of TRPC channels could be a common downstream event of many signaling pathways that contribute to seizure and excitotoxicity, such as N-methyl-D-aspartate (NMDA receptor-mediated Ca2+ influx, or metabotropic glutamate receptor activation. Recent studies with genetic ablation of various TRPC family members have demonstrated that TRPC channels, in particular heteromeric TRPC1/4 channels and homomeric TRPC5 channels, play a critical role in both pilocarpine-induced acute seizures and neuronal cell death. However, exact underlying mechanisms remain to be fully elucidated, and selective TRPC modulators and antibodies with better specificity are urgently needed for future research.

  14. Can {alpha}-tocopherol and {beta}-carotene supplementation reduce adverse radiation effects on salivary glands?

    Energy Technology Data Exchange (ETDEWEB)

    Funegaard, U.; Johansson, I.; Ericson, T. [Umeaa Univ. (Sweden). Dept. of Cariology; Malmer, B.; Henriksson, R. [Umeaa Univ. (Sweden). Dept. of Oncology

    1995-12-31

    In this study, we evaluated whether supplementation with antioxidant vitamins can reduce the adverse effects of irradiation on the salivary glands in the rat. Four groups of adult Sprague-Dawley rats were given a basic diet providing 0.6 mg {alpha}-tocopherol and no {beta}-carotene per day. In two groups the basic diet was supplemented with 3.4 mg {alpha}-tocopherol and 6 mg {beta}-carotene per day from 14 days before irradiation until 12 days after complete irradiation. One group of rats given basic diet and one group given supplemented diet were irradiated with 7 Gy daily for five consecutive days. Isoproterenol and pilocarpine-stimulated whole saliva was collected from all rats 2, 4 and 26 weeks after irradiation. Vitamin-supplemented irradiated rats had higher secretion rates on all three occasions compared with those of irradiated rats given basic diet. The changes in saliva composition seen in irradiated rats were less accentuated in vitamin-supplemented, irradiated rats. The proportions of acinar cells were significantly decreased both in parotid and submandibular glands 26 weeks after irradiation. Supplementation with {alpha}-tocopherol and {beta}-carotene did not alter the morphology of the glands. (author).

  15. Autonomic thermoregulatory dysfunction in neurofibromatosis type 1

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    Luciana G Madeira

    Full Text Available ABSTRACT Objective Neurofibromatosis type 1 (NF1 causes neural and cutaneous disorders and reduced exercise capacity. Exercise/heat exposure increasing internal temperature must be compensated by eccrine sweat function and warmed skin vasodilation. We suspected NF1 could adversely affect eccrine sweat function and/or vascular thermoregulatory responses (VTR. Methods The eccrine sweat function and VTR of 25 NF1 volunteers (14 males, 11 females; 16–57 years old were compared with 23 non-NF1 controls matched by sex, age, height and weight (CG. Sweating was induced by 1 pilocarpine 1% iontophoresis (PILO; and 2 by passive heating (HEAT via the lower third of the legs being immersed in 42°C water for one hour. Previously established eccrine sweat function and VTR protocols were used. Results The NF1 group showed: a lower sweat rate than the CG group during PILO; b a smaller diastolic pressure decrease; and c higher tympanic temperatures than controls during HEAT (p < 0.05. Conclusion Reduced sweating and vascular thermoregulatory responses suggest autonomic dysfunction in NF1 individuals.

  16. Radiation-induced xerostomia: pathophysiology, clinical course and supportive treatment.

    Science.gov (United States)

    Guchelaar, H J; Vermes, A; Meerwaldt, J H

    1997-07-01

    Xerostomia, or oral dryness, is one of the most common complaints experienced by patients who have had radiotherapy of the oral cavity and neck region. The hallmarks of radiation-induced damage are acinar atrophy and chronic inflammation of the salivary glands. The early response, resulting in atrophy of the secretory cells without inflammation might be due to radiation-induced apoptosis. In contrast, the late response with inflammation could be a result of radiation-induced necrosis. The subjective complaint of a dry mouth appears to be poorly correlated with objective findings of salivary gland dysfunction. Xerostomia, with secondary symptoms of increased dental caries, difficulty in chewing, swallowing and speaking, and an increased incidence of oral candidiasis, can have a significant effect on the quality of life. At present there is no causal treatment for radiation-induced xerostomia. Temporary symptomatic relief can be offered by moistening agents and saliva substitutes, and is the only option for patients without residual salivary function. In patients with residual salivary function, oral administration of pilocarpine 5-10 mg three times a day is effective in increasing salivary flow and improving the symptoms of xerostomia, and this therapy should be considered as the treatment of choice. Effectiveness of sialogogue treatment requires residual salivary function, which emphasizes the potential benefit from sparing normal tissue during irradiation. The hypothesis concerning the existence of early apoptotic and late necrotic effects of irradiation on the salivary glands theoretically offers a way of achieving this goal.

  17. Evaluation of drug-muscarinic receptor affinities using cell membrane chromatography and radioligand binding assay in guinea pig jejunum membrane

    Institute of Scientific and Technical Information of China (English)

    Bing-xiang YUAN; Jin HOU; Lang-chong HE; Guang-de YANG

    2005-01-01

    Aim: To study if cell membrane chromatography (CMC) could reflect drug-receptor interaction and evaluate the affinity and competitive binding to muscarinic acetylcholine receptor (mAChR). Methods: The cell membrane stationary phase(CMSP) was prepared by immobilizing guinea pig jejunum cell membrane on the surface of a silica carrier, and was used for the rapid on-line chromatographic evaluation of ligand binding affinities to mAChR. The affinity to mAChR was also evaluated from radioligand binding assays (RBA) using the same jejunum membrane preparation. Results: The capacity factor (k') profiles in guinea pig jejunum CMSP were: (-)QNB (15.4)>(+)QNB (11.5)>atropine (5.35)>pirenzepine(5.26)>4-DAMP (4.45)>AF-DX 116 (4.18)>pilocarpine (3.93)>acetylcholine(1.31). These results compared with the affinity rank orders obtained from radioligand binding assays indicated that there wasa positive correlation (r2=0.8525, P<0.0001) between both data sets. Conclusion: The CMC method can be used to evaluate drug-receptor affinities for drug candidates.

  18. Inherent vulnerabilities in monoaminergic pathways predict the emergence of depressive impairments in an animal model of chronic epilepsy.

    Science.gov (United States)

    Medel-Matus, Jesús-Servando; Shin, Don; Sankar, Raman; Mazarati, Andrey

    2017-08-01

    The objective was to determine whether the depression comorbid with epilepsy could be predicted based on inherent premorbid patterns of monoaminergic transmission. In male Wistar rats, despair-like and anhedonia-like behaviors were examined using forced swimming and taste preference tests, respectively. Serotonergic raphe nucleus (RN)-prefrontal cortex (PFC) and dopaminergic ventral tegmental area (VTA)-nucleus accumbens (NAcc) pathways were interrogated by fast scan cyclic voltammetry (FSCV). The assays were performed before and 2 months after pilocarpine status epilepticus. In a subset of naive rats, FSCV, coupled with the intensity-dependent stimulation paradigm, detected specific deviations in each pathway (six rats for RN-PFC and seven rats for VTA-NAcc, with overlap in two, of 19 total subjects) in the absence of behavioral impairments. During epilepsy, animals with preexisting deviations in RN-PFC invariably developed despair, and rats with deviations in VTA-NAcc developed anhedonia. Serotonergic and dopaminergic pathways, respectively, showed signs of explicit deterioration. We suggest that epilepsy triggers decompensations in the already vulnerable depression-relevant neuronal circuits, which culminate in depression. The established connection between the identified specific signatures in monoamine transmission in naive rats and specific symptoms of epilepsy-associated depression may help in understanding causes of comorbidity and in developing its early biomarkers. Wiley Periodicals, Inc. © 2017 International League Against Epilepsy.

  19. Holmes-Adie syndrome, autoimmune hepatitis and celiac disease: A case report

    Institute of Scientific and Technical Information of China (English)

    Timea Csak; Aniko Folhoffer; Andrea Horvath; Judit Halász; Csaba Diczházi; Zsuzsa Schaff; Ferenc Szalay

    2006-01-01

    A 35-year-old female patient presented with the following symptoms of Holmes-Adie syndrome: photophobia,enlargement of the left pupil unresponsive to light,Achilles areflexia. The pilocarpine test was positive. No tumor or other neurological abnormality was found. She had a 19-year history of autoimmune hepatitis. Flares up were observed following each 3 deliveries. At age of 31she presented with diarrhea and weight loss. Abdominal tumor was detected by ultrasound. The surgically removed tumor was histologically a benign mesenteric multicystic lymphangioma. Simultaneously, celiac disease was diagnosed. Gluten-free diet resulted in a significant improvement of celiac disease, but not of autoimmune hepatitis. Autonomic neuropathy was proven by standard cardiovascular tests. The patient was a homozygous carrier for HLA DQ2 antigen characteristic for celiac disease and heterozygous for HLA DR3 B8 frequent in autoimmune liver diseases. Our novel observation on association of Holmes-Adie syndrome with autoimmune hepatitis and celiac disease is suggestive for a common immunological background for all three entities present in a patient with mesenteric multicystic lymphangioma.

  20. Different microRNA profiles in chronic epilepsy versus acute seizure mouse models.

    Science.gov (United States)

    Kretschmann, Anita; Danis, Benedicte; Andonovic, Lidija; Abnaof, Khalid; van Rikxoort, Marijke; Siegel, Franziska; Mazzuferi, Manuela; Godard, Patrice; Hanon, Etienne; Fröhlich, Holger; Kaminski, Rafal M; Foerch, Patrik; Pfeifer, Alexander

    2015-02-01

    Epilepsy affects around 50 million people worldwide, and in about 65% of patients, the etiology of disease is unknown. MicroRNAs are small non-coding RNAs that have been suggested to play a role in the pathophysiology of epilepsy. Here, we compared microRNA expression patterns in the hippocampus using two chronic models of epilepsy characterised by recurrent spontaneous seizures (pilocarpine and self-sustained status epilepticus (SSSE)) and an acute 6-Hz seizure model. The vast majority of microRNAs deregulated in the acute model exhibited increased expression with 146 microRNAs up-regulated within 6 h after a single seizure. In contrast, in the chronic models, the number of up-regulated microRNAs was similar to the number of down-regulated microRNAs. Three microRNAs-miR-142-5p, miR-331-3p and miR-30a-5p-were commonly deregulated in all three models. However, there is a clear overlap of differentially expressed microRNAs within the chronic models with 36 and 15 microRNAs co-regulated at 24 h and at 28 days following status epilepticus, respectively. Pathway analysis revealed that the altered microRNAs are associated with inflammation, innate immunity and cell cycle regulation. Taken together, the identified microRNAs and the pathways they modulate might represent candidates for novel molecular approaches for the treatment of patients with epilepsy.

  1. A new futuristic glaucoma therapeutic management paradigm

    Directory of Open Access Journals (Sweden)

    Anusuya Gehlot

    2015-04-01

    Full Text Available Glaucoma is a group of diseases, characterized by a progressive form of optic nerve damage. Current studies indicate more selective pathophysiological involvement, thereby targeted therapies are warranted. Although both the prostaglandin analogs and beta blockers are still, most commonly used drugs for glaucoma, due to their efficacy, lack of adverse effects. In addition, a stepped care approach is the corner stone for its management. In addition, attempts have been made to enhance patient compliance and ocular delivery of already available anti-glaucoma drugs such as pilocarpine and timolol maleate. Notable among futuristic treatment options are; novel delivery systems, benzalkonium chloride-free drugs, various glaucoma drainage devices, new targeted therapies and prompt diagnosis plus aggressive treatment, in patients with primary angle closure glaucoma. Promising new focus on vision sparing, greater patient safety and tolerability will provide improved treatment options and long-term preservation of vision and quality of life. [Int J Basic Clin Pharmacol 2015; 4(2.000: 195-197

  2. Intersegmental coordination of walking movements in stick insects.

    Science.gov (United States)

    Ludwar, Björn Ch; Göritz, Marie L; Schmidt, Joachim

    2005-03-01

    Locomotion requires the coordination of movements across body segments, which in walking animals is expressed as gaits. We studied the underlying neural mechanisms of this coordination in a semi-intact walking preparation of the stick insect Carausius morosus. During walking of a single front leg on a treadmill, leg motoneuron (MN) activity tonically increased and became rhythmically modulated in the ipsilateral deafferented and deefferented mesothoracic (middle leg) ganglion. The pattern of modulation was correlated with the front leg cycle and specific for a given MN pool, although it was not consistent with functional leg movements for all MN pools. In an isolated preparation of a pair of ganglia, where one ganglion was made rhythmically active by application of pilocarpine, we found no evidence for coupling between segmental central pattern generators (CPGs) that could account for the modulation of MN activity observed in the semi-intact walking preparation. However, a third preparation provided evidence that signals from the front leg's femoral chordotonal organ (fCO) influenced activity of ipsilateral MNs in the adjacent mesothoracic ganglion. These intersegmental signals could be partially responsible for the observed MN activity modulation during front leg walking. While afferent signals from a single walking front leg modulate the activity of MNs in the adjacent segment, additional afferent signals, local or from contralateral or posterior legs, might be necessary to produce the functional motor pattern observed in freely walking animals.

  3. Melatonin Alleviates the Epilepsy-Associated Impairments in Hippocampal LTP and Spatial Learning Through Rescue of Surface GluR2 Expression at Hippocampal CA1 Synapses.

    Science.gov (United States)

    Ma, Yue; Sun, Xiaolong; Li, Juan; Jia, Ruihua; Yuan, Fang; Wei, Dong; Jiang, Wen

    2017-02-18

    Epilepsy-associated cognitive impairment is common, and negatively impacts patients' quality of life. However, most antiepileptic drugs focus on the suppression of seizures, and fewer emphasize treatment of cognitive dysfunction. Melatonin, an indolamine synthesized primarily in the pineal grand, is reported to be neuroprotective against several central nervous system disorders. In this study, we investigated whether melatonin could reverse cognitive dysfunction in lithium-pilocarpine treated rats. Chronic treatment with melatonin (8 mg/kg daily for 15 days) after induction of status epilepticus significantly alleviated seizure severity, reduced neuronal death in the CA1 region of the hippocampus, improved spatial learning (as measured by the Morris water maze test), and reversed LTP impairments, compared to vehicle treatment. Furthermore, we found that melatonin rescued the decreased surface levels of GluR2 in the CA1 region observed in epilepsy, which might be the underlying mechanism of the neuroprotective and synapse-modulating function of melatonin. Our study provides experimental evidence for the possible clinical utility of melatonin as an adjunctive therapy to prevent epilepsy-associated cognitive impairments.

  4. Down-regulation of Pin1 in Temporal Lobe Epilepsy Patients and Mouse Model.

    Science.gov (United States)

    Tang, Lan; Zhang, Yanke; Chen, Guojun; Xiong, Yan; Wang, Xuefeng; Zhu, Binglin

    2017-02-27

    Peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1) is a unique PPIase belonging to the parvulin family, and it isomerizes peptide bond between phospho-(Ser/Thr) and Pro. Pin1 has been linked to the pathogenesis of various human diseases; however, its exact biological functions remain unclear. The aim of the present study is to explore the expression pattern of Pin1 in patients with refractory epilepsy and in a chronic pilocarpine-induced epileptic mouse model. Using Western blot, immunofluorescence and immunoprecipitation analysis, we found that Pin1 protein was mainly distributed in neurons, demonstrated by colocalization with the dendritic marker, MAP2. However, the expression of Pin1 decreased remarkably in epileptic patients and experimental mice. Furthermore, the reciprocal coimmunoprecipitation analysis showed that Pin1 interacted with NR2A and NR2B-containing NMDA receptors not AMPA receptors in epileptic mouse models. Our results are the first to indicate that the expression of Pin1 in epileptic brain tissue could play important roles in epilepsy.

  5. Treatment of xerostomia and hyposalivation in the elderly: A systematic review

    Science.gov (United States)

    Gil-Montoya, José-Antonio; Silvestre, Francisco-Javier; Barrios, Rocío

    2016-01-01

    Background Therapeutic strategies for xerostomia, regardless of etiology, have so far not had definitive or clearly effective results. Objectives. To systematically revise the latest scientific evidence available regarding the treatment of dry mouth, regardless of the cause of the problem. Material and Methods The literature search was conducted in March 2015, using the Medline and Embase databases. The “Clinical Trial”, from 2006 to March 2015, was carried out in English and only on human cases. The draft of the systematic review and assessment of the methodological quality of the trials was carried out following the criteria of PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) and the “Oxford Quality Scale”. Results Finally, a total of 26 trials were identified that met the previously defined selection and quality criteria; 14 related to drug treatments for dry mouth, 10 with non-pharmacological treatment and 2 with alternative treatments. Conclusions Pilocarpine continues to be the best performing sialogogue drug for subjects with xerostomia due to radiation on head and neck cancer or diseases such as Sjogren’s Syndrome. For patients with dry mouth caused solely by medication, there are some positive indications from the use of malic acid, along with other elements that counteract the harmful effect on dental enamel. In general, lubrication of oral mucous membrane reduces the symptoms, although the effects are short-lived. Key words:Systematic review, xerostomia, clinical trial, hyposalivation. PMID:27031061

  6. UNILATERAL MYDRIASIS WITH CHOLINERGIC SUPER SENSITIVITY: A DIAGNOSTIC DILEMMA - A CASE SERIES REPORT

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    Sandhya

    2014-05-01

    Full Text Available Pupillary abnormalities are a common feature of general ophthalmic practice. It often causes confusion as they can be manifestations of local and/ or systemic diseases. These diseases may range from vision threatening to life endangering to innocuous ones. A keen observational and clinical skill can help the ophthalmologist in diagnosis & timely referral when necessary. We report 3 cases of acquired mydriasis, with cholinergic supersensitivity. The short history poses a diagnostic dilemma as to whether it is Adie’s Tonic pupil or a harbinger of a serious neurological problem. 2 of the patients with mydriasis were younger, 32 & 35 years of age, presenting with recent onset of blurring of Vision for distance and difficulty in reading. The 3rd patient was a 45 year old presbyope who presented with sudden drop in near vision in one eye. Our cases raise several important question regarding so-called “benign pupillary dilation of the young” and its relationship with Adie‘s tonic pupil. Demonstration of probable transient parasympathetic dysfunction suggests that pharmacologic testing with dilute pilocarpine should be considered in patients reporting with near vision problems with isolated unilateral recent onset mydriasis which is probably intermittent. Thorough history and basic clinical neurological examination are mandatory. The importance of timely referral to neurologist must be borne in mind always in such cases.

  7. TRPC6在匹罗卡品致痫大鼠海马苔藓纤维出芽中的作用%Effect of TRPC6 on mossy fiber sprouting in hippocampus of rats with pilocapine-induced epilepsy

    Institute of Scientific and Technical Information of China (English)

    唐薇婷; 曾畅; 李国良; 冯莉; 陈锶; 肖波

    2012-01-01

    Objective To observe the dynamic changes of canonical transient receptor potential channels 6 (TRPC6) expression in the rat hippocampus with pilocarpine-induced epilepsy, and to investigate its effect on mossy fiber sprouting in hippocampus. Methods Seventy-two healthy male Sprague-Dawley (SD) rats were divided randomly into experimental group (n =60) and control group (n - 12), The temporal lobe epilepsy model was established by intraperitoneal injection of lithium and pilocarpine, while the controls were injected with equal dose of saline (NS). The experimental rats were equally divided into 5 subgroups at time points 1 day、7 days、15 days、30 days and 60 days after status epileptics (SE). Each subgroup and the control rats were subsequently divided into 2 panels for the following tests respectively: ①expression of TRPC6 and Synaptophysin protein in rats' hippocampus by Western blot; ②Timm staining and score. Results The expression of TRPC6 was markedly increased and reached its peak on 1 days after SE (P <0. 01) , and it was higher than the control at all the other time points (P <0, 01). Compared with the control, the expression of Synaptophysin was markedly up-regulated on 15 days、30 days 、60 days after SE (P<0.05 or P <0. 01 ) , and reached the peak on 30 days after SE. Timm granules appeared in molecular layer of gyrus dentatus in the hippocampus of experimental rats since 7 days after SE, and then gradually increased. Conclusions TRPC6 may play a potential role in mossy fiber sprouting, in which BDNF may involve.%目的 观察传统型瞬时受体电位通道6(TRPC6)蛋白在匹罗卡品致痫大鼠海马中的表达变化,探讨其在海马苔藓纤维出芽中的作用.方法 72只SD大鼠随机分为实验组(n=60)和对照组(n=12).实验组采用氯化锂-匹罗卡品腹腔注射法建立颞叶癫痫模型;对照组腹腔注射等量无菌生理盐水.实验组按癫痫持续状态(SE)后1d、7d、15d、30 d和60 d分为5

  8. 丘脑前核在三叉神经电刺激减轻癫痫发作和海马神经元损伤中的作用%Role of anterior thalamic nucleus in trigeminal nerve electrostimulation-induced reduction of seizures and hippocampal damage

    Institute of Scientific and Technical Information of China (English)

    王先红; 田苗苗; 潘晴晴; 鲁亚楠; 王玉

    2014-01-01

    [ ABSTRACT] AIM: To investigate the role of anterior thalamic nucleus in trigeminal nerve electrostimulation ( TNS)-induced effects on seizures and hippocampal damage.METHODS: The rats were intraperitoneally injected with pilocarpine to induce chronic epilepsy, and then received sham treatment, TNS treatment and TNS treatment after stereotac-tic lesion to the anterior thalamic nucleus.The TNS treatment lasted for 1 month in each group.Another injection of pilo-carpine was conducted to induce seizures, and the severity and duration of seizures were quantitatively evaluated.TUNEL, Fluoro-Jade B ( FJB) and Nissl staining were applied to determine in situ apoptosis, neuronal degeneration and neuron loss in the hippocampal CA1 area respectively.RESULTS:Compared with TNS group, TNS treatment after stereotactic lesion to the anterior thalamic nucleus significantly increased the severity and duration of seizures (P<0.05), and the numbers of TUNEL positive cells, FJB positive cells and lost neurons in the hippocampal CA1 area (P<0.01).CONCLUSION:Anterior thalamic nucleus plays a role in TNS-induced reduction of seizures and hippocampal damage.The mechanisms might be due to the chronically activation of the cortex through anterior thalamic nucleus pathway induced by TNS, resulting in the down-regulation of neuronal excitatory susceptibility.%目的:探讨丘脑前核在三叉神经电刺激( TNS)减轻癫痫发作和海马神经元损伤中的作用。方法:大鼠经腹腔注射匹罗卡品建立慢性癫痫模型,模型大鼠分别给予假刺激、三叉神经电刺激和立体定向毁损丘脑前核预处理后三叉神经电刺激1月,再次诱导癫痫发作,观察大鼠的癫痫行为表现,并通过TUNEL、Fluoro-Jade B ( FJB)染色和Nissl染色观察大鼠海马CA1区神经元的凋亡、变性及脱失情况。结果:与未经毁损丘脑前核的TNS处理大鼠相比,毁损丘脑前核后的TNS处理大鼠癫痫发作的级别分数

  9. 人参皂甙Rd对匹罗卡品致(癎)大鼠模型中netrin-1表达及神经元凋亡的影响%Effects of ginsenoside-Rd on expression levels of netrin-1 and neuron apoptosis in epilepsy model

    Institute of Scientific and Technical Information of China (English)

    郑佳丽; 杨金升; 刘学娟; 马亚杰

    2011-01-01

    目的 探讨人参皂甙Rd(GSRd)对大鼠癫(癎)持续状态后齿状回netrin-1表达及海马神经元凋亡的影响.方法 健康Wistar大鼠30只,随机分为颞叶癫(癎)组、GSRd组和对照组,每组10只.氯化锂-匹罗卡品建立大鼠颞叶癫(癎)模型,采用免疫组织化学法及TUNEL法观察颞叶癫(癎)组、GSRd组和对照组齿状回netrin-1蛋白表达及海马神经元凋亡细胞数情况.结果 与对照组比较,颞叶癫(癎)组大鼠30 d齿状回netrin-1蛋白表达明显增高,海马CA3区TUNEL阳性细胞数在癫(癎)持续状态后7d明显增高,差异有统计学意义(P< 0.05);与颞叶癫(癎)组比较,GSRd组大鼠30 d齿状回netrin-1蛋白表达明显降低,海马CA3区TUNEL阳性细胞数在癫(癎)持续状态后7d明显降低,差异有统计学意义(P< 0.05).结论 GSRd可能通过下调netrin-1的表达,并使神经元凋亡数目减少,从而发挥对神经元的保护作用.%Objective To investigate the effects of ginsenoside-Rd(GSRd) on the expression levels of netrin-1 and neuron apoptosis in the rat hippocampus after lithium-pilocarpine induced epilepsy.Methods Thirty Wistar rats were randomly divided into the NS control group,temporal lobe epilepsy(TLE) group and GSRd group.The TLE animal model was established using lithium-pilocarpine.Netrin-1 proteins were detected with immunohistochemical method.The neuron apoptosis was observed with TdT-mediated dUTP nick end labeling(TUNEL) method.Results The expression of netrin-1 in GSRd group was notably lower than that of TLE group (P < 0.05).The TUNEL positive cells in hippocampus CA3 of TLE group were more than those of NS control group (P < 0.05).TUNEL positive cells in TLE group were significantly more than those in NS control group (P < 0.05).TUNEL positive cells in GSRd treated group significantly decreased as compared with TLE group (P < 0.05).Conclusions GSRd may serve as an effective agent for curing the brain damage after TLE in vivo.The mechanism

  10. Olomoucine在匹罗卡品所致颞叶癫(癎)中的作用及机制的研究%Effects and mechanisms of olomoucine on temporal lobe epilepsy

    Institute of Scientific and Technical Information of China (English)

    殷亚萍; 刘希金; 杨志勇; 邓学军

    2013-01-01

    目的 研究细胞周期依赖激酶(CDK)抑制剂在氯化锂-匹罗卡品(Licl-Pilocarpine)所致颞叶癫(癎)大鼠癎性发作中的作用及其相关机制.方法 用Licl-Pilocarpine制作大鼠颞叶癫(癎)模型;用CDK抑制剂Olomoucine在大鼠癫(癎)持续状态后第3、4、5d进行干预,并将参与实验的大鼠分为正常组、模型组、干预组和对照组;用视频监测各组大鼠的癫(癎)发作情况,记录距离第1次慢性自发性发作的潜伏期、慢性自发性发作的频率及贯穿于整个过程中大鼠的病死率;用脑电图监测各组出现慢性自发性发作大鼠的脑电活动的变化,记录发作间期棘波发放的情况;用免疫组化检测GFAP(胶质纤维酸性蛋白,一种星形胶质细胞的标记物)和ADK(腺苷激酶)的表达水平.结果 干预组较模型组距第1次发作的潜伏期明显延长,临床发作次数明显减少,棘波发作的频率和持续时间明显缩短(P<0.05);GFAP和ADK的表达量明显下降(P<0.05);模型组较正常组GFAP和ADK的表达量明显增加;对照组较模型组无明显变化.结论 CDK抑制剂在一定程度上能够减少癫(癎)大鼠的癎性发作次数和脑电波的棘波发放,其机制可能为细胞周期依赖激酶抑制剂能够抑制星形胶质细胞的增生,减少ADK的表达,间接的增加了内源性止癎剂腺苷的含量.%Objective To investigate the correlation between temporal lobe epilepsy (TLE) and cell cycle inhibition through pilocarpine-induced rat models of temporal lobe epilepsy and surgical samples of temporal neocortex from intractable epilepsy patients.Methods Olomoucine (a CDK inhibitor) was used at an acute stage of induced TLE.Behavioral changes and electroencephalogram (EEG) were interpreted after intervention.We also used immunohistochemistry (IHC) to detect the expression of glial fibrillary acidic protein (GFAP) and adenosine kinase (ADK).Results We found that the frequency of seizures and the

  11. Combined role of seizure-induced dendritic morphology alterations and spine loss in newborn granule cells with mossy fiber sprouting on the hyperexcitability of a computer model of the dentate gyrus.

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    Tejada, Julian; Garcia-Cairasco, Norberto; Roque, Antonio C

    2014-05-01

    Temporal lobe epilepsy strongly affects hippocampal dentate gyrus granule cells morphology. These cells exhibit seizure-induced anatomical alterations including mossy fiber sprouting, changes in the apical and basal dendritic tree and suffer substantial dendritic spine loss. The effect of some of these changes on the hyperexcitability of the dentate gyrus has been widely studied. For example, mossy fiber sprouting increases the excitability of the circuit while dendritic spine loss may have the opposite effect. However, the effect of the interplay of these different morphological alterations on the hyperexcitability of the dentate gyrus is still unknown. Here we adapted an existing computational model of the dentate gyrus by replacing the reduced granule cell models with morphologically detailed models coming from three-dimensional reconstructions of mature cells. The model simulates a network with 10% of the mossy fiber sprouting observed in the pilocarpine (PILO) model of epilepsy. Different fractions of the mature granule cell models were replaced by morphologically reconstructed models of newborn dentate granule cells from animals with PILO-induced Status Epilepticus, which have apical dendritic alterations and spine loss, and control animals, which do not have these alterations. This complex arrangement of cells and processes allowed us to study the combined effect of mossy fiber sprouting, altered apical dendritic tree and dendritic spine loss in newborn granule cells on the excitability of the dentate gyrus model. Our simulations suggest that alterations in the apical dendritic tree and dendritic spine loss in newborn granule cells have opposing effects on the excitability of the dentate gyrus after Status Epilepticus. Apical dendritic alterations potentiate the increase of excitability provoked by mossy fiber sprouting while spine loss curtails this increase.

  12. Preictal activity of subicular, CA1, and dentate gyrus principal neurons in the dorsal hippocampus before spontaneous seizures in a rat model of temporal lobe epilepsy.

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    Fujita, Satoshi; Toyoda, Izumi; Thamattoor, Ajoy K; Buckmaster, Paul S

    2014-12-10

    Previous studies suggest that spontaneous seizures in patients with temporal lobe epilepsy might be preceded by increased action potential firing of hippocampal neurons. Preictal activity is potentially important because it might provide new opportunities for predicting when a seizure is about to occur and insight into how spontaneous seizures are generated. We evaluated local field potentials and unit activity of single, putative excitatory neurons in the subiculum, CA1, CA3, and dentate gyrus of the dorsal hippocampus in epileptic pilocarpine-treated rats as they experienced spontaneous seizures. Average action potential firing rates of neurons in the subiculum, CA1, and dentate gyrus, but not CA3, increased significantly and progressively beginning 2-4 min before locally recorded spontaneous seizures. In the subiculum, CA1, and dentate gyrus, but not CA3, 41-57% of neurons displayed increased preictal activity with significant consistency across multiple seizures. Much of the increased preictal firing of neurons in the subiculum and CA1 correlated with preictal theta activity, whereas preictal firing of neurons in the dentate gyrus was independent of theta. In addition, some CA1 and dentate gyrus neurons displayed reduced firing rates preictally. These results reveal that different hippocampal subregions exhibit differences in the extent and potential underlying mechanisms of preictal activity. The finding of robust and significantly consistent preictal activity of subicular, CA1, and dentate neurons in the dorsal hippocampus, despite the likelihood that many seizures initiated in other brain regions, suggests the existence of a broader neuronal network whose activity changes minutes before spontaneous seizures initiate. Copyright © 2014 the authors 0270-6474/14/3416671-17$15.00/0.

  13. GABAergic transmission facilitates ictogenesis and synchrony between CA3, hilus, and dentate gyrus in slices from epileptic rats

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    Gafurov, Boris

    2013-01-01

    The impact of regional hippocampal interactions and GABAergic transmission on ictogenesis remain unclear. Cortico-hippocampal slices from pilocarpine-treated epileptic rats were compared with controls to investigate associations between seizurelike events (SLE), GABAergic transmission, and neuronal synchrony within and between cortico-hippocampal regions. Multielectrode array recordings revealed more prevalent hippocampal SLE in epileptic tissue when excitatory transmission was enhanced and GABAergic transmission was intact [removal of Mg2+ (0Mg)] than when GABAergic transmission was blocked [removal of Mg2+ + bicuculline methiodide (0Mg+BMI)]. When activity within individual regions was analyzed, spectral and temporal slow oscillation/SLE correlations and cross-correlations were highest within the hilus of epileptic tissue during SLE but were similar in 0Mg and 0Mg+BMI. GABAergic facilitation of spectral “slow” oscillation and ripple correlations was most prominent within CA3 of epileptic tissue during SLE. When activity between regions was analyzed, slow oscillation and ripple coherence was highest between the hilus and dentate gyrus as well as between the hilus and CA3 of epileptic tissue during SLE and was significantly higher in 0Mg than 0Mg+BMI. High 0Mg-induced SLE cross-correlations between the hilus and dentate gyrus as well as between the hilus and CA3 were reduced or abolished in 0Mg+BMI. SLE cross-correlation lag measurements provided evidence for a monosynaptic connection from the hilus to the dentate gyrus during SLE. Findings implicate the hilus as an oscillation generator, whose impact on other cortico-hippocampal regions is mediated by GABAergic transmission. Data also suggest that GABAA receptor-mediated transmission facilitates back-propagation from CA3/hilus to the dentate gyrus and that this back-propagation augments SLE in epileptic hippocampus. PMID:23615549

  14. Whole transcriptome analysis of the hippocampus: toward a molecular portrait of epileptogenesis

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    Scorza Fúlvio A

    2010-04-01

    Full Text Available Abstract Background Uncovering the molecular mechanisms involved in epileptogenesis is critical to better understand the physiopathology of epilepsies and to help develop new therapeutic strategies for this prevalent and severe neurological condition that affects millions of people worldwide. Results Changes in the transcriptome of hippocampal cells from rats subjected to the pilocarpine model of epilepsy were evaluated by microarrays covering 34,000 transcripts representing all annotated rat genes to date. Using such genome-wide approach, differential expression of nearly 1,400 genes was detected during the course of epileptogenesis, from the early events post status epilepticus (SE to the onset of recurrent spontaneous seizures. Most of these genes are novel and displayed an up-regulation after SE. Noteworthy, a group of 128 genes was found consistently hyper-expressed throughout epileptogenesis, indicating stable modulation of the p38MAPK, Jak-STAT, PI3K, and mTOR signaling pathways. In particular, up-regulation of genes from the TGF-beta and IGF-1 signaling pathways, with opposite effects on neurogenesis, correlate with the physiopathological changes reported in humans. Conclusions A consistent regulation of genes functioning in intracellular signal transduction regulating neurogenesis have been identified during epileptogenesis, some of which with parallel expression patterns reported in patients with epilepsy, strengthening the link between these processes and development of epilepsy. These findings reveal dynamic molecular changes occurring in the hippocampus that may serve as a starting point for designing alternative therapeutic strategies to prevent the development of epilepsy after acquired brain insults.

  15. Correlation between IL-10 and microRNA-187 expression in epileptic rat hippocampus and patients with temporal lobe epilepsy

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    Walid A. Alsharafi

    2015-12-01

    Full Text Available Accumulating evidence is emerging that microRNAs (miRs are key regulators controlling neuroinflammatory processes, which are known to play a potential role in the pathogenesis of temporal lobe epilepsy (TLE. The aim of the present study was to investigate the dynamic expression pattern of interleukin (IL–10 as an anti-inflammatory cytokine and miR-187 and post-transcriptional inflammation-related miRNA in the hippocampus of a rat model of status epilepticus (SE and patients with TLE. We performed a real-time quantitative PCR and western blot on rat hippocampus (2 hours, 7 days, 21 days and 60 days following pilocarpine-induced SE, and on hippocampus obtained from TLE patients and normal controls. To detect the relationship between IL-10 and miR-187 on neurons, lipopolysaccharide (LPS and IL-10-stimulated neurons were prepared. Furthermore, we identified the effect of antagonizing of miR-187 by its antagomir on IL-10 secretion. Here we reported that that IL-10 secretion and miR-187 expression levels are inversely correlated after SE.. In patients with TLE, the expression levels of IL-10 was also significantly upregulated, whereas miR-187 expression was significantly downregulated. Moreover, miR-187 expression was significantly reduced following IL-10 stimulation in an IL-10–dependent manner. On the other hand, antagonizing miR-187 reduced the production of IL-10 in hippocampal tissues of rat model of SE. Our findings demonstrate a critical role of miR-187 in the physiological regulation of IL-10 anti-inflammatory responses and elucidate the role of neuro-inflammation in the pathogenesis of TLE. Therefore, modulation of the IL-10 / miR-187 axis may be a new therapeutic approach for TLE.

  16. Corneal healing after riboflavin ultraviolet-A collagen cross-linking determined by confocal laser scanning microscopy in vivo: early and late modifications.

    Science.gov (United States)

    Mazzotta, Cosimo; Traversi, Claudio; Baiocchi, Stefano; Caporossi, Orsola; Bovone, Cristina; Sparano, Maria Caterina; Balestrazzi, Angelo; Caporossi, Aldo

    2008-10-01

    To assess early and late micromorphological modifications of cross-linked corneas in vivo by means of Heidelberg Retinal Tomography (HRT) II confocal microscopy. Prospective nonrandomized open trial. Micromorphological examination of 44 cross-linked keratoconic corneas was performed in vivo by HRT II confocal laser scanning microscopy. Riboflavin ultraviolet (UV)-A-induced corneal collagen cross-linking (CXL) was performed according to the Siena protocol: pilocarpin 1% drops 30 minutes before, topical anesthesia with lidocaine 4% drops 15 minutes before irradiation, mechanical scraping of epithelium (9-mm-diameter area), preirradiation soaking for 10 minutes in riboflavin solution 0.1% (Ricrolin, Sooft, Italy) applied every 2.5 minutes for 30 minutes, 30 minutes exposure to solid-state UVA illuminator (Caporossi; Baiocchi; Mazzotta, X-linker, CSO, Italy), 8-mm-diameter irradiated area, energy delivered 3 mW/cm(2). All patients were examined by confocal scans preoperatively and at the following times after treatment: one, three, and six months, and one, two, and three years. No damage to the limbal region was observed. Epithelial regrowth was complete after four days of soft contact lens bandage. The anatomy of the subepithelial plexus was restored one year after the operation with full corneal sensitivity. Increased density of extracellular matrix in late postoperative period indicated cross-linked collagen to a depth of 340 microm expressed by a late demarcation line. In vivo confocal microscopy showed early and late modification of corneal microstructure after the treatment. The three-year stability of CXL recorded could be related to increased cross-links formation, synthesis of well-structured collagen and new lamellar interconnections.

  17. Corneal collagen cross-linking to stop corneal ectasia exacerbated by radial keratotomy.

    Science.gov (United States)

    Mazzotta, Cosimo; Baiocchi, Stefano; Denaro, Rosario; Tosi, Gian Marco; Caporossi, Tomaso

    2011-02-01

    To assess the efficacy of riboflavin ultraviolet A (UV-A) corneal collagen cross-linking in the management of keratoconic corneal ectasia exacerbated by radial keratotomy (RK). A patient with progressive corneal ectasia and hyperopic shift, occurring 10 years after RK performed in the left eye, was treated with riboflavin UV-A corneal collagen cross-linking according to the Siena protocol: Pilocarpin 0.1% drop (1 hour before), lidocaine 4% drops 15 minutes before, mechanical scraping of epithelium (9-mm-diameter area), preirradiation stromal soaking for 10 minutes in riboflavin 0.1%-dextrane 20% (Ricrolin; Sooft Italy) applied every 2 minutes, and 30 minutes of total exposure (6 steps of 5 minutes) to solid-state UV-A illuminator (Caporossi, Baiocchi, Mazzotta Vega X linker; CSO Opthalmics, Florence, Italy), energy delivered 3 mW/cm, and irradiated area 9 mm in diameter. After the operation, uncorrected visual acuity and best spectacle-corrected visual acuity improved from 0.2 to 0.6 and from 0.3 to 0.8 Snellen lines, respectively, in a 12-month follow-up. Improved topographical K readings and corneal symmetry index were also recorded starting from the first postoperative month and continuing thereafter. No adverse effects were recorded after treatment. Riboflavin UV-A-induced corneal cross-linking seems to be a promising surgical option in the management of unstable corneal ectasia exacerbated by RK, particularly in eyes with preexisting keratoconus. A large cohort and longer follow-up are needed to determine its long-term efficacy in this clinical setting.

  18. Unit Activity of Hippocampal Interneurons before Spontaneous Seizures in an Animal Model of Temporal Lobe Epilepsy.

    Science.gov (United States)

    Toyoda, Izumi; Fujita, Satoshi; Thamattoor, Ajoy K; Buckmaster, Paul S

    2015-04-22

    Mechanisms of seizure initiation are unclear. To evaluate the possible roles of inhibitory neurons, unit recordings were obtained in the dentate gyrus, CA3, CA1, and subiculum of epileptic pilocarpine-treated rats as they experienced spontaneous seizures. Most interneurons in the dentate gyrus, CA1, and subiculum increased their firing rate before seizures, and did so with significant consistency from seizure to seizure. Identification of CA1 interneuron subtypes based on firing characteristics during theta and sharp waves suggested that a parvalbumin-positive basket cell and putative bistratified cells, but not oriens lacunosum moleculare cells, were activated preictally. Preictal changes occurred much earlier than those described by most previous in vitro studies. Preictal activation of interneurons began earliest (>4 min before seizure onset), increased most, was most prevalent in the subiculum, and was minimal in CA3. Preictal inactivation of interneurons was most common in CA1 (27% of interneurons) and included a putative ivy cell and parvalbumin-positive basket cell. Increased or decreased preictal activity correlated with whether interneurons fired faster or slower, respectively, during theta activity. Theta waves were more likely to occur before seizure onset, and increased preictal firing of subicular interneurons correlated with theta activity. Preictal changes by other hippocampal interneurons were largely independent of theta waves. Within seconds of seizure onset, many interneurons displayed a brief pause in firing and a later, longer drop that was associated with reduced action potential amplitude. These findings suggest that many interneurons inactivate during seizures, most increase their activity preictally, but some fail to do so at the critical time before seizure onset.

  19. Genetic testing for cystic fibrosis in adult patients

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    Marina Mencinger

    2006-02-01

    Full Text Available Background: Cystic fibrosis (CF is an autosomal recessive disease caused by mutations in gene encoding cystic fibrosis transmembrane regulator (CFTR protein. Over 1400 mutations found in the gene contribute to the complexity of the CF phenotypes ranging from a classic multiorgan disease commonly involving respiratory, gastrointestinal and reproductive tract to mild and monosymptomatic presentations. Pilocarpine iontophoresis is considered as standard diagnostic test for CF, but it often fails in atypical forms of CF.Methods: In order to provide an additional diagnostic test to assure the diagnosis and provide patients with a proper medical care, we performed a genetic testing on 16 adults suspected to have atypical form of CF. Following counselling, parents of patients with possible homozygote variant of mutations were tested. On a personal request testing was also performed in an adult sibling of a patient with two known mutations to investigate possible carrier hood. The allele specific polymerase chain reaction method (PCR was used to detect 29 most common mutations in the cftr gene.Results: The diagnosis was proved in 3 individuals, a homozygote for Δ F508, and two compound heterozygotes Δ F508/R1162X and Δ F508/3849+10kbC>T. In three cases only one mutation was found: I148T, 2789+5G>A and Δ F508 in a heterozygote form.Conclusions: The genetic testing for CF is a valuable diagnostic tool in atypical forms of CF. Exclusion of possible differential diagnosis is warranted because of a variable CF phenotype. In cases where only one or no mutation was detected a necessity of whole gene sequencing is indicated to exclude rare mutations and polymorphisms that could be implicated in the pathogenesis of atypical CF.

  20. Effects of cobalt 60 irradiation on rat salivary glands; Correlation of early changes with irradiation dose

    Energy Technology Data Exchange (ETDEWEB)

    Shigematsu, Yoshiaki (Meikai Univ., Sakado, Saitama (Japan). School of Dentistry)

    1991-11-01

    A single irradiation of Co-60 was given to the head and neck region in male Sprague-Dawley rats. According to the time, dose and fractionation factor (TDF), the experimental animals were equally divided into the group of 11.36 Gy (TDF 50), the group of 17.82 Gy (TDF 100), and the group of 27.97 Gy (TDF 200). Parotid, submandibular, and sublingual salivary glands were examined by light microscopy, transmission electron microscopy, and biochemically one, 3, and 7 days after irradiation. Parotid saliva samples were also collected after stimulation with pilocarpine, and the flow rate, concentration of total salivary protein, and {alpha}-amylase activity were assessed. The parotid gland showed the most significant decrease in {alpha}-amylase activity at 3 days in all groups. {alpha}-amylase activity in the submandibular and sublingual glands was much lower than in the parotid gland. Non-lysosome enzyme, acid p-nitrophenyl phosphatase activity in the salivary glands was increased. Similarly, a lysosomal enzyme, {beta}-D-glucuronidase in the salivary glands was increased. Alkaline phosphatase activity was increased in the parotid gland and decreased in the submandibular and sublingual glands. Salivary flow rate in the parotid gland was decreased one, 3, and 7 days after irradiation, markedly depending on irradiation doses. Light microscopy of the salivary glands revealed atrophy, vacuolization, and degranulation in acinar cells, especially in granular tubule cells. Electron microscopy revealed vacuolization and degeneration of mitochondria, secretory granules, and other organelles in the cytoplasm of acinar cells. Morphological changes of secretory granules were dose-dependent. Not only morphological but also biochemical changes in the parotid gland occurred synchronously. (N.K.) 56 refs.

  1. Salivary gland hypofunction in tyrosylprotein sulfotransferase-2 knockout mice is due to primary hypothyroidism.

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    Andrew D Westmuckett

    Full Text Available BACKGROUND: Protein-tyrosine sulfation is a post-translational modification of an unknown number of secreted and membrane proteins mediated by two known Golgi tyrosylprotein sulfotransferases (TPST-1 and TPST-2. We reported that Tpst2-/- mice have mild-moderate primary hypothyroidism, whereas Tpst1-/- mice are euthyroid. While using magnetic resonance imaging (MRI to look at the thyroid gland we noticed that the salivary glands in Tpst2-/- mice appeared smaller than in wild type mice. This prompted a detailed analysis to compare salivary gland structure and function in wild type, Tpst1-/-, and Tpst2 -/- mice. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative MRI imaging documented that salivary glands in Tpst2-/- females were (≈ 30% smaller than wild type or Tpst1-/- mice and that the granular convoluted tubules in Tpst2-/- submandibular glands were less prominent and were almost completely devoid of exocrine secretory granules compared to glands from wild type or Tpst1-/- mice. In addition, pilocarpine-induced salivary flow and salivary α-amylase activity in Tpst2-/- mice of both sexes was substantially lower than in wild type and Tpst1-/- mice. Anti-sulfotyrosine Western blots of salivary gland extracts and saliva showed no differences between wild type, Tpst1-/-, and Tpst2-/- mice, suggesting that the salivary gland hypofunction is due to factor(s extrinsic to the salivary glands. Finally, we found that all indicators of hypothyroidism (serum T4, body weight and salivary gland hypofunction (salivary flow, salivary α-amylase activity, histological changes were restored to normal or near normal by thyroid hormone supplementation. CONCLUSIONS/SIGNIFICANCE: Our findings conclusively demonstrate that low body weight and salivary gland hypofunction in Tpst2-/- mice is due solely to primary hypothyroidism.

  2. Effects of split-dose X irradiation on rat salivary gland function

    Energy Technology Data Exchange (ETDEWEB)

    Vissink, A.; s-Gravenmade, E.J.; Ligeon, E.E.; Konings, A.W. (Department of Radiobiology, University of Groningen (Netherlands))

    1991-07-01

    The effect of a single local dose of 15 Gy on salivary gland function in male Albino Wistar rats was compared with the effect of two doses of 7.5 Gy. The intervals chosen were 0-24 h and 1 week. Before and 1-30 days after the last radiation dose, samples of parotid and submandibular saliva were collected simultaneously after stimulation of the glands with pilocarpine. Irradiation with the single dose resulted in an increased lag phase and potassium concentration, and a decreased flow rate and sodium concentration. The rate of secretion of amylase was decreased during Days 1-6, increased at Day 10, and was decreased again at Day 30. With two dose fractions, substantial dose-sparing effects on lag phase, flow rate, and secretion of amylase were observed for both the very early (0-6 days postirradiation) and later (6-30 days postirradiation) effects. These effects were maximal when the interval between the fractions was 6 h. A significant dose-sparing effect on electrolytes was observed for the later effects only, again with a maximum for the 6-h interval. The dose-sparing observed for the very early effects cannot be explained satisfactorily by repair of sublethal damage (SLD), redistribution of cells over the cell cycle, or repopulation of salivary gland tissue between the doses. In contrast to the earlier dose-sparing effects, the split-dose recovery seen for later damage may be attributed, in part, to SLD repair in providing for greater reproductive survival of intercalated ductal cells and enhanced tissue regeneration.

  3. Vascular endothelial growth factor is up-regulated after status epilepticus and protects against seizure-induced neuronal loss in hippocampus.

    Science.gov (United States)

    Nicoletti, J N; Shah, S K; McCloskey, D P; Goodman, J H; Elkady, A; Atassi, H; Hylton, D; Rudge, J S; Scharfman, H E; Croll, S D

    2008-01-02

    Vascular endothelial growth factor (VEGF) is a protein factor which has been found to play a significant role in both normal and pathological states. Its role as an angiogenic factor is well-established. More recently, VEGF has been shown to protect neurons from cell death both in vivo and in vitro. While VEGF's potential as a protective factor has been demonstrated in hypoxia-ischemia, in vitro excitotoxicity, and motor neuron degeneration, its role in seizure-induced cell loss has received little attention. A potential role in seizures is suggested by Newton et al.'s [Newton SS, Collier EF, Hunsberger J, Adams D, Terwilliger R, Selvanayagam E, Duman RS (2003) Gene profile of electroconvulsive seizures: Induction of neurotrophic and angiogenic factors. J Neurosci 23:10841-10851] finding that VEGF mRNA increases in areas of the brain that are susceptible to cell loss after electroconvulsive-shock induced seizures. Because a linear relationship does not always exist between expression of mRNA and protein, we investigated whether VEGF protein expression increased after pilocarpine-induced status epilepticus. In addition, we administered exogenous VEGF in one experiment and blocked endogenous VEGF in another to determine whether VEGF exerts a neuroprotective effect against status epilepticus-induced cell loss in one vulnerable brain region, the rat hippocampus. Our data revealed that VEGF is dramatically up-regulated in neurons and glia in hippocampus, thalamus, amygdala, and neocortex 24 h after status epilepticus. VEGF induced significant preservation of hippocampal neurons, suggesting that VEGF may play a neuroprotective role following status epilepticus.

  4. Seizure induces activation of multiple subtypes of neural progenitors and growth factors in hippocampus with neuronal maturation confined to dentate gyrus

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    Indulekha, Chandrasekharan L.; Sanalkumar, Rajendran [Neuro Stem Cell Biology Laboratory, Department of Neurobiology, Rajiv Gandhi Center for Biotechnology, Thiruvananthapuram, Kerala 695 014 (India); Thekkuveettil, Anoopkumar [Molecular Medicine, Biomedical Technology Wing, Sree Chitra Thirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala (India); James, Jackson, E-mail: jjames@rgcb.res.in [Neuro Stem Cell Biology Laboratory, Department of Neurobiology, Rajiv Gandhi Center for Biotechnology, Thiruvananthapuram, Kerala 695 014 (India)

    2010-03-19

    Adult hippocampal neurogenesis is altered in response to different physiological and pathological stimuli. GFAP{sup +ve}/nestin{sup +ve} radial glial like Type-1 progenitors are considered to be the resident stem cell population in adult hippocampus. During neurogenesis these Type-1 progenitors matures to GFAP{sup -ve}/nestin{sup +ve} Type-2 progenitors and then to Type-3 neuroblasts and finally differentiates into granule cell neurons. In our study, using pilocarpine-induced seizure model, we showed that seizure initiated activation of multiple progenitors in the entire hippocampal area such as DG, CA1 and CA3. Seizure induction resulted in activation of two subtypes of Type-1 progenitors, Type-1a (GFAP{sup +ve}/nestin{sup +ve}/BrdU{sup +ve}) and Type-1b (GFAP{sup +ve}/nestin{sup +ve}/BrdU{sup -ve}). We showed that majority of Type-1b progenitors were undergoing only a transition from a state of dormancy to activated form immediately after seizures rather than proliferating, whereas Type-1a showed maximum proliferation by 3 days post-seizure induction. Type-2 (GFAP{sup -ve}/nestin{sup +ve}/BrdU{sup +ve}) progenitors were few compared to Type-1. Type-3 (DCX{sup +ve}) progenitors showed increased expression of immature neurons only in DG region by 3 days after seizure induction indicating maturation of progenitors happens only in microenvironment of DG even though progenitors are activated in CA1 and CA3 regions of hippocampus. Also parallel increase in growth factors expression after seizure induction suggests that microenvironmental niche has a profound effect on stimulation of adult neural progenitors.

  5. Structure-Function Studies of Naphthalene, Phenanthrene, Biphenyl, and Their Derivatives in Interaction with and Oxidation by Cytochromes P450 2A13 and 2A6.

    Science.gov (United States)

    Shimada, Tsutomu; Takenaka, Shigeo; Kakimoto, Kensaku; Murayama, Norie; Lim, Young-Ran; Kim, Donghak; Foroozesh, Maryam K; Yamazaki, Hiroshi; Guengerich, F Peter; Komori, Masayuki

    2016-06-20

    Naphthalene, phenanthrene, biphenyl, and their derivatives having different ethynyl, propynyl, butynyl, and propargyl ether substitutions were examined for their interaction with and oxidation by cytochromes P450 (P450) 2A13 and 2A6. Spectral interaction studies suggested that most of these chemicals interacted with P450 2A13 to induce Type I binding spectra more readily than with P450 2A6. Among the various substituted derivatives examined, 2-ethynylnaphthalene, 2-naphthalene propargyl ether, 3-ethynylphenanthrene, and 4-biphenyl propargyl ether had larger ΔAmax/Ks values in inducing Type I binding spectra with P450 2A13 than their parent compounds. P450 2A13 was found to oxidize naphthalene, phenanthrene, and biphenyl to 1-naphthol, 9-hydroxyphenanthrene, and 2- and/or 4-hydroxybiphenyl, respectively, at much higher rates than P450 2A6. Other human P450 enzymes including P450s 1A1, 1A2, 1B1, 2C9, and 3A4 had lower rates of oxidation of naphthalene, phenanthrene, and biphenyl than P450s 2A13 and 2A6. Those alkynylated derivatives that strongly induced Type I binding spectra with P450s 2A13 and 2A6 were extensively oxidized by these enzymes upon analysis with HPLC. Molecular docking studies supported the hypothesis that ligand-interaction energies (U values) obtained with reported crystal structures of P450 2A13 and 2A6 bound to 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, indole, pilocarpine, nicotine, and coumarin are of use in understanding the basis of possible molecular interactions of these xenobiotic chemicals with the active sites of P450 2A13 and 2A6 enzymes. In fact, the ligand-interaction energies with P450 2A13 4EJG bound to these chemicals were found to relate to their induction of Type I binding spectra.

  6. Effect of levetiracetam and carbenoxolone on epilepsy model rats and related molecular mechanism

    Institute of Scientific and Technical Information of China (English)

    Yi Jia; Xiao-Jie Dai; Li Zhang; Jian-Jun Liu

    2016-01-01

    Objective:To study the effect of levetiracetam and carbenoxolone on epileptic seizures, neuron apoptosis and connexin expression in epilepsy model rats.Methods:Male SD rats were selected and divided into control group, epilepsy model group, carbenoxolone group and levetiracetam group; epilepsy model group, carbenoxolone group and levetiracetam group received intraperitoneal injection of lithium chloride-pilocarpine to establish epilepsy models and were given saline, carbenoxolone and levetiracetam treatment respectively. The epileptic seizures of the four groups of rats were observed, and the cell apoptosis as well as Cx26, Cx30, Cx43, Bcl-2, Bax and Caspase-3 expression levels in brain hippocampus was determined.Results: Epileptic seizure incubation of levetiracetam group and carbenoxolone group were significantly longer than that of epilepsy model group and the frequency of seizures were significantly less than that of epilepsy model group, epileptic seizure incubation of levetiracetam group was significantly longer than that of carbenoxolone group and the frequency of seizures was significantly less than that of carbenoxolone group. Cx26, Cx30, Cx43, Bax and Caspase-3 protein levels in hippocampus tissue of levetiracetam group and carbenoxolone group were significantly lower than those of epilepsy model group, Bcl-2 protein levels and positive Nissl staining cell number were significantly higher than that of epilepsy model group, positive TUNNEL staining cell number were significantly less than that of epilepsy model group; Cx26, Cx30, Cx43, Bax and Caspase-3 protein levels and positive Nissl staining cell number in hippocampus tissue of levetiracetam group were significantly lower than those of carbenoxolone group, Bcl-2 protein level was significantly higher than that of carbenoxolone group, positive TUNNEL staining cell number was significantly less than that of carbenoxolone group.Conclusions: Levetiracetam has better effect on inhibiting the epileptic

  7. [Pharmacological effects of N-acetyl-L-cysteine on the respiratory tract. (I). Quantitative and qualitative changes in respiratory tract fluid and sputum (author's transl)].

    Science.gov (United States)

    Kogi, K; Saito, T; Kasé, Y; Hitoshi, T

    1981-06-01

    The following three experiments were performed to determine the effects of N-acetyl-L-cysteine (NAC) on the quantity and quality of respiratory tract fluid (RTF) and sputum. All drugs used were administered into the stomach through a gastric tube. 1) Indirect measurement of bronchial secretion in rats, which was expressed by the amounts of dye excreted into the respiratory tract, was carried out according the the Sakuno's method, with some modification. Some expectorants of the secretomotor type, such as bromhexine and pilocarpine, significantly increased the secretion, even at low doses. On the other hand, mucolytic agents such as NAC augmented the secretion only in doses of 500 to 1500 mg/kg. 2)As a direct method of measurements, Kasé's modification of Perry and Boyd's method was used to collect RTF, quantitatively, from rabbits. The RTF of healthy rabbits was colorless and watery. The administration of NAC in doses of 500 to 1500 mg/kg augmented the output volume and RTF became slightly turbid, probably due to an increase in the viscous mucus. 3) Rabbits with subacute bronchitis were prepared by long-term exposure to air contaminated with SO2 gas and sputa were collected before and after administration of NAC, respectively, according to the Kase's method. The sputa were opalescent and viscous gel included nodular masses. The administration of NAC, 1000 and 1500 mg/kg resulted in a dose dependent decrease in the relative viscosity. The percent-decreased in viscosity with NAC was statistically correlated with that in amounts of dry matter, those in protein and polysaccharide in the sputa. From the results described above, it was concluded that NAC given into the stomach can liquefy sputum by splitting mucoprotein disulphide linkages, that is, altering the rheological characteristics of sputum to facilitate expectoration.

  8. Actions of brain-derived neurotrophic factor in slices from rats with spontaneous seizures and mossy fiber sprouting in the dentate gyrus.

    Science.gov (United States)

    Scharfman, H E; Goodman, J H; Sollas, A L

    1999-07-01

    This study examined the acute actions of brain-derived neurotrophic factor (BDNF) in the rat dentate gyrus after seizures, because previous studies have shown that BDNF has acute effects on dentate granule cell synaptic transmission, and other studies have demonstrated that BDNF expression increases in granule cells after seizures. Pilocarpine-treated rats were studied because they not only have seizures and increased BDNF expression in granule cells, but they also have reorganization of granule cell "mossy fiber" axons. This reorganization, referred to as "sprouting," involves collaterals that grow into novel areas, i.e., the inner molecular layer, where granule cell and interneuron dendrites are located. Thus, this animal model allowed us to address the effects of BDNF in the dentate gyrus after seizures, as well as the actions of BDNF on mossy fiber transmission after reorganization. In slices with sprouting, BDNF bath application enhanced responses recorded in the inner molecular layer to mossy fiber stimulation. Spontaneous bursts of granule cells occurred, and these were apparently generated at the site of the sprouted axon plexus. These effects were not accompanied by major changes in perforant path-evoked responses or paired-pulse inhibition, occurred only after prolonged (30-60 min) exposure to BDNF, and were blocked by K252a. The results suggest a preferential action of BDNF at mossy fiber synapses, even after substantial changes in the dentate gyrus network. Moreover, the results suggest that activation of trkB receptors could contribute to the hyperexcitability observed in animals with sprouting. Because human granule cells also express increased BDNF mRNA after seizures, and sprouting can occur in temporal lobe epileptics, the results may have implications for understanding temporal lobe epilepsy.

  9. Deficits in avoidance responding after paradoxical sleep deprivation are not associated with altered [3H]pirenzepine binding to M1 muscarinic receptors in rat brain.

    Science.gov (United States)

    Moreira, Karin M; Hipólide, Débora C; Nobrega, José N; Bueno, Orlando F A; Tufik, Sergio; Oliveira, Maria Gabriela M

    2003-07-04

    Previous work had indicated that animals that were sleep-deprived and then trained on a passive avoidance task show poor retention of the task 24 h later after being allowed to sleep freely again. Cholinergic involvement is suggested by the fact that this effect is prevented by treatment with the muscarinic agonist pilocarpine during sleep deprivation. The observation that similar deficits are observed in non-deprived rats after treatment with M1-selective antagonist compounds such as dicyclomine or pirenzepine cause similar impairments, and gave rise to the hypothesis that sleep deprivation might induce significant reductions in M1 binding in brain areas involved in learning and memory processes. Rats were deprived of sleep for 96 h and then either immediately killed, or allowed to recover sleep for 24 h before being killed. [3H]pirenzepine binding to M1 sites was examined by quantitative autoradiography in 39 different brain areas in cage controls, sleep-deprived and sleep-recovered animals (N=8 per group). No significant differences among groups were found in any brain region. A separate group of animals was subjected to the sleep deprivation procedure and then trained in a simple avoidance task. Animals were then allowed to sleep and retested 24 h later. This group showed a significant impairment in the avoidance task compared to cage controls, in agreement with previous observations. These data suggest that proactive learning/memory deficits induced by sleep deprivation cannot be attributed to altered M1 binding either immediately after deprivation (when avoidance training occurs) or after sleep has recovered (when acquisition/retention are tested). The possibility remains that alterations in M1 function occur at post-membrane second messenger systems.

  10. /sup 3/H)pirenzepine and (-)-(/sup 3/H)quinuclidinyl benzilate binding to rat cerebral cortical and cardiac muscarinic cholinergic sites. I. Characterization and regulation of agonist binding to putative muscarinic subtypes

    Energy Technology Data Exchange (ETDEWEB)

    Watson, M.; Yamamura, H.I.; Roeske, W.R.

    1986-05-01

    The binding and regulation of selected muscarinic agonists to putative subtypes in rat cerebral cortex and heart were studied. Parallel inhibition studies of (/sup 3/H)pirenzepine ((/sup 3/H)PZ) and (-)-(/sup 3/H)quinuclidinylbenzilate ((-)-(/sup 3/H)QNB)-labeled membranes were done with and without 30 microM guanyl-5'-yl imidodiphosphate (Gpp(NH)p) at 25 degrees C in 10 mM Na-K-phosphate buffer which enhances PZ binding affinity and in modified Krebs-phosphate buffer, which mimics physiological conditions. Classical agonists such as carbachol, oxotremorine and acetylcholine inhibited (-)-(/sup 3/H)QNB binding to membranes with shallow Hill values (nH less than 1), were better fit to a 2-state model, were Gpp(NH)p-regulated and showed lower affinity in modified Krebs-phosphate buffer than in 10 mM Na-K-phosphate buffer. Some agonists were not significantly better fit to a 2-state model in (/sup 3/H)PZ-labeled cortical membranes, especially in 10 mM Na-K-phosphate buffer. Whereas putative M1 and M2 binding sites distinguished by PZ possessed multiple agonist affinity states, as judged by carbachol, and agonist binding to (/sup 3/H)PZ-labeled sites were Gpp(NH)p modulated, the partial agonist pilocarpine and nonclassical agonist McN-A-343 (3-(m-chlorophenylcarbamoyloxy)-2-butynyl trimethylammonium chloride) showed little Gpp(NH)p-induced shift in (/sup 3/H)PZ-labeled cortical membranes in physiological conditions. Agonist binding to (-)-(/sup 3/H)QNB-labeled putative M2 cardiac sites was more sensitive to Gpp(NH)p than (-)-(/sup 3/H)QNB-labeled cortical sites. Carbachol and acetylcholine showed significant selectivity for putative M2 sites.

  11. The contribution of oxazolidinone frame to the biological activity of pharmaceutical drugs and natural products.

    Science.gov (United States)

    Zappia, Giovanni; Menendez, Pilar; Monache, Giuliano Delle; Misiti, Domenico; Nevola, Laura; Botta, Bruno

    2007-04-01

    The development of resistance by the antibiotics in the Gram-positive pathogenic bacteria over the last twenty years and continuing today has created a need for new antibiotic classes, which may be unaffected by existing bacterial resistance. The oxazolidin-2-ones represent not only a new class with a novel mechanism of action, but also satisfy the requirement for overcoming the resistance mechanisms. Both linezolid and eperozolid, the first chemical candidates, arose from the piperazine subclass, with the first one being chosen further development because of its enhanced pharmacokinetic properties. The main attractive traits of the oxazolidinone series has encouraged further work in the area, and the patent literature reveals that extensive chemical investigation is currently being made. The unexpected early resistance development emphasizes the need for further exploration of features of the oxazolidinone to eliminate these deficiencies. Recently, several changes, involving the C5 side chain as well the N-phenyl heterocyclic ring, give promise for such improvement. Oxazolidinone antibacterial agents comprise also ketolides, derivatives of macrolides, such as erythromycin A, with a newly formed carbamate cycle, with a largely unexplored potential. The oxazolidinone nucleus does not appear only in the structures of antimicrobial drugs, but a number of biological activities are connected with frameworks including the oxazolidinone ring. A partial list of these activities comprises enzyme inhibitors, agonists and antagonists, with a particular citation for a new generation of selective monoamino oxidase inhibitors (befloxatone). The oxazolidinone moiety was found in the structure of few biologically active natural products, such as (-)-cytoxazone and streptazolin. Moreover, in some cases the oxazolidinone ring has been chosen for the preparation of isosteric aza analogues of natural compounds (podophyllotoxin, pilocarpine) that can be more easily synthesised and more

  12. Change of MicroRNA-134, CREB and p-CREB expression in epileptic rat

    Institute of Scientific and Technical Information of China (English)

    Yan Zhu; Cheng-Shan Li; Yuan-Ye Wang; Sheng-Nian Zhou

    2015-01-01

    Objective: To To investigate the changes of MicroRNA-134, CREB and p-CREB expression in epileptic rat brains in order to elucidate the molecular mechanisms of epilepsy, providing new ideas for clinical treatment. Methods: Sixty-four Spraque-Dawley (SD) rats were divided into groups randomly, including control group, six hours after seizure group, 24-hour group, three-day group, one-week group, two-week group, four-week group, and eight-week group. All groups were placed under a pilocarpine-induced epilepsy model except the control group, and all rats were decapitated in different points of time. Brain specimens were taken for quantitative PCR experiments, immunohistochemistry and Western blot experiments. The results of the epilepsy model groups and the control group were compared. Results: There were no significant differences between the six hours after seizure group, the 24-hour group and the control group about the MicroRNA-134 levels. MicroRNA-134 in the hippocampus tissue of the three-day group significantly reduced compared with the control group; same result was observed with the one-week, two-week, four-week and eight-week groups. The CREB and p-CREB levels in the three-day group’s rat hippocampus significantly increased compared with the control group; and the high levels of CREB and p-CREB were constantly maintained in the one-week, two-week, four-week and eight-week groups. Conclusions: The MicroRNA-134 level of the epileptic rat hippocampus is significantly lower than normal after three days, and continues to maintain a low level; while CREB and p-CREB levels are rsignificantly increased after three days, and continue to remain at a high level. MicroRNA-134 plays a role in inhibiting synaptic plasticity by inhibiting CREB and p-CREB expressions.

  13. Altered anxiety-related and abnormal social behaviors in rats exposed to early life seizures

    Directory of Open Access Journals (Sweden)

    Adelisandra S. S. Castelhano

    2013-05-01

    Full Text Available Neonatal seizures are the most common manifestation of neurological dysfunction in the neonate. The prognosis of neonatal seizures is highly variable, and the controversy remains whether the severity, duration or frequency of seizures may contribute to brain damage independently of its etiology. Animal data indicates that seizures during development are associated with a high probability of long-term adverse effects such as learning and memory impairment, behavioral changes and even epilepsy, which is strongly age dependent, as well as the severity, duration and frequency of seizures. In preliminary studies, we demonstrated that adolescent male rats exposed to one-single neonatal status epilepticus (SE episode showed social behavior impairment, and we proposed the model as relevant for studies of developmental disorders. Based on these facts, the goal of this study was to verify the existence of a persistent deficit and if the anxiety-related behavior could be associated with that impairment. To do so, male Wistar rats at 9 days postnatal were submitted to a single episode of status epilepticus (SE by pilocarpine injection (380 mg/kg, i.p. and control animals received saline (0.9 %, 0,1mL/10 g. It was possible to demonstrate that in adulthood, animals exposed to neonatal SE displayed low preference for social novelty, anxiety-related behavior and increased stereotyped behavior in anxiogenic environment with no locomotor activity changes. On the balance, these data suggests that neonatal status epilepticus in rodents leads to altered anxiety-related and abnormal social behaviors.

  14. Muscarinic and nicotinic modulation of thalamo-prefrontal cortex synaptic plasticity [corrected] in vivo.

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    Lezio Soares Bueno-Junior

    Full Text Available The mediodorsal nucleus of the thalamus (MD is a rich source of afferents to the medial prefrontal cortex (mPFC. Dysfunctions in the thalamo-prefrontal connections can impair networks implicated in working memory, some of which are affected in Alzheimer disease and schizophrenia. Considering the importance of the cholinergic system to cortical functioning, our study aimed to investigate the effects of global cholinergic activation of the brain on MD-mPFC synaptic plasticity by measuring the dynamics of long-term potentiation (LTP and depression (LTD in vivo. Therefore, rats received intraventricular injections either of the muscarinic agonist pilocarpine (PILO; 40 nmol/µL, the nicotinic agonist nicotine (NIC; 320 nmol/µL, or vehicle. The injections were administered prior to either thalamic high-frequency (HFS or low-frequency stimulation (LFS. Test pulses were applied to MD for 30 min during baseline and 240 min after HFS or LFS, while field postsynaptic potentials were recorded in the mPFC. The transient oscillatory effects of PILO and NIC were monitored through recording of thalamic and cortical local field potentials. Our results show that HFS did not affect mPFC responses in vehicle-injected rats, but induced a delayed-onset LTP with distinct effects when applied following PILO or NIC. Conversely, LFS induced a stable LTD in control subjects, but was unable to induce LTD when applied after PILO or NIC. Taken together, our findings show distinct modulatory effects of each cholinergic brain activation on MD-mPFC plasticity following HFS and LFS. The LTP-inducing action and long-lasting suppression of cortical LTD induced by PILO and NIC might implicate differential modulation of thalamo-prefrontal functions under low and high input drive.

  15. Heterogeneous effects of antiepileptic drugs in an in vitro epilepsy model--a functional multineuron calcium imaging study.

    Science.gov (United States)

    Hongo, Yoshie; Takasu, Keiko; Ikegaya, Yuji; Hasegawa, Minoru; Sakaguchi, Gaku; Ogawa, Koichi

    2015-07-01

    Epilepsy is a chronic brain disease characterised by recurrent seizures. Many studies of this disease have focused on local neuronal activity, such as local field potentials in the brain. In addition, several recent studies have elucidated the collective behavior of individual neurons in a neuronal network that emits epileptic activity. However, little is known about the effects of antiepileptic drugs on neuronal networks during seizure-like events (SLEs) at single-cell resolution. Using functional multineuron Ca(2+) imaging (fMCI), we monitored the activities of multiple neurons in the rat hippocampal CA1 region on treatment with the proconvulsant bicuculline under Mg(2+) -free conditions. Bicuculline induced recurrent synchronous Ca(2+) influx, and the events were correlated with SLEs. Other proconvulsants, such as 4-aminopyridine, pentetrazol, and pilocarpine, also induced synchronous Ca(2+) influx. We found that the antiepileptic drugs phenytoin, flupirtine, and ethosuximide, which have different mechanisms of action, exerted heterogeneous effects on bicuculline-induced synchronous Ca(2+) influx. Phenytoin and flupirtine significantly decreased the peak, the amount of Ca(2+) influx and the duration of synchronous events in parallel with the duration of SLEs, whereas they did not abolish the synchronous events themselves. Ethosuximide increased the duration of synchronous Ca(2+) influx and SLEs. Furthermore, the magnitude of the inhibitory effect of phenytoin on the peak synchronous Ca(2+) influx level differed according to the peak amplitude of the synchronous event in each individual cell. Evaluation of the collective behavior of individual neurons by fMCI seems to be a powerful tool for elucidating the profiles of antiepileptic drugs.

  16. The Journal of Biological Chemistry, Volume 203, 1953: Enzyme secretion and the incorporation of P32 into phospholipides of pancreas slices.

    Science.gov (United States)

    Hokin, M R; Hokin, L E

    1989-06-01

    1. When enzyme secretion was stimulated by carbamylcholine or acetylcholine (with eserine) in slices of pigeon pancreas, the incorporation of P32 into the phospholipide fraction of the stimulated slices was, after 2 hours, 4.8 to 8.7 (average, 7.0) times greater than the incorporation of P32 into the phospholipides of control slices. Neither respiration nor the incorporation of P32 into acid-soluble phosphate esters was increased. 2. Pilocarpine, which on a weight for weight basis was much less effective than carbamylcholine or acetylcholine in stimulating enzyme secretion in pancreas slices, was also much less effective in stimulating the uptake of P32 into phospholipides. 3. The stimulatory effects of carbamylcholine on both enzyme secretion and the incorporation of P32 into phospholipides were abolished by atropine. 4. The specific activity of the phospholipides from slices incubated anaerobically was less than 5 per cent of that observed aerobically. Anaerobically, carbamylcholine did not stimulate the incorporation of P32 into phospholipides to any significant extent. The specific activity of the acid-soluble phosphate esters after anaerobic incubation was 34 per cent of that found aerobically. 5. Cholinergic drugs had little or no effect on the incorporation of P32 into the phospholipides of the following tissue slices: pigeon and guinea pig liver, guinea pig heart ventricle, pigeon gizzard (smooth muscle), and guinea pig kidney cortex. A relatively slight stimulation of P32 uptake into phospholipides was observed in slices of pigeon brain (65 per cent) and guinea pig brain cortex (40 per cent). 6. Stimulation of amylase synthesis in slices of pigeon pancreas by the addition of a mixture of amino acids had no effect on the incorporation of P32 into phospholipides.

  17. Animal models of epilepsy: use and limitations

    Directory of Open Access Journals (Sweden)

    Kandratavicius L

    2014-09-01

    Full Text Available Ludmyla Kandratavicius,1 Priscila Alves Balista,1 Cleiton Lopes-Aguiar,1 Rafael Naime Ruggiero,1 Eduardo Henrique Umeoka,2 Norberto Garcia-Cairasco,2 Lezio Soares Bueno-Junior,1 Joao Pereira Leite11Department of Neurosciences and Behavior, 2Department of Physiology, Ribeirao Preto School of Medicine, University of Sao Paulo, Ribeirao Preto, BrazilAbstract: Epilepsy is a chronic neurological condition characterized by recurrent seizures that affects millions of people worldwide. Comprehension of the complex mechanisms underlying epileptogenesis and seizure generation in temporal lobe epilepsy and other forms of epilepsy cannot be fully acquired in clinical studies with humans. As a result, the use of appropriate animal models is essential. Some of these models replicate the natural history of symptomatic focal epilepsy with an initial epileptogenic insult, which is followed by an apparent latent period and by a subsequent period of chronic spontaneous seizures. Seizures are a combination of electrical and behavioral events that are able to induce chemical, molecular, and anatomic alterations. In this review, we summarize the most frequently used models of chronic epilepsy and models of acute seizures induced by chemoconvulsants, traumatic brain injury, and electrical or sound stimuli. Genetic models of absence seizures and models of seizures and status epilepticus in the immature brain were also examined. Major uses and limitations were highlighted, and neuropathological, behavioral, and neurophysiological similarities and differences between the model and the human equivalent were considered. The quest for seizure mechanisms can provide insights into overall brain functions and consciousness, and animal models of epilepsy will continue to promote the progress of both epilepsy and neurophysiology research.Keywords: epilepsy, animal model, pilocarpine, kindling, neurodevelopment

  18. Bumetanide is not capable of terminating status epilepticus but enhances phenobarbital efficacy in different rat models.

    Science.gov (United States)

    Töllner, Kathrin; Brandt, Claudia; Erker, Thomas; Löscher, Wolfgang

    2015-01-05

    In about 20-40% of patients, status epilepticus (SE) is refractory to standard treatment with benzodiazepines, necessitating second- and third-line treatments that are not always successful, resulting in increased mortality. Rat models of refractory SE are instrumental in studying the changes underlying refractoriness and to develop more effective treatments for this severe medical emergency. Failure of GABAergic inhibition is a likely cause of the development of benzodiazepine resistance during SE. In addition to changes in GABAA receptor expression, trafficking, and function, alterations in Cl(-) homeostasis with increased intraneuronal Cl(-) levels may be involved. Bumetanide, which reduces intraneuronal Cl(-) by inhibiting the Cl(-) intruding Na(+), K(+), Cl(-) cotransporter NKCC1, has been reported to interrupt SE induced by kainate in urethane-anesthetized rats, indicating t