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Sample records for a-beta drg neurons

  1. Delayed onset of changes in soma action potential genesis in nociceptive A-beta DRG neurons in vivo in a rat model of osteoarthritis

    Henry James L

    2009-09-01

    Full Text Available Abstract Background Clinical data on osteoarthritis (OA suggest widespread changes in sensory function that vary during the progression of OA. In previous studies on a surgically-induced animal model of OA we have observed that changes in structure and gene expression follow a variable trajectory over the initial days and weeks. To investigate mechanisms underlying changes in sensory function in this model, the present electrophysiological study compared properties of primary sensory nociceptive neurons at one and two months after model induction with properties in naïve control animals. Pilot data indicated no difference in C- or Aδ-fiber associated neurons and therefore the focus is on Aβ-fiber nociceptive neurons. Results At one month after unilateral derangement of the knee by cutting the anterior cruciate ligament and removing the medial meniscus, the only changes observed in Aβ-fiber dorsal root ganglion (DRG neurons were in nociceptor-like unresponsive neurons bearing a hump on the repolarization phase; these changes consisted of longer half width, reflecting slowed dynamics of AP genesis, a depolarized Vm and an increased AP amplitude. At two months, changes observed were in Aβ-fiber high threshold mechanoreceptors, which exhibited shorter AP duration at base and half width, shorter rise time and fall time, and faster maximum rising rate/maximum falling rate, reflecting accelerated dynamics of AP genesis. Conclusion These data indicate that Aβ nociceptive neurons undergo significant changes that vary in time and occur later than changes in structure and in nociceptive scores in this surgically induced OA model. Thus, if changes in Aβ-fiber nociceptive neurons in this model reflect a role in OA pain, they may relate to mechanisms underlying pain associated with advanced OA.

  2. Daidzein induces neuritogenesis in DRG neuronal cultures

    Yang Shih-Hung

    2012-08-01

    Full Text Available Absract Background Daidzein, a phytoestrogen found in isoflavone, is known to exert neurotrophic and neuroprotective effects on the nervous system. Using primary rat dorsal root ganglion (DRG neuronal cultures, we have examined the potential neurite outgrowth effect of daidzein. Methods Dissociated dorsal root ganglia (DRG cultures were used to study the signaling mechanism of daidzein-induced neuritogenesis by immunocytochemistry and Western blotting. Results In response to daidzein treatment, DRG neurons showed a significant increase in total neurite length and in tip number per neuron. The neuritogenic effect of daidzein was significantly hampered by specific blockers for Src, protein kinase C delta (PKCδ and mitogen-activated protein kinase/extracellular signal-regulated kinase kinases (MEK/ERK, but not by those for estrogen receptor (ER. Moreover, daidzein induced phosphorylation of Src, PKCδ and ERK. The activation of PKCδ by daidzein was attenuated in the presence of a Src kinase inhibitor, and that of ERK by daidzein was diminished in the presence of either a Src or PKCδ inhibitor. Conclusion Daidzein may stimulate neurite outgrowth of DRG neurons depending on Src kinase, PKCδ and ERK signaling pathway.

  3. Comparative functional expression of nAChR subtypes in rodent DRG neurons

    J. Michael McIntosh

    2013-11-01

    Full Text Available We investigated the functional expression of nicotinic acetylcholine receptors (nAChRs in heterogeneous populations of dissociated rat and mouse lumbar dorsal root ganglion (DRG neurons by calcium imaging. By this experimental approach, it is possible to investigate the functional expression of multiple receptor and ion-channel subtypes across more than 100 neuronal and glial cells simultaneously. Based on nAChR expression, DRG neurons could be divided into four subclasses: 1 neurons that express predominantly alpha3beta4 and alpha6beta4 nAChRs; 2 neurons that express predominantly alpha7 nAChRs; 3 neurons that express a combination of alpha3beta4/alpha6beta4 and alpha7 nAChRs; and 4 neurons that do not express nAChRs. In this comparative study, the same four neuronal subclasses were observed in mouse and rat DRG. However, the expression frequency differed between species: substantially more rat DRG neurons were in the first three subclasses than mouse DRG neurons, at all developmental time points tested in our study. Approximately 70-80% of rat DRG neurons expressed functional nAChRs, in contrast to only ~15-30% of mouse DRG neurons. Our study also demonstrated functional coupling between nAChRs, voltage-gated calcium channels and mitochondrial Ca2+ transport in discrete subsets of DRG neurons. In contrast to the expression of nAChRs in DRG neurons, we demonstrated that a subset of non-neuronal DRG cells expressed muscarinic acetylcholine receptors (mAChRs and not nAChRs. The general approach to comparative cellular neurobiology outlined in this paper has the potential to better integrate molecular and systems neuroscience by uncovering the spectrum of neuronal subclasses present in a given cell population and the functionally integrated signaling components expressed in each subclass.

  4. Metabolic changes of cultured DRG neurons induced by adenosine using confocal microscopy imaging

    Zheng, Liqin; Huang, Yimei; Chen, Jiangxu; Wang, Yuhua; Yang, Hongqin; Zhang, Yanding; Xie, Shusen

    2012-12-01

    Adenosine exerts multiple effects on pain transmission in the peripheral nervous system. This study was performed to use confocal microscopy to evaluate whether adenosine could affect dorsal root ganglia (DRG) neurons in vitro and test which adenosine receptor mediates the effect of adenosine on DRG neurons. After adding adenosine with different concentration, we compared the metabolic changes by the real time imaging of calcium and mitochondria membrane potential using confocal microscopy. The results showed that the effect of 500 μM adenosine on the metabolic changes of DRG neurons was more significant than others. Furthermore, four different adenosine receptor antagonists were used to study which receptor mediated the influences of adenosine on the cultured DRG neurons. All adenosine receptor antagonists especially A1 receptor antagonist (DPCPX) had effect on the Ca2+ and mitochondria membrane potential dynamics of DRG neurons. The above studies demonstrated that the effect of adenosine which may be involved in the signal transmission on the sensory neurons was dose-dependent, and all the four adenosine receptors especially the A1R may mediate the transmission.

  5. Sensory discrimination between innocuous and noxious cold by TRPM8-expressing DRG neurons of rats

    Sarria Ignacio

    2012-10-01

    Full Text Available Abstract The TRPM8 channel is a principal cold transducer that is expressed on some primary afferents of the somatic and cranial sensory systems. However, it is uncertain whether TRPM8-expressing afferent neurons have the ability to convey innocuous and noxious cold stimuli with sensory discrimination between the two sub-modalities. Using rat dorsal root ganglion (DRG neurons and the patch-clamp recording technique, we characterized membrane and action potential properties of TRPM8-expressing DRG neurons at 24°C and 10°C. TRPM8-expressing neurons could be classified into TTX-sensitive (TTXs/TRPM8 and TTX-resistant (TTXr/TRPM8 subtypes based on the sensitivity to tetrodotoxin (TTX block of their action potentials. These two subtypes of cold-sensing cells displayed different membrane and action potential properties. Voltage-activated inward Na+ currents were highly susceptible to cooling temperature and abolished by ~95% at 10°C in TTXs/TRPM8 DRG neurons, but remained substantially large at 10°C in TTXr/TRPM8 cells. In both TTXs/TRPM8 and TTXr/TRPM8 cells, voltage-activated outward K+ currents were substantially inhibited at 10°C, and the cooling-sensitive outward currents resembled A-type K+ currents. TTXs/TRPM8 neurons and TTXr/TRPM8 neurons were shown to fire action potentials at innocuous and noxious cold temperatures respectively, demonstrating sensory discrimination between innocuous and noxious cold by the two subpopulations of cold-sensing DRG neurons. The effects of cooling temperatures on voltage-gated Na+ channels and A-type K+ currents are likely to be contributing factors to sensory discrimination of cold by TTXs/TRPM8 and TTXr/TRPM8 afferent neurons.

  6. Reactive oxygen species mediate TNFR1 increase after TRPV1 activation in mouse DRG neurons

    Westlund Karin N

    2009-06-01

    Full Text Available Abstract Background Transient receptor potential vanilloid subtype 1 (TRPV1 is activated by low pH/protons and is well known to be involved in hyperalgesia during inflammation. Tumor necrosis factor α (TNF-α, a proinflammatory cytokine, is involved in nociceptive responses causing hyperalgesia through TNF receptor type 1 (TNFR1 activation. Reactive oxygen species (ROS production is also prominently increased in inflamed tissue. The present study investigated TNFR1 receptors in primary cultured mouse dorsal root ganglion (DRG neurons after TRPV1 activation and the involvement of ROS. C57BL/6 mice, both TRPV1 knockout and wild type, were used for immunofluorescent and live cell imaging. The L4 and L5 DRGs were dissected bilaterally and cultured overnight. TRPV1 was stimulated with capsaicin or its potent analog, resiniferatoxin. ROS production was measured with live cell imaging and TNFR1 was detected with immunofluorescence in DRG primary cultures. The TRPV1 knockout mice, TRPV1 antagonist, capsazepine, and ROS scavenger, N-tert-Butyl-α-phenylnitrone (PBN, were employed to explore the functional relationship among TRPV1, ROS and TNFR1 in these studies. Results The results demonstrate that TRPV1 activation increases TNFR1 receptors and ROS generation in primary cultures of mouse DRG neurons. Activated increases in TNFR1 receptors and ROS production are absent in TRPV1 deficient mice. The PBN blocks increases in TNFR1 and ROS production induced by capsaicin/resiniferatoxin. Conclusion TRPV1 activation increases TNFR1 in cultured mouse DRG neurons through a ROS signaling pathway, a novel sensitization mechanism in DRG neurons.

  7. Oxaliplatin-induced loss of phosphorylated heavy neurofilament subunit neuronal immunoreactivity in rat DRG tissue

    Connor Bronwen

    2009-11-01

    Full Text Available Abstract Background Oxaliplatin and related chemotherapeutic drugs cause painful chronic peripheral neuropathies in cancer patients. We investigated changes in neuronal size profiles and neurofilament immunoreactivity in L5 dorsal root ganglion (DRG tissue of adult female Wistar rats after multiple-dose treatment with oxaliplatin, cisplatin, carboplatin or paclitaxel. Results After treatment with oxaliplatin, phosphorylated neurofilament heavy subunit (pNF-H immunoreactivity was reduced in neuronal cell bodies, but unchanged in nerve fibres, of the L5 DRG. Morphometric analysis confirmed significant changes in the number (-75%; P P P = 0.82, NF-M (-1%, P = 0.96 or NF-H (0%; P = 0.93 after oxaliplatin treatment, although the sizes of parvalbumin (-29%, P = 0.047, NF-M (-11%, P = 0.038 and NF-H (-28%; P = 0.0033 immunoreactive neurons were reduced. In an independent comparison of different chemotherapeutic agents, the number of pNF-H-immunoreactive neurons was significantly altered by oxaliplatin (-77.2%; P P = 0.03 but not by carboplatin or paclitaxel, and their mean cell body area was significantly changed by oxaliplatin (-31.1%; P = 0.008 but not by cisplatin, carboplatin or paclitaxel. Conclusion This study has demonstrated a specific pattern of loss of pNF-H immunoreactivity in rat DRG tissue that corresponds with the relative neurotoxicity of oxaliplatin, cisplatin and carboplatin. Loss of pNF-H may be mechanistically linked to oxaliplatin-induced neuronal atrophy, and serves as a readily measureable endpoint of its neurotoxicity in the rat model.

  8. Neurofilament protein synthesis in DRG neurons decreases more after peripheral axotomy than after central axotomy

    Cytoskeletal protein synthesis was studied in DRG neurons after transecting either their peripheral or their central branch axons. Specifically, the axons were transected 5-10 mm from the lumbar-5 ganglion on one side of the animal; the DRGs from the transected side and contralateral control side were labeled with radiolabeled amino acids in vitro; radiolabeled proteins were separated by 2-dimensional (2D) PAGE; and the amounts of radiolabel in certain proteins of the experimental and control ganglia were quantified and compared. We focused on the neurofilament proteins because they are neuron-specific. If either the peripheral or central axons were cut, the amounts of radiolabeled neurofilament protein synthesized by the DRG neurons decreased between 1 and 10 d after transection. Neurofilament protein labeling decreased more after transection of the peripheral axons than after transection of the central axons. In contrast to axonal transections, sham operations or heat shock did not decrease the radiolabeling of the neurofilament proteins, and these procedures also affected the labeling of actin, tubulin, and the heat-shock proteins differently from transection. These results and others indicate that axonal transection leads to specific changes in the synthesis of cytoskeletal proteins of DRG neurons, and that these changes differ from those produced by stress to the animal or ganglia. Studies of the changes in neurofilament protein synthesis from 1 to 40 d after axonal transection indicate that the amounts of radiolabeled neurofilament protein synthesis were decreased during axonal elongation, but that they returned toward control levels when the axons reached cells that stopped elongation

  9. Sensitization of capsaicin and icilin responses in oxaliplatin treated adult rat DRG neurons

    Anand Praveen

    2010-11-01

    Full Text Available Abstract Background Oxaliplatin chemotherapy induced neuropathy is a dose related cumulative toxicity that manifests as tingling, numbness, and chronic pain, compromising the quality of life and leading to discontinued chemotherapy. Patients report marked hypersensitivity to cold stimuli at early stages of treatment, when sensory testing reveals cold and heat hyperalgesia. This study examined the morphological and functional effects of oxaliplatin treatment in cultured adult rat DRG neurons. Results 48 hour exposure to oxaliplatin resulted in dose related reduction in neurite length, density, and number of neurons compared to vehicle treated controls, using Gap43 immunostaining. Neurons treated acutely with 20 μg/ml oxaliplatin showed significantly higher signal intensity for cyclic AMP immunofluorescence (160.5 ± 13 a.u., n = 3, P Conclusions Oxaliplatin treatment induces TRP sensitization mediated by increased intracellular cAMP, which may cause neuronal damage. These effects may be mitigated by co-treatment with adenylyl cyclase inhibitors, like CB2 agonists, to alleviate the neurotoxic effects of oxaliplatin.

  10. Single-cell RNA-seq reveals distinct injury responses in different types of DRG sensory neurons

    Hu, Ganlu; Huang, Kevin; Hu, Youjin; Du, Guizhen; Xue, Zhigang; Zhu, Xianmin; Fan, Guoping

    2016-01-01

    Peripheral nerve injury leads to various injury-induced responses in sensory neurons including physiological pain, neuronal cell death, and nerve regeneration. In this study, we performed single-cell RNA-sequencing (scRNA-seq) analysis of mouse nonpeptidergic nociceptors (NP), peptidergic nociceptors (PEP), and large myelinated sensory neurons (LM) under both control and injury conditions at 3 days after sciatic nerve transection (SNT). After performing principle component and weighted gene co-expression network analysis, we categorized dorsal root ganglion (DRG) neurons into different subtypes and discovered co-regulated injury-response genes including novel regeneration associated genes (RAGs) in association with neuronal development, protein translation and cytoplasm transportation. In addition, we found significant up-regulation of the genes associated with cell death such as Pdcd2 in a subset of NP neurons after axotomy, implicating their actions in neuronal cell death upon nerve injury. Our study revealed the distinctive and sustained heterogeneity of transcriptomic responses to injury at single neuron level, implicating the involvement of different gene regulatory networks in nerve regeneration, neuronal cell death and neuropathy in different population of DRG neurons. PMID:27558660

  11. Role of laser fluence in protein synthesis of cultured DRG neurons following low-level laser irradiation

    Zheng, Liqin; Qiu, Caimin; Wang, Yuhua; Zeng, Yixiu; Yang, Hongqin; Zhang, Yanding; Xie, Shusen

    2014-11-01

    Low-level lasers have been used to relieve pain in clinical for many years. But the mechanism is not fully clear. In animal models, nitric oxide (NO) has been reported involving in the transmission and modulation of nociceptive signals. So the objective of this study was to establish whether low-level laser with different fluence could stimulate the production of nitric oxide synthese (NOS), which produces NO in cultured primary dorsal root ganglion neurons (DRG neurons). The primary DRG neurons were isolated from healthy Sprague Dawley rats (8-12 weeks of age) and spread on 35 mm culture dishes specially used for confocal microscopy. 24 hours after spreading, cells were irradiated with 658 nm laser for two consecutive days at the energy density of 20, 40, 60 and 80 mJ·cm-2 respectively. Control groups were not exposed to the laser, but were kept under the same conditions as the irradiated ones. The synthesis of NOS after laser irradiation was detected by immunofluorescence assay, and the changes of NOS were evaluated using confocal microscopy and Image J software. The results showed that all the laser fluence could promote the production of NOS in DRG neurons, especially the 60 mJ·cm-2 . These results demonstrated that low-level laser irradiation could modify protein synthesis in a dose- or fluence- dependent manner, and indicated that low-level laser irradiation might achieve the analgesic effect through modulation of NO production.

  12. Effects of ranolazine on wild-type and mutant hNav1.7 channels and on DRG neuron excitability

    Estacion Mark

    2010-06-01

    Full Text Available Abstract Background A direct role of sodium channels in pain has recently been confirmed by establishing a monogenic link between SCN9A, the gene which encodes sodium channel Nav1.7, and pain disorders in humans, with gain-of-function mutations causing severe pain syndromes, and loss-of-function mutations causing congenital indifference to pain. Expression of sodium channel Nav1.8 in DRG neurons has also been shown to be essential for the manifestation of mutant Nav1.7-induced neuronal hyperexcitability. These findings have confirmed key roles of Nav1.7 and Nav1.8 in pain and identify these channels as novel targets for pain therapeutic development. Ranolazine preferentially blocks cardiac late sodium currents at concentrations that do not significantly reduce peak sodium current. Ranolazine also blocks wild-type Nav1.7 and Nav1.8 channels in a use-dependent manner. However, ranolazine's effects on gain-of-function mutations of Nav1.7 and on DRG neuron excitability have not been investigated. We used voltage- and current-clamp recordings to evaluate the hypothesis that ranolazine may be effective in regulating Nav1.7-induced DRG neuron hyperexcitability. Results We show that ranolazine produces comparable block of peak and ramp currents of wild-type Nav1.7 and mutant Nav1.7 channels linked to Inherited Erythromelalgia and Paroxysmal Extreme Pain Disorder. We also show that ranolazine, at a clinically-relevant concentration, blocks high-frequency firing of DRG neurons expressing wild-type but not mutant channels. Conclusions Our data suggest that ranalozine can attenuate hyperexcitability of DRG neurons over-expressing wild-type Nav1.7 channels, as occurs in acquired neuropathic and inflammatory pain, and thus merits further study as an alternative to existing non-selective sodium channel blockers.

  13. Paroxysmal extreme pain disorder M1627K mutation in human Nav1.7 renders DRG neurons hyperexcitable

    Tyrrell Lynda

    2008-09-01

    Full Text Available Abstract Background Paroxysmal extreme pain disorder (PEPD is an autosomal dominant painful neuropathy with many, but not all, cases linked to gain-of-function mutations in SCN9A which encodes voltage-gated sodium channel Nav1.7. Severe pain episodes and skin flushing start in infancy and are induced by perianal probing or bowl movement, and pain progresses to ocular and mandibular areas with age. Carbamazepine has been effective in relieving symptoms, while other drugs including other anti-epileptics are less effective. Results Sequencing of SCN9A coding exons from an English patient, diagnosed with PEPD, has identified a methionine 1627 to lysine (M1627K substitution in the linker joining segments S4 and S5 in domain IV. We confirm that M1627K depolarizes the voltage-dependence of fast-inactivation without substantially altering activation or slow-inactivation, and inactivates from the open state with slower kinetics. We show here that M1627K does not alter development of closed-state inactivation, and that M1627K channels recover from fast-inactivation faster than wild type channels, and produce larger currents in response to a slow ramp stimulus. Using current-clamp recordings, we also show that the M1627K mutant channel reduces the threshold for single action potentials in DRG neurons and increases the number of action potentials in response to graded stimuli. Conclusion M1627K mutation was previously identified in a sporadic case of PEPD from France, and we now report it in an English family. We confirm the initial characterization of mutant M1627K effect on fast-inactivation of Nav1.7 and extend the analysis to other gating properties of the channel. We also show that M1627K mutant channels render DRG neurons hyperexcitable. Our new data provide a link between altered channel biophysics and pain in PEPD patients.

  14. Expression of Nav1.7 in DRG neurons extends from peripheral terminals in the skin to central preterminal branches and terminals in the dorsal horn

    Black Joel A

    2012-11-01

    Full Text Available Abstract Background Sodium channel Nav1.7 has emerged as a target of considerable interest in pain research, since loss-of-function mutations in SCN9A, the gene that encodes Nav1.7, are associated with a syndrome of congenital insensitivity to pain, gain-of-function mutations are linked to the debiliting chronic pain conditions erythromelalgia and paroxysmal extreme pain disorder, and upregulated expression of Nav1.7 accompanies pain in diabetes and inflammation. Since Nav1.7 has been implicated as playing a critical role in pain pathways, we examined by immunocytochemical methods the expression and distribution of Nav1.7 in rat dorsal root ganglia neurons, from peripheral terminals in the skin to central terminals in the spinal cord dorsal horn. Results Nav1.7 is robustly expressed within the somata of peptidergic and non-peptidergic DRG neurons, and along the peripherally- and centrally-directed C-fibers of these cells. Nav1.7 is also expressed at nodes of Ranvier in a subpopulation of Aδ-fibers within sciatic nerve and dorsal root. The peripheral terminals of DRG neurons within skin, intraepidermal nerve fibers (IENF, exhibit robust Nav1.7 immunolabeling. The central projections of DRG neurons in the superficial lamina of spinal cord dorsal horn also display Nav1.7 immunoreactivity which extends to presynaptic terminals. Conclusions The expression of Nav1.7 in DRG neurons extends from peripheral terminals in the skin to preterminal central branches and terminals in the dorsal horn. These data support a major contribution for Nav1.7 in pain pathways, including action potential electrogenesis, conduction along axonal trunks and depolarization/invasion of presynaptic axons. The findings presented here may be important for pharmaceutical development, where target engagement in the right compartment is essential.

  15. Effects of ranolazine on wild-type and mutant hNav1.7 channels and on DRG neuron excitability

    Estacion Mark; Waxman Stephen G; Dib-Hajj Sulayman D

    2010-01-01

    Abstract Background A direct role of sodium channels in pain has recently been confirmed by establishing a monogenic link between SCN9A, the gene which encodes sodium channel Nav1.7, and pain disorders in humans, with gain-of-function mutations causing severe pain syndromes, and loss-of-function mutations causing congenital indifference to pain. Expression of sodium channel Nav1.8 in DRG neurons has also been shown to be essential for the manifestation of mutant Nav1.7-induced neuronal hypere...

  16. Expression of mRNAs for PPT, CGRP, NF-200, and MAP-2 in cocultures of dissociated DRG neurons and skeletal muscle cells in administration of NGF or NT-3

    Weiwei Zhang

    2012-07-01

    Full Text Available Both neurotrophins (NTs and target skeletal muscle (SKM cells are essential for the maintenance of the function of neurons and nerve-muscle communication. However, much less is known about the association of target SKM cells with distinct NTs on the expression of mRNAs for preprotachykinin (PPT, calcitonin-gene related peptide (CGRP, neurofilament 200 (NF-200, and microtubule associated protein 2 (MAP-2 in dorsal root ganglion (DRG sensory neurons. In the present study, a neuromuscular coculture model of dissociated dorsal root ganglion (DRG neurons and SKM cells was established. The morphology of DRG neurons and SKM cells in coculture was observed with an inverted phase contrast microscope. The effects of nerve growth factor (NGF or neurotrophin-3 (NT-3 on the expression of mRNAs for PPT, CGRP, NF-200, and MAP-2 was analyzed by real time-PCR assay. The morphology of DRG neuronal cell bodies and SKM cells in neuromuscular coculture at different conditions was similar. The neurons presented evidence of dense neurite outgrowth in the presence of distinct NTs in neuromuscular cocultures. NGF and NT-3 increased mRNA levels of PPT, CGRP, and NF-200, but not MAP-2, in neuromuscular cocultures. These results offer new clues towards a better understanding of the association of target SKM cells with distinct NTs on the expression of mRNAs for PPT, CGRP, NF-200 and MAP-2, and implicate the association of target SKM cells and NTs with DRG sensory neuronal phenotypes.

  17. Modulation of Extracellular ATP Content of Mast Cells and DRG Neurons by Irradiation: Studies on Underlying Mechanism of Low-Level-Laser Therapy

    Lina Wang

    2015-01-01

    Full Text Available Low-level-laser therapy (LLLT is an effective complementary treatment, especially for anti-inflammation and wound healing in which dermis or mucus mast cells (MCs are involved. In periphery, MCs crosstalk with neurons via purinergic signals and participate in various physiological and pathophysiological processes. Whether extracellular ATP, an important purine in purinergic signaling, of MCs and neurons could be modulated by irradiation remains unknown. In this study, effects of red-laser irradiation on extracellular ATP content of MCs and dorsal root ganglia (DRG neurons were investigated and underlying mechanisms were explored in vitro. Our results show that irradiation led to elevation of extracellular ATP level in the human mast cell line HMC-1 in a dose-dependent manner, which was accompanied by elevation of intracellular ATP content, an indicator for ATP synthesis, together with [Ca2+]i elevation, a trigger signal for exocytotic ATP release. In contrast to MCs, irradiation attenuated the extracellular ATP content of neurons, which could be abolished by ARL 67156, a nonspecific ecto-ATPases inhibitor. Our results suggest that irradiation potentiates extracellular ATP of MCs by promoting ATP synthesis and release and attenuates extracellular ATP of neurons by upregulating ecto-ATPase activity. The opposite responses of these two cell types indicate complex mechanisms underlying LLLT.

  18. The inhibition of subchondral bone lesions significantly reversed the weight-bearing deficit and the overexpression of CGRP in DRG neurons, GFAP and Iba-1 in the spinal dorsal horn in the monosodium iodoacetate induced model of osteoarthritis pain.

    Degang Yu

    Full Text Available BACKGROUND: Chronic pain is the most prominent and disabling symptom of osteoarthritis (OA. Clinical data suggest that subchondral bone lesions contribute to the occurrence of joint pain. The present study investigated the effect of the inhibition of subchondral bone lesions on joint pain. METHODS: Osteoarthritic pain was induced by an injection of monosodium iodoacetate (MIA into the rat knee joint. Zoledronic acid (ZOL, a third generation of bisphosphonate, was used to inhibit subchondral bone lesions. Joint histomorphology was evaluated using X-ray micro computed tomography scanning and hematoxylin-eosin staining. The activity of osteoclast in subchondral bone was evaluated using tartrate-resistant acid phosphatase staining. Joint pain was evaluated using weight-bearing asymmetry, the expression of calcitonin gene-related peptide (CGRP in the dorsal root ganglion (DRG, and spinal glial activation status using glial fibrillary acidic protein (GFAP and ionized calcium binding adaptor molecule-1 (Iba-1 immunofluorescence. Afferent neurons in the DRGs that innervated the joints were identified using retrograde fluorogold labeling. RESULTS: MIA injections induced significant histomorphological alterations and joint pain. The inhibition of subchondral bone lesions by ZOL significantly reduced the MIA-induced weight-bearing deficit and overexpression of CGRP in DRG neurons, GFAP and Iba-1 in the spinal dorsal horn at 3 and 6 weeks after MIA injection; however, joint swelling and synovial reaction were unaffected. CONCLUSIONS: The inhibition of subchondral bone lesions alleviated joint pain. Subchondral bone lesions should be a key target in the management of osteoarthritic joint pain.

  19. Comparison of the Force Exerted by Hippocampal and DRG Growth Cones

    Amin, Ladan; Ercolini, Erika; Ban, Jelena; Torre, Vincent

    2013-01-01

    Mechanical properties such as force generation are fundamental for neuronal motility, development and regeneration. We used optical tweezers to compare the force exerted by growth cones (GCs) of neurons from the Peripheral Nervous System (PNS), such as Dorsal Root Ganglia (DRG) neurons, and from the Central Nervous System (CNS) such as hippocampal neurons. Developing GCs from dissociated DRG and hippocampal neurons were obtained from P1-P2 and P10-P12 rats. Comparing their morphology, we obse...

  20. Mouse DRG Cell Line with Properties of Nociceptors.

    Ciara Doran

    Full Text Available In vitro cell lines from DRG neurons aid drug discovery because they can be used for early stage, high-throughput screens for drugs targeting pain pathways, with minimal dependence on animals. We have established a conditionally immortal DRG cell line from the Immortomouse. Using immunocytochemistry, RT-PCR and calcium microfluorimetry, we demonstrate that the cell line MED17.11 expresses markers of cells committed to the sensory neuron lineage. Within a few hours under differentiating conditions, MED17.11 cells extend processes and following seven days of differentiation, express markers of more mature DRG neurons, such as NaV1.7 and Piezo2. However, at least at this time-point, the nociceptive marker NaV1.8 is not expressed, but the cells respond to compounds known to excite nociceptors, including the TRPV1 agonist capsaicin, the purinergic receptor agonist ATP and the voltage gated sodium channel agonist, veratridine. Robust calcium transients are observed in the presence of the inflammatory mediators bradykinin, histamine and norepinephrine. MED17.11 cells have the potential to replace or reduce the use of primary DRG culture in sensory, pain and developmental research by providing a simple model to study acute nociception, neurite outgrowth and the developmental specification of DRG neurons.

  1. Digital Raster Graphics (DRG) Lambert

    Kansas Data Access and Support Center — The Digital Raster Graphic-Lambert (DRG-Lam) is a raster image of a scanned USGS topographic map with the collar information clipped out, georeferenced to the...

  2. Modulation of Voltage-Gated Sodium Channels by Activation of Tumor Necrosis Factor Receptor-1 and Receptor-2 in Small DRG Neurons of Rats

    M. Leo

    2015-01-01

    Full Text Available Tumor necrosis factor- (TNF- α is a proinflammatory cytokine involved in the development and maintenance of inflammatory and neuropathic pain. Its effects are mediated by two receptors, TNF receptor-1 (TNFR-1 and TNF receptor-2 (TNFR-2. These receptors play a crucial role in the sensitization of voltage-gated sodium channels (VGSCs, a key mechanism in the pathogenesis of chronic pain. Using the whole-cell patch-clamp technique, we examined the influence of TNFR-1 and TNFR-2 on VGSCs and TTX-resistant NaV1.8 channels in isolated rat dorsal root ganglion neurons by using selective TNFR agonists. The TNFR-1 agonist R32W (10 pg/mL caused an increase in the VGSC current (INa(V by 27.2 ± 5.1%, while the TNFR-2 agonist D145 (10 pg/mL increased the current by 44.9 ± 2.6%. This effect was dose dependent. Treating isolated NaV1.8 with R32W (100 pg/mL resulted in an increase in INaV(1.8 by 18.9 ± 1.6%, while treatment with D145 (100 pg/mL increased the current by 14.5 ± 3.7%. Based on the current-voltage relationship, 10 pg of R32W or D145 led to an increase in INa(V in a bell-shaped, voltage-dependent manner with a maximum effect at −30 mV. The effects of TNFR activation on VGSCs promote excitation in primary afferent neurons and this might explain the sensitization mechanisms associated with neuropathic and inflammatory pain.

  3. Effects of serum immunoglobulins from patients with complex regional pain syndrome (CRPS) on depolarisation-induced calcium transients in isolated dorsal root ganglion (DRG) neurons.

    Reilly, Joanne M; Dharmalingam, Backialakshmi; Marsh, Stephen J; Thompson, Victoria; Goebel, Andreas; Brown, David A

    2016-03-01

    Complex regional pain syndrome (CRPS) is thought to have an auto-immune component. One such target recently proposed from the effects of auto-immune IgGs on Ca(2+) transients in cardiac myocytes and cell lines is the α1-adrenoceptor. We have tested whether such IgGs exerted comparable effects on nociceptive sensory neurons isolated from rat dorsal root ganglia. Depolarisation-induced [Ca(2+)]i transients were generated by applying 30mM KCl for 2min and monitored by Fura-2 fluorescence imaging. No IgGs tested (including 3 from CRPS patients) had any significant effect on these [Ca(2+)]i transients. However, IgG from one CRPS patient consistently and significantly reduced the K(+)-induced response of cells that had been pre-incubated for 24h with a mixture of inflammatory mediators (1μM histamine, 5-hydroxytryptamine, bradykinin and PGE2). Since this pre-incubation also appeared to induce a comparable inhibitory response to the α1-agonist phenylephrine, this is compatible with the α1-adrenoceptor as a target for CRPS auto-immunity. A mechanism whereby this might enhance pain is suggested. PMID:26708558

  4. Metabotropic glutamate receptor activation and intracellular cyclic ADP-ribose release Ca2+ from the same store in cultured DRG neurones.

    Pollock, J; Crawford, J H; Wootton, J F; Seabrook, G R; Scott, R H

    1999-01-01

    The whole cell patch clamp technique has been used to record Ca(2+)-activated cation and chloride conductances evoked by release of Ca2+ from intracellular stores of cultured neonatal dorsal root ganglion neurones. The aim of this study was to investigate metabotropic glutamate receptor (mGluR) mechanisms and evaluate a possible role for cyclic ADP-ribose as an intracellular signalling molecule. Glutamate and the metabotropic glutamate receptor agonist (1S, 3R)-ACPD-evoked transient depolarizations, Ca(2+)-activated inward currents and rises in intracellular Ca2+. The (1S, 3R)-ACPD-activated currents were insensitive to InsP3 signalling inhibitors, heparin and pentosan polysulphate. Intracellular application of ryanodine alone activated currents in this study and proved a difficult tool to use as a potential inhibitor of cyclic ADP-ribose-mediated responses. However, intracellular dantrolene did attenuate both (1S, 3R)-ACPD and cyclic ADP-ribose responses. Intracellular photo-release of cGMP and cyclic ADP-ribose mimicked the responses to mGluR receptor activation. Intracellular application of nicotinamide and W7 inhibited the responses to photo-released cGMP but did not prevent responses to mGluR activation. The cyclic ADP-ribose receptor antagonist 8-amino cyclic ADP-ribose attenuated responses to (1S, 3R)-ACPD, cGMP and cyclic ADP-ribose, but some Ca(2+)-activated inward currents were still observed in the presence of this antagonist. In conclusion, mGluR receptor activation, cGMP and cyclic ADP-ribose release Ca2+ from intracellular stores. Some evidence suggests that pharmacologically related pathways are involved. PMID:10598278

  5. An Optimized Culture Method of Rat Dorsal Root Ganglion Neurons

    LIUYin; CHENJing-Hong; GONGZe-Hui

    2004-01-01

    AIM: To establish a primary culture technique of acutely isolated dorsal root ganglion (DRG) neurons, and provide a simple & useful in vitro model for study of analgesia. Methods: Acutely isolated dorsal root ganglion (DRG) neurons were planted and cultured; the configuration and growth characters of DRG neurons were observed through inverted microscope.

  6. Effects of Jinmaitong on ultrastructure of DRG neurons in STZ-DM rats%中药筋脉通对糖尿病大鼠背根神经节神经元超微结构影响的研究

    杨丹; 梁晓春; 屈岭; 吴群励; 吴亚楠; 石玥; 戴威

    2016-01-01

    Objective To study the effects of Jinmaitong on ultrastructure of dorsal root ganglion ( DRG) neurons in STZ-induced diabetic rats. Methods The STZ-induced diabetic rats were randomly divided into 4 groups including normal control group (Con), Diabetic group (DM), JMT group (JMT) and Taurine group ( Tau) . Six rats were in each group. All rats were given intragastric administration for 16 weeks and then culled. The mechanical withdrawal threshold was tested by electronic Von Frey instrument before death. The DRG of the rats was collected and used for transmission electron microscopy. Results Compared with the control group, the mechanical pain threshold of the DM group was significantly decreased (P<0. 01). The ultrastructure of mitochondria in DRG neurons of the DM group was severely damaged. The number of mitochondria in neurons was significantly decreased (P<0. 01), and the rate of swelling mitochondria and mitochondria vacuolar degeneration in DRG neurons was significantly increased of the DM group (P<0. 01). The mechanical pain threshold and the number of mitochondria of two treatment groups were significantly improved compared with the DM group. The rate of swelling mitochondria and mitochondria vacuolar degeneration of two treatment groups were decreased strikingly (P<0. 01), and the JMT group was more effective than the Tau group (P<0. 01). Conclusion JMT could ameliorate the peripheral neuropathy by improving the pathological morphology of mitochondria in DRG neurons.%目的:观察中药筋脉通对链尿佐菌素( streptozotocin,STZ)诱导的糖尿病( diabetes mellitus,DM)大鼠背根神经节(dorsal root ganglia,DRG)感觉神经元超微结构的影响。方法雄性SD大鼠18只,腹腔内注射STZ诱导建立糖尿病大鼠模型,随机分为糖尿病组、筋脉通组与牛磺酸组,每组6只大鼠,并设正常对照组6只。成模后每天1次灌胃给药,持续16周。药物干预16周后,用电子Von Frey仪检测机械痛阈值,取大鼠背

  7. 42 CFR 412.60 - DRG classification and weighting factors.

    2010-10-01

    ... 42 Public Health 2 2010-10-01 2010-10-01 false DRG classification and weighting factors. 412.60... Determining Prospective Payment Federal Rates for Inpatient Operating Costs § 412.60 DRG classification and... discharges by Diagnosis-Related Groups (DRGs). (b) DRG weighting factors. CMS assigns, for each DRG,...

  8. 42 CFR 412.515 - LTC-DRG weighting factors.

    2010-10-01

    ... 42 Public Health 2 2010-10-01 2010-10-01 false LTC-DRG weighting factors. 412.515 Section 412.515 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES MEDICARE...-Term Care Hospitals § 412.515 LTC-DRG weighting factors. For each LTC-DRG, CMS assigns an...

  9. Phosphoinositide signaling in somatosensory neurons.

    Rohacs, Tibor

    2016-05-01

    Somatosensory neurons of the dorsal root ganglia (DRG) and trigeminal ganglia (TG) are responsible for detecting thermal and tactile stimuli. They are also the primary neurons mediating pain and itch. A large number of cell surface receptors in these neurons couple to phospholipase C (PLC) enzymes leading to the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and the generation of downstream signaling molecules. These neurons also express many different ion channels, several of which are regulated by phosphoinositides. This review will summarize the knowledge on phosphoinositide signaling in DRG neurons, with special focus on effects on sensory and other ion channels. PMID:26724974

  10. Oligomerization and toxicity of A{beta} fusion proteins

    Caine, Joanne M., E-mail: Jo.Caine@csiro.au [CSIRO Materials Science and Engineering and the Preventive Health Flagship, Parkville, Victoria (Australia); Bharadwaj, Prashant R. [CSIRO Materials Science and Engineering and the Preventive Health Flagship, Parkville, Victoria (Australia); Centre for Excellence for Alzheimer' s Disease Research and Care, School of Exercise, Biomedical and Health Sciences, Edith Cowan University, Western Australia (Australia); Sankovich, Sonia E. [CSIRO Materials Science and Engineering and the Preventive Health Flagship, Parkville, Victoria (Australia); Ciccotosto, Giuseppe D. [The Department of Pathology and Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Victoria 3010 (Australia); Streltsov, Victor A.; Varghese, Jose [CSIRO Materials Science and Engineering and the Preventive Health Flagship, Parkville, Victoria (Australia)

    2011-06-10

    Highlights: {yields} We expressed amyloid-{beta} (A{beta}) peptide as a soluble maltose binding protein fusion (MBP-A{beta}42 and MBP-A{beta}16). {yields} The full length A{beta} peptide fusion, MBP-A{beta}42, forms oligomeric species as determined by SDS-PAGE gels, gel filtration and DLS. {yields} The MBP-A{beta}42, but not MBP-A{beta}16 or MBP alone, is toxic to both yeast and mammalian cells as determined by toxicity assays. -- Abstract: This study has found that the Maltose binding protein A{beta}42 fusion protein (MBP-A{beta}42) forms soluble oligomers while the shorter MBP-A{beta}16 fusion and control MBP did not. MBP-A{beta}42, but neither MBP-A{beta}16 nor control MBP, was toxic in a dose-dependent manner in both yeast and primary cortical neuronal cells. This study demonstrates the potential utility of MBP-A{beta}42 as a reagent for drug screening assays in yeast and neuronal cell cultures and as a candidate for further A{beta}42 characterization.

  11. Amitriptyline Activates TrkA to Aid Neuronal Growth and Attenuate Anesthesia-Induced Neurodegeneration in Rat Dorsal Root Ganglion Neurons.

    Zheng, Xiaochun; Chen, Feng; Zheng, Ting; Huang, Fengyi; Chen, Jianghu; Tu, Wenshao

    2016-05-01

    Tricyclic antidepressant amitriptyline (AM) has been shown to exert neurotrophic activity on neurons. We thus explored whether AM may aid the neuronal development and protect anesthesia-induced neuro-injury in young spinal cord dorsal root ganglion (DRG) neurons.The DRG explants were prepared from 1-day-old rats. The effect of AM on aiding DRG neural development was examined by immunohistochemistry at dose-dependent manner. AM-induced changes in gene and protein expressions, and also phosphorylation states of tyrosine kinases receptor A (TrkA) and B (TrkB) in DRG, were examined by quantitative real-time polymerase chain reaction and western blot. The effect of AM on attenuating lidocaine-induced DRG neurodegeneration was examined by immunohistochemistry, and small interfering RNA (siRNA)-mediated TrkA/B down-regulation.Amitriptyline stimulated DRG neuronal development in dose-dependent manner, but exerted toxic effect at concentrations higher than 10 M. AM activated TrkA in DRG through phosphorylation, whereas it had little effect on TrkB-signaling pathway. AM reduced lidocaine-induced DRG neurodegeneration by regenerating neurites and growth cones. Moreover, the neuroprotection of AM on lidocaine-injured neurodegeneration was blocked by siRNA-mediated TrkA down-regulation, but not by TrkB down-regulation.Amitriptyline facilitated neuronal development and had protective effect on lidocaine-induced neurodegeneration, very likely through the activation of TrkA-signaling pathway in DRG. PMID:27149473

  12. Characteristics of dorsal root ganglia neurons sensitive to Substance P

    Moraes, Eder Ricardo; Kushmerick, Christopher; Naves, Ligia Araujo

    2014-01-01

    Background Substance P modulates ion channels and the excitability of sensory neurons in pain pathways. Within the heterogeneous population of Dorsal Root Ganglia (DRG) primary sensory neurons, the properties of cells that are sensitive to Substance P are poorly characterized. To define this population better, dissociated rat DRG neurons were tested for their responsiveness to capsaicin, ATP and acid. Responses to ATP were classified according to the kinetics of current activation and desensi...

  13. Digital Raster Graphics (DRG) UTM NAD 83 Clipped

    Kansas Data Access and Support Center — The Digital Raster Graphic (DRG) is a raster image of a scanned USGS topographic map including the collar information, georeferenced to the UTM grid. For display...

  14. Digital Raster Graphics (DRG) UTM NAD 83 County

    Kansas Data Access and Support Center — The Digital Raster Graphic (DRG) is a raster image of a scanned USGS topographic map including the collar information, georeferenced to the UTM grid and tiled by...

  15. Radioiodine therapy within the German DRG-system 2005

    With introduction of a diagnosis-related groups system (DRG-system) in Germany the previous duration of stay based refunding is also replaced for the radioiodine therapies by a performance oriented reimbursement system. Since the at first optional start of the DRG-system in 2003 the adaptations which take place every year should lead, up to the planned end of the convergence phase in 2009, to a transparent, fair and economical financing system of the stationary hospital service. The physician is responsible for the right and complete coding of the diagnoses and procedures, which serve as essential parameters for the determination of the diagnosis related group (DRG) of a hospital case. In the actual version of the year 2005 the DRG-system still supplies for radioiodine therapy of thyroid carcinoma some unclarity in the coding of the diagnosis, as well as clear inadequacy with the fair mapping of the therapy costs. (orig.)

  16. Digital Raster Graphics (DRG) UTM NAD 27 Clipped

    Kansas Data Access and Support Center — The Digital Raster Graphic (DRG) is a raster image of a scanned USGS topographic map including the collar information, georeferenced to the UTM grid. For display...

  17. USGS DRG Mosaic For Appomattox Court House National Historical Park

    National Park Service, Department of the Interior — This Digital Raster Graphic (DRG) is a mosaic of the USGS topographic map for Vera and Appomattox with the collar information clipped and georeferenced to the UTM...

  18. Critical appraisal of the DRG system: problem areas for DRG reimbursement in the USA.

    Harper, D L

    1985-03-01

    One may conclude that the prospective payment system known as Diagnosis Related Groups is the initial thrust of the government to reduce the rate of expenditures for Medicare patients in the acute hospital setting. More will come and probably soon. Among the criticism and concern, one must not ignore the non-inclusion of the physician provider in this system. Cost shifting to the private payer will probably result in DRGs in that area also. Concern about the ability to provide the technology necessary to render quality care is utmost on the minds of providers. The shift of patients into the alternate health care field will leave hospitals with fewer patients who are overall much sicker and the hospital will have fewer dollars from which to provide that care. There is specific criticism of some of the DRG groupings and concern with the way in which physicians will be brought under the system. The issue of bad debts and care for the indigent must be addressed. A great deal of attention must be paid in the manner in which capital costs are reimbursed. Medical education costs must be satisfactorily addressed on a national basis. MEDPAR data used to provide data base for determining the DRG rates, must be upgraded to bring fairness to the system. Now medical technology must be examined quickly and the costs of those desirable technological advances incorporated into the DRG rate so as not to impede their use. Perhaps the greatest concern relates to the factors which will be incorporated into the rate basis and those which affect the allowable rate increases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3929757

  19. G protein-coupled inwardly rectifying potassium channels in dorsal root ganglion neurons

    Xiao-fei GAO; Hai-lin ZHANG; Zhen-dong YOU; Chang-lin LU; Cheng HE

    2007-01-01

    Aim: G protein-coupled inwardly rectifying potassium channels (GIRK) are important for neuronal signaling and membrane excitability. In the present study, we intend to find whether GIRK channels express functionally in adult rat dorsal root ganglion (DRG) neurons. Methods: We used RT-PCR to detect mRNA for4 subunits of GIRK in the adult DRG. The whole-cell patch clamp recording was used to confirm GIRK channels functionally expressed. Results: The mRNA for the 4 subunits of GIRK were detected in the adult DRG. GTPγS enhanced inwardly rectifying potassium (K+) currents of the DRG neurons, while Ba2+inhibited such currents. Furthermore, the GIRK channels were shown to be coupled to the GABAB receptor, a member of the G protein-coupled receptor family, as baclofen increased the inwardly rectifying K+ currents. Conclusion: GIRK channels are expressed and functionally coupled with GABAB receptors in adult rat DRG neurons.

  20. Postembryonic neuronal addition in Zebrafish dorsal root ganglia is regulated by Notch signaling

    McGraw Hillary

    2012-06-01

    Full Text Available Abstract Background The sensory neurons and glia of the dorsal root ganglia (DRG arise from neural crest cells in the developing vertebrate embryo. In mouse and chick, DRG formation is completed during embryogenesis. In contrast, zebrafish continue to add neurons and glia to the DRG into adulthood, long after neural crest migration is complete. The molecular and cellular regulation of late DRG growth in the zebrafish remains to be characterized. Results In the present study, we use transgenic zebrafish lines to examine neuronal addition during postembryonic DRG growth. Neuronal addition is continuous over the period of larval development. Fate-mapping experiments support the hypothesis that new neurons are added from a population of resident, neural crest-derived progenitor cells. Conditional inhibition of Notch signaling was used to assess the role of this signaling pathway in neuronal addition. An increase in the number of DRG neurons is seen when Notch signaling is inhibited during both early and late larval development. Conclusions Postembryonic growth of the zebrafish DRG comes about, in part, by addition of new neurons from a resident progenitor population, a process regulated by Notch signaling.

  1. Implementation of DRG Payment in France: issues and recent developments.

    Or, Zeynep

    2014-08-01

    In France, a DRG-based payment system was introduced in 2004/2005 for funding acute services in all hospitals with the objectives of improving hospital efficiency, transparency and fairness in payments to public and private hospitals. Despite the initial consensus on the necessity of the reform, providers have become increasingly critical of the system because of the problems encountered during the implementation. In 2012 the government announced its intention to modify the payment model to better deal with its adverse effects. The paper reports on the issues raised by the DRG-based payment in the French hospital sector and provides an overview of the main problems with the French DRG payment model. It also summarises the evidence on its impact and presents recent developments for reforming the current model. DRG-based payment addressed some of the chronic problems inherent in the French hospital market and improved accountability and productivity of health-care facilities. However, it has also created new problems for controlling hospital activity and ensuring that care provided is medically appropriate. In order to alter its adverse effects the French DRG model needs to better align greater efficiency with the objectives of better quality and effectiveness of care. PMID:24962536

  2. Oxytocin-induced membrane hyperpolarization in pain-sensitive dorsal root ganglia neurons mediated by Ca(2+)/nNOS/NO/KATP pathway.

    Gong, L; Gao, F; Li, J; Li, J; Yu, X; Ma, X; Zheng, W; Cui, S; Liu, K; Zhang, M; Kunze, W; Liu, C Y

    2015-03-19

    Oxytocin (OT) plays an important role in pain modulation and antinociception in the central nervous system. However, little is known about its peripheral effects. This study was conducted to investigate the effect of OT on the electrical properties of neurons in the dorsal root ganglia (DRG) and the underlying mechanisms. DRG neurons from adult rats were acutely dissociated and cultured. Intracellular Ca(2+) was determined by fluorescent microscopy using an indicator dye. The electrical properties of DRG neurons were tested by patch-clamp recording. The oxytocin receptor (OTR) and neuronal nitric oxide synthase (nNOS) on DRG neurons were assessed with immunofluorescence assays. OTR co-localized with nNOS in most of Isolectin B4 (IB4)-binding cultured DRG neurons in rats. OT decreased the excitability, increased the outward current, and evoked the membrane hyperpolarization in cultured DRG neurons. Sodium nitroprusside (SNP), the donor of nitric oxide (NO), exerted similar effects as OT on the membrane potential of cultured DRG neurons. OT increased the production of NO in DRGs and cultured DRG neurons. Pre-treatment of the OTR antagonist atosiban or the selective nNOS inhibitor N-Propyl-l-arginine (NPLA) significantly attenuated the hyperpolarization effect evoked by OT. OT produced a concentration-dependent increase in intracellular Ca(2+) in DRG neurons that responds to capsaicin, which can be attenuated by atosiban, but not by NPLA. OT-evoked membrane hyperpolarization and increase of outward current were distinctly attenuated by glibenclamide, a blocker of ATP-sensitive K(+) (KATP) channel. OT might be an endogenous antinociceptive agent and the peripheral antinociceptive effects of OT are mediated by activation of the Ca(2+)/nNOS/NO/KATP pathway in DRG neurons. PMID:25617653

  3. 97. German Roentgen congress of the DRG. Program with abstracts

    The Volume with program and abstracts of the 97. German Roentgen congress of the DRG covers the following issues: The future of radiology - where are we heading? Cardiovascular imaging - the future; brain imaging - the essentials; chest imaging - not only nodules; abdominal imaging - the liver and beyond; muscoskeletal imaging - joints and bones; head and neck radiology - made easy.

  4. 42 CFR 476.84 - Changes as a result of DRG validation.

    2010-10-01

    ... 42 Public Health 4 2010-10-01 2010-10-01 false Changes as a result of DRG validation. 476.84... § 476.84 Changes as a result of DRG validation. A provider or practitioner may obtain a review by a QIO... in DRG assignment as a result of QIO validation activities....

  5. 42 CFR 412.517 - Revision of LTC-DRG group classifications and weighting factors.

    2010-10-01

    ... 42 Public Health 2 2010-10-01 2010-10-01 false Revision of LTC-DRG group classifications and... SERVICES Prospective Payment System for Long-Term Care Hospitals § 412.517 Revision of LTC-DRG group... the LTC-DRG classifications and recalibration of the weighting factors described in paragraph (a)...

  6. 42 CFR 412.10 - Changes in the DRG classification system.

    2010-10-01

    ... 42 Public Health 2 2010-10-01 2010-10-01 false Changes in the DRG classification system. 412.10... § 412.10 Changes in the DRG classification system. (a) General rule. CMS issues changes in the DRG classification system in a Federal Register notice at least annually. Except as specified in paragraphs (c)...

  7. 97. German Roentgen congress of the DRG. Program with abstracts; 97. Deutscher Roentgenkongress der DRG. Vollstaendiges Programm mit Abstracts

    NONE

    2016-05-15

    The Volume with program and abstracts of the 97. German Roentgen congress of the DRG covers the following issues: The future of radiology - where are we heading? Cardiovascular imaging - the future; brain imaging - the essentials; chest imaging - not only nodules; abdominal imaging - the liver and beyond; muscoskeletal imaging - joints and bones; head and neck radiology - made easy.

  8. Physicians' Patient Load per DRG, the Consumption of Hospital Resources, and the Incentives of the DRG Prospective Payment System.

    Munoz, Eric; And Others

    1990-01-01

    The relationship between numbers (high or low) of patients per diagnosis-related group (DRG) treated by individual physicians and hospital resource consumption of the patients at a large academic medical center was studied for the period 1985-87. The findings, although a result of many varied factors, suggest a relationship between the two…

  9. Growth Cone Biomechanics in Peripheral and Central Nervous System Neurons

    Urbach, Jeffrey; Koch, Daniel; Rosoff, Will; Geller, Herbert

    2012-02-01

    The growth cone, a highly motile structure at the tip of an axon, integrates information about the local environment and modulates outgrowth and guidance, but little is known about effects of external mechanical cues and internal mechanical forces on growth-cone mediated guidance. We have investigated neurite outgrowth, traction forces and cytoskeletal substrate coupling on soft elastic substrates for dorsal root ganglion (DRG) neurons (from the peripheral nervous system) and hippocampal neurons (from the central) to see how the mechanics of the microenvironment affect different populations. We find that the biomechanics of DRG neurons are dramatically different from hippocampal, with DRG neurons displaying relatively large, steady traction forces and maximal outgrowth and forces on substrates of intermediate stiffness, while hippocampal neurons display weak, intermittent forces and limited dependence of outgrowth and forces on substrate stiffness. DRG growth cones have slower rates of retrograde actin flow and higher density of localized paxillin (a protein associated with substrate adhesion complexes) compared to hippocampal neurons, suggesting that the difference in force generation is due to stronger adhesions and therefore stronger substrate coupling in DRG growth cones.

  10. Diagnosis-Related Groups (DRG and Hospital Business Performance Management

    Szynkiewicz Piotr

    2014-12-01

    Full Text Available The goal of this article is to present the possibility of using Diagnosis- Related Groups (DRG in the hospital management process and to analyse the need for business performance management on the part of hospital management staff. The following research methods were used: literature analysis, case studies, and poll analysis. It is not possible to increase the effectiveness of operation of healthcare entities without increasing the importance of IT systems and using DRG more effectively in the management process. Training users in IT and the use of DRGs is important to achieving hospital effectiveness. The increased importance of analyses and planning in a hospital should be reflected in the organisational structure of service providers. Hospital controllers should have a similar role to those present in most companies in other industries.

  11. 96. German Roentgen congress of the DRG. Program with abstracts

    The volume contains the program and the abstracts of the 96th German Roentgen congress of the DRG covering the following issues: profession and environment caused diseases, medical imaging - image processing - software, experimental radiology, whole-body diagnostics, gastro- and abdomen diagnostics, vascular diagnostics, device technology, heart diagnostics, interventional radiology (vascular diseases, oncology, others), pediatric radiology, contrast agents, head- and neck diagnostics, molecular imaging, musco-skeletal diagnostics, neuroradiology, emergency diagnostics - intensive care, oncologic imaging, quality management, radiation protection, thorax diagnostics.

  12. [G-DRG version 2007: a short overview].

    Müller, Marcel L; Forschner, Andrea; Luger, Thomas A; Rompel, Rainer; Roeder, Norbert; Hensen, Peter

    2007-09-01

    Just as in the year before, the structure and regulations of the new GDRG version 2007 was successfully agreed on a mutual basis by the national self-governing bodies in the German health care system. Although some problems in high-specialized medicine or day clinic care will remain, the current developments demonstrate once more the learning aptitude of the G-DRG-system. Some beneficial and major changes have been made in 2007, but most of them do not touch dermatology. Additional procedure-based payments have been introduced in 2007 including the parenteral administration of such expensive agents as etanercept and itraconazole. A statistical analysis of cost weights of the year 2006 versus 2007 for two university clinics suggests that in dermatology, the increasing complexity of the G-DRG system partly leads to lower cost weights. Overall in 2007 a remarkable increase of complexity and differentiation throughout the DRG-system can be identified as well as a careful expansion of procedure-based payments. PMID:17760899

  13. FPGA platform for MEMS Disc Resonance Gyroscope (DRG) control

    Keymeulen, Didier; Peay, Chris; Foor, David; Trung, Tran; Bakhshi, Alireza; Withington, Phil; Yee, Karl; Terrile, Rich

    2008-04-01

    Inertial navigation systems based upon optical gyroscopes tend to be expensive, large, power consumptive, and are not long lived. Micro-Electromechanical Systems (MEMS) based gyros do not have these shortcomings; however, until recently, the performance of MEMS based gyros had been below navigation grade. Boeing and JPL have been cooperating since 1997 to develop high performance MEMS gyroscopes for miniature, low power space Inertial Reference Unit applications. The efforts resulted in demonstration of a Post Resonator Gyroscope (PRG). This experience led to the more compact Disc Resonator Gyroscope (DRG) for further reduced size and power with potentially increased performance. Currently, the mass, volume and power of the DRG are dominated by the size of the electronics. This paper will detail the FPGA based digital electronics architecture and its implementation for the DRG which will allow reduction of size and power and will increase performance through a reduction in electronics noise. Using the digital control based on FPGA, we can program and modify in real-time the control loop to adapt to the specificity of each particular gyro and the change of the mechanical characteristic of the gyro during its life time.

  14. Association of Differentiation-Related Gene-1 (DRG1) with Breast Cancer Survival and in Vitro Impact of DRG1 Suppression

    Differentiation-related gene-1, DRG1, is a metastasis suppressor gene whose expression has been shown to be dysregulated in a number of malignancies. The current study examines the expression of DRG1 in a clinical breast cohort and its association with a number of clinical pathological factors using quantitative polymerase chain reaction. Additionally, DRG1 expression is targeted in vitro using ribozyme transgene technology to explore the function of DRG1 in two human breast cancer cell lines. Low levels of DRG1 were found in patients who developed metastasis (p = 0.036) and who died of breast cancer (p = 0.0048) compared to disease free patients. Knockdown of DRG1 also resulted in significantly increased invasion and motility, but decreased matrix-adhesion in MCF7 cells. Knockdown of DRG1 seemed to have minimal impact on the cellular functions of the MDA-MB-231 breast cancer cell line causing no significant differences in cell growth, invasion, motility or matrix-adhesion. Thus, DRG1 appears to be linked to development of metastasis and death in patients who died as a result of breast cancer and may be useful as a prognostic factor as its knockdown appears to be linked with increased invasion and motility and decreased adhesion in MCF7 breast cancer cells

  15. Association of Differentiation-Related Gene-1 (DRG1) with Breast Cancer Survival and in Vitro Impact of DRG1 Suppression

    Baig, Ruqia Mehmood [Metastasis and Angiogenesis Research Group, Institute of Cancer and Genetics, Cardiff University School of Medicine, Cardiff CF14 4XN (United Kingdom); Cancer Genetics Lab, COMSATS Institute of Information Technology, Islamabad 44000 (Pakistan); Sanders, Andrew J. [Metastasis and Angiogenesis Research Group, Institute of Cancer and Genetics, Cardiff University School of Medicine, Cardiff CF14 4XN (United Kingdom); Kayani, Mahmood Akhtar [Cancer Genetics Lab, COMSATS Institute of Information Technology, Islamabad 44000 (Pakistan); Jiang, Wen G., E-mail: jiangw@cf.ac.uk [Metastasis and Angiogenesis Research Group, Institute of Cancer and Genetics, Cardiff University School of Medicine, Cardiff CF14 4XN (United Kingdom)

    2012-07-10

    Differentiation-related gene-1, DRG1, is a metastasis suppressor gene whose expression has been shown to be dysregulated in a number of malignancies. The current study examines the expression of DRG1 in a clinical breast cohort and its association with a number of clinical pathological factors using quantitative polymerase chain reaction. Additionally, DRG1 expression is targeted in vitro using ribozyme transgene technology to explore the function of DRG1 in two human breast cancer cell lines. Low levels of DRG1 were found in patients who developed metastasis (p = 0.036) and who died of breast cancer (p = 0.0048) compared to disease free patients. Knockdown of DRG1 also resulted in significantly increased invasion and motility, but decreased matrix-adhesion in MCF7 cells. Knockdown of DRG1 seemed to have minimal impact on the cellular functions of the MDA-MB-231 breast cancer cell line causing no significant differences in cell growth, invasion, motility or matrix-adhesion. Thus, DRG1 appears to be linked to development of metastasis and death in patients who died as a result of breast cancer and may be useful as a prognostic factor as its knockdown appears to be linked with increased invasion and motility and decreased adhesion in MCF7 breast cancer cells.

  16. Association of Differentiation-Related Gene-1 (DRG1 with Breast Cancer Survival and in Vitro Impact of DRG1 Suppression

    Mahmood Akhtar Kayani

    2012-07-01

    Full Text Available Differentiation-related gene-1, DRG1, is a metastasis suppressor gene whose expression has been shown to be dysregulated in a number of malignancies. The current study examines the expression of DRG1 in a clinical breast cohort and its association with a number of clinical pathological factors using quantitative polymerase chain reaction. Additionally, DRG1 expression is targeted in vitro using ribozyme transgene technology to explore the function of DRG1 in two human breast cancer cell lines. Low levels of DRG1 were found in patients who developed metastasis (p = 0.036 and who died of breast cancer (p = 0.0048 compared to disease free patients. Knockdown of DRG1 also resulted in significantly increased invasion and motility, but decreased matrix-adhesion in MCF7 cells. Knockdown of DRG1 seemed to have minimal impact on the cellular functions of the MDA-MB-231 breast cancer cell line causing no significant differences in cell growth, invasion, motility or matrix-adhesion. Thus, DRG1 appears to be linked to development of metastasis and death in patients who died as a result of breast cancer and may be useful as a prognostic factor as its knockdown appears to be linked with increased invasion and motility and decreased adhesion in MCF7 breast cancer cells.

  17. Cannabinoid inhibition of the capsaicin-induced calcium response in rat dorsal root ganglion neurones

    Millns, Paul J; Chapman, Victoria; Kendall, David A.

    2001-01-01

    Cannabinoids have marked inhibitory effects on somatosensory processing, which may arise from actions at both peripheral and central cannabinoid receptors. Here, the effect of a synthetic cannabinoid agonist HU210 on capsaicin-evoked responses in adult rat dorsal root ganglion (DRG) neurones was studied. The vanilloid capsaicin produced a concentration-related increase in intracellular calcium in DRG neurones, which was significantly inhibited by HU210 (1 μM). The cannabinoid CB1 receptor ant...

  18. 96. German Roentgen congress of the DRG. Program with abstracts; 96. Deutscher Roentgenkongress der DRG. Vollstaendiges Program mit Abstracts

    NONE

    2015-05-15

    The volume contains the program and the abstracts of the 96th German Roentgen congress of the DRG covering the following issues: profession and environment caused diseases, medical imaging - image processing - software, experimental radiology, whole-body diagnostics, gastro- and abdomen diagnostics, vascular diagnostics, device technology, heart diagnostics, interventional radiology (vascular diseases, oncology, others), pediatric radiology, contrast agents, head- and neck diagnostics, molecular imaging, musco-skeletal diagnostics, neuroradiology, emergency diagnostics - intensive care, oncologic imaging, quality management, radiation protection, thorax diagnostics.

  19. Neuronal Subtype and Satellite Cell Tropism Are Determinants of Varicella-Zoster Virus Virulence in Human Dorsal Root Ganglia Xenografts In Vivo.

    Leigh Zerboni

    2015-06-01

    Full Text Available Varicella zoster virus (VZV, a human alphaherpesvirus, causes varicella during primary infection. VZV reactivation from neuronal latency may cause herpes zoster, post herpetic neuralgia (PHN and other neurologic syndromes. To investigate VZV neuropathogenesis, we developed a model using human dorsal root ganglia (DRG xenografts in immunodeficient (SCID mice. The SCID DRG model provides an opportunity to examine characteristics of VZV infection that occur in the context of the specialized architecture of DRG, in which nerve cell bodies are ensheathed by satellite glial cells (SGC which support neuronal homeostasis. We hypothesized that VZV exhibits neuron-subtype specific tropism and that VZV tropism for SGC contributes to VZV-related ganglionopathy. Based on quantitative analyses of viral and cell protein expression in DRG tissue sections, we demonstrated that, whereas DRG neurons had an immature neuronal phenotype prior to implantation, subtype heterogeneity was observed within 20 weeks and SGC retained the capacity to maintain neuronal homeostasis longterm. Profiling VZV protein expression in DRG neurons showed that VZV enters peripherin+ nociceptive and RT97+ mechanoreceptive neurons by both axonal transport and contiguous spread from SGC, but replication in RT97+ neurons is blocked. Restriction occurs even when the SGC surrounding the neuronal cell body were infected and after entry and ORF61 expression, but before IE62 or IE63 protein expression. Notably, although contiguous VZV spread with loss of SGC support would be predicted to affect survival of both nociceptive and mechanoreceptive neurons, RT97+ neurons showed selective loss relative to peripherin+ neurons at later times in DRG infection. Profiling cell factors that were upregulated in VZV-infected DRG indicated that VZV infection induced marked pro-inflammatory responses, as well as proteins of the interferon pathway and neuroprotective responses. These neuropathologic changes

  20. Neuronal Subtype and Satellite Cell Tropism Are Determinants of Varicella-Zoster Virus Virulence in Human Dorsal Root Ganglia Xenografts In Vivo.

    Zerboni, Leigh; Arvin, Ann

    2015-06-01

    Varicella zoster virus (VZV), a human alphaherpesvirus, causes varicella during primary infection. VZV reactivation from neuronal latency may cause herpes zoster, post herpetic neuralgia (PHN) and other neurologic syndromes. To investigate VZV neuropathogenesis, we developed a model using human dorsal root ganglia (DRG) xenografts in immunodeficient (SCID) mice. The SCID DRG model provides an opportunity to examine characteristics of VZV infection that occur in the context of the specialized architecture of DRG, in which nerve cell bodies are ensheathed by satellite glial cells (SGC) which support neuronal homeostasis. We hypothesized that VZV exhibits neuron-subtype specific tropism and that VZV tropism for SGC contributes to VZV-related ganglionopathy. Based on quantitative analyses of viral and cell protein expression in DRG tissue sections, we demonstrated that, whereas DRG neurons had an immature neuronal phenotype prior to implantation, subtype heterogeneity was observed within 20 weeks and SGC retained the capacity to maintain neuronal homeostasis longterm. Profiling VZV protein expression in DRG neurons showed that VZV enters peripherin+ nociceptive and RT97+ mechanoreceptive neurons by both axonal transport and contiguous spread from SGC, but replication in RT97+ neurons is blocked. Restriction occurs even when the SGC surrounding the neuronal cell body were infected and after entry and ORF61 expression, but before IE62 or IE63 protein expression. Notably, although contiguous VZV spread with loss of SGC support would be predicted to affect survival of both nociceptive and mechanoreceptive neurons, RT97+ neurons showed selective loss relative to peripherin+ neurons at later times in DRG infection. Profiling cell factors that were upregulated in VZV-infected DRG indicated that VZV infection induced marked pro-inflammatory responses, as well as proteins of the interferon pathway and neuroprotective responses. These neuropathologic changes observed in sensory

  1. Diabetes does not accelerate neuronal loss following nerve injury

    Severinsen, Kaare; Jakobsen, Johannes

    2007-01-01

    To determine the resistance of neuronal dorsal root ganglion (DRG) cells in experimental diabetes, we studied the neuronal cell loss after severe axonal injury in streptozotocin (STZ) diabetic rats with unilateral transection of the L5 spinal nerve for 12 weeks. Fifty 18-week-old inbred male Wist...

  2. Hyperexcitable neurons and altered non-neuronal cells in the compressed spinal ganglion

    Robert H. LaMotte; Chao MA

    2008-01-01

    The cell body or soma in the dosal root ganglion (DRG) is normally excitable and this excitability can increase and persist after an injury of peripheral sensory neurons. In a rat model of radicular pain, an intraforaminal implantation of a rod that chronically compressed the lumbar DRG ("CCD" model) resulted in neuronal somal hyperexcitability and spontaneous activity that was accom-panied by hyperalgesia in the ipsilateral hind paw. By the 5th day after onset of CCD, there was a novel upregulation in neuronal expression of the chemokine, monocyte chemoattractant protein-1 (MCP- 1 or CCL2) and also its receptor, CCR2. The neurons developed, in response to topically applied MCP-1, an excitatory response that they normally do not have. CCD also activated non-neuronal cells including, for example, the endothelial cells as evidenced by angiogenesis in the form of an increased number of capillaries in the DRG after 7 days. A working hypothesis is that the CCD induced changes in neurons and non-neuronal cells that may act together to promote the survival of the injured tissue. The release of ligands such as CCL2, in addition to possibly activating nociceptive neurons (maintaining the pain), may also act to preserve injured cells in the face of ischemia and hypoxia, for example, by promoting angiogenesis. Thus, somal hyperexcitability, as often said of inflammation, may represent a double edged sword.

  3. Six1 is a key regulator of the developmental and evolutionary architecture of sensory neurons in craniates

    Yajima, Hiroshi; SUZUKI, MAKOTO; Ochi, Haruki; Ikeda, Keiko; Sato, Shigeru; Yamamura, Ken-ichi; Ogino, Hajime; Ueno, Naoto; Kawakami, Kiyoshi

    2014-01-01

    Background Various senses and sensory nerve architectures of animals have evolved during adaptation to exploit diverse environments. In craniates, the trunk sensory system has evolved from simple mechanosensory neurons inside the spinal cord (intramedullary), called Rohon-Beard (RB) cells, to multimodal sensory neurons of dorsal root ganglia (DRG) outside the spinal cord (extramedullary). The fish and amphibian trunk sensory systems switch from RB cells to DRG during development, while amniot...

  4. Differentiation patterns of mouse embryonic stem cells and induced pluripotent stem cells into neurons.

    Nakamura, Mai; Kamishibahara, Yu; Kitazawa, Ayako; Kawaguchi, Hideo; Shimizu, Norio

    2016-05-01

    Mouse embryonic stem (ES) cells and induced pluripotent stem (iPS) cells have the ability to differentiate in vitro into various cell lineages including neurons. The differentiation of these cells into neurons has potential applications in regenerative medicine. Previously, we reported that a chick dorsal root ganglion (DRG)-conditioned medium (CM) promoted the differentiation of mouse ES and iPS cells into neurons. Here, we used real-time PCR to investigate the differentiation patterns of ES and iPS cells into neurons when DRG-CM was added. DRG-CM promoted the expression levels of βIII-tubulin gene (a marker of postmitotic neurons) in ES and iPS cells. ES cells differentiated into neurons faster than iPS cells, and the maximum peaks of gene expression involved in motor, sensory, and dopaminergic neurons were different. Rho kinase (ROCK) inhibitors could be very valuable at numerous stages in the production and use of stem cells in basic research and eventual cell-based therapies. Thus, we investigated whether the addition of a ROCK inhibitor Y-27632 and DRG-CM on the basis of the differentiation patterns promotes the neuronal differentiation of ES cells. When the ROCK inhibitor was added to the culture medium at the initial stages of cultivation, it stimulated the neuronal differentiation of ES cells more strongly than that stimulated by DRG-CM. Moreover, the combination of the ROCK inhibitor and DRG-CM promoted the neuronal differentiation of ES cells when the ROCK inhibitor was added to the culture medium at day 3. The ROCK inhibitor may be useful for promoting neuronal differentiation of ES cells. PMID:25354731

  5. Hospital Coding Practice, Data Quality, and DRG-Based Reimbursement under the Thai Universal Coverage Scheme

    Pongpirul, Krit

    2011-01-01

    In the Thai Universal Coverage scheme, hospital providers are paid for their inpatient care using Diagnosis Related Group (DRG) reimbursement. Questionable quality of the submitted DRG codes has been of concern whereas knowledge about hospital coding practice has been lacking. The objectives of this thesis are (1) To explore hospital coding…

  6. 42 CFR 478.15 - QIO review of changes resulting from DRG validation.

    2010-10-01

    ... 42 Public Health 4 2010-10-01 2010-10-01 false QIO review of changes resulting from DRG validation... review of changes resulting from DRG validation. (a) General rules. (1) A provider or practitioner dissatisfied with a change to the diagnostic or procedural coding information made by a QIO as a result of...

  7. Caspase-Mediated Apoptosis in Sensory Neurons of Cultured Dorsal Root Ganglia in Adult Mouse

    Hamid Reza Momeni

    2013-01-01

    Full Text Available Objective: Sensory neurons in dorsal root ganglia (DRG undergo apoptosis after peripheral nerve injury. The aim of this study was to investigate sensory neuron death and the mechanism involved in the death of these neurons in cultured DRG.Materials and Methods: In this experimental study, L5 DRG from adult mouse were dissected and incubated in culture medium for 24, 48, 72 and 96 hours. Freshly dissected and cultured DRG were then fixed and sectioned using a cryostat. Morphological and biochemical features of apoptosis were investigated using fluorescent staining (Propidium iodide and Hoechst 33342 and the terminal Deoxynucleotide transferase dUTP nick end labeling (TUNEL method respectively. To study the role of caspases, general caspase inhibitor (Z-VAD.fmk, 100 μM and immunohistochemistry for activated caspase-3 were used.Results: After 24, 48, 72 and 96 hours in culture, sensory neurons not only displayed morphological features of apoptosis but also they appeared TUNEL positive. The application of Z-VAD.fmk inhibited apoptosis in these neurons over the same time period. In addition, intense activated caspase-3 immunoreactivity was found both in the cytoplasm and the nuclei of these neurons after 24 and 48 hours.Conclusion: Results of the present study show caspase-dependent apoptosis in the sensory neurons of cultured DRG from adult mouse.

  8. 42 CFR 476.93 - Opportunity to discuss proposed initial denial determination and changes as a result of a DRG...

    2010-10-01

    ... determination and changes as a result of a DRG validation. 476.93 Section 476.93 Public Health CENTERS FOR... initial denial determination and changes as a result of a DRG validation. Before a QIO reaches an initial denial determination or makes a change as a result of a DRG validation, it must— (a) Promptly notify...

  9. 42 CFR 476.85 - Conclusive effect of QIO initial denial determinations and changes as a result of DRG validations.

    2010-10-01

    ... determinations and changes as a result of DRG validations. 476.85 Section 476.85 Public Health CENTERS FOR... denial determinations and changes as a result of DRG validations. A QIO initial denial determination or change as a result of DRG validation is final and binding unless, in accordance with the procedures...

  10. 42 CFR 476.94 - Notice of QIO initial denial determination and changes as a result of a DRG validation.

    2010-10-01

    ... changes as a result of a DRG validation. 476.94 Section 476.94 Public Health CENTERS FOR MEDICARE... changes as a result of a DRG validation. (a) Notice of initial denial determination—(1) Parties to be... retrospective review, (excluding DRG validation and post procedure review), within 3 working days of the...

  11. 42 CFR 476.96 - Review period and reopening of initial denial determinations and changes as a result of DRG...

    2010-10-01

    ... determinations and changes as a result of DRG validations. 476.96 Section 476.96 Public Health CENTERS FOR... initial denial determinations and changes as a result of DRG validations. (a) General timeframe. A QIO or... initial denial determination or a change as a result of a DRG validation. (b) Extended timeframes. (1)...

  12. Intrathecal administration of rapamycin inhibits the phosphorylation of DRG Nav1.8 and attenuates STZ-induced painful diabetic neuropathy in rats.

    He, Wan-You; Zhang, Bin; Xiong, Qing-Ming; Yang, Cheng-Xiang; Zhao, Wei-Cheng; He, Jian; Zhou, Jun; Wang, Han-Bing

    2016-04-21

    The mammalian target of rapamycin (mTOR) is a key regulator of mRNA translation and protein synthesis, and it is specifically inhibited by rapamycin. In chronic pain conditions, mTOR-mediated local protein synthesis is crucial for neuronal hyperexcitability and synaptic plasticity. The tetrodotoxin-resistant (TTX-R) sodium channel Nav1.8 plays a major role in action potential initiation and propagation and cellular excitability in DRG (dorsal root ganglion) neurons. In this study, we investigated if mTOR modulates the phosphorylation of Nav1.8 that is associated with neuronal hyperexcitability and behavioral hypersensitivity in STZ-induced diabetic rats. Painful diabetic neuropathy (PDN) was induced in Sprague-Dawley rats by intraperitoneal injection with streptozotocin (STZ) at 60mg/kg. After the onset of PDN, the rats received daily intrathecal administrations of rapamycin (1μg, 3μg, or 10μg/day) for 7 days; other diabetic rats received the same volumes of dimethyl sulfoxide (DMSO). Herein, we demonstrate a marked increase in protein expression of total mTOR and phospho-mTOR (p-mTOR) together with the up-regulation of phosphor-Nav1.8 (p-Nav1.8) prior to the mechanical withdrawal threshold reaching a significant reduction in dorsal root ganglions (DRGs). Furthermore, the intrathecal administration of rapamycin, inhibiting the activity of mTOR, suppressed the phosphorylation of DRG Nav1.8, reduced the TTX-R current density, heightened the voltage threshold for activation and lowered the voltage threshold for inactivation and relieved mechanical hypersensitivity in diabetic rats. An intrathecal injection (i.t.) of rapamycin inhibited the phosphorylation and enhanced the functional availability of DRG Nav1.8 attenuated STZ-induced hyperalgesia. These results suggest that rapamycin is a potential therapeutic intervention for clinical PDN. PMID:26946108

  13. srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons by inactivating RAC1

    Quan-Peng Zhang; Hai-Ying Zhang; Xian-Fang Zhang; Jiu-Hong Zhao; Zhi-Jian Ma; Dan Zhao; Xi-Nan Yi

    2014-01-01

    Objective:To explore effect of srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons.Methods:In this study, expression ofSlit1 was observed predominantly in the glia, while expression ofRobo2 and srGAP3 was detected in sensory neurons of postnatal rat cultured dorsal root ganglion(DRG).Furthermore, upregulation of srGAP3 following sciatic nerve transection was detected by immunohistochemistry andWestern blotting.Results:It was observed that inhibition of neurite outgrowth in cultured adultDRG neurons following treatment with anti-srGAP3 or anti-Robo2 was more effectively(1.5-fold higher) than that following treatment with an anti-BDNF positive control antibody.It demonstrated that srGAP3 interacted withRobo2 andSlit1 protein to decreaseRac1-GTP activity in cultured adult ratDRG neurons and the opposite effect onRac1-GTP activity was detected by co-immunoprecipitation and immunoblotting analyses following treatment with anti-Robo2 or anti-srGAP3.These data demonstrated a role for srGAP3 in neurite outgrowth ofDRG sensory neurons.Conclusions:Our observations suggest that srGAP3 promotes neurite outgrowth and filopodial growth cones by interacting withRobo2 to inactivateRac1 in mammalianDRG neurons.

  14. [The DRG law--planning gone wrong? The conversion from the physician's point of view].

    Rochell, Bernhard; Roeder, Norbert

    2002-08-01

    With the Health Care Reform Act of 2000 the German government initiated the introduction of a new hospital funding system based on an internationally used Diagnosis Related Group (DRG) system. In June 2000, the medical self-governing bodies (consisting of representatives of the German Hospital Federation, the German Statutory Health Insurance Funds and the Association of Private Health Insurances) commissioned for the execution of this project decided to use the Australian Refined DRG system, version 4.1 (AR DRG) as the basis for the future German (Refined) DRG system (G-DRG). It is planned for voluntary hospitals to replace the previous German hospital reimbursement system by the new DRG-based hospital funding system in January 2003. From January 2004, the change of the reimbursement system will be mandatory for all hospitals with the exception of psychiatric, psychosomatic and psychotherapeutic hospitals or units. The new reimbursement system is not only intended to cover acute hospital care but also parts of early rehabilitation, palliative and sub-acute care. Due to its economic incentives the effects of introducing the DRG system in Germany will not only affect the hospital sector but ambulatory care, nursing and rehabilitation as well. PMID:12244873

  15. Sympathectomy attenuates excitability of dorsal root ganglion neurons and pain behaviour in a lumbar radiculopathy model

    Iwase, T; Takebayashi, T; Tanimoto, K; Terashima, Y.; Miyakawa, T.; Kobayashi, T.; Tohse, N.; Yamashita, T

    2012-01-01

    Objectives In order to elucidate the influence of sympathetic nerves on lumbar radiculopathy, we investigated whether sympathectomy attenuated pain behaviour and altered the electrical properties of the dorsal root ganglion (DRG) neurons in a rat model of lumbar root constriction. Methods Sprague-Dawley rats were divided into three experimental groups. In the root constriction group, the left L5 spinal nerve root was ligated proximal to the DRG as a lumbar radiculopathy model. In the root con...

  16. Long-Term Activation of Group I Metabotropic Glutamate Receptors Increases Functional TRPV1-Expressing Neurons in Mouse Dorsal Root Ganglia

    Masuoka, Takayoshi; Kudo, Makiko; Yoshida, Junko; Ishibashi, Takaharu; Muramatsu, Ikunobu; Kato, Nobuo; Imaizumi, Noriko; Nishio, Matomo

    2016-01-01

    Damaged tissues release glutamate and other chemical mediators for several hours. These chemical mediators contribute to modulation of pruritus and pain. Herein, we investigated the effects of long-term activation of excitatory glutamate receptors on functional expression of transient receptor potential vaniloid type 1 (TRPV1) in dorsal root ganglion (DRG) neurons and then on thermal pain behavior. In order to detect the TRPV1-mediated responses in cultured DRG neurons, we monitored intracell...

  17. A comparison of peripheral and central axotomy effects on neurofilament and tubulin gene expression in rat dorsal root ganglion neurons

    The expression of major cytoskeletal protein mRNAs was studied in adult rat dorsal root ganglion (DRG) neurons after crushing either their central or peripheral branch axons. mRNA levels in DRG neurons were examined by quantitative in situ hybridization with radiolabeled cDNA probes specific for the low-molecular-weight neurofilament protein (NF-L) and beta-tubulin. The large-sized (greater than 1000 microns 2) neurons which give rise to myelinated axons in lumbar ganglia (L4 and L5) were studied 1 d through 8 weeks after either dorsal root or sciatic nerve crush. NF-L and beta-tubulin mRNA levels in axotomized DRG neurons were compared to those in contralateral control DRG neurons, as well as to those in normal (completely untreated) DRG cells. In the case of NF-L mRNA, changes were observed after central as well as peripheral branch axotomy and the time course and magnitude of changes were similar after both types of axotomy. NF-L mRNA levels initially decreased (first 2 weeks after crush) and then began to return towards control levels at longer survival times. Similar, but less pronounced, changes in NF-L mRNA levels also occurred in contralateral DRG neurons (which were uninjured); the changes in contralateral neurons were not simply a result of surgical stress since no changes in NF-L mRNA levels were observed in sham-operated DRG neurons. In the case of tubulin mRNA, changes were observed after central as well as peripheral branch axotomy by in situ hybridization, but the time course and magnitude of changes were different after each type of axotomy

  18. Somatosensory neuron types identified by high-coverage single-cell RNA-sequencing and functional heterogeneity.

    Li, Chang-Lin; Li, Kai-Cheng; Wu, Dan; Chen, Yan; Luo, Hao; Zhao, Jing-Rong; Wang, Sa-Shuang; Sun, Ming-Ming; Lu, Ying-Jin; Zhong, Yan-Qing; Hu, Xu-Ye; Hou, Rui; Zhou, Bei-Bei; Bao, Lan; Xiao, Hua-Sheng; Zhang, Xu

    2016-01-01

    Sensory neurons are distinguished by distinct signaling networks and receptive characteristics. Thus, sensory neuron types can be defined by linking transcriptome-based neuron typing with the sensory phenotypes. Here we classify somatosensory neurons of the mouse dorsal root ganglion (DRG) by high-coverage single-cell RNA-sequencing (10 950 ± 1 218 genes per neuron) and neuron size-based hierarchical clustering. Moreover, single DRG neurons responding to cutaneous stimuli are recorded using an in vivo whole-cell patch clamp technique and classified by neuron-type genetic markers. Small diameter DRG neurons are classified into one type of low-threshold mechanoreceptor and five types of mechanoheat nociceptors (MHNs). Each of the MHN types is further categorized into two subtypes. Large DRG neurons are categorized into four types, including neurexophilin 1-expressing MHNs and mechanical nociceptors (MNs) expressing BAI1-associated protein 2-like 1 (Baiap2l1). Mechanoreceptors expressing trafficking protein particle complex 3-like and Baiap2l1-marked MNs are subdivided into two subtypes each. These results provide a new system for cataloging somatosensory neurons and their transcriptome databases. PMID:26691752

  19. Beta-amyloid peptide blocks the fast-inactivating K+ current in rat hippocampal neurons.

    Good, T A; Smith, D. O.; Murphy, R M

    1996-01-01

    Deposition of beta-amyloid peptide (A beta) in senile plaques is a hallmark of Alzheimer disease neuropathology. Chronic exposure of neuronal cultures to synthetic A beta is directly toxic, or enhances neuronal susceptibility to excitotoxins. Exposure to A beta may cause a loss of cellular calcium homeostasis, but the mechanism by which this occurs is uncertain. In this work, the acute response of rat hippocampal neurons to applications of synthetic A beta was measured using whole-cell voltag...

  20. Topographic Digital Raster Graphics, USGS DRG's; reprojected to state plane, Published in 2006, Washoe County.

    NSGIC GIS Inventory (aka Ramona) — This Topographic Digital Raster Graphics dataset, was produced all or in part from Hardcopy Maps information as of 2006. It is described as 'USGS DRG's; reprojected...

  1. Indications for CT and MR arthrography. Recommendations of the Musculoskeletal Workgroup of the DRG; Indikationen der MR- und CT-Arthrografie. Empfehlungen der AG Muskuloskelettale Radiologie der DRG

    Fischer, W. [Hessingpark Clinic, Radiologengemeinschaft Augsburg (Germany); Bohndorf, K.; Zentner, J. [Zentralklinikum Augsburg (Germany). Klinik fuer Diagnostische Radiologie und Neuroradiologie; Kreitner, K.F. [Universitaetsklinikum Mainz (Germany). Klinik und Poliklinik fuer Radiologie; Schmitt, R. [Herz- und Gefaessklinik GmbH, Bad Neustadt an der Saale (Germany). Inst. fuer Diagnostische und Interventionelle Radiologie; Woertler, K. [Klinikum rechts der Isar, Muenchen (Germany). Inst. fuer Roentgendiagnostik

    2009-05-15

    The ongoing discussion about CT and MR arthrography is at least in part due to the lack of definite guidelines. The intention of the musculoskeletal workgroup of the DRG (Deutsche Roentgengesellschaft) was the establishment of recommendations for general guidance. After review of the recent literature, the indications for arthrographic examinations were discussed during a consensus meeting. Since the published data are insufficient and partially contradictory, no precise statements could be extracted from the literature. Therefore, the proposed recommendations are mainly based on expert opinions. In this review the main statements of the published literature are summarized and the recommendations of the musculoskeletal workgroup of the DRG are presented. (orig.)

  2. The majority of dorsal spinal cord gastrin releasing peptide is synthesized locally whereas neuromedin B is highly expressed in pain- and itch-sensing somatosensory neurons

    Fleming Michael S

    2012-07-01

    Full Text Available Abstract Background Itch is one of the major somatosensory modalities. Some recent findings have proposed that gastrin releasing peptide (Grp is expressed in a subset of dorsal root ganglion (DRG neurons and functions as a selective neurotransmitter for transferring itch information to spinal cord interneurons. However, expression data from public databases and earlier literatures indicate that Grp mRNA is only detected in dorsal spinal cord (dSC whereas its family member neuromedin B (Nmb is highly expressed in DRG neurons. These contradictory results argue that a thorough characterization of the expression of Grp and Nmb is warranted. Findings Grp mRNA is highly expressed in dSC but is barely detectable in DRGs of juvenile and adult mice. Anti-bombesin serum specifically recognizes Grp but not Nmb. Grp is present in a small number of small-diameter DRG neurons and in abundance in layers I and II of the spinal cord. The reduction of dSC Grp after dorsal root rhizotomy is significantly different from those of DRG derived markers but similar to that of a spinal cord neuronal marker. Double fluorescent in situ of Nmb and other molecular markers indicate that Nmb is highly and selectively expressed in nociceptive and itch-sensitive DRG neurons. Conclusion The majority of dSC Grp is synthesized locally in dorsal spinal cord neurons. On the other hand, Nmb is highly expressed in pain- and itch-sensing DRG neurons. Our findings provide direct anatomic evidence that Grp could function locally in the dorsal spinal cord in addition to its roles in DRG neurons and that Nmb has potential roles in nociceptive and itch-sensitive neurons. These results will improve our understanding about roles of Grp and Nmb in mediating itch sensation.

  3. Correlation of the electrophysiological profiles and sodium channel transcripts of individual rat dorsal root ganglia neurons

    Mohamed Chahine

    2014-09-01

    We used the patch-clamp technique in the whole-cell configuration to record Na+ currents and action potentials from acutely dissociated small diameter DRG neurons (<30 µM from adult rats. We also performed single cell qPCR on the same neurons. Our results revealed that there is a strong correlation between Na+ currents and mRNA transcripts in individual neurons. A cluster analysis showed that subgroups formed by Na+ channel transcripts by mRNA quantification have different biophysical properties. In addition, the firing frequency of the neurons was not affected by the relative populations of Na+ channel. The synergy between populations of Na+ channel in individual small diameter DRG neurons gives each neuron a unique electrophysiological profile. The Na+ channel remodeling that occurs in different pathological pain states may be responsible for the sensitization of the neurons.

  4. Mechanical Compression and Nucleus Pulposus Application on Dorsal Root Ganglia Differentially Modify Evoked Neuronal Activity in the Thalamus

    Nilsson, Elin; Brisby, Helena; Rask, Katarina; Hammar, Ingela

    2013-01-01

    Abstract A combination of mechanical compression caused by a protruding disc and leakage of nucleus pulposus (NP) from the disc core is presumed to contribute to intervertebral disc hernia-related pain. Experimental models of disc hernia including both components have resulted in changes in neuronal activity at the level of the dorsal root ganglion (DRG) and spinal cord, but changes within the brain have been less well studied. However, acute application of NP to a DRG without mechanical comp...

  5. The role of matrix metalloproteinases in regulating neuronal and nonneuronal cell invasion into PEGylated fibrinogen hydrogels.

    Sarig-Nadir, Offra; Seliktar, Dror

    2010-09-01

    Injured peripheral nerve tissue could benefit from biomaterial nerve guidance conduits (NGCs) that are designed to promote neuronal regeneration. Nerve regeneration is a complex multi-step process that involves the remodeling of the ECM surrounding the regenerating neural tissue. Hydrogel biomaterials have been used as provisional matrices to regulate this regeneration process by providing the desired physical properties and controllable degradation characteristics. The purpose of this investigation was to understand the mechanism by which nerve cells penetrate into a hydrogel made from PEGylated fibrinogen. In this context, the dorsal root ganglion (DRG) assay was used as an in vitro model to study the cellular invasion behavior of both neural and nonneuronal cells. Our hypothesis stipulated that DRG cells employ matrix metalloproteinases (MMPs) in order to degrade the dense hydrogel matrix and penetrate the biomaterial. Three dimensional (3D) DRG-hydrogel constructs were cultured with MMP inhibitors (MMPi) and the effect of the inhibitors on DRG cell outgrowth was investigated. We also examined the effect of inhibitors on two dimensional (2D) DRG cell outgrowth on PEGylated fibrinogen hydrogels and on tissue culture polystyrene (TCP). Our results demonstrate that DRG cell outgrowth into and onto PEGylated fibrinogen hydrogels was inhibited by MMPi and that the outgrowth characteristics was dependent on the type of inhibitor and its concentration. MMP-3i and MMP-8i decreased both neuronal and nonneuronal outgrowth, where MMP-3i had a stronger inhibitory effect on nonneuronal cells. MMP-2/9i, on the other hand, affected the neuronal outgrowth much more than the others. We concluded that MMPs play a central role in the process of DRG cell penetration into PEGylated fibrinogen hydrogels and may also regulate the adhesion, migration and elongation of neuronal cells on the surface of these hydrogel biomaterials. PMID:20537384

  6. Inflammatory mediator bradykinin increases population of sensory neurons expressing functional T-type Ca2+ channels

    Huang, Dongyang; Liang, Ce; Zhang, Fan; Men, Hongchao; Du, Xiaona; Gamper, Nikita; Zhang, Hailin

    2016-01-01

    T-type Ca2+ channels are important regulators of peripheral sensory neuron excitability. Accordingly, T-type Ca2+ currents are often increased in various pathological pain conditions, such as inflammation or nerve injury. Here we investigated effects of inflammation on functional expression of T-type Ca2+ channels in small-diameter cultured dorsal root ganglion (DRG) neurons. We found that overnight treatment of DRG cultures with a cocktail of inflammatory mediators bradykinin (BK), adenosine triphosphate (ATP), norepinephrine (NE) and prostaglandin E2 (PGE2) strongly increased the population size of the small-diameter neurons displaying low-voltage activated (LVA, T-type) Ca2+ currents while having no effect on the peak LVA current amplitude. When applied individually, BK and ATP also increased the population size of LVA-positive neurons while NE and PGE2 had no effect. The PLC inhibitor U-73122 and B2 receptor antagonist, Hoe-140, both abolished the increase of the population of LVA-positive DRG neurons. Inflammatory treatment did not affect CaV3.2 mRNA or protein levels in DRG cultures. Furthermore, an ubiquitination inhibitor, MG132, did not increase the population of LVA-positive neurons. Our data suggest that inflammatory mediators BK and ATP increase the abundance of LVA-positive DRG neurons in total neuronal population by stimulating the recruitment of a ‘reserve pool’ of CaV3.2 channels, particularly in neurons that do not display measurable LVA currents under control conditions. PMID:26944020

  7. Inflammatory mediator bradykinin increases population of sensory neurons expressing functional T-type Ca(2+) channels.

    Huang, Dongyang; Liang, Ce; Zhang, Fan; Men, Hongchao; Du, Xiaona; Gamper, Nikita; Zhang, Hailin

    2016-04-29

    T-type Ca(2+) channels are important regulators of peripheral sensory neuron excitability. Accordingly, T-type Ca(2+) currents are often increased in various pathological pain conditions, such as inflammation or nerve injury. Here we investigated effects of inflammation on functional expression of T-type Ca(2+) channels in small-diameter cultured dorsal root ganglion (DRG) neurons. We found that overnight treatment of DRG cultures with a cocktail of inflammatory mediators bradykinin (BK), adenosine triphosphate (ATP), norepinephrine (NE) and prostaglandin E2 (PGE2) strongly increased the population size of the small-diameter neurons displaying low-voltage activated (LVA, T-type) Ca(2+) currents while having no effect on the peak LVA current amplitude. When applied individually, BK and ATP also increased the population size of LVA-positive neurons while NE and PGE2 had no effect. The PLC inhibitor U-73122 and B2 receptor antagonist, Hoe-140, both abolished the increase of the population of LVA-positive DRG neurons. Inflammatory treatment did not affect CaV3.2 mRNA or protein levels in DRG cultures. Furthermore, an ubiquitination inhibitor, MG132, did not increase the population of LVA-positive neurons. Our data suggest that inflammatory mediators BK and ATP increase the abundance of LVA-positive DRG neurons in total neuronal population by stimulating the recruitment of a 'reserve pool' of CaV3.2 channels, particularly in neurons that do not display measurable LVA currents under control conditions. PMID:26944020

  8. Nesfatin-1 increases intracellular calcium concentration by protein kinase C activation in cultured rat dorsal root ganglion neurons.

    Ozcan, Mete; Gok, Zeynep Betul; Kacar, Emine; Serhatlioglu, Ihsan; Kelestimur, Haluk

    2016-04-21

    Nesfatin-1 is a recently identified anorexigenic hypothalamic polypeptide derived from the posttranslational processing of nucleobindin 2 (NUCB2). Several studies have indicated that this neuropeptide may be participated in somatosensory and visceral transmission including pain signals in addition to energy metabolism. The aim of this study was to explore the possible role of nesfatin-1 in the transmission of peripheral neural signals by investigating the effects of nesfatin-1 on intracellular free calcium levels ([Ca(2+)]i) in cultured neonatal rat dorsal root ganglion (DRG) neurons. The effects of nesfatin-1 on [Ca(2+)]i in DRG neurons were investigated by using an in vitro calcium imaging system. DRG neurons were grown in primary culture following enzymatic and mechanical dissociation of ganglia from 1-or 2-day-old neonatal Wistar rats. Using the fura-2-based calcium imaging technique, the effects of nesfatin-1 on [Ca(2+)]i and role of the protein kinase C (PKC)-mediated pathway in nesfatin-1 effect were assessed. Nesfatin-1 elevated [Ca(2+)]i in cultured DRG neurons. The response was prevented by pretreating the cells with pertussis toxin. The protein kinase C inhibitor chelerythrine chloride suppressed nesfatin-1-induced rise in [Ca(2+)]i. The result shows that nesfatin-1 interacts with a G protein-coupled receptor, leading to an increase of [Ca(2+)]i, which is linked to protein kinase C activation in cultured rat DRG neurons. PMID:26975784

  9. Policy trends and reforms in the German DRG-based hospital payment system.

    Klein-Hitpaß, Uwe; Scheller-Kreinsen, David

    2015-03-01

    A central structural point in all DRG-based hospital payment systems is the conversion of relative weights into actual payments. In this context policy makers need to address (amongst other things) (a) how the price level of DRG-payments from one period to the following period is changed and (b) whether and how hospital payments based on DRGs are to be differentiated beyond patient characteristics, e.g. by organizational, regional or state-level factors. Both policy problems can be and in international comparison often are empirically addressed. In Germany relative weights are derived from a highly sophisticated empirical cost calculation, whereas the annual changes of DRG-based payments (base rates) as well as the differentiation of DRG-based hospital payments beyond patient characteristics are not empirically addressed. Rather a complex set of regulations and quasi-market negotiations are applied. There were over the last decade also timid attempts to foster the use of empirical data to address these points. However, these reforms failed to increase the fairness, transparency and rationality of the mechanism to convert relative weights into actual DRG-based hospital payments. PMID:25638648

  10. The Endoplasmic Reticulum of Dorsal Root Ganglion Neurons Contains Functional TRPV1 Channels*

    Gallego-Sandín, Sonia; Rodríguez-García, Arancha; Alonso, María Teresa; García-Sancho, Javier

    2009-01-01

    Transient receptor potential vanilloid type 1 (TRPV1) is a plasma membrane Ca2+ channel involved in transduction of painful stimuli. Dorsal root ganglion (DRG) neurons express ectopic but functional TRPV1 channels in the endoplasmic reticulum (ER) (TRPV1ER). We have studied the properties of TRPV1ER in DRG neurons and HEK293T cells expressing TRPV1. Activation of TRPV1ER with capsaicin or other vanilloids produced an increase of cytosolic Ca2+ due to Ca2+ release from the ER. The decrease of ...

  11. Evaluation of the productivity of Brazilian hospitals by the methodology of diagnosis related group (DRG)

    Filho, Jose Carlos Serufo; Grillo, Tania Moreira

    2014-01-01

    The management requires a hospital organization to provision their costs/expenses with tools that approximate reality. The task of measuring productivity can be complex and uncertain, several methods are tested and the use of the DRG has been efficient, being used to assess the productivity through clinical outcomes. Cross-sectional study evaluated 145.710 hospitalizations in the period 2012-2014, using the DRG methodology for measuring productivity from the median length of hospitalization. When we group all hospitalizations in clinical (37.6%) and surgical (62.4%), multiple analyzes could be made according to this criterion. The DRG as a tool for prediction of hospital days is an effective alternative, thereby contributing to the control of productivity that directly influences the costs of hospital expenses and product and service quality.

  12. Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity

    Chiu, Isaac M; Barrett, Lee B.; Williams, Erika K.; Strochlic, David E; Lee, Seungkyu; Weyer, Andy D.; Lou, Shan; Bryman, Gregory S; Roberson, David P.; Ghasemlou, Nader; Piccoli, Cara; Ahat, Ezgi; Wang, Victor; Cobos del Moral, Enrique Jos??; Cheryl L. Stucky

    2014-01-01

    The somatosensory nervous system is critical for the organism's ability to respond to mechanical, thermal, and nociceptive stimuli. Somatosensory neurons are functionally and anatomically diverse but their molecular profiles are not well-defined. Here, we used transcriptional profiling to analyze the detailed molecular signatures of dorsal root ganglion (DRG) sensory neurons. We used two mouse reporter lines and surface IB4 labeling to purify three major non-overlapping classes of neurons: 1)...

  13. Regulation of Piezo2 Mechanotransduction by Static Plasma Membrane Tension in Primary Afferent Neurons.

    Jia, Zhanfeng; Ikeda, Ryo; Ling, Jennifer; Viatchenko-Karpinski, Viacheslav; Gu, Jianguo G

    2016-04-22

    The Piezo2 channel is a newly identified mammalian mechanical transducer that confers rapidly adapting mechanically activated (RA-MA) currents in primary afferent neurons. The Piezo2 channels sense rapid membrane displacement, but it is not clear whether they are sensitive to osmotic swelling, which slowly increases static plasma membrane tension (SPMT). Here, we show that SPMT exerts a profound impact on the mechanical sensitivity of RA-MA channels in primary afferent neurons. RA-MA currents are greatly enhanced, and the mechanical threshold was reduced in both primary afferent neurons of rat dorsal root ganglia (DRG) and HEK293 cells heterologously expressing Piezo2 when these cells undergo osmotic swelling to increase SPMT. Osmotic swelling switches the kinetics of RA-MA currents to the slowly adapting type in both cultured DRG neurons and HEK293 cells heterologously expressing Piezo2. The potentiation of RA-MA currents is abolished when cultured DRG neurons are treated with cytochalasin D, an actin filament disruptor that prevents SPMT of cultured DRG neurons from an increase by osmotic swelling. Osmotic swelling significantly increases DRG neuron mechano-excitability such that a subthreshold mechanical stimulus can result in action potential firing. Behaviorally, the mechanical hind paw withdrawal threshold in rats is reduced following the injection of a hypotonic solution, but this osmotic effect is abolished when cytochalasin D or Gd(3+) is co-administered with the hypo-osmotic solution. Taken together, our findings suggest that Piezo2-mediated mechanotransduction is regulated by SPMT in primary afferent neurons. Because SPMT can be changed by multiple biological factors, our findings may have broad implications in mechanical sensitivity under physiological and pathological conditions. PMID:26929410

  14. Indications for CT and MR arthrography. Recommendations of the Musculoskeletal Workgroup of the DRG

    The ongoing discussion about CT and MR arthrography is at least in part due to the lack of definite guidelines. The intention of the musculoskeletal workgroup of the DRG (Deutsche Roentgengesellschaft) was the establishment of recommendations for general guidance. After review of the recent literature, the indications for arthrographic examinations were discussed during a consensus meeting. Since the published data are insufficient and partially contradictory, no precise statements could be extracted from the literature. Therefore, the proposed recommendations are mainly based on expert opinions. In this review the main statements of the published literature are summarized and the recommendations of the musculoskeletal workgroup of the DRG are presented. (orig.)

  15. CC chemokine ligand 2 upregulates the current density and expression of TRPV1 channels and Nav1.8 sodium channels in dorsal root ganglion neurons

    Kao Der-Jang

    2012-08-01

    Full Text Available Abstract Background Inflammation or nerve injury-induced upregulation and release of chemokine CC chemokine ligand 2 (CCL2 within the dorsal root ganglion (DRG is believed to enhance the activity of DRG nociceptive neurons and cause hyperalgesia. Transient receptor potential vanilloid receptor 1 (TRPV1 and tetrodotoxin (TTX-resistant Nav1.8 sodium channels play an essential role in regulating the excitability and pain transmission of DRG nociceptive neurons. We therefore tested the hypothesis that CCL2 causes peripheral sensitization of nociceptive DRG neurons by upregulating the function and expression of TRPV1 and Nav1.8 channels. Methods DRG neuronal culture was prepared from 3-week-old Sprague–Dawley rats and incubated with various concentrations of CCL2 for 24 to 36 hours. Whole-cell voltage-clamp recordings were performed to record TRPV1 agonist capsaicin-evoked inward currents or TTX-insensitive Na+ currents from control or CCL2-treated small DRG sensory neurons. The CCL2 effect on the mRNA expression of TRPV1 or Nav1.8 was measured by real-time quantitative RT-PCR assay. Results Pretreatment of CCL2 for 24 to 36 hours dose-dependently (EC50 value = 0.6 ± 0.05 nM increased the density of capsaicin-induced currents in small putative DRG nociceptive neurons. TRPV1 mRNA expression was greatly upregulated in DRG neurons preincubated with 5 nM CCL2. Pretreating small DRG sensory neurons with CCL2 also increased the density of TTX-resistant Na+ currents with a concentration-dependent manner (EC50 value = 0.7 ± 0.06 nM. The Nav1.8 mRNA level was significantly increased in DRG neurons pretreated with CCL2. In contrast, CCL2 preincubation failed to affect the mRNA level of TTX-resistant Nav1.9. In the presence of the specific phosphatidylinositol-3 kinase (PI3K inhibitor LY294002 or Akt inhibitor IV, CCL2 pretreatment failed to increase the current density of capsaicin-evoked inward currents or TTX-insensitive Na+ currents and

  16. Reimbursement of radiologically guided vascular interventions within the DRG-System: What wil change?; Verguetung radiologischer Gefaessinterventionen im DRG-System: Was wird sich aendern?

    Strotzer, M. [Krankenhaus Hohe Warte, Bayreuth (Germany). Abt. fuer Radiologie; Feuerbach, S. [Klinikum der Univ. Regensburg (Germany). Inst. fuer Diagnostische Radiologie; Voelk, M. [Bundeswehrkrankenhaus Ulm (Germany). Abt. Radiologie

    2004-09-01

    Purpose: To evaluate reimbursement within the DRG-system ('diagnosis-related groups') compared with traditional reimbursement for interventional therapy of hospitalized patients. Materials and Methods: Reimbursement calculation was prospectively analyzed in two respects for 30 consecutive patients who underwent percutaneous transluminal angioplasty (PTA) of the lower extremity arteries: (1) based on the DRG-system; (2) based on the traditional system. Additional evaluation was performed for five further, typical vascular procedures on the basis of real documentation and calculation data (stenting of the carotid artery, fibrinolytic therapy of basilar artery occlusion, stenting of renal artery stenosis, angioplasty of hemodialysis-shunt stenosis and aspiration thrombectomy of an infrapopliteal arterial occlusion). Results: In our hospital, the introduction of the DRG system would reduce reimbursement by approximately 1100 Euro per PTA patient. However, the other vascular radiological procedures can be expected to increase the payments by up to 4500 Euro. Conclusion: To minimize imminent reduction of reimbursement for patients with peripheral PTA, complete documentation and economical patient management is mandatory. Payment may increase significantly for patients with the other reported vascular interventional procedures. (orig.)

  17. Exogenous brain-derived neurotrophic factor relieves pain symptoms of diabetic rats by reducing excitability of dorsal root ganglion neurons.

    Li, Lei; Yu, Ting; Yu, Liling; Li, Haijun; Liu, Yongjuan; Wang, Dongqin

    2016-08-01

    Diabetic peripheral neuropathy (DPN) is a common complication of diabetes lacking of effective treatments. Enhanced excitability of dorsal root ganglion (DRG) neuron plays a crucial role in the progression of diabetic neuropathic hyperalgesia. Brain-derived neurotrophic factor (BDNF) is known as a neuromodulator of nociception, but whether and how BDNF modulates the excitability of DRG neurons in the development of DPN remain to be clarified. This study investigated the role of exogenous BDNF and its high-affinity tropomyosin receptor kinase B (TrkB) in rats with streptozotocin-induced diabetic neuropathic pain. The results showed that continued intrathecal administration of BDNF to diabetic rats dramatically alleviated mechanical and thermal hyperalgesia, as well as inhibited hyperexcitability of DRG neurons. These effects were blocked by pretreatment with TrkB Fc (a synthetic fusion protein consisting of the extracellular ligand-binding domain of the TrkB receptor). The expression of BDNF and TrkB was upregulated in the DRG of diabetic rats. Intrathecal administration of BDNF did not affect this upregulation. These data provide novel information that exogenous BDNF relieved pain symptoms of diabetic rats by reducing hyperexcitability of DRG neurons and might be the potential treatment of painful diabetic neuropathy. PMID:26441011

  18. Modulating nitric oxide levels in dorsal root ganglion neurons of rat with low-level laser therapy

    Zheng, Li-qin; Wang, Yu-hua; He, Yi-peng; Zhou, Jie; Yang, Hong-qin; Zhang, Yan-ding; Xie, Shu-sen

    2015-05-01

    Nitric oxide (NO) and nitric oxide synthase (NOS) have an important role in pain signaling transmission in animal models. Low-level laser therapy (LLLT) is known to have an analgesic effect, but the mechanism is unclear. The aim of the study is to investigate the influence of LLLT on NO release and NOS synthesis in dorsal root ganglion (DRG) neurons, in order to find whether LLLI can ameliorate pain through modulating NO production at the cellular level. The results show that in stress conditions, the laser irradiation at 658 nm can modulate NO production in DRG neurons with soma diameter of about 20 μm in a short time after illumination, and affect NOS synthesis in a dose-dependent manner. It is demonstrated that LLLT might treat pain by altering NO release directly and indirectly in DRG neurons.

  19. The up-regulation of IL-6 in DRG and spinal dorsal horn contributes to neuropathic pain following L5 ventral root transection.

    Wei, Xu-Hong; Na, Xiao-Dong; Liao, Guang-Jie; Chen, Qiu-Ying; Cui, Yu; Chen, Feng-Ying; Li, Yong-Yong; Zang, Ying; Liu, Xian-Guo

    2013-03-01

    Our previous works have shown that pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α) plays an important role in neuropathic pain produced by lumber 5 ventral root transection (L5-VRT). In the present work we evaluate the role of interleukin-6 (IL-6), another key inflammatory cytokine, in the L5-VRT model. We found that IL-6 was up-regulated in the ipsilateral L4 and L5 dorsal root ganglian (DRG) neurons and in bilateral lumbar spinal cord following L5-VRT. Double immunofluorescence stainings revealed that in DRGs the increased immunoreactivity (IR) of IL-6 was almost restricted in neuronal cells, while in the spinal dorsal horn IL-6-IR up-regulated in both glial cells (astrocyte and microglia) and neurons. Intrathecal administration of IL-6 neutralizing antibody significantly delayed the induction of mechanical allodynia in bilateral hindpaws after L5-VRT. Furthermore, inhibition of TNF-α synthesis by intraperitoneal thalidomide prevented both mechanical allodynia and the up-regulation of IL-6 in DRGs following L5-VRT. These data suggested that the increased IL-6 in afferent neurons and spinal cord contribute to the development of neuropathic pain following motor fiber injury, and that TNF-α is responsible for the up-regulation of IL-6. PMID:23261764

  20. 7.5-MINUTE DIGITAL RASTER GRAPHICS (DRG), VA PORTION OF ALBE

    7.5-minute DRG for Virginia portion of Albemarle-Pamlico Estuary watershed. Files obtained from www.runet.edu:8800/~geoserve/Virginia.html at Radford University.Digital Raster Graphics (DRGs) come from the US Geological Survey in Universal Trans Mercator (UTM), North American...

  1. Enhanced excitability of small dorsal root ganglion neurons in rats with bone cancer pain

    Zheng Qin

    2012-04-01

    Full Text Available Abstract Background Primary and metastatic cancers that affect bone are frequently associated with severe and intractable pain. The mechanisms underlying the development of bone cancer pain are largely unknown. The aim of this study was to determine whether enhanced excitability of primary sensory neurons contributed to peripheral sensitization and tumor-induced hyperalgesia during cancer condition. In this study, using techniques of whole-cell patch-clamp recording associated with immunofluorescent staining, single-cell reverse-transcriptase PCR and behavioral test, we investigated whether the intrinsic membrane properties and the excitability of small-sized dorsal root ganglion (DRG neurons altered in a rat model of bone cancer pain, and whether suppression of DRG neurons activity inhibited the bone cancer-induced pain. Results Our present study showed that implantation of MRMT-1 tumor cells into the tibial canal in rats produced significant mechanical and thermal hyperalgesia in the ipsilateral hind paw. Moreover, implantation of tumor cells provoked spontaneous discharges and tonic excitatory discharges evoked by a depolarizing current pulse in small-sized DRG neurons. In line with these findings, alterations in intrinsic membrane properties that reflect the enhanced neuronal excitability were observed in small DRG neurons in bone cancer rats, of which including: 1 depolarized resting membrane potential (RMP; 2 decreased input resistance (Rin; 3 a marked reduction in current threshold (CT and voltage threshold (TP of action potential (AP; 4 a dramatic decrease in amplitude, overshot, and duration of evoked action potentials as well as in amplitude and duration of afterhyperpolarization (AHP; and 5 a significant increase in the firing frequency of evoked action potentials. Here, the decreased AP threshold and increased firing frequency of evoked action potentials implicate the occurrence of hyperexcitability in small-sized DRG neurons in bone

  2. [OR minute myth : Guidelines for calculation of DRG revenues per OR minute].

    Waeschle, R M; Hinz, J; Bleeker, F; Sliwa, B; Popov, A; Schmidt, C E; Bauer, M

    2016-02-01

    The economic situation in German Hospitals is tense and needs the implementation of differentiated controlling instruments. Accordingly, parameters of revenue development of different organizational units within a hospital are needed. This is particularly necessary in the revenue and cost-intensive operating theater field. So far there are only barely established productivity data for the control of operating room (OR) revenues during the year available. This article describes a valid method for the calculation of case-related revenues per OR minute conform to the diagnosis-related groups (DRG).For this purpose the relevant datasets from the OR information system and the § 21 productivity report (DRG grouping) of the University Medical Center Göttingen were combined. The revenues defined in the DRG browser of the Institute for Hospital Reimbursement (InEK) were assigned to the corresponding process times--incision-suture time (SNZ), operative preparation time and anesthesiology time--according to the InEK system. All full time stationary DRG cases treated within the OR were included and differentiated according to the surgical department responsible. The cost centers "OR section" and "anesthesia" were isolated to calculate the revenues of the operating theater. SNZ clusters and cost type groups were formed to demonstrate their impact on the revenues per OR minute. A surgical personal simultaneity factor (GZF) was calculated by division of the revenues for surgeons and anesthesiologists. This factor resembles the maximum DRG financed personnel deployment for surgeons in German hospitals.The revenue per OR minute including all cost types and DRG was 16.63 €/min. The revenues ranged from 10.45 to 24.34 €/min depending on the surgical field. The revenues were stable when SNZ clusters were analyzed. The differentiation of cost type groups revealed a revenue reduction especially after exclusion of revenues for implants and infrastructure. The calculated GZF over

  3. Expression and Distribution of LRRN3 Membrance Protein in Human Embryo Dorsal Root Ganglion (DRG)%LRRN3膜蛋白在人胚脊神经节的分布和表达

    李明; 吴海平; 贺旭; 潘爱华

    2011-01-01

    Objective: To explore the expression and localization of membrance protein LRRN3 of dorsal root ganglion (DRG) in human embryo at the mRNA and protein levels. Methods: mRNA and protein of membrance protein LRRN3 were separated from DRG, and RT-PCR, Western Blotting, immunocytochemistry staining and Immunofluorescence were used to detect expression of membrance protein LRRN3. Results: RT-PCR production cDNA of LRRN3 C-terminal domain protein were detected in DRG at each gestational age, and the length is about 500 bp; Western blot analysis showed membrance protein LRRN3 were expressed and the relative molecular mass is about 78 kD; immunocytochemistry staining and Immunofiuorescence detection showed all membrance protein LRRN3 positively expressed cells were sensory neurons in DRG, some weak-positively expressed and some not expressed. Conclusion: The expression of LRRN3 in DRG suggests that LRRN3 is closely associated with embryonic development, morphogenesis and repair following injury of sensory neurons in Dorsal root ganglion.%目的:探讨神经系统富亮氨酸重复超家族成员LRRN3膜蛋白在人胚脊神经节的表达和分布.方法:从人胚脊神经节分离mRNA和蛋白质,用RT-PCR、Western印记杂交法和免疫组织化学方法检测LRRN3膜蛋白表达.结果:LRRN3膜蛋白C端序列RT-PCR扩增产物cDNA在各胎龄脊神经节均有表达,长度约500bp; Western印记杂交结果显示LRRN3膜蛋白存在,分子质量约78 kD;免疫组织化学和免疫荧光组织化学结果表明LRRN3阳性表达细胞均为脊神经节感觉神经细胞,部分神经细胞弱阳性表达,部分未表达.结论:LRRN3膜蛋白在脊神经节神经元的表达,推断与脊神经节感觉神经元的发育、形态构建及损伤后修复有密切的关系.

  4. Differential expression of ATP-gated P2X receptors in DRG between chronic neuropathic pain and visceralgia rat models.

    Chen, Lin; Liu, Yu-Wei; Yue, Kai; Ru, Qin; Xiong, Qi; Ma, Bao-Miao; Tian, Xiang; Li, Chao-Ying

    2016-03-01

    There are divergences between neuropathic pain and visceralgia in terms of the duration, location, and character of hyperalgesia. It is generally recognized that nociceptive receptors, including P2X receptors, may play different roles in nociceptive mechanisms. The different roles of P2X1-7 receptors have not been fully understood both in neuropathic pain and visceral hyperalgesia. In order to explore the different expressions of P2X1-7 receptors in these two hyperalgesia models, the lumbosacral dorsal root ganglion (DRG) neurons from rat sciatic nerve chronic constriction injury (CCI) model and neonatal colorectal distention (NCRD) model were studied (both the primary nociceptive neuron afferents of those two models projected to the same segment of spinal cord). Both immunohistochemistry (IHC) technique and real-time fluorescence quantitative polymerase chain reaction (RT-PCR) technology were applied to analyze the protein expression levels and nucleic acid of P2X1-7 receptors. We found that except P2X2 and P2X3, the expression levels of P2X1 and P2X5 receptors increased in neuropathic pain while those expression levels of P2X4, P2X6, and P2X7 receptors increased in visceral pain. Our results also suggested that in addition to P2X2/3 heteromeric, other P2X subunits may also involved in generation heteromeric such as P2X1/5 and/or P2X2/5 in neuropathic pain and P2X4/6 and/or P2X4/7 in visceral pain. PMID:26531254

  5. Kv4 channels underlie the subthreshold-operating A-type K+-current in nociceptive dorsal root ganglion neurons

    Thanawath R Na Phuket

    2009-07-01

    Full Text Available The dorsal root ganglion (DRG contains heterogeneous populations of sensory neurons including primary nociceptive neurons and C-fibers implicated in pain signaling.  Recent studies have demonstrated DRG hyperexcitability associated with downregulation of A-type K+ channels; however, the molecular correlate of the corresponding A-type K+ current (IA has remained hypothetical.  Kv4 channels may underlie the IA in DRG neurons.  We combined electrophysiology, molecular biology (whole-tissue and single-cell RT-PCR and immunohistochemistry to investigate the molecular basis of the IA in acutely dissociated DRG neurons from 7-8 day-old rats.  Whole-cell recordings demonstrate a robust tetraethylammonium-resistant (20 mM and 4-aminopyridine-sensitive (5 mM IA.  Matching Kv4 channel properties, activation and inactivation of this IA occur in the subthreshold range of membrane potentials and the rate of recovery from inactivation is rapid and voltage-dependent.  Among Kv4 transcripts, the DRG expresses significant levels of Kv4.1 and Kv4.3 mRNAs.  Also, single small-medium diameter DRG neurons (~30 mm exhibit correlated frequent expression of mRNAs encoding Kv4.1 and Nav1.8, a known nociceptor marker.  In contrast, the expressions of Kv1.4 and Kv4.2 mRNAs at the whole-tissue and single-cell levels are relatively low and infrequent.  Kv4 protein expression in nociceptive DRG neurons was confirmed by immunohistochemistry, which demonstrates colocalization of Kv4.3 and Nav1.8, and negligible expression of Kv4.2.  Furthermore, specific dominant-negative suppression and overexpression strategies confirmed the contribution of Kv4 channels to IA in DRG neurons.  Contrasting the expression patterns of Kv4 channels in the central and peripheral nervous systems, we discuss possible functional roles of these channels in primary sensory neurons.

  6. Enhancement of GABAA Receptor-Mediated Conductances Induced by Nerve Injury in a Subclass of Sensory Neurons

    OYELESE, ADETOKUNBO A.; ENG, DOUGLAS L.; Richerson, George B.; Kocsis, Jeffery D.

    1995-01-01

    The effects of axotomy on the electrophysiologic properties of adult rat dorsal root ganglion (DRG) neurons were studied to understand the changes in excitability induced by traumatic nerve injury. Nerve injury was induced in vivo by sciatic nerve ligation with distal nerve transection. Two to four weeks after nerve ligation, a time when a neuroma forms, lumbar (L4 and L5) DRG neurons were removed and placed in short-term tissue culture. Whole cell patch-clamp recordings were made 5–24 h afte...

  7. Etanercept decreases HMGB1 expression in dorsal root ganglion neuron cells in a rat chronic constriction injury model

    WANG, RUI-KE; Zhang, Qin-Qin; PAN, YUN-DAN; Guo, Qu-Lian

    2012-01-01

    In the present study, we examined the effect of etanercept on high mobility group box 1 (HMGB1) expression in dorsal root ganglion (DRG) neuron cells in a rat model of chronic constriction injury (CCI) of the sciatic nerve, with the aim of exploring the molecular mechanism underlying the therapeutic effect of etanercept on sciatica-related nociception and the potential interaction between tumor necrosis factor-α (TNF-α) and HMGB1 in DRG neuron cells. A rat CCI model was employed and the anima...

  8. DRG-related prices applied in a public health care system--can Finland learn from Norway and Sweden?

    Mikkola, Hennamari; Keskimäki, Ilmo; Häkkinen, Unto

    2002-01-01

    In the early 1990s, DRG based hospital financing was introduced into some hospital districts in Finland. The 1993 state subsidy reform decentralising all hospital financing to municipalities, and the aim of improving productivity, were the driving forces for introducing DRG. This study addresses the pros and cons of DRG in hospital financing in the Finnish health care system and puts forward several solutions to avoid potential problems. We consider the objectives and optimal features of hospital financing systems in the context of the public health care system, where the public sector owns and finances hospitals. We analyse impacts of introducing different types of DRG based hospital financing systems, taking into account earlier experiences in countries such as Sweden and Norway, as well as Finnish system specific features. DRG could assist the Finnish municipalities to compare quality, costs and prices of services between hospitals, and related cost information might help them budget expenditure more accurately. System specific features mean that traditional uses of DRG in hospital pricing are not feasible in Finland. But some benefits of DRG could be exploited, for instance in the controlled contracts between municipalities and hospitals. PMID:11786173

  9. Ellagic acid promotes A{beta}42 fibrillization and inhibits A{beta}42-induced neurotoxicity

    Feng, Ying [Department of Histology and Embryology, College of Basic Medical Science, China Medical University, Shenyang 110001 (China); Tsinghua University School of Medicine, Haidian District, Beijing 100084 (China); Yang, Shi-gao; Du, Xue-ting; Zhang, Xi; Sun, Xiao-xia; Zhao, Min [Tsinghua University School of Medicine, Haidian District, Beijing 100084 (China); Sun, Gui-yuan, E-mail: sungy2004@sohu.com [Department of Histology and Embryology, College of Basic Medical Science, China Medical University, Shenyang 110001 (China); Liu, Rui-tian, E-mail: rtliu@tsinghua.edu.cn [Tsinghua University School of Medicine, Haidian District, Beijing 100084 (China)

    2009-12-25

    Smaller, soluble oligomers of {beta}-amyloid (A{beta}) play a critical role in the pathogenesis of Alzheimer's disease (AD). Selective inhibition of A{beta} oligomer formation provides an optimum target for AD therapy. Some polyphenols have potent anti-amyloidogenic activities and protect against A{beta} neurotoxicity. Here, we tested the effects of ellagic acid (EA), a polyphenolic compound, on A{beta}42 aggregation and neurotoxicity in vitro. EA promoted A{beta} fibril formation and significant oligomer loss, contrary to previous results that polyphenols inhibited A{beta} aggregation. The results of transmission electron microscopy (TEM) and Western blot displayed more fibrils in A{beta}42 samples co-incubated with EA in earlier phases of aggregation. Consistent with the hypothesis that plaque formation may represent a protective mechanism in which the body sequesters toxic A{beta} aggregates to render them harmless, our MTT results showed that EA could significantly reduce A{beta}42-induced neurotoxicity toward SH-SY5Y cells. Taken together, our results suggest that EA, an active ingredient in many fruits and nuts, may have therapeutic potential in AD.

  10. Glutaminase Increases in Rat Dorsal Root Ganglion Neurons after Unilateral Adjuvant-Induced Hind Paw Inflammation

    E. Matthew Hoffman

    2016-01-01

    Full Text Available Glutamate is a neurotransmitter used at both the peripheral and central terminals of nociceptive primary sensory neurons, yet little is known concerning regulation of glutamate metabolism during peripheral inflammation. Glutaminase (GLS is an enzyme of the glutamate-glutamine cycle that converts glutamine into glutamate for neurotransmission and is implicated in producing elevated levels of glutamate in central and peripheral terminals. A potential mechanism for increased levels of glutamate is an elevation in GLS expression. We assessed GLS expression after unilateral hind paw inflammation by measuring GLS immunoreactivity (ir with quantitative image analysis of L4 dorsal root ganglion (DRG neurons after one, two, four, and eight days of adjuvant-induced arthritis (AIA compared to saline injected controls. No significant elevation in GLS-ir occurred in the DRG ipsilateral to the inflamed hind paw after one or two days of AIA. After four days AIA, GLS-ir was elevated significantly in all sizes of DRG neurons. After eight days AIA, GLS-ir remained elevated in small (<400 µm2, presumably nociceptive neurons. Western blot analysis of the L4 DRG at day four AIA confirmed the elevated GLS-ir. The present study indicates that GLS expression is increased in the chronic stage of inflammation and may be a target for chronic pain therapy.

  11. Glutaminase Increases in Rat Dorsal Root Ganglion Neurons after Unilateral Adjuvant-Induced Hind Paw Inflammation.

    Hoffman, E Matthew; Zhang, Zijia; Schechter, Ruben; Miller, Kenneth E

    2016-01-01

    Glutamate is a neurotransmitter used at both the peripheral and central terminals of nociceptive primary sensory neurons, yet little is known concerning regulation of glutamate metabolism during peripheral inflammation. Glutaminase (GLS) is an enzyme of the glutamate-glutamine cycle that converts glutamine into glutamate for neurotransmission and is implicated in producing elevated levels of glutamate in central and peripheral terminals. A potential mechanism for increased levels of glutamate is an elevation in GLS expression. We assessed GLS expression after unilateral hind paw inflammation by measuring GLS immunoreactivity (ir) with quantitative image analysis of L4 dorsal root ganglion (DRG) neurons after one, two, four, and eight days of adjuvant-induced arthritis (AIA) compared to saline injected controls. No significant elevation in GLS-ir occurred in the DRG ipsilateral to the inflamed hind paw after one or two days of AIA. After four days AIA, GLS-ir was elevated significantly in all sizes of DRG neurons. After eight days AIA, GLS-ir remained elevated in small (<400 µm²), presumably nociceptive neurons. Western blot analysis of the L4 DRG at day four AIA confirmed the elevated GLS-ir. The present study indicates that GLS expression is increased in the chronic stage of inflammation and may be a target for chronic pain therapy. PMID:26771651

  12. [Introduction of the DRG system from the point of view of private health insurers].

    Fritze, Jürgen; Miebach, Jürgen; Hüdig, Wolfgang

    2002-08-01

    For the first time, there has been a worldwide attempt to fund all hospital services almost completely by a DRG system supplemented by additional charges, rebates, and procedural rates. In the interest of the efficiency and transparency of hospital services the introduction of a German DRG system settling the current implausible price differences would be welcome. The system selected by the medical self-governing bodies in Germany is based upon the Australian AR-DRG classification. In contrast to other systems, the latter provides the best medical plausibility, the highest transparency of the assignment algorithm and the highest potential for flexibility and adaptations to changing morbidity patterns and medical progress. The adaptation to the conditions of the German health care system requires considerable efforts on the part of hospitals as well as sickness funds and health insurers. Hospitals need to establish a cost unit accounting system satisfying the rules of Applied Economics to allow, among other things, the calculation of relative cost weights. The self-governing bodies will have to consent on a complex regulation system. The German Hospital Federation declared the break down of negotiations concerning a provisional DRG system to be optionally available to hospitals in 2003. The Federal Ministry of Health will now have to decide whether to implement the system through executive fiat. The comprehensive DRG system will introduce new risks. The economic risks of the individual hospital, though not the individual insurer's risks, will be partially compensated for in the introductory phase by revenue balance mechanisms, for example. In particular, both the privately insured and civil servants will face a rise in costs as they will no longer benefit from a shorter length of hospital stay. To end this discrimination against private health insurers, the double counting of the costs associated with medical treatment (included in the DRG price and additionally

  13. La valutazione dei 43 DRG a rischio di inappropriatezza nella regione Abruzzo

    V. Cofini

    2003-05-01

    Full Text Available

    Obiettivi: l’obiettivo dello studio è rappresentato dalla valutazione dei ricoveri relativi ai 43 DRG ospedalieri, individuati nell’allegato 2C del DPCM 29 novembre 2001 (ad alto rischio di inappropriatezza nelle strutture di ricovero pubbliche e private, della Regione Abruzzo.

    Metodi: sono stati elaborati i dati forniti dalla Regione Abruzzo relativi ai ricoveri ed alle giornate di degenza, DRG specifici, per gli anni 2000 e 2001, in regime di degenza ordinaria e day hospital. È stata effettuata un’analisi comparativa degli indicatori dell’attività ospedaliera e della spesa sanitaria legati al sistema DRG, tra i presidi oggetto di studio, ed un confronto temporale degli stessi indicatori, per ogni singolo presidio. I dati disponibili su supporto informatico sono stati elaborati attraverso il pacchetto statistico sas/stat.

    Risultati: nei due anni considerati il rapporto tra i ricoveri ordinari ed i ricoveri in regime di day hospital è passato dal 78% del 2000 al 74% del 2001. È stato registrato un decremento del 1.5% nei ricoveri in regime di degenza ordinaria ed un importante incremento del 21% per i ricoveri in day hospital. Le giornate di degenza prodotte dai rispettivi DRG analizzati, hanno subito un decremento del 6% in regime di degenza ordinaria e poco più del 5% in regime di day hospital. Dal confronto tra le strutture pubbliche con più di 500 posti letto, sono stati evidenziati un decremento dell’1% dei ricoveri ordinari ed un incremento del 38% dei ricoveri in day hospital.

    Conclusioni: dall’analisi è stato evidenziato un aumento del numero dei ricoveri “inappropriati”, seguita da un decremento delle giornate di degenza prodotte dagli stessi DRG. Uno degli effetti più rilevanti dell’analisi è stata la crescita della quota della spesa sanitaria in corrispondenza dei 43 DRG valutati. Tale considerazione rende necessaria

  14. Role of neurotrophin signalling in the differentiation of neurons from dorsal root ganglia and sympathetic ganglia.

    Ernsberger, Uwe

    2009-06-01

    Manipulation of neurotrophin (NT) signalling by administration or depletion of NTs, by transgenic overexpression or by deletion of genes coding for NTs and their receptors has demonstrated the importance of NT signalling for the survival and differentiation of neurons in sympathetic and dorsal root ganglia (DRG). Combination with mutation of the proapoptotic Bax gene allows the separation of survival and differentiation effects. These studies together with cell culture analysis suggest that NT signalling directly regulates the differentiation of neuron subpopulations and their integration into neural networks. The high-affinity NT receptors trkA, trkB and trkC are restricted to subpopulations of mature neurons, whereas their expression at early developmental stages largely overlaps. trkC is expressed throughout sympathetic ganglia and DRG early after ganglion formation but becomes restricted to small neuron subpopulations during embryogenesis when trkA is turned on. The temporal relationship between trkA and trkC expression is conserved between sympathetic ganglia and DRG. In DRG, NGF signalling is required not only for survival, but also for the differentiation of nociceptors. Expression of neuropeptides calcitonin gene-related peptide and substance P, which specify peptidergic nociceptors, depends on nerve growth factor (NGF) signalling. ret expression indicative of non-peptidergic nociceptors is also promoted by the NGF-signalling pathway. Regulation of TRP channels by NGF signalling might specify the temperature sensitivity of afferent neurons embryonically. The manipulation of NGF levels "tunes" heat sensitivity in nociceptors at postnatal and adult stages. Brain-derived neurotrophic factor signalling is required for subpopulations of DRG neurons that are not fully characterized; it affects mechanical sensitivity in slowly adapting, low-threshold mechanoreceptors and might involve the regulation of DEG/ENaC ion channels. NT3 signalling is required for the

  15. Wirtschaftlichkeitsanalyse von kieferorthopädischen Korrektureingriffen unter DRG-Bedingungen an einem Zentrum der Maximalversorgung

    Tschakert, Robert Julian

    2015-01-01

    Orthognathic surgical procedures comprise a major portion of all oral and maxillofacial surgeries. In 2007 and 2008, 106 patients at the Department of Oral and Maxillofacial Surgery and Clinical Navigation at Charité Campus Virchow-Klinikum Universitätsmedizin Berlin (WMKG) underwent orthognathic surgery. This doctoral thesis evaluates the economics of these corrective procedures in consideration of the German Diagnosis Related Groups (DRG) system, a billing system whereby inpatient proce...

  16. Reimbursement of radiologically guided vascular interventions within the DRG-System: What wil change?

    Purpose: To evaluate reimbursement within the DRG-system ('diagnosis-related groups') compared with traditional reimbursement for interventional therapy of hospitalized patients. Materials and Methods: Reimbursement calculation was prospectively analyzed in two respects for 30 consecutive patients who underwent percutaneous transluminal angioplasty (PTA) of the lower extremity arteries: (1) based on the DRG-system; (2) based on the traditional system. Additional evaluation was performed for five further, typical vascular procedures on the basis of real documentation and calculation data (stenting of the carotid artery, fibrinolytic therapy of basilar artery occlusion, stenting of renal artery stenosis, angioplasty of hemodialysis-shunt stenosis and aspiration thrombectomy of an infrapopliteal arterial occlusion). Results: In our hospital, the introduction of the DRG system would reduce reimbursement by approximately 1100 Euro per PTA patient. However, the other vascular radiological procedures can be expected to increase the payments by up to 4500 Euro. Conclusion: To minimize imminent reduction of reimbursement for patients with peripheral PTA, complete documentation and economical patient management is mandatory. Payment may increase significantly for patients with the other reported vascular interventional procedures. (orig.)

  17. Heterogeneity in Patient Recruitment Between Public and Private Hospitals Within DRG (GHM 164

    Allemand H

    2001-12-01

    Full Text Available Aim: The generalization of the medical program for information systems (PMSI to the two hospital sectors (public and private has offered the possibility for cost comparisons: the costs of a synthetic activity index point (point ISA, cost according to DRG (GHM… However, these analyses assume that the contents of a GHM are homogenous in both the public and private sector. The aim of this study was to test this hypothesis by using one GHM (n° 164 whose heading seems to describe a very homogenous group: implantation of a permanent cardiac pacemaker in the absence of myocardial infarction or congestive heart failure. Method: We used the comprehensive national data from both public and private institutions emanating from all the anonymous discharge summaries (RSA during the last six months of 1997. Results: A comparison of hospital stays for DRG 164 between both hospital sectors showed disparities in type of apparatus implanted, patients’ ages and length of stay. Differences in length of stay were not due to a greater propensity for the public sector to keep patients longer but rather to the recruitment of clinically different patients. Conclusion: Cost comparisons between the public and private sectors using DRGs should verify the homogeneity of hospital stays within each DRG when analyzing identical expenditures. This verification should include the procedure performed, the patients’ ages, the primary diagnoses and the number of summaries from medical units (RUM contained in the RSA. For a valid comparison, all these elements must be taken into account.

  18. The effects of neurotrophic substances on primary sensory neurons following peripheral nerve injury

    Eriksson, Petri

    1997-01-01

    Sensory impulses from the periphery is transmitted by primary sensory neurons located in dorsal root and cranial nerve ganglia. The primary sensory neurons are phenotypically diverse with regard to their expression of various chemical components. Thus, various subpopulations of dorsal root ganglion (DRG) cells express the peptides substance P (SP) and calcitonin gene related peptide (CGRP), the enzyme fluoride-resistant acid phosphatase (FRAP) and bind the lectin Gr...

  19. Prostatic Acid Phosphatase Is Expressed in Peptidergic and Nonpeptidergic Nociceptive Neurons of Mice and Rats

    Taylor-Blake, Bonnie; Zylka, Mark J.

    2010-01-01

    Thiamine monophosphatase (TMPase, also known as Fluoride-resistant acid phosphatase or FRAP) is a classic histochemical marker of small- to medium-diameter dorsal root ganglia (DRG) neurons and has primarily been studied in the rat. Previously, we found that TMPase was molecularly identical to Prostatic acid phosphatase (PAP) using mice. In addition, PAP was expressed in a majority of nonpeptidergic, isolectin B4-binding (IB4+) nociceptive neurons and a subset of peptidergic, calcitonin gene-...

  20. [A DRG based future hospital funding scheme: state of implementation and impact in the field of rehabilitation].

    Rochell, B; Roeder, N

    2002-02-01

    With the statutory health insurance reform act 2000 the German government started to introduce a new hospital funding system based on an internationally used diagnosis related groups (DRGs) system. In June 2000 the German self-administration board (consisting of the German hospital federation, the German statutory health insurance funds and the association of private health insurers), which is in charge of realizing this project, decided to develop the future German (Refined) DRG System (G-DRG) with reference to the Australian Refined DRG System (AR-DRG) Version 4.1. Replacement of the previous German hospital reimbursement system by the new DRG-based hospital funding system is planned for January 2003 on a voluntary basis. From January 2004 on, the change of the reimbursement system is to become mandatory for all hospitals with the exception of psychiatry. The new reimbursement system is intended to not only cover acute hospital care but also parts of early rehabilitation, palliative and sub-acute care. Because of its economic incentives the effects of DRG introduction in Germany will not only be limited to the hospital scene but will also affect rehabilitation. PMID:11830787

  1. The sensitivity of neurons with non-periodic activity to sympathetic stimulation in rat injured dorsal root ganglion

    Hong-Jun YANG; San-Jue HU; Pu-Lin GONG; Jian-Hong DUAN

    2006-01-01

    Objective The relationship between firing pattern and sensitivity of neurons was studied in chronically compressed dorsal root ganglion (DRG) neurons and the Hindmarsh-Rose (HR) neuronal model. Methods Spontaneous activities from single fibers of chronically compressed DRG neurons in rats were recorded, and divided into periodic and non-periodic firing patterns. The sensitivity of the two kinds of firing pattern neuron to sympathetic stimulation (SS)was compared. Result It was found that 27.3% of periodic firing neurons and 93.2% of non-periodic firing neurons responded to SS respectively ( periodic vs non-periodic, P < 0.01 ). The responses to SS with different stimulation time were greater non-periodic firing neurons than periodic firing neurons (P < 0.01 ). The non-periodic firing neurons obviously responded to SS. After the firing pattern of these neurons transformed to periodic firing pattern, their responses to SS disappeared or decreased obviously. The HR neuronal model exhibited a significantly greater response to perturbation in non-periodic (chaotic) firing pattern than in periodic firing pattern. Conclusion The non-periodic firing neurons with deterministic chaos are more sensitive to external stimuli than the periodic firing neurons.

  2. Dramatic Co-Activation of WWOX/WOX1 with CREB and NF-κB in Delayed Loss of Small Dorsal Root Ganglion Neurons upon Sciatic Nerve Transection in Rats

    Li, Meng-Yen; Lai, Feng-Jie; Hsu, Li-Jin; Lo, Chen-Peng; Cheng, Ching-Li; Lin, Sing-Ru; Lee, Ming-Hui; Chang, Jean-Yun; Subhan, Dudekula; Tsai, Ming-Shu; Sze, Chun-I; Pugazhenthi, Subbiah; Chang, Nan-Shan; Chen, Shur-Tzu

    2009-01-01

    Background Tumor suppressor WOX1 (also named WWOX or FOR) is known to participate in neuronal apoptosis in vivo. Here, we investigated the functional role of WOX1 and transcription factors in the delayed loss of axotomized neurons in dorsal root ganglia (DRG) in rats. Methodology/Principal Findings Sciatic nerve transection in rats rapidly induced JNK1 activation and upregulation of mRNA and protein expression of WOX1 in the injured DRG neurons in 30 min. Accumulation of p-WOX1, p-JNK1, p-CRE...

  3. Demethylation regulation of BDNF gene expression in dorsal root ganglion neurons is implicated in opioid-induced pain hypersensitivity in rats.

    Chao, Yu-Chieh; Xie, Fang; Li, Xueyang; Guo, Ruijuan; Yang, Ning; Zhang, Chen; Shi, Rong; Guan, Yun; Yue, Yun; Wang, Yun

    2016-07-01

    Repeated administration of morphine may result in opioid-induced hypersensitivity (OIH), which involves altered expression of numerous genes, including brain-derived neurotrophic factor (BDNF) in dorsal root ganglion (DRG) neurons. Yet, it remains unclear how BDNF expression is increased in DRG neurons after repeated morphine treatment. DNA methylation is an important mechanism of epigenetic control of gene expression. In the current study, we hypothesized that the demethylation regulation of certain BDNF gene promoters in DRG neurons may contribute to the development of OIH. Real-time RT-PCR was used to assess changes in the mRNA transcription levels of major BDNF exons including exon I, II, IV, VI, as well as total BDNF mRNA in DRGs from rats after repeated morphine administration. The levels of exon IV and total BDNF mRNA were significantly upregulated by repeated morphine administration, as compared to that in saline control group. Further, ELISA array and immunocytochemistry study revealed a robust upregulation of BDNF protein expression in DRG neurons after repeated morphine exposure. Correspondingly, the methylation levels of BDNF exon IV promoter showed a significant downregulation by morphine treatment. Importantly, intrathecal administration of a BDNF antibody, but not control IgG, significantly inhibited mechanical hypersensitivity that developed in rats after repeated morphine treatment. Conversely, intrathecal administration of an inhibitor of DNA methylation, 5-aza-2'-deoxycytidine (5-aza-dC) markedly upregulated the BDNF protein expression in DRG neurons and enhanced the mechanical allodynia after repeated morphine exposure. Together, our findings suggest that demethylation regulation of BDNF gene promoter may be implicated in the development of OIH through epigenetic control of BDNF expression in DRG neurons. PMID:26970395

  4. Low-frequency electrical stimulation improves neurite outgrowth of dorsal root ganglion neurons in vitro via upregulating Ca2+-mediated brain-derived neurotrophic factor expression

    Lidan Wan; Rong Xia; Wenlong Ding

    2010-01-01

    Short-term,low-frequency electrical stimulation of neural tissues significantly enhances axonal regeneration of peripheral nerves following injury.However,little is known about the mechanisms of electrical stimulation to induce neurite outgrowth.In the present study,short-term,low-frequency electrical stimulation,using identical stimulation parameters of in vivo experiments,was administered to in vitro dorsal root ganglion(DRG)neurons.Enhanced neurite outgrowth,as well as synthesis and release of brain-derived neurotrophic factor(BDNF),were examined in electrical stimulation-treated DRG neuronal cultures.Because the effects of electrical stimulation on neuronal intracellular signaling molecules are less reported,classic calcium intracellular signals are directly or indirectly involved in electrical stimulation effects on neurons.Cultured DRG neurons were pretreated with the calcium channel blocker nifedipine,followed by electrical stimulation.Results suggested that electrical stimulation not only promoted in vitro neurite outgrowth,but also enhanced BDNF expression.However,nifedipine reduced electrical stimulation-enhanced neurite outgrowth and BDNF biosynthesis.These results suggest that the promoting effects of electrical stimulation on DRG neurite outgrowth could be associated with altered calcium influx,which is involved induction of neuronal BDNF expression and secretion.

  5. Acrylamide Retards the Slow Axonal Transport of Neurofilaments in Rat Cultured Dorsal Root Ganglia Neurons and the Corresponding Mechanisms.

    An, Lihong; Li, Guozhen; Si, Jiliang; Zhang, Cuili; Han, Xiaoying; Wang, Shuo; Jiang, Lulu; Xie, Keqin

    2016-05-01

    Chronic acrylamide (ACR) exposure induces peripheral-central axonopathy in occupational workers and laboratory animals, but the underlying mechanisms remain unclear. In this study, we first investigated the effects of ACR on slow axonal transport of neurofilaments in cultured rat dorsal root ganglia (DRG) neurons through live-cell imaging approach. Then for the underlying mechanisms exploration, the protein level of neurofilament subunits, motor proteins kinesin and dynein, and dynamitin subunit of dynactin in DRG neurons were assessed by western blotting and the concentrations of ATP was detected using ATP Assay Kit. The results showed that ACR treatment results in a dose-dependent decrease of slow axonal transport of neurofilaments. Furthermore, ACR intoxication significantly increases the protein levels of the three neurofilament subunits (NF-L, NF-M, NF-H), kinesin, dynein, and dynamitin subunit of dynactin in DRG neurons. In addition, ATP level decreased significantly in ACR-treated DRG neurons. Our findings indicate that ACR exposure retards slow axonal transport of NF-M, and suggest that the increase of neurofilament cargoes, motor proteins, dynamitin of dynactin, and the inadequate ATP supply contribute to the ACR-induced retardation of slow axonal transport. PMID:26721510

  6. Curcumin protects against intracellular amyloid toxicity in rat primary neurons

    Ye, Jelina; Zhang, Yan

    2012-01-01

    To investigate whether curcumin is protective against intracellular amyloid beta (A beta) toxicity, different concentrations of curcumin were applied to with intracellular A beta in rat primary hippocampal neurons in culture. We find that at low dosages, curcumin effectively inhibits intracellular A

  7. Differential adeno-associated virus mediated gene transfer to sensory neurons following intrathecal delivery by direct lumbar puncture

    Kitto Kelley F

    2010-05-01

    Full Text Available Abstract Background Neuronal transduction by adeno-associated viral (AAV vectors has been demonstrated in cortex, brainstem, cerebellum, and sensory ganglia. Intrathecal delivery of AAV serotypes that transduce neurons in dorsal root ganglia (DRG and spinal cord offers substantial opportunities to 1 further study mechanisms underlying chronic pain, and 2 develop novel gene-based therapies for the treatment and management of chronic pain using a non-invasive delivery route with established safety margins. In this study we have compared expression patterns of AAV serotype 5 (AAV5- and AAV serotype 8 (AAV8-mediated gene transfer to sensory neurons following intrathecal delivery by direct lumbar puncture. Results Intravenous mannitol pre-treatment significantly enhanced transduction of primary sensory neurons after direct lumbar puncture injection of AAV5 (rAAV5-GFP or AAV8 (rAAV8-GFP carrying the green fluorescent protein (GFP gene. The presence of GFP in DRG neurons was consistent with the following evidence for primary afferent origin of the majority of GFP-positive fibers in spinal cord: 1 GFP-positive axons were evident in both dorsal roots and dorsal columns; and 2 dorsal rhizotomy, which severs the primary afferent input to spinal cord, abolished the majority of GFP labeling in dorsal horn. We found that both rAAV5-GFP and rAAV8-GFP appear to preferentially target large-diameter DRG neurons, while excluding the isolectin-B4 (IB4 -binding population of small diameter neurons. In addition, a larger proportion of CGRP-positive cells was transduced by rAAV5-GFP, compared to rAAV8-GFP. Conclusions The present study demonstrates the feasibility of minimally invasive gene transfer to sensory neurons using direct lumbar puncture and provides evidence for differential targeting of subtypes of DRG neurons by AAV vectors.

  8. Distinct functional and temporal requirements for zebrafish Hdac1 during neural crest-derived craniofacial and peripheral neuron development.

    Myron S Ignatius

    Full Text Available The regulation of gene expression is accomplished by both genetic and epigenetic means and is required for the precise control of the development of the neural crest. In hdac1(b382 mutants, craniofacial cartilage development is defective in two distinct ways. First, fewer hoxb3a, dlx2 and dlx3-expressing posterior branchial arch precursors are specified and many of those that are consequently undergo apoptosis. Second, in contrast, normal numbers of progenitors are present in the anterior mandibular and hyoid arches, but chondrocyte precursors fail to terminally differentiate. In the peripheral nervous system, there is a disruption of enteric, DRG and sympathetic neuron differentiation in hdac1(b382 mutants compared to wildtype embryos. Specifically, enteric and DRG-precursors differentiate into neurons in the anterior gut and trunk respectively, while enteric and DRG neurons are rarely present in the posterior gut and tail. Sympathetic neuron precursors are specified in hdac1(b382 mutants and they undergo generic neuronal differentiation but fail to undergo noradrenergic differentiation. Using the HDAC inhibitor TSA, we isolated enzyme activity and temporal requirements for HDAC function that reproduce hdac1(b382 defects in craniofacial and sympathetic neuron development. Our study reveals distinct functional and temporal requirements for zebrafish hdac1 during neural crest-derived craniofacial and peripheral neuron development.

  9. 游离NgR促进新生大鼠背根神经节突起体外生长%Role of soluble NgR in promotion of DRG neurite sprouting in vitro in rats

    马永刚; 刘世清; 范里; 陶海鹰; 卫爱林

    2009-01-01

    目的 观察生物合成的游离NgR对新生大鼠原代背根神经节(DRG)神经元突起生长的影响.方法 以ViraPowerTM慢病毒将NgR(310)ecto基因转染人培养的骨髓基质细胞,收集培养上清获得游离NgR.分离新生大鼠DRG神经元并分为两组,对照组直接将神经元种植到经含中枢神经系统(CNS)髓鞘蛋白包被的培养皿中培养;实验组先将骨髓基质细胞培养上清加入含CNS髓鞘蛋白的培养皿中,孵育4 h后再植入DRG神经元.两组细胞均在培养24 h后固定,进行βⅢ-tubulin免疫组织化学染色,图像观察神经元及其突起生长情况.结果 基因修饰的骨髓基质细胞,转染后48 h间接免疫荧光的方法检测可见胞浆内荧光表达.种植24 h后对照组DRG细胞很少有突起长出;而实验组DRG细胞中60%的神经元长出突起,且突起较长.结论 游离NgR能够竞争性结合脊髓损伤后局部分泌的轴突生长抑制因子,从而保护生理性NgR,可能是促轴突再生和脊髓功能恢复的一种新策略.%Objective To observe the effect of soluble NgR on rat DRG neurite sprouting in vitro. Methods Gene encoding NgR(310)ecto was transduced into bone marrow stromal cells (BM-SCs). Soluble NgR was obtained from the supernatant of BMSCs culture. DRG neurons of neonate rat were isolated and allocated into two groups, ie, control group (DRG neurons were cultured in slides coa-ted with myelin protein of spinal cord) and experiment group (the slides were pre-incubated with the su-pernatant of BMSCs for four hours). BMSCs were immobilized after 24 hours of culture. Immunohis-tostaining with βⅢ-tubulin and fluorescence microscope were employed to analyze the neurite growth.Results Positive staining of intracytoplasm was found with indirect fluorescence microscope in the BM-SCs 48 hours following transduction. Little neurite was found sprout to DRG neurons in the control group 24 hours after transduction. In contrast, longer neurite was observed

  10. Heterogeneity in Patient Recruitment Between Public and Private Hospitals Within DRG (GHM) 164

    Allemand H; Fender P; Vincke B

    2001-01-01

    Aim: The generalization of the medical program for information systems (PMSI) to the two hospital sectors (public and private) has offered the possibility for cost comparisons: the costs of a synthetic activity index point (point ISA), cost according to DRG (GHM)… However, these analyses assume that the contents of a GHM are homogenous in both the public and private sector. The aim of this study was to test this hypothesis by using one GHM (n° 164) whose heading seems to describe a very homog...

  11. TRESK channel contribution to nociceptive sensory neurons excitability: modulation by nerve injury

    Serra Jordi

    2011-04-01

    Full Text Available Abstract Background Neuronal hyperexcitability is a crucial phenomenon underlying spontaneous and evoked pain. In invertebrate nociceptors, the S-type leak K+ channel (analogous to TREK-1 in mammals plays a critical role of in determining neuronal excitability following nerve injury. Few data are available on the role of leak K2P channels after peripheral axotomy in mammals. Results Here we describe that rat sciatic nerve axotomy induces hyperexcitability of L4-L5 DRG sensory neurons and decreases TRESK (K2P18.1 expression, a channel with a major contribution to total leak current in DRGs. While the expression of other channels from the same family did not significantly change, injury markers ATF3 and Cacna2d1 were highly upregulated. Similarly, acute sensory neuron dissociation (in vitro axotomy produced marked hyperexcitability and similar total background currents compared with neurons injured in vivo. In addition, the sanshool derivative IBA, which blocked TRESK currents in transfected HEK293 cells and DRGs, increased intracellular calcium in 49% of DRG neurons in culture. Most IBA-responding neurons (71% also responded to the TRPV1 agonist capsaicin, indicating that they were nociceptors. Additional evidence of a biological role of TRESK channels was provided by behavioral evidence of pain (flinching and licking, in vivo electrophysiological evidence of C-nociceptor activation following IBA injection in the rat hindpaw, and increased sensitivity to painful pressure after TRESK knockdown in vivo. Conclusions In summary, our results clearly support an important role of TRESK channels in determining neuronal excitability in specific DRG neurons subpopulations, and show that axonal injury down-regulates TRESK channels, therefore contributing to neuronal hyperexcitability.

  12. Neuregulin-1β Regulates the migration of Different Neurochemical Phenotypic Neurons from Organotypically Cultured Dorsal Root Ganglion Explants.

    Li, Yunfeng; Liu, Guixiang; Li, Hao; Bi, Yanwen

    2016-01-01

    Neuregulin-1β (NRG-1β) has multiple roles in the development and function in the nervous system and exhibits potent neuroprotective properties. In the present study, organotypically cultured dorsal root ganglion (DRG) explants were used to evaluate the effects of NRG-1β on migration of two major phenotypic classes of DRG neurons. The signaling pathways involved in these effects were also determined. Organotypically cultured DRG explants were exposed to NRG-1β (20 nmol/L), the phosphatidylinositol 3-kinase inhibitor LY294002 (10 μmol/L) plus NRG-1β (20 nmol/L), the extracellular signal-regulated protein kinase (ERK1/2) inhibitor PD98059 (10 μmol/L) plus NRG-1β (20 nmol/L), and LY294002 (10 μmol/L) plus PD98059 (10 μmol/L) plus NRG-1β (20 nmol/L), respectively, for 3 days. The DRG explants were continuously exposed to culture media as a control. After that, all above cultures were processed for detecting the mRNA levels of calcitonin gene-related peptide (CGRP) and neurofilament-200 (NF-200) by real-time PCR analysis. CGRP and NF-200 expression in situ was determined by fluorescent labeling technique. The results showed that NRG-1β elevated the mRNA and protein levels of CGRP and NF-200. NRG-1β also increased the number and the percentage of CGRP-immunoreactive (IR) migrating neurons and NF-200-IR migrating neurons. Inhibitors (LY294002, PD98059) either alone or in combination blocked the effects of NRG-1β. The contribution of NRG-1β on modulating distinct neurochemical phenotypic plasticity of DRG neurons suggested that NRG-1β signaling system might play an important role on the biological effects of primary sensory neurons. PMID:26093851

  13. Extracellular Nm23H1 stimulates neurite outgrowth from dorsal root ganglia neurons in vitro independently of nerve growth factor supplementation or its nucleoside diphosphate kinase activity

    Wright, K.T. [Keele University at the RJAH Orthopaedic Hospital, Oswestry, Shropshire (United Kingdom); Seabright, R.; Logan, A. [Neuropharmacology and Neurobiology, School of Clinical and Experimental Medicine, Birmingham University, Birmingham (United Kingdom); Lilly, A.J.; Khanim, F.; Bunce, C.M. [Biosciences, Birmingham University, Birmingham (United Kingdom); Johnson, W.E.B., E-mail: w.e.johnson@aston.ac.uk [Life and Health Sciences, Aston University, Birmingham (United Kingdom)

    2010-07-16

    Research highlights: {yields} Extracellular Nm23H1 stimulates nerve growth. {yields} Extracellular Nm23H1 provides pathfinding cues to growth cones. {yields} The neurotrophic activity of Nm23H1 is independent of NDP kinase activity. {yields} The neurotrophic activity of Nm23H1 is independent of NGF. -- Abstract: The nucleoside diphosphate (NDP) kinase, Nm23H1, is a highly expressed during neuronal development, whilst induced over-expression in neuronal cells results in increased neurite outgrowth. Extracellular Nm23H1 affects the survival, proliferation and differentiation of non-neuronal cells. Therefore, this study has examined whether extracellular Nm23H1 regulates nerve growth. We have immobilised recombinant Nm23H1 proteins to defined locations of culture plates, which were then seeded with explants of embryonic chick dorsal root ganglia (DRG) or dissociated adult rat DRG neurons. The substratum-bound extracellular Nm23H1 was stimulatory for neurite outgrowth from chick DRG explants in a concentration-dependent manner. On high concentrations of Nm23H1, chick DRG neurite outgrowth was extensive and effectively limited to the location of the Nm23H1, i.e. neuronal growth cones turned away from adjacent collagen-coated substrata. Nm23H1-coated substrata also significantly enhanced rat DRG neuronal cell adhesion and neurite outgrowth in comparison to collagen-coated substrata. These effects were independent of NGF supplementation. Recombinant Nm23H1 (H118F), which does not possess NDP kinase activity, exhibited the same activity as the wild-type protein. Hence, a novel neuro-stimulatory activity for extracellular Nm23H1 has been identified in vitro, which may function in developing neuronal systems.

  14. Effects of complete Freund's adjuvant on immunohistochemical distribution of IL-1β and IL-1R I in neurons and glia cells of dorsal root ganglion

    Man LI; Chuan-you LIN; Xin-min GUAN; Jing SHI; Jun-rui TANG; Di CHEN; Bo AI; Jun CHEN; Li-na WANG; Fu-yuan CAO; Ling-li LI

    2005-01-01

    Aim: To investigate the effects of complete Freund's adjuvant (CFA) on inflammatory hyperalgesia and morphological change of the coexsistence of interleukin- 1 beta (IL-1β) and type Ⅰ IL-1 receptor (IL-1RI) in neurons and glia cells of rat dorsal root ganglion (DRG). Methods: The pain-related parameters and the expression of IL- 1RI and IL-1β positive neurons and glia cells of DRG in normal saline (NS) and adjuvant-induced arthritic (AA) group were examined with pain behavior assessment methods and immunohistochemical assay, respectively. Results: Five hours,1 d, and 2 d after intra-articular injection of 50 μL CFA, tactile hyperalgesia in duced by CFA was observed in the foot flexion and extension scores of the ipsilateral hindpaw of AA group. Three days after injection, the distribution of IL-1RI/ IL-1β double-stained coexisted neurons and glia cells were observed in ipsilateral DRG of both groups. The number of IL-1β positive neurons, IL-1RI positive neurons, IL-1 β/IL-1RI double-stained neurons, and IL-1RI positive glia cells in ipsilateral DRG of the AA group were higher than that of NS group (P<0.05 or P< 0.01). Conclusion: The coexsistence of IL- 1 β and IL- 1 RI in neurons and nonneuronal cells suggests an as yet unknown autocrine and/or paracrine function of IL-1 β in the DRG. The function was enhanced in articular arthritis induced by CFA and could play an important role in hyperalgesia under inflammatory conditions.

  15. Extracellular Nm23H1 stimulates neurite outgrowth from dorsal root ganglia neurons in vitro independently of nerve growth factor supplementation or its nucleoside diphosphate kinase activity

    Research highlights: → Extracellular Nm23H1 stimulates nerve growth. → Extracellular Nm23H1 provides pathfinding cues to growth cones. → The neurotrophic activity of Nm23H1 is independent of NDP kinase activity. → The neurotrophic activity of Nm23H1 is independent of NGF. -- Abstract: The nucleoside diphosphate (NDP) kinase, Nm23H1, is a highly expressed during neuronal development, whilst induced over-expression in neuronal cells results in increased neurite outgrowth. Extracellular Nm23H1 affects the survival, proliferation and differentiation of non-neuronal cells. Therefore, this study has examined whether extracellular Nm23H1 regulates nerve growth. We have immobilised recombinant Nm23H1 proteins to defined locations of culture plates, which were then seeded with explants of embryonic chick dorsal root ganglia (DRG) or dissociated adult rat DRG neurons. The substratum-bound extracellular Nm23H1 was stimulatory for neurite outgrowth from chick DRG explants in a concentration-dependent manner. On high concentrations of Nm23H1, chick DRG neurite outgrowth was extensive and effectively limited to the location of the Nm23H1, i.e. neuronal growth cones turned away from adjacent collagen-coated substrata. Nm23H1-coated substrata also significantly enhanced rat DRG neuronal cell adhesion and neurite outgrowth in comparison to collagen-coated substrata. These effects were independent of NGF supplementation. Recombinant Nm23H1 (H118F), which does not possess NDP kinase activity, exhibited the same activity as the wild-type protein. Hence, a novel neuro-stimulatory activity for extracellular Nm23H1 has been identified in vitro, which may function in developing neuronal systems.

  16. Sigma-1 Receptor Antagonism Restores Injury-Induced Decrease of Voltage-Gated Ca2+ Current in Sensory Neurons

    Pan, Bin; Guo, Yuan; Kwok, Wai-Meng; Hogan, Quinn; Wu, Hsiang-en

    2014-01-01

    Sigma-1 receptor (σ1R), an endoplasmic reticulum–chaperone protein, can modulate painful response after peripheral nerve injury. We have demonstrated that voltage-gated calcium current is inhibited in axotomized sensory neurons. We examined whether σ1R contributes to the sensory dysfunction of voltage-gated calcium channel (VGCC) after peripheral nerve injury through electrophysiological approach in dissociated rat dorsal root ganglion (DRG) neurons. Animals received either skin incision (Con...

  17. [Potential uses of AP-DRG to describe the health care profile in hospital units].

    Noronha, Marina Ferreira de; Portela, Margareth Crisóstomo; Lebrão, Maria Lúcia

    2004-01-01

    The All Patient Diagnosis Related Groups (AP-DRG) provide a classification system for general hospital inpatients, aggregating hospitalizations based on resource use and clinical criteria. The different versions of the AP-DRG have been applied to inpatient care management and reimbursement. This paper aimed to describe the classification and explore the potential for generating information on inpatient care management based on data from the Ribeirão Preto region (Sao Paulo State, Brazil) in 1997, including public, private, and charity hospitals. We compared average length of stay related to DRGs in the Ribeirão Preto region and the United States. Using the relative cost weights of the AP-DRGs constructed for New York State, we verified the profile of inpatient care provided by 30 hospitals in the Ribeirão Preto region, and reimbursement for hospital care provided to patients referred from other municipalities and covered by the Unified National Health System (SUS). Our findings indicate the applicability of the classification to inform the decision-making process on inpatient care regionalization and organization in levels of complexity, as well as for improvement of inpatient care monitoring and reimbursement. PMID:15608938

  18. Cost Analysis of Integrative Inpatient Treatment Based on DRG Data: The Example of Anthroposophic Medicine

    Jürgen Heinz

    2013-01-01

    Full Text Available Background. Much work has been done to evaluate the outcome of integrative inpatient treatment but scarcely the costs. This paper evaluates the costs for inpatient treatment in three anthroposophic hospitals (AHs. Material and Methods. Cost and performance data from a total of 23,180 cases were analyzed and compared to national reference data. Subgroup analysis was performed between the cases with and without anthroposophic medical complex (AMC treatment. Results. Costs and length of stay in the cases without AMC displayed no relevant differences compared to the national reference data. In contrast the inlier cases with AMC caused an average of € 1,394 more costs. However costs per diem were not higher than those in the national reference data. Hence, the delivery of AMC was associated with a prolonged length of stay. 46.6% of the cases with AMC were high outliers. Only 10.6% of the inlier cases with AMC were discharged before reaching the mean length of stay of each DRG. Discussion. Treatment in an AH is not generally associated with an increased use of resources. However, the provision of AMC leads to a prolonged length of stay and cannot be adequately reimbursed by the current G-DRG system. Due to the heterogeneity of the patient population, an additional payment should be negotiated individually.

  19. Cost unit accounting based on a clinical pathway: a practical tool for DRG implementation.

    Feyrer, R; Rösch, J; Weyand, M; Kunzmann, U

    2005-10-01

    Setting up a reliable cost unit accounting system in a hospital is a fundamental necessity for economic survival, given the current general conditions in the healthcare system. Definition of a suitable cost unit is a crucial factor for success. We present here the development and use of a clinical pathway as a cost unit as an alternative to the DRG. Elective coronary artery bypass grafting was selected as an example. Development of the clinical pathway was conducted according to a modular concept that mirrored all the treatment processes across various levels and modules. Using service records and analyses the process algorithms of the clinical pathway were developed and visualized with CorelTM iGrafix Process 2003. A detailed process cost record constituted the basis of the pathway costing, in which financial evaluation of the treatment processes was performed. The result of this study was a structured clinical pathway for coronary artery bypass grafting together with a cost calculation in the form of cost unit accounting. The use of a clinical pathway as a cost unit offers considerable advantages compared to the DRG or clinical case. The variance in the diagnoses and procedures within a pathway is minimal, so the consumption of resources is homogeneous. This leads to a considerable improvement in the value of cost unit accounting as a strategic control instrument in hospitals. PMID:16208610

  20. Vestibular Neuronitis

    ... Prevent Painful Swimmer's Ear Additional Content Medical News Vestibular Neuronitis By Lawrence R. Lustig, MD NOTE: This ... Drugs Herpes Zoster Oticus Meniere Disease Purulent Labyrinthitis Vestibular Neuronitis Vestibular neuronitis is a disorder characterized by ...

  1. A SAGE-based screen for genes expressed in sub-populations of neurons in the mouse dorsal root ganglion

    Garces Alain

    2007-11-01

    Full Text Available Abstract Background The different sensory modalities temperature, pain, touch and muscle proprioception are carried by somatosensory neurons of the dorsal root ganglia. Study of this system is hampered by the lack of molecular markers for many of these neuronal sub-types. In order to detect genes expressed in sub-populations of somatosensory neurons, gene profiling was carried out on wild-type and TrkA mutant neonatal dorsal root ganglia (DRG using SAGE (serial analysis of gene expression methodology. Thermo-nociceptors constitute up to 80 % of the neurons in the DRG. In TrkA mutant DRGs, the nociceptor sub-class of sensory neurons is lost due to absence of nerve growth factor survival signaling through its receptor TrkA. Thus, comparison of wild-type and TrkA mutants allows the identification of transcripts preferentially expressed in the nociceptor or mechano-proprioceptor subclasses, respectively. Results Our comparison revealed 240 genes differentially expressed between the two tissues (P Conclusion We have identified and characterized the detailed expression patterns of three genes in the developing DRG, placing them in the context of the known major neuronal sub-types defined by molecular markers. Further analysis of differentially expressed genes in this tissue promises to extend our knowledge of the molecular diversity of different cell types and forms the basis for understanding their particular functional specificities.

  2. Vorschläge der DGPP zur Abbildung phoniatrisch-pädaudiologischer Leistungen im G-DRG-System

    Matulat, P; Lamprecht-Dinnesen, A; Roeder, N; Gross, M.

    2003-01-01

    Im Rahmen des Vorschlagsverfahrens zur Einbindung des medizinischen, wissenschaftlichen und weiteren Sachverstandes bei der Weiterentwicklung des G-DRG-Systems hat die DGPP dem Institut für das Entgeltsystem im Krankenhaus (InEk) und dem Deutschen Institut für Medizinische Dokumentation und Information (DIMDI) Vorschläge für das Jahr 2004 unterbreitet. Auf der Grundlage von Behandlungs- und Kostendaten aus den Kliniken in Erlangen, Mainz und Münster wurde dem InEK für die Revision des G-DRG ...

  3. Nociceptin/orphanin FQ receptor expression in clinical pain disorders and functional effects in cultured neurons.

    Anand, Praveen; Yiangou, Yiangos; Anand, Uma; Mukerji, Gaurav; Sinisi, Marco; Fox, Michael; McQuillan, Anthony; Quick, Tom; Korchev, Yuri E; Hein, Peter

    2016-09-01

    The nociceptin/orphanin FQ peptide receptor (NOP), activated by its endogenous peptide ligand nociceptin/orphanin FQ (N/OFQ), exerts several effects including modulation of pain signalling. We have examined, for the first time, the tissue distribution of the NOP receptor in clinical visceral and somatic pain disorders by immunohistochemistry and assessed functional effects of NOP and μ-opioid receptor activation in cultured human and rat dorsal root ganglion (DRG) neurons. Quantification of NOP-positive nerve fibres within the bladder suburothelium revealed a remarkable several-fold increase in detrusor overactivity (P numbers. NOP immunoreactivity was significantly decreased in injured peripheral nerves (P = 0.0004), and also in painful neuromas (P = 0.025). Calcium-imaging studies in cultured rat DRG neurons demonstrated dose-dependent inhibition of capsaicin responses in the presence of N/OFQ, with an IC50 of 8.6 pM. In cultured human DRG neurons, 32% inhibition of capsaicin responses was observed in the presence of 1 pM N/OFQ (P < 0.001). The maximum inhibition of capsaicin responses was greater with N/OFQ than μ-opioid receptor agonist DAMGO. Our findings highlight the potential of NOP agonists, particularly in urinary bladder overactivity and pain syndromes. The regulation of NOP expression in visceral and somatic sensory neurons by target-derived neurotrophic factors deserves further study, and the efficacy of NOP selective agonists in clinical trials. PMID:27127846

  4. Stimulation of neuronal neurite outgrowth using functionalized carbon nanotubes

    Low concentrations (0.11-1.7 μg ml-1) of functionalized carbon nanotubes (CNTs), which are multi-walled CNTs modified by amino groups, when added with nerve growth factor (NGF), promoted outgrowth of neuronal neurites in dorsal root ganglion (DRG) neurons and rat pheochromocytoma cell line PC12h cells in culture media. The quantity of active extracellular signal-regulated kinase (ERK) was higher after the addition of both 0.85 μg ml-1 CNTs and NGF than that with NGF alone. CNTs increased the number of cells with neurite outgrowth in DRG neurons and PC12h cells after the inhibition of the ERK signaling pathway using a mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor. Active ERK proteins were detected in MEK inhibitor-treated neurons after the addition of CNTs to the culture medium. These results demonstrate that CNTs may stimulate neurite outgrowth by activation of the ERK signaling pathway. Thus, CNTs are biocompatible and are promising candidates for biological applications and devices.

  5. Stimulation of neuronal neurite outgrowth using functionalized carbon nanotubes

    Matsumoto, K; Sato, C; Shimizu, N [Graduate School of Life Sciences, Toyo University, 1-1-1 Izumino, Itakura-machi, Ora-gun, Gunma 374-0193 (Japan); Naka, Y [Bio-Nano Electronics Research Center, Toyo University, 2100 Kujirai, Kawagoe-shi, Saitama 350-8585 (Japan); Whitby, R, E-mail: shimizu@toyonet.toyo.ac.jp [School of Pharmacy and Biomolecular Sciences, University of Brighton, Cockroft Building, Lewes Road, Brighton BN2 4GJ (United Kingdom)

    2010-03-19

    Low concentrations (0.11-1.7 {mu}g ml{sup -1}) of functionalized carbon nanotubes (CNTs), which are multi-walled CNTs modified by amino groups, when added with nerve growth factor (NGF), promoted outgrowth of neuronal neurites in dorsal root ganglion (DRG) neurons and rat pheochromocytoma cell line PC12h cells in culture media. The quantity of active extracellular signal-regulated kinase (ERK) was higher after the addition of both 0.85 {mu}g ml{sup -1} CNTs and NGF than that with NGF alone. CNTs increased the number of cells with neurite outgrowth in DRG neurons and PC12h cells after the inhibition of the ERK signaling pathway using a mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor. Active ERK proteins were detected in MEK inhibitor-treated neurons after the addition of CNTs to the culture medium. These results demonstrate that CNTs may stimulate neurite outgrowth by activation of the ERK signaling pathway. Thus, CNTs are biocompatible and are promising candidates for biological applications and devices.

  6. Block of voltage-gated potassium channels by Pacific ciguatoxin-1 contributes to increased neuronal excitability in rat sensory neurons

    The present study investigated the actions of the polyether marine toxin Pacific ciguatoxin-1 (P-CTX-1) on neuronal excitability in rat dorsal root ganglion (DRG) neurons using patch-clamp recording techniques. Under current-clamp conditions, bath application of 2-20 nM P-CTX-1 caused a rapid, concentration-dependent depolarization of the resting membrane potential in neurons expressing tetrodotoxin (TTX)-sensitive voltage-gated sodium (Nav) channels. This action was completely suppressed by the addition of 200 nM TTX to the external solution, indicating that this effect was mediated through TTX-sensitive Nav channels. In addition, P-CTX-1 also prolonged action potential and afterhyperpolarization (AHP) duration. In a subpopulation of neurons, P-CTX-1 also produced tonic action potential firing, an effect that was not accompanied by significant oscillation of the resting membrane potential. Conversely, in neurons expressing TTX-resistant Nav currents, P-CTX-1 failed to alter any parameter of neuronal excitability examined in this study. Under voltage-clamp conditions in rat DRG neurons, P-CTX-1 inhibited both delayed-rectifier and 'A-type' potassium currents in a dose-dependent manner, actions that occurred in the absence of alterations to the voltage dependence of activation. These actions appear to underlie the prolongation of the action potential and AHP, and contribute to repetitive firing. These data indicate that a block of potassium channels contributes to the increase in neuronal excitability, associated with a modulation of Nav channel gating, observed clinically in response to ciguatera poisoning

  7. Calcium activity of upper thoracic dorsal root ganglion neurons in zucker diabetic Fatty rats

    Ghorbani, Marie Louise; Nyborg, Niels C B; Fjalland, Bjarne;

    2013-01-01

    The aim of the present study was to examine the calcium activity of C8-T5 dorsal root ganglion (DRG) neurons from Zucker diabetic fatty rats. In total, 8 diabetic ZDF fatty animals and 8 age-matched control ZDF lean rats were employed in the study. C8-T5 dorsal root ganglia were isolated bilatera......The aim of the present study was to examine the calcium activity of C8-T5 dorsal root ganglion (DRG) neurons from Zucker diabetic fatty rats. In total, 8 diabetic ZDF fatty animals and 8 age-matched control ZDF lean rats were employed in the study. C8-T5 dorsal root ganglia were isolated...

  8. [Diseases of the digestive system and a system for patient classification: diagnostic-related groups (DRG)].

    Poves Martínez, E

    1996-10-01

    In the last years different classification systems for hospitalized patients have been developed in the United States, that allow to evaluate different aspects of any Hospital: The quality, the efficacy and the efficiency. The classification of "The Diagnosis Related Groups" (DRG) relates the hospitalized patients with the expenses that they generate, and it is the most used in Europe. In the version DRG-All Patient, all the diagnoses and procedures of the hospitalized patients, using the International Classification of Diseases, are divided in to 25 Major Diagnostic Categories (MDC) where each one excludes all the others. We describe the behavior with respect to the age, sex and stay of all the GDR integrated in the CDM 6 and 7: diseases and disorders of the digestive tract, and diseases and disorders of the hepatobiliary system and the pancreas. The study has been carried out with a national data base of two millions of cases seen from 214 acute-care Hospitals. The MDC number 6 is the most frequent, with an important number of patients in the pediatric age; it is more frequent in males; the average stay is 8.59 days and 19 GDR (36.5%) have a variation coefficient greater than one. The other MDC, number 7 is less frequent, the majority of the patients are between 60-80 years of age, the average stay is 12.2 days with a coefficient of variation greater than one in 4 GDR (17.4%). Clinical Services should know the characteristics and behavior of their patients, as well as the comparison with the national data bases. This may allow a control of the quality and the costs by using a common "language" with the managers. PMID:8983307

  9. Nerve Injury Diminishes Opioid Analgesia through Lysine Methyltransferase-mediated Transcriptional Repression of μ-Opioid Receptors in Primary Sensory Neurons.

    Zhang, Yuhao; Chen, Shao-Rui; Laumet, Geoffroy; Chen, Hong; Pan, Hui-Lin

    2016-04-15

    The μ-opioid receptor (MOR, encoded by Oprm1) agonists are the mainstay analgesics for treating moderate to severe pain. Nerve injury causes down-regulation of MORs in the dorsal root ganglion (DRG) and diminishes the opioid effect on neuropathic pain. However, the epigenetic mechanisms underlying the diminished MOR expression caused by nerve injury are not clear. G9a (encoded by Ehmt2), a histone 3 at lysine 9 methyltransferase, is a key chromatin regulator responsible for gene silencing. In this study, we determined the role of G9a in diminished MOR expression and opioid analgesic effects in animal models of neuropathic pain. We found that nerve injury in rats induced a long-lasting reduction in the expression level of MORs in the DRG but not in the spinal cord. Nerve injury consistently increased the enrichment of the G9a product histone 3 at lysine 9 dimethylation in the promoter of Oprm1 in the DRG. G9a inhibition or siRNA knockdown fully reversed MOR expression in the injured DRG and potentiated the morphine effect on pain hypersensitivity induced by nerve injury. In mice lacking Ehmt2 in DRG neurons, nerve injury failed to reduce the expression level of MORs and the morphine effect. In addition, G9a inhibition or Ehmt2 knockout in DRG neurons normalized nerve injury-induced reduction in the inhibitory effect of the opioid on synaptic glutamate release from primary afferent nerves. Our findings indicate that G9a contributes critically to transcriptional repression of MORs in primary sensory neurons in neuropathic pain. G9a inhibitors may be used to enhance the opioid analgesic effect in the treatment of chronic neuropathic pain. PMID:26917724

  10. Functional up-regulation of Nav1.8 sodium channel on dorsal root ganglia neurons contributes to the induction of scorpion sting pain.

    Ye, Pin; Hua, Liming; Jiao, Yunlu; Li, Zhenwei; Qin, Shichao; Fu, Jin; Jiang, Feng; Liu, Tong; Ji, Yonghua

    2016-02-01

    BmK I, purified from the venom of scorpion Buthus martensi Karsch (BmK), is a receptor site-3-specific modulator of voltage-gated sodium channels (VGSCs) and can induce pain-related behaviors in rats. The tetrodotoxin-resistant (TTX-R) sodium channel Nav1.8 contributes to most of the sodium current underlying the action potential upstroke in dorsal root ganglia (DRG) neurons and may serve as a critical ion channel targeted by BmK I. Herein, using electrophysiological, molecular, and behavioral approaches, we investigated whether the aberrant expression of Nav1.8 in DRG contributes to generation of pain induced by BmK I. The expression of Nav1.8 was found to be significantly increased at both mRNA and protein levels following intraplantar injection of BmK I in rats. In addition, the current density of TTX-R Nav1.8 sodium channel is significantly increased and the gating kinetics of Nav1.8 is also altered in DRG neurons from BmK I-treated rats. Furthermore, spontaneous pain and mechanical allodynia, but not thermal hyperalgesia induced by BmK I, are significantly alleviated through either blockade of the Nav1.8 sodium channel by its selective blocker A-803467 or knockdown of the Nav1.8 expression in DRG by antisense oligodeoxynucleotide (AS-ODN) targeting Nav1.8 in rats. Finally, BmK I was shown to induce enhanced pain behaviors in complete freund's adjuvant (CFA)-inflamed rats, which was partly due to the over-expression of Nav1.8 in DRG. Our results suggest that functional up-regulation of Nav1.8 channel on DRG neurons contributes to the development of BmK I-induced pain in rats. PMID:26764239

  11. Autoimmune neurological disorders associated with group-A beta-hemolytic streptococcal infection.

    Hachiya, Yasuo; Miyata, Rie; Tanuma, Naoyuki; Hongou, Kazuhisa; Tanaka, Keiko; Shimoda, Konomi; Kanda, Sachiko; Hoshino, Ai; Hanafusa, Yukiko; Kumada, Satoko; Kurihara, Eiji; Hayashi, Masaharu

    2013-08-01

    Although central nervous system (CNS) disorders associated with group-A beta-hemolytic streptococcal (GABHS) infection occur only rarely, Sydenham's chorea is a well-recognized disease that can arise following infection. Children may develop a tic, obsessive compulsive disorder (OCD), and extrapyramidal movement subsequent to GABHS infection. These disorders have been termed pediatric autoimmune neuropsychiatric disorders associated with streptococci (PANDAS). Herein we report one case each of acute disseminated encephalomyelitis (ADEM), PANDAS and subacute encephalitis associated with GABHS infection. To evaluate the pathogenesis of the CNS disorders associated with GABHS infection, we measured levels of neurotransmitters, cytokines, anti-neuronal autoantibodies, and performed immunohistochemistry using patient sera to stain human brain sections. All three cases showed psychiatric behavioral disorders. Immunotherapy was effective, and homovanillic acid levels in the cerebrospinal fluid (CSF) were elevated at the acute stage in all three cases. In each case of ADEM and PANDAS, immunohistochemistry demonstrated neuronal impairment in the basal ganglia during the acute stage. Neuronal immunoreactivity was visualized in the cerebral cortex at the acute stage in the case of subacute encephalitis. There was no direct correlation between immunoreactivity of patient sera on the brain sections and positivity of anti-neuronal autoantibodies or CSF biomarkers. The results suggest that autoimmune responses may modulate neurotransmission, and the use of patient serum for immunohistochemistry is a sensitive screening method for the detection of anti-neuronal autoantibodies in CNS disorders associated with GABHS infection. PMID:23142103

  12. Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity.

    Chiu, Isaac M; Barrett, Lee B; Williams, Erika K; Strochlic, David E; Lee, Seungkyu; Weyer, Andy D; Lou, Shan; Bryman, Gregory S; Roberson, David P; Ghasemlou, Nader; Piccoli, Cara; Ahat, Ezgi; Wang, Victor; Cobos, Enrique J; Stucky, Cheryl L; Ma, Qiufu; Liberles, Stephen D; Woolf, Clifford J

    2014-01-01

    The somatosensory nervous system is critical for the organism's ability to respond to mechanical, thermal, and nociceptive stimuli. Somatosensory neurons are functionally and anatomically diverse but their molecular profiles are not well-defined. Here, we used transcriptional profiling to analyze the detailed molecular signatures of dorsal root ganglion (DRG) sensory neurons. We used two mouse reporter lines and surface IB4 labeling to purify three major non-overlapping classes of neurons: 1) IB4(+)SNS-Cre/TdTomato(+), 2) IB4(-)SNS-Cre/TdTomato(+), and 3) Parv-Cre/TdTomato(+) cells, encompassing the majority of nociceptive, pruriceptive, and proprioceptive neurons. These neurons displayed distinct expression patterns of ion channels, transcription factors, and GPCRs. Highly parallel qRT-PCR analysis of 334 single neurons selected by membership of the three populations demonstrated further diversity, with unbiased clustering analysis identifying six distinct subgroups. These data significantly increase our knowledge of the molecular identities of known DRG populations and uncover potentially novel subsets, revealing the complexity and diversity of those neurons underlying somatosensation. PMID:25525749

  13. Shaping the System - The DRG Evaluation Project of the German Society for Gynaecology and Obstetrics (Deutsche Gesellschaft für Gynäkologie und Geburtshilfe, DGGG).

    Fiori, W; Renner, S P; Siam, K; Babapirali, J; Roeder, N; Dausch, E; Hildebrandt, T; Hillemanns, P; Nehmzow, M; Zygmunt, M; Piroth, D; Schem, C; Schwenzer, T; Friese, K; Wallwiener, D; Beckmann, M W

    2013-08-01

    Introduction: The German DRG system is annually adapted to the changing services provided. For the further development, the self-governing body and its DRG Institute (InEK) depend on participation of the users. Methods: For one of the DRG evaluation projects initiated by DGGG, cost and performance data for the year 2011 from 16 hospitals were available. After plausibility checks and corrections, analyses for service and cost homogeneity were performed. In cases of inadequate DRG-representation attributes were sought that would make an appropriate reimbursement possible. Conspicuities and potential solutions were checked for clinical plausibility. Results: 44 concrete modification proposals for further development of the G-DRG system were formulated and submitted in due time to the InEK. In addition, 3 modification proposals were addressed to the German Institute for Medical Documentation and Information (Deutsches Institut für Medizinische Dokumentation und Information, DIMDI) for further development of the diagnosis classification ICD-10-GM. For all modification proposals care was taken to minimise misdirected incentives and to reduce the potential for disputes with the cost bearers and their auditors services in settlements. Discussion: The publication of the G-DRG system 2014 shows which modification proposals have been realised. Essentially, an appropriate redistribution of the resources among the gynaecological and obstetrics departments is to be expected. The financial pressure that is caused by the generally inadequate financing of hospitals will not be reduced by a further development of the G-DRG system. PMID:24771931

  14. Activation of Six1 Expression in Vertebrate Sensory Neurons.

    Shigeru Sato

    Full Text Available SIX1 homeodomain protein is one of the essential key regulators of sensory organ development. Six1-deficient mice lack the olfactory epithelium, vomeronasal organs, cochlea, vestibule and vestibuloacoustic ganglion, and also show poor neural differentiation in the distal part of the cranial ganglia. Simultaneous loss of both Six1 and Six4 leads to additional abnormalities such as small trigeminal ganglion and abnormal dorsal root ganglia (DRG. The aim of this study was to understand the molecular mechanism that controls Six1 expression in sensory organs, particularly in the trigeminal ganglion and DRG. To this end, we focused on the sensory ganglia-specific Six1 enhancer (Six1-8 conserved between chick and mouse. In vivo reporter assays using both animals identified an important core region comprising binding consensus sequences for several transcription factors including nuclear hormone receptors, TCF/LEF, SMAD, POU homeodomain and basic-helix-loop-helix proteins. The results provided information on upstream factors and signals potentially relevant to Six1 regulation in sensory neurons. We also report the establishment of a new transgenic mouse line (mSix1-8-NLSCre that expresses Cre recombinase under the control of mouse Six1-8. Cre-mediated recombination was detected specifically in ISL1/2-positive sensory neurons of Six1-positive cranial sensory ganglia and DRG. The unique features of the mSix1-8-NLSCre line are the absence of Cre-mediated recombination in SOX10-positive glial cells and central nervous system and ability to induce recombination in a subset of neurons derived from the olfactory placode/epithelium. This mouse model can be potentially used to advance research on sensory development.

  15. Activation of Six1 Expression in Vertebrate Sensory Neurons.

    Sato, Shigeru; Yajima, Hiroshi; Furuta, Yasuhide; Ikeda, Keiko; Kawakami, Kiyoshi

    2015-01-01

    SIX1 homeodomain protein is one of the essential key regulators of sensory organ development. Six1-deficient mice lack the olfactory epithelium, vomeronasal organs, cochlea, vestibule and vestibuloacoustic ganglion, and also show poor neural differentiation in the distal part of the cranial ganglia. Simultaneous loss of both Six1 and Six4 leads to additional abnormalities such as small trigeminal ganglion and abnormal dorsal root ganglia (DRG). The aim of this study was to understand the molecular mechanism that controls Six1 expression in sensory organs, particularly in the trigeminal ganglion and DRG. To this end, we focused on the sensory ganglia-specific Six1 enhancer (Six1-8) conserved between chick and mouse. In vivo reporter assays using both animals identified an important core region comprising binding consensus sequences for several transcription factors including nuclear hormone receptors, TCF/LEF, SMAD, POU homeodomain and basic-helix-loop-helix proteins. The results provided information on upstream factors and signals potentially relevant to Six1 regulation in sensory neurons. We also report the establishment of a new transgenic mouse line (mSix1-8-NLSCre) that expresses Cre recombinase under the control of mouse Six1-8. Cre-mediated recombination was detected specifically in ISL1/2-positive sensory neurons of Six1-positive cranial sensory ganglia and DRG. The unique features of the mSix1-8-NLSCre line are the absence of Cre-mediated recombination in SOX10-positive glial cells and central nervous system and ability to induce recombination in a subset of neurons derived from the olfactory placode/epithelium. This mouse model can be potentially used to advance research on sensory development. PMID:26313368

  16. TRPA1 modulates mechanotransduction in cutaneous sensory neurons

    Kwan, Kelvin Y.; Glazer, Joshua M.; David P Corey; Rice, Frank L.; Stucky, Cheryl L.

    2009-01-01

    TRPA1 is expressed by nociceptive neurons of the dorsal root ganglia (DRG) and trigeminal ganglia, but its roles in cold and mechanotransduction are controversial. To determine the contribution of TRPA1 to cold and mechanotransduction in cutaneous primary afferent terminals, we used the ex-vivo skin-nerve preparation from Trpa1+/+, Trpa1+/− or Trpa1−/− adult mouse littermates. Cutaneous fibers from TRPA1-deficient mice showed no deficits in acute cold sensitivity, but they displayed striking ...

  17. Topographic Digital Raster Graphics, DRG - 1:250,000 Topo - Quads (7.5 Minute), Published in 1996, Arizona State Land Department.

    NSGIC GIS Inventory (aka Ramona) — This Topographic Digital Raster Graphics dataset, was produced all or in part from Hardcopy Maps information as of 1996. It is described as 'DRG - 1:250,000 Topo -...

  18. Topographic Digital Raster Graphics, DRG - 1:24,000 Topo - Quads (7.5 Minute), Published in 1996, Arizona State Land Department.

    NSGIC GIS Inventory (aka Ramona) — This Topographic Digital Raster Graphics dataset, was produced all or in part from Hardcopy Maps information as of 1996. It is described as 'DRG - 1:24,000 Topo -...

  19. Topographic Digital Raster Graphics, DRG - 1:100,000 Topo - Quads (7.5 Minute), Published in 1996, Arizona State Land Department.

    NSGIC GIS Inventory (aka Ramona) — This Topographic Digital Raster Graphics dataset, was produced all or in part from Hardcopy Maps information as of 1996. It is described as 'DRG - 1:100,000 Topo -...

  20. Impact of a DRG-based hospital financing system on quality and outcomes of care in Italy.

    Louis, D Z; Yuen, E J; Braga, M.; A. Cicchetti; Rabinowitz, C; Laine, C; Gonnella, J S

    1999-01-01

    OBJECTIVE: To examine potential changes in quality of care associated with a recent financing system implementation in Italy: in 1995, hospital financing reform implemented in Italy included the introduction of a DRG-based hospital financing system with the goals of controlling the growth of hospital costs and making hospitals more accountable for their productivity. DATA SOURCES: Hospital discharge abstract data from 1993 through 1996 for all hospitals (N=32) in the Friuli-Venezia-Giulia reg...

  1. Effects of DRG-based hospital payment in Poland on treatment of patients with stroke.

    Bystrov, Victor; Staszewska-Bystrova, Anna; Rutkowski, Daniel; Hermanowski, Tomasz

    2015-08-01

    A prospective payment system based on Diagnosis Related Groups (DRGs) presents strong financial incentives to healthcare providers. These incentives may have intended as well as unintended consequences for the healthcare system. In this paper we use administrative data on stroke admissions to Polish hospitals in order to demonstrate the response of hospitals to the incentives embedded in the design of stroke-related groups in Poland. The design was intended to motivate hospitals for the development of specialized stroke units by paying significantly higher tariffs for treatment of patients in these units. As a result, an extensive network of stroke units has emerged. However, as it is shown in the paper, there is no evidence that outcomes in hospitals with stroke units are significantly different from outcomes in hospitals without stroke units. It is also demonstrated that the reliance on the length of stay as a major grouping variable provides incentives for regrouping patients into more expensive groups by extending their length of stay in stroke units. The results of the study are limited by the incompleteness of the casemix data. There is a need to develop information and audit systems which would further inform a revision of the DRG system aimed to reduce the risk of regrouping and up-coding. PMID:26008985

  2. Optimizing diagnostic workup in the DRG environment: Dynamic algorithms and minimizing radiologic costs may cost your hospital money

    In certain diagnosis-related group (DRG) categories, the availability of sufficient CT scanners or of new equipment, such as MR equipment, can expedite the definitive workup. This will reduce the average length of stay and hospital cost. We analyzed the total hospital and radiologic charges by DRG category for all patients admitted to our hospital in 1985 and 1986. Although the cost per procedure is relatively high, the radiologic component is a small percentage of total hospital costs (median, 3%; maximum, <10%). The authors developed alternative diagnostic algorithms for radiologic-intensive DRG categories. Different diagnostic algorithms proposed for the same clinical problems were compared analytically in terms of impact on the hospital (cost, equipment availability, and length of stay). An example is the workup for FUO. Traditional approach uses plain x-rays and gallium scans and only uses CT when localizing symptoms are present. An alternative approach is to perform CT only. Although more CT examinations would be required, there is considerable reduction in the length of hospital stay and in overall charges. Neurologic and thoracic workups will be given as examples of classes or problems that can be addressed analytically: sequencing of the workup; prevalence; patient population; resource of allocation; and introduction of new imaging modality

  3. Modulation of chloride homeostasis by inflammatory mediators in dorsal root ganglion neurons

    Möhrlen Frank

    2008-08-01

    Full Text Available Abstract Background Chloride currents in peripheral nociceptive neurons have been implicated in the generation of afferent nociceptive signals, as Cl- accumulation in sensory endings establishes the driving force for depolarizing, and even excitatory, Cl- currents. The intracellular Cl- concentration can, however, vary considerably between individual DRG neurons. This raises the question, whether the contribution of Cl- currents to signal generation differs between individual afferent neurons, and whether the specific Cl- levels in these neurons are subject to modulation. Based on the hypothesis that modulation of the peripheral Cl- homeostasis is involved in the generation of inflammatory hyperalgesia, we examined the effects of inflammatory mediators on intracellular Cl- concentrations and on the expression levels of Cl- transporters in rat DRG neurons. Results We developed an in vitro assay for testing how inflammatory mediators influence Cl- concentration and the expression of Cl- transporters. Intact DRGs were treated with 100 ng/ml NGF, 1.8 μM ATP, 0.9 μM bradykinin, and 1.4 μM PGE2 for 1–3 hours. Two-photon fluorescence lifetime imaging with the Cl--sensitive dye MQAE revealed an increase of the intracellular Cl- concentration within 2 hours of treatment. This effect coincided with enhanced phosphorylation of the Na+-K+-2Cl- cotransporter NKCC1, suggesting that an increased activity of that transporter caused the early rise of intracellular Cl- levels. Immunohistochemistry of NKCC1 and KCC2, the main neuronal Cl- importer and exporter, respectively, exposed an inverse regulation by the inflammatory mediators. While the NKCC1 immunosignal increased, that of KCC2 declined after 3 hours of treatment. In contrast, the mRNA levels of the two transporters did not change markedly during this time. These data demonstrate a fundamental transition in Cl- homeostasis toward a state of augmented Cl- accumulation, which is induced by a 1–3 hour

  4. GABAB receptors inhibit low-voltage activated and high-voltage activated Ca(2+) channels in sensory neurons via distinct mechanisms.

    Huang, Dongyang; Huang, Sha; Peers, Chris; Du, Xiaona; Zhang, Hailin; Gamper, Nikita

    2015-09-18

    Growing evidence suggests that mammalian peripheral somatosensory neurons express functional receptors for gamma-aminobutyric acid, GABAA and GABAB. Moreover, local release of GABA by pain-sensing (nociceptive) nerve fibres has also been suggested. Yet, the functional significance of GABA receptor triggering in nociceptive neurons is not fully understood. Here we used patch-clamp recordings from small-diameter cultured DRG neurons to investigate effects of GABAB receptor agonist baclofen on voltage-gated Ca(2+) currents. We found that baclofen inhibited both low-voltage activated (LVA, T-type) and high-voltage activated (HVA) Ca(2+) currents in a proportion of DRG neurons by 22% and 32% respectively; both effects were sensitive to Gi/o inhibitor pertussis toxin. Inhibitory effect of baclofen on both current types was about twice less efficacious as compared to that of the μ-opioid receptor agonist DAMGO. Surprisingly, only HVA but not LVA current modulation by baclofen was partially prevented by G protein inhibitor GDP-β-S. In contrast, only LVA but not HVA current modulation was reversed by the application of a reducing agent dithiothreitol (DTT). Inhibition of T-type Ca(2+) current by baclofen and the recovery of such inhibition by DTT were successfully reconstituted in the expression system. Our data suggest that inhibition of LVA current in DRG neurons by baclofen is partially mediated by an unconventional signaling pathway that involves a redox mechanism. These findings reinforce the idea of targeting peripheral GABA receptors for pain relief. PMID:26239659

  5. Assembly of juxtaparanodes in myelinating DRG culture: Differential clustering of the Kv1/Caspr2 complex and scaffolding protein 4.1B.

    Hivert, Bruno; Pinatel, Delphine; Labasque, Marilyne; Tricaud, Nicolas; Goutebroze, Laurence; Faivre-Sarrailh, Catherine

    2016-05-01

    The precise distribution of ion channels at the nodes of Ranvier is essential for the efficient propagation of action potentials along myelinated axons. The voltage-gated potassium channels Kv1.1/1.2 are clustered at the juxtaparanodes in association with the cell adhesion molecules, Caspr2 and TAG-1 and the scaffolding protein 4.1B. In the present study, we set up myelinating cultures of DRG neurons and Schwann cells to look through the formation of juxtaparanodes in vitro. We showed that the Kv1.1/Kv1.2 channels were first enriched at paranodes before being restricted to distal paranodes and juxtaparanodes. In addition, the Kv1 channels displayed an asymmetric expression enriched at the distal juxtaparanodes. Caspr2 was strongly co-localized with Kv1.2 whereas the scaffolding protein 4.1B was preferentially recruited at paranodes while being present at juxtaparanodes too. Kv1.2/Caspr2 but not 4.1B, also transiently accumulated within the nodal region both in myelinated cultures and developing sciatic nerves. Studying cultures and sciatic nerves from 4.1B KO mice, we further showed that 4.1B is required for the proper targeting of Caspr2 early during myelination. Moreover, using adenoviral-mediated expression of Caspr-GFP and photobleaching experiments, we analyzed the stability of paranodal junctions and showed that the lateral stability of paranodal Caspr was not altered in 4.1B KO mice indicating that 4.1B is not required for the assembly and stability of the paranodal junctions. Thus, developing an adapted culture paradigm, we provide new insights into the dynamic and differential distribution of Kv1 channels and associated proteins during myelination. GLIA 2016;64:840-852. PMID:26840208

  6. Ultrafast Unzipping of a Beta-Hairpin Peptide

    Zinth, W.; Schrader, T.E.; Schreier, W.J.; Koller, F.O.; Cordes, T.; Babitzki, G.; Denschlag, R.; Tavan, P.; Löweneck, M.; Dong, Shou-Liang; Moroder, L.; Renner, C.; Corkum, Paul; Jonas, David M.; Miller, R.J. Dwayne; Weiner, Andrew M.

    2007-01-01

    Light induced switching of a beta-hairpin structure is investigated by femtosecond IR spectroscopy. While the unzipping process comprises ultrafast kinetics and is finished within 1 ns, the folding into the hairpin structure is a much slower process.

  7. Reduced thermal sensitivity and Nav1.8 and TRPV1 channel expression in sensory neurons of aged mice

    Wang, Shuying; Davis, Brian M.; Zwick, Melissa; Waxman, Stephen G.; Albers, Kathryn M.

    2005-01-01

    Sensory neurons in aging mammals undergo changes in anatomy, physiology and gene expression that correlate with reduced sensory perception. In this study we compared young and aged mice to identify proteins that might contribute to this loss of sensation. We first show using behavioral testing that thermal sensitivity in aged male and female mice is reduced. Expression of sodium channel (Nav1.8 and Nav1.9) and transient receptor potential vanilloid (TRPV) channels in DRG and peripheral nerves...

  8. Increase of TRPV1-Immunoreactivity in Dorsal Root Ganglia Neurons Innervating the Femur in a Rat Model of Osteoporosis

    Yoshino, Kensuke; Suzuki, Miyako; Kawarai, Yuya; Sakuma, Yoshihiro; Inoue, Gen; Orita, Sumihisa; Yamauchi, Kazuyo; Aoki, Yasuchika; Ishikawa, Tetsuhiro; Miyagi, Masayuki; Kamoda, Hiroto; Kubota, Gou; Oikawa, Yasuhiro; Inage, Kazuhide; Sainoh, Takeshi

    2014-01-01

    Purpose Transient receptor potential vanilloid 1 (TRPV1) is a ligand-gated nonselective cation channel, which can be activated by capsaicin and other noxious stimuli. Recently, an association between bone pain and TRPV1 has been reported. However, the influence of osteoporosis on TRPV1 in the sensory system innervating the femur has not been reported. Materials and Methods TRPV1-immunoreactive (ir) in dorsal root ganglia (DRG) neurons labeled with neurotracer [Fluoro-Gold (FG)] innervating th...

  9. Prostaglandin E2 potentiation of P2X3 receptor mediated currents in dorsal root ganglion neurons

    Huang Li-Yen

    2007-08-01

    Full Text Available Abstract Prostaglandin E2 (PGE2 is a well-known inflammatory mediator that enhances the excitability of DRG neurons. Homomeric P2X3 and heteromeric P2X2/3 receptors are abundantly expressed in dorsal root ganglia (DRG neurons and participate in the transmission of nociceptive signals. The interaction between PGE2 and P2X3 receptors has not been well delineated. We studied the actions of PGE2 on ATP-activated currents in dissociated DRG neurons under voltage-clamp conditions. PGE2 had no effects on P2X2/3 receptor-mediated responses, but significantly potentiated fast-inactivating ATP currents mediated by homomeric P2X3 receptors. PGE2 exerted its action by activating EP3 receptors. To study the mechanism underlying the action of PGE2, we found that the adenylyl cyclase activator, forskolin and the membrane-permeable cAMP analogue, 8-Br-cAMP increased ATP currents, mimicking the effect of PGE2. In addition, forskolin occluded the enhancement produced by PGE2. The protein kinase A (PKA inhibitors, H89 and PKA-I blocked the PGE2 effect. In contrast, the PKC inhibitor, bisindolymaleimide (Bis did not change the potentiating action of PGE2. We further showed that PGE2 enhanced α,β-meATP-induced allodynia and hyperalgesia and the enhancement was blocked by H89. These observations suggest that PGE2 binds to EP3 receptors, resulting in the activation of cAMP/PKA signaling pathway and leading to an enhancement of P2X3 homomeric receptor-mediated ATP responses in DRG neurons.

  10. AMPA and NMDA glutamate receptors are found in both peptidergic and non-peptidergic primary afferent neurons in the rat

    Willcockson, Helen; Valtschanoff, Juli

    2008-01-01

    Two distinct classes of nociceptive primary afferents, peptidergic and non-peptidergic, respond similarly to acute noxious stimulation; however the peptidergic afferents are more likely to play a role in inflammatory pain, while the non-peptidergic afferents may be more characteristically involved in neuropathic pain. Using multiple immunofluorescence, we determined the proportions of neurons in the rat L4 dorsal root ganglion (DRG) that co-express AMPA or NMDA glutamate receptors and markers...

  11. Expression and Regulation of Cav3.2 T-Type Calcium Channels during Inflammatory Hyperalgesia in Mouse Dorsal Root Ganglion Neurons.

    Masaya Watanabe

    Full Text Available The Cav3.2 isoform of the T-type calcium channel is expressed in primary sensory neurons of the dorsal root ganglion (DRG, and these channels contribute to nociceptive and neuropathic pain in rats. However, there are conflicting reports on the roles of these channels in pain processing in rats and mice. In addition, the function of T-type channels in persistent inflammatory hyperalgesia is poorly understood. We performed behavioral and comprehensive histochemical analyses to characterize Cav3.2-expressing DRG neurons and examined the regulation of T-type channels in DRGs from C57BL/6 mice with carrageenan-induced inflammatory hyperalgesia. We show that approximately 20% of mouse DRG neurons express Cav3.2 mRNA and protein. The size of the majority of Cav3.2-positive DRG neurons (69 ± 8% ranged from 300 to 700 μm2 in cross-sectional area and 20 to 30 μm in estimated diameter. These channels co-localized with either neurofilament-H (NF-H or peripherin. The peripherin-positive cells also overlapped with neurons that were positive for isolectin B4 (IB4 and calcitonin gene-related peptide (CGRP but were distinct from transient receptor potential vanilloid 1 (TRPV1-positive neurons during normal mouse states. In mice with carrageenan-induced inflammatory hyperalgesia, Cav3.2 channels, but not Cav3.1 or Cav3.3 channels, were upregulated in ipsilateral DRG neurons during the sub-acute phase. The increased Cav3.2 expression partially resulted from an increased number of Cav3.2-immunoreactive neurons; this increase in number was particularly significant for TRPV1-positive neurons. Finally, preceding and periodic intraplantar treatment with the T-type calcium channel blockers mibefradil and NNC 55-0396 markedly reduced and reversed mechanical hyperalgesia during the acute and sub-acute phases, respectively, in mice. These data suggest that Cav3.2 T-type channels participate in the development of inflammatory hyperalgesia, and this channel might play an

  12. SPARCL1-containing neurons in the human brainstem and sensory ganglion.

    Hashimoto, Naoya; Sato, Tadasu; Yajima, Takehiro; Fujita, Masatoshi; Sato, Ayumi; Shimizu, Yoshinaka; Shimada, Yusuke; Shoji, Noriaki; Sasano, Takashi; Ichikawa, Hiroyuki

    2016-06-01

    Secreted protein, acidic and rich in cysteine-like 1 (SPARCL1) is a member of the osteonectin family of proteins. In this study, immunohistochemistry for SPARCL1 was performed to obtain its distribution in the human brainstem, cervical spinal cord, and sensory ganglion. SPARCL1-immunoreactivity was detected in neuronal cell bodies including perikarya and proximal dendrites, and the neuropil. The motor nuclei of the IIIrd, Vth, VIth, VIIth, IXth, Xth, XIth, and XIIth cranial nerves and spinal nerves contained many SPARCL1-immunoreactive (-IR) neurons with medium-sized to large cell bodies. Small and medium-sized SPARCL1-IR neurons were distributed in sensory nuclei of the Vth, VIIth, VIIIth, IXth, and Xth cranial nerves. In the medulla oblongata, the dorsal column nuclei also had small to medium-sized SPARCL1-IR neurons. In addition, SPARCL1-IR neurons were detected in the nucleus of the trapezoid body and pontine nucleus within the pons and the arcuate nucleus in the medulla oblongata. In the cervical spinal cord, the ventral horn contained some SPARCL1-IR neurons with large cell bodies. These findings suggest that SPARCL1-containing neurons function to relay and regulate motor and sensory signals in the human brainstem. In the dorsal root (DRG) and trigeminal ganglia (TG), primary sensory neurons contained SPARCL1-immunoreactivity. The proportion of SPARCL1-IR neurons in the TG (mean ± SD, 39.9 ± 2.4%) was higher than in the DRG (30.6 ± 2.1%). SPARCL1-IR neurons were mostly medium-sized to large (mean ± SD, 1494.5 ± 708.3 μm(2); range, 320.4-4353.4 μm(2)) in the DRG, whereas such neurons were of various cell body sizes in the TG (mean ± SD, 1291.2 ± 532.8 μm(2); range, 209.3-4326.4 μm(2)). There appears to be a SPARCL1-containing sensory pathway in the ganglion and brainstem of the spinal and trigeminal nervous systems. PMID:27357901

  13. Quantitative Analysis of Rat Dorsal Root Ganglion Neurons Cultured on Microelectrode Arrays Based on Fluorescence Microscopy Image Processing.

    Mari, João Fernando; Saito, José Hiroki; Neves, Amanda Ferreira; Lotufo, Celina Monteiro da Cruz; Destro-Filho, João-Batista; Nicoletti, Maria do Carmo

    2015-12-01

    Microelectrode Arrays (MEA) are devices for long term electrophysiological recording of extracellular spontaneous or evocated activities on in vitro neuron culture. This work proposes and develops a framework for quantitative and morphological analysis of neuron cultures on MEAs, by processing their corresponding images, acquired by fluorescence microscopy. The neurons are segmented from the fluorescence channel images using a combination of segmentation by thresholding, watershed transform, and object classification. The positioning of microelectrodes is obtained from the transmitted light channel images using the circular Hough transform. The proposed method was applied to images of dissociated culture of rat dorsal root ganglion (DRG) neuronal cells. The morphological and topological quantitative analysis carried out produced information regarding the state of culture, such as population count, neuron-to-neuron and neuron-to-microelectrode distances, soma morphologies, neuron sizes, neuron and microelectrode spatial distributions. Most of the analysis of microscopy images taken from neuronal cultures on MEA only consider simple qualitative analysis. Also, the proposed framework aims to standardize the image processing and to compute quantitative useful measures for integrated image-signal studies and further computational simulations. As results show, the implemented microelectrode identification method is robust and so are the implemented neuron segmentation and classification one (with a correct segmentation rate up to 84%). The quantitative information retrieved by the method is highly relevant to assist the integrated signal-image study of recorded electrophysiological signals as well as the physical aspects of the neuron culture on MEA. Although the experiments deal with DRG cell images, cortical and hippocampal cell images could also be processed with small adjustments in the image processing parameter estimation. PMID:26510475

  14. Shaping the System – The DRG Evaluation Project of the German Society for Gynaecology and Obstetrics (Deutsche Gesellschaft für Gynäkologie und Geburtshilfe, DGGG)

    Fiori, W; Renner, S. P.; Siam, K.; Babapirali, J.; Roeder, N; Dausch, E.; Hildebrandt, T.; Hillemanns, P; Nehmzow, M.; Zygmunt, M.; Piroth, D.; Schem, C; Schwenzer, T.; Friese, K.; Wallwiener, D.

    2013-01-01

    Introduction: The German DRG system is annually adapted to the changing services provided. For the further development, the self-governing body and its DRG Institute (InEK) depend on participation of the users. Methods: For one of the DRG evaluation projects initiated by DGGG, cost and performance data for the year 2011 from 16 hospitals were available. After plausibility checks and corrections, analyses for service and cost homogeneity were performed. In cases of inadequate...

  15. 95. German Roentgen congress and 7. joint congress of the DRG and OeRG. Program with abstracts; 95. Deutscher Roentgenkongress und 7. Gemeinsamer Kongress von DRG und OeRG. Vollstaendiges Programm mit Abstracts

    Diederich, Stefan; Lammer, Johannes (eds.)

    2014-05-15

    The volume contains the program and the abstracts of the 95th German Roentgen congress and the 7th joint congress of the DRG and OeRG. The radiological focal points of the congress were thorax radiology (pneumology: lung fibrosis, emphysema); oncological radiology: skeletal carcinoma, lung carcinoma, kidneys, lung metastases, primary liver carcinoma, liver metastases. Further topics included radiology in hospitals and medical centers, tele-radiology, ambulant health care, legal issues, financial accounting and management issues. Several courses an specific radiological issues and radiological techniques were offered, including radiation protection and legal aspects.

  16. The mast cell degranulator compound 48/80 directly activates neurons.

    Michael Schemann

    Full Text Available BACKGROUND: Compound 48/80 is widely used in animal and tissue models as a "selective" mast cell activator. With this study we demonstrate that compound 48/80 also directly activates enteric neurons and visceral afferents. METHODOLOGY/PRINCIPAL FINDINGS: We used in vivo recordings from extrinsic intestinal afferents together with Ca(++ imaging from primary cultures of DRG and nodose neurons. Enteric neuronal activation was examined by Ca(++ and voltage sensitive dye imaging in isolated gut preparations and primary cultures of enteric neurons. Intraluminal application of compound 48/80 evoked marked afferent firing which desensitized on subsequent administration. In egg albumen-sensitized animals, intraluminal antigen evoked a similar pattern of afferent activation which also desensitized on subsequent exposure to antigen. In cross-desensitization experiments prior administration of compound 48/80 failed to influence the mast cell mediated response. Application of 1 and 10 µg/ml compound 48/80 evoked spike discharge and Ca(++ transients in enteric neurons. The same nerve activating effect was observed in primary cultures of DRG and nodose ganglion cells. Enteric neuron cultures were devoid of mast cells confirmed by negative staining for c-kit or toluidine blue. In addition, in cultured enteric neurons the excitatory action of compound 48/80 was preserved in the presence of histamine H(1 and H(2 antagonists. The mast cell stabilizer cromolyn attenuated compound 48/80 and nicotine evoked Ca(++ transients in mast cell-free enteric neuron cultures. CONCLUSIONS/SIGNIFICANCE: The results showed direct excitatory action of compound 48/80 on enteric neurons and visceral afferents. Therefore, functional changes measured in tissue or animal models may involve a mast cell independent effect of compound 48/80 and cromolyn.

  17. Using DRG to analyze hospital production: a re-classification model based on a linear tree-network topology

    Achille Lanzarini

    2014-09-01

    Full Text Available Background: Hospital discharge records are widely classified through the Diagnosis Related Group (DRG system; the version currently used in Italy counts 538 different codes, including thousands of diagnosis and procedures. These numbers reflect the considerable effort of simplification, yet the current classification system is of little use to evaluate hospital production and performance.Methods: As the case-mix of a given Hospital Unit (HU is driven by its physicians’ specializations, a grouping of DRGs into a specialization-driven classification system has been conceived through the analysis of HUs discharging and the ICD-9-CM codes. We propose a three-folded classification, based on the analysis of 1,670,755 Hospital Discharge Cards (HDCs produced by Lombardy Hospitals in 2010; it consists of 32 specializations (e.g. Neurosurgery, 124 sub-specialization (e.g. skull surgery and 337 sub-sub-specialization (e.g. craniotomy.Results: We give a practical application of the three-layered approach, based on the production of a Neurosurgical HU; we observe synthetically the profile of production (1,305 hospital discharges for 79 different DRG codes of 16 different MDC are grouped in few groups of homogeneous DRG codes, a more informative production comparison (through process-specific comparisons, rather than crude or case-mix standardized comparisons and a potentially more adequate production planning (considering the Neurosurgical HUs of the same city, those produce a limited quote of the whole neurosurgical production, because the same activity can be realized by non-Neurosugical HUs.Conclusion: Our work may help to evaluate the hospital production for a rational planning of available resources, blunting information asymmetries between physicians and managers. 

  18. The expression of ELK transcription factors in adult DRG: novel isoforms, antisense transcripts and upregulation by nerve damage

    Kerr, Niall; Pintzas, Alexander; Holmes, Fiona; Hobson, Sally-Ann; Pope, Robert; Wallace, Mark; Wasylyk, Christine; Wasylyk, Bohdan; Wynick, David

    2010-01-01

    ELK transcription factors are expressed in brain, but it is unknown whether they are expressed in the peripheral nervous system. We show by RT-PCR that the previously described Elk1, Elk3/Elk3b/Elk3c and Elk4 mRNAs are expressed in adult dorsal root ganglia (DRG), together with the novel alternatively spliced isoforms Elk1b, Elk3d and Elk4c/Elk4d/Elk4e. These isoforms are also expressed in brain, heart, kidney and testis. In contrast to Elk3 protein, the novel Elk3d isoform is cytoplasmic, fa...

  19. Monitoring the impact of the DRG payment system on nursing service context factors in Swiss acute care hospitals: Study protocol

    Spirig, Rebecca

    2014-03-01

    Full Text Available [english] Aims: With this study protocol, a research program is introduced. Its overall aim is to prepare the instruments and to conduct the first monitoring of nursing service context factors at three university and two cantonal hospitals in Switzerland prior to the introduction of the reimbursement system based on Diagnosis Related Groups (DRG and to further develop a theoretical model as well as a methodology for future monitoring following the introduction of DRGs.Background: DRG was introduced to all acute care hospitals in Switzerland in 2012. In other countries, DRG introduction led to rationing and subsequently to a reduction in nursing care. As result, nursing-sensitive patient outcomes were seriously jeopardised. Switzerland has the opportunity to learn from the consequences experienced by other countries when they introduced DRGs. Their experiences highlight that DRGs influence nursing service context factors such as complexity of nursing care or leadership, which in turn influence nursing-sensitive patient outcomes. For this reason, the monitoring of nursing service context factors needs to be an integral part of the introduction of DRGs. However, most acute care hospitals in Switzerland do not monitor nursing service context data. Nursing managers and hospital executive boards will be in need of this data in the future, in order to distribute resources effectively. Methods/Design: A mixed methods design in the form of a sequential explanatory strategy was chosen. During the preparation phase, starting in spring 2011, instruments were selected and prepared, and the access to patient and nursing data in the hospitals was organized. Following this, online collection of quantitative data was conducted in fall 2011. In summer 2012, qualitative data was gathered using focus group interviews, which helped to describe the processes in more detail. During 2013 and 2014, an integration process is being conducted involving complementing

  20. 95. German Roentgen congress and 7. joint congress of the DRG and OeRG. Program with abstracts

    The volume contains the program and the abstracts of the 95th German Roentgen congress and the 7th joint congress of the DRG and OeRG. The radiological focal points of the congress were thorax radiology (pneumology: lung fibrosis, emphysema); oncological radiology: skeletal carcinoma, lung carcinoma, kidneys, lung metastases, primary liver carcinoma, liver metastases. Further topics included radiology in hospitals and medical centers, tele-radiology, ambulant health care, legal issues, financial accounting and management issues. Several courses an specific radiological issues and radiological techniques were offered, including radiation protection and legal aspects.

  1. The pathophysiology of chronic pain--increased sensitivity to low threshold A beta-fibre inputs.

    Woolf, C J; Doubell, T P

    1994-08-01

    Chronic pain is characterized by abnormal sensitivity, which is due to the generation of pain in response to the activation of the low-threshold mechanoreceptive A beta fibres that normally generate innocuous sensations. Three different processes in the spinal cord can account for this dramatic alteration in sensory processing in the somatosensory system: increased excitability, decreased inhibition and structural reorganization. All have been shown to occur and each may contribute separately or together to the wide range of chronic inflammatory and neuropathic pain disorders. The unravelling of the cellular mechanisms involved both offers the potential for developing novel therapeutic strategies, which reduce functional synaptic plasticity and prevent central atrophic and regenerative responses in injured neurones, and illustrates the capacity of the adult nervous system for maladaptive modification. PMID:7812141

  2. Segmental distribution and morphometric features of primary sensory neurons projecting to the tibial periosteum in the rat.

    Tadeusz Cichocki

    2004-07-01

    Full Text Available Previous reports have demonstrated very rich innervation pattern in the periosteum. Most of the periosteal fibers were found to be sensory in nature. The aim of this study was to identify the primary sensory neurons that innervate the tibial periosteum in the adult rat and to describe the morphometric features of their perikarya. To this end, an axonal fluorescent carbocyanine tracer, DiI, was injected into the periosteum on the medial surface of the tibia. The perikarya of the sensory fibers were traced back in the dorsal root ganglia (DRG L1-L6 by means of fluorescent microscopy on cryosections. DiI-containing neurons were counted in each section and their segmental distribution was determined. Using PC-assisted image analysis system, the size and shape of the traced perikarya were analyzed. DiI-labeled sensory neurons innervating the periosteum of the tibia were located in the DRG ipsilateral to the injection site, with the highest distribution in L3 and L4 (57% and 23%, respectively. The majority of the traced neurons were of small size (area < 850 microm2, which is consistent with the size distribution of CGRP- and SP-containing cells, regarded as primary sensory neurons responsible for perception of pain and temperature. A small proportion of labeled cells had large perikarya and probably supplied corpuscular sense receptors observed in the periosteum. No differences were found in the shape distribution of neurons belonging to different size classes.

  3. The expression of Toll-like receptor 4, 7 and co-receptors in neurochemical sub-populations of rat trigeminal ganglion sensory neurons.

    Helley, M P; Abate, W; Jackson, S K; Bennett, J H; Thompson, S W N

    2015-12-01

    The recent discovery that mammalian nociceptors express Toll-like receptors (TLRs) has raised the possibility that these cells directly detect and respond to pathogens with implications for either direct nociceptor activation or sensitization. A range of neuronal TLRs have been identified, however a detailed description regarding the distribution of expression of these receptors within sub-populations of sensory neurons is lacking. There is also some debate as to the composition of the TLR4 receptor complex on sensory neurons. Here we use a range of techniques to quantify the expression of TLR4, TLR7 and some associated molecules within neurochemically-identified sub-populations of trigeminal (TG) and dorsal root (DRG) ganglion sensory neurons. We also detail the pattern of expression and co-expression of two isoforms of lysophosphatidylcholine acyltransferase (LPCAT), a phospholipid remodeling enzyme previously shown to be involved in the lipopolysaccharide-dependent TLR4 response in monocytes, within sensory ganglia. Immunohistochemistry shows that both TLR4 and TLR7 preferentially co-localize with transient receptor potential vallinoid 1 (TRPV1) and purinergic receptor P2X ligand-gated ion channel 3 (P2X3), markers of nociceptor populations, within both TG and DRG. A gene expression profile shows that TG sensory neurons express a range of TLR-associated molecules. LPCAT1 is expressed by a proportion of both nociceptors and non-nociceptive neurons. LPCAT2 immunostaining is absent from neuronal profiles within both TG and DRG and is confined to non-neuronal cell types under naïve conditions. Together, our results show that nociceptors express the molecular machinery required to directly respond to pathogenic challenge independently from the innate immune system. PMID:26434622

  4. Predictors of High Profit and High Deficit Outliers under SwissDRG of a Tertiary Care Center

    Mehra, Tarun; Müller, Christian Thomas Benedikt; Volbracht, Jörk; Seifert, Burkhardt; Moos, Rudolf

    2015-01-01

    Principles Case weights of Diagnosis Related Groups (DRGs) are determined by the average cost of cases from a previous billing period. However, a significant amount of cases are largely over- or underfunded. We therefore decided to analyze earning outliers of our hospital as to search for predictors enabling a better grouping under SwissDRG. Methods 28,893 inpatient cases without additional private insurance discharged from our hospital in 2012 were included in our analysis. Outliers were defined by the interquartile range method. Predictors for deficit and profit outliers were determined with logistic regressions. Predictors were shortlisted with the LASSO regularized logistic regression method and compared to results of Random forest analysis. 10 of these parameters were selected for quantile regression analysis as to quantify their impact on earnings. Results Psychiatric diagnosis and admission as an emergency case were significant predictors for higher deficit with negative regression coefficients for all analyzed quantiles (pDRG split criteria as they predict large, consistent and significant losses. PMID:26517545

  5. The expression of ELK transcription factors in adult DRG: Novel isoforms, antisense transcripts and upregulation by nerve damage.

    Kerr, Niall; Pintzas, Alexander; Holmes, Fiona; Hobson, Sally-Ann; Pope, Robert; Wallace, Mark; Wasylyk, Christine; Wasylyk, Bohdan; Wynick, David

    2010-06-01

    ELK transcription factors are known to be expressed in a number of regions in the nervous system. We show by RT-PCR that the previously described Elk1, Elk3/Elk3b/Elk3c and Elk4 mRNAs are expressed in adult dorsal root ganglia (DRG), together with the novel alternatively spliced isoforms Elk1b, Elk3d and Elk4c/Elk4d/Elk4e. These isoforms are also expressed in brain, heart, kidney and testis. In contrast to Elk3 protein, the novel Elk3d isoform is cytoplasmic, fails to bind ETS binding sites and yet can activate transcription by an indirect mechanism. The Elk3 and Elk4 genes are overlapped by co-expressed Pctk2 (Cdk17) and Mfsd4 genes, respectively, with the potential formation of Elk3/Pctaire2 and Elk4/Mfsd4 sense-antisense mRNA heteroduplexes. After peripheral nerve injury the Elk3 mRNA isoforms are each upregulated approximately 2.3-fold in DRG (P<0.005), whereas the natural antisense Pctaire2 isoforms show only a small increase (21%, P<0.01) and Elk1 and Elk4 mRNAs are unchanged. PMID:20304071

  6. Has DRG payment influenced the technical efficiency and productivity of diagnostic technologies in Portuguese public hospitals? An empirical analysis using parametric and non-parametric methods.

    Dismuke, C E; Sena, V

    1999-05-01

    The use of Diagnosis Related Groups (DRG) as a mechanism for hospital financing is a currently debated topic in Portugal. The DRG system was scheduled to be initiated by the Health Ministry of Portugal on January 1, 1990 as an instrument for the allocation of public hospital budgets funded by the National Health Service (NHS), and as a method of payment for other third party payers (e.g., Public Employees (ADSE), private insurers, etc.). Based on experience from other countries such as the United States, it was expected that implementation of this system would result in more efficient hospital resource utilisation and a more equitable distribution of hospital budgets. However, in order to minimise the potentially adverse financial impact on hospitals, the Portuguese Health Ministry decided to gradually phase in the use of the DRG system for budget allocation by using blended hospital-specific and national DRG case-mix rates. Since implementation in 1990, the percentage of each hospital's budget based on hospital specific costs was to decrease, while the percentage based on DRG case-mix was to increase. This was scheduled to continue until 1995 when the plan called for allocating yearly budgets on a 50% national and 50% hospital-specific cost basis. While all other non-NHS third party payers are currently paying based on DRGs, the adoption of DRG case-mix as a National Health Service budget setting tool has been slower than anticipated. There is now some argument in both the political and academic communities as to the appropriateness of DRGs as a budget setting criterion as well as to their impact on hospital efficiency in Portugal. This paper uses a two-stage procedure to assess the impact of actual DRG payment on the productivity (through its components, i.e., technological change and technical efficiency change) of diagnostic technology in Portuguese hospitals during the years 1992-1994, using both parametric and non-parametric frontier models. We find evidence

  7. The majority of dorsal spinal cord gastrin releasing peptide is synthesized locally whereas neuromedin B is highly expressed in pain- and itch-sensing somatosensory neurons

    Fleming, Michael S; Ramos, Daniel; Han, Seung Baek; Zhao, JianYuan; Son, Young-Jin; Luo, Wenqin

    2012-01-01

    Background Itch is one of the major somatosensory modalities. Some recent findings have proposed that gastrin releasing peptide (Grp) is expressed in a subset of dorsal root ganglion (DRG) neurons and functions as a selective neurotransmitter for transferring itch information to spinal cord interneurons. However, expression data from public databases and earlier literatures indicate that Grp mRNA is only detected in dorsal spinal cord (dSC) whereas its family member neuromedin B (Nmb) is high...

  8. The majority of dorsal spinal cord gastrin releasing peptide is synthesized locally whereas neuromedin B is highly expressed in pain- and itch-sensing somatosensory neurons

    Fleming Michael S; Ramos Daniel; Han Seung; Zhao Jianyuan; Son Young-Jin; Luo Wenqin

    2012-01-01

    Abstract Background Itch is one of the major somatosensory modalities. Some recent findings have proposed that gastrin releasing peptide (Grp) is expressed in a subset of dorsal root ganglion (DRG) neurons and functions as a selective neurotransmitter for transferring itch information to spinal cord interneurons. However, expression data from public databases and earlier literatures indicate that Grp mRNA is only detected in dorsal spinal cord (dSC) whereas its family member neuromedin B (Nmb...

  9. Prokaryotic expression of recombinant human p75NTR-Fc fusion protein and its effect on the neurite outgrowth of dorsal root ganglia neuron

    Zhu Feng; Wang Yongtang; Lu Xiumin; Zeng Lin; Wu Yamin

    2009-01-01

    Objective: To clone, express, and identify the extracellular domain gene of human p75 neurotrophin receptor with IgG-Fc (hp75NTR-Fc) in prokaryotic expression system, and investigate the effect of the recombinant protein on dorsal root ganglia (DRG) neuron neurites. Methods: The hp75NTR-Fc coding sequence was amplified from pcDNA-hp75NTR-Fc by polymerase chain reaction (PCR) and subcloned into vector pET30a (+), in which hp75NTR-Fc expression was controlled under the T7 promoter. The recombinant vectors were amplified in E. coli DH5a and identified by PCR, enzyme digestion and sequencing, and then transformed into E. coli BL21 (DE3). The expression product was analyzed with SDS-PAGE and Western blot. Then after the recombinant protein purified with Protein A affinity chromatograph, and renaturated with dialysis, respectively, the effect of the recombinant protein on DRG neuron neuritis was further investigated. Results: The results of PCR, enzyme digestion, and sequencing demonstrated the success of inserting the hp75NTR-Fc fragment into vector pET30a (+). SDS-PAGE and Western blot showed a positive protein band with molecular weight about 50 kD in the expression product, which is accordant with the interest protein, and this band could be specifically recognized by rabbit anti-NGFRp75 antibody. The purified infusion protein following dialysis could promote neurite outgrowth of DRG neurons cultured with myelin-associated glycoprotein (MAG). Conclusion: The hp75NTR-Fc coding sequence was subcloned into the expression vector pET30a (+) correctly and expressed successfully in the prokaryotic expression system. The infusion protein could promote neurite outgrowth of DRG neurons cultured with MAG.

  10. Modulation of dragon's blood on tetrodotoxin-resistant sodium currents in dorsal root ganglion neurons and identification of its material basis for efficacy

    LIU Xiangming; CHEN Su; ZHANG Yuxia; ZHANG Fan

    2006-01-01

    To clarify the modulation of dragon's blood on the tetrodotoxin-resistant (TTX-R) sodium currents in dorsal root ganglion (DRG) neurons and explore its corresponding material basis for the efficacy, using whole-cell patch clamp technique, the effects of dragon's blood and the combined effects of three components (cochinchinenin A, cochinchinenin B, and loureirin B) extracted from dragon's blood on the TTX-R sodium currents in acute-isolated DRG neurons of rats were observed. According to the operational definition of material basis for the efficacy of TCM established, the material basis of the modulation on the TTX-R sodium currents in DRG neurons of dragon's blood was judged from the experimental results. The drug interaction equation of Greco et al. was used to assess the interaction of the three components extracted from dragon's blood. This investigation demonstrated that dragon's blood suppressed the peak TTX-R sodium currents in a dose-dependent way and affected the activations of TTX-R sodium currents. The effects of the combination of cochinchinenin A, cochinchinenin B, and loureirin B were in good agreement with those of dragon's blood. Although the three components used alone could modulate TTX-R sodium currents, the concentrations of the three components used alone were respectively higher than those used in combination when the inhibition rates on the TTX-R sodium currents of them used alone and in combination were the same. The combined effects of the three components were synergistic. These results suggested that the interference with pain messages caused by the modulation of dragon's blood on TTX-R sodium currents in DRG neurons may explain some of the analgesic effect of dragon's blood and the corresponding material basis for the efficacy is the combination of cochinchinenin A, cochinchinenin B, and loureirin B.

  11. Cold shock induces apoptosis of dorsal root ganglion neurons plated on infrared windows.

    Aboualizadeh, Ebrahim; Mattson, Eric C; O'Hara, Crystal L; Smith, Amanda K; Stucky, Cheryl L; Hirschmugl, Carol J

    2015-06-21

    The chemical status of live sensory neurons is accessible with infrared microspectroscopy of appropriately prepared cells. In this paper, individual dorsal root ganglion (DRG) neurons have been prepared with two different protocols, and plated on glass cover slips, BaF2 and CaF2 substrates. The first protocol exposes the intact DRGs to 4 °C for between 20-30 minutes before dissociating individual neurons and plating 2 hours later. The second protocol maintains the neurons at 23 °C for the entire duration of the sample preparation. The visual appearance of the neurons is similar. The viability was assessed by means of trypan blue exclusion method to determine the viability of the neurons. The neurons prepared under the first protocol (cold exposure) and plated on BaF2 reveal a distinct chemical signature and chemical distribution that is different from the other sample preparations described in the paper. Importantly, results for other sample preparation methods, using various substrates and temperature protocols, when compared across the overlapping spectral bandwidth, present normal chemical distribution within the neurons. The unusual chemically specific spatial variation is dominated by a lack of protein and carbohydrates in the center of the neurons and signatures of unraveling DNA are detected. We suggest that cold shock leads to apoptosis of DRGs, followed by osmotic stress originating from ion gradients across the cell membrane leading to cell lysis. PMID:26000346

  12. Thy1.2 YFP-16 transgenic mouse labels a subset of large-diameter sensory neurons that lack TRPV1 expression.

    Thomas E Taylor-Clark

    Full Text Available The Thy1.2 YFP-16 mouse expresses yellow fluorescent protein (YFP in specific subsets of peripheral and central neurons. The original characterization of this model suggested that YFP was expressed in all sensory neurons, and this model has been subsequently used to study sensory nerve structure and function. Here, we have characterized the expression of YFP in the sensory ganglia (DRG, trigeminal and vagal of the Thy1.2 YFP-16 mouse, using biochemical, functional and anatomical analyses. Despite previous reports, we found that YFP was only expressed in approximately half of DRG and trigeminal neurons and less than 10% of vagal neurons. YFP-expression was only found in medium and large-diameter neurons that expressed neurofilament but not TRPV1. YFP-expressing neurons failed to respond to selective agonists for TRPV1, P2X(2/3 and TRPM8 channels in Ca2+ imaging assays. Confocal analysis of glabrous skin, hairy skin of the back and ear and skeletal muscle indicated that YFP was expressed in some peripheral terminals with structures consistent with their presumed non-nociceptive nature. In summary, the Thy1.2 YFP-16 mouse expresses robust YFP expression in only a subset of sensory neurons. But this mouse model is not suitable for the study of nociceptive nerves or the function of such nerves in pain and neuropathies.

  13. Endomorphin-2 is released from newborn rat primary sensory neurons in a frequency- and calcium-dependent manner.

    Scanlin, Heather L; Carroll, Elizabeth A; Jenkins, Victoria K; Balkowiec, Agnieszka

    2008-05-01

    Recent evidence indicates that endomorphins, endogenous mu-opioid receptor (MOR) agonists, modulate synaptic transmission in both somatic and visceral sensory pathways. Here we show that endomorphin-2 (END-2) is expressed in newborn rat dorsal root ganglion (DRG) and nodose-petrosal ganglion complex (NPG) neurons, and rarely co-localizes with brain-derived neurotrophic factor (BDNF). In order to examine activity-dependent release of END-2 from neurons, we established a model using dispersed cultures of DRG and NPG cells activated by patterned electrical field stimulation. To detect release of END-2, we developed a novel rapid capture enzyme-linked immunosorbent assay (ELISA), in which END-2 capture antibody was added to neuronal cultures shortly before their electrical stimulation. The conventional assay was effective at reliably detecting END-2 only when the cells were stimulated in the presence of CTAP, a MOR-selective antagonist. This suggests that the strength of the novel assay is related primarily to rapid capture of released END-2 before it binds to endogenous MORs. Using the rapid capture ELISA, we found that stimulation protocols known to induce plastic changes at sensory synapses were highly effective at releasing END-2. Removal of extracellular calcium or blocking voltage-activated calcium channels significantly reduced the release. Together, our data provide the first evidence that END-2 is expressed by newborn DRG neurons of all sizes found in this age group, and can be released from these, as well as from NPG neurons, in an activity-dependent manner. These results point to END-2 as a likely mediator of activity-dependent plasticity in sensory pathways. PMID:18513316

  14. Potenciais usos dos AP-DRG para discriminar o perfil da assistência de unidades hospitalares Potential uses of AP-DRG to describe the health care profile in hospital units

    Marina Ferreira de Noronha

    2004-01-01

    Full Text Available Os All Patient Diagnosis Related Groups (AP-DRG constituem-se em um sistema de classificação de pacientes internados em hospitais gerais, que reúne pacientes em grupos, considerando diversas características com impacto no consumo de recursos hospitalares e critérios clínicos. O AP-DRG tem suas diversas versões utilizadas para a gerência e como unidade de pagamento a hospitais. O objetivo deste artigo foi descrever a classificação, explorando suas potencialidades para gerar informações para a gestão de hospitais. Foi utilizada a base de dados da região de Ribeirão Preto, São Paulo Brasil, de 1997, que inclui unidades hospitalares públicas, privadas e filantrópicas. Foram realizadas comparações entre Ribeirão Preto e Estados Unidos quanto ao tempo médio de permanência hospitalar de AP-DRGs selecionados. Utilizando-se os pesos relacionados ao custo relativo dos DRGs, construídos para o Estado de Nova York, Estados Unidos, foi possível verificar o perfil da atenção prestada por trinta hospitais da região de Ribeirão Preto e também o perfil dos pacientes pagos pelo SUS procedentes de outras localidades. Avalia-se que a classificação fornece informações acerca do perfil de complexidade da atenção hospitalar, tanto no que diz respeito à regionalização e sua organização em níveis de complexidade, como para subsidiar as decisões de composição dessa rede de assistência, auxiliar no seu monitoramento e contribuir para o aprimoramento do pagamento aos hospitais.The All Patient Diagnosis Related Groups (AP-DRG provide a classification system for general hospital inpatients, aggregating hospitalizations based on resource use and clinical criteria. The different versions of the AP-DRG have been applied to inpatient care management and reimbursement. This paper aimed to describe the classification and explore the potential for generating information on inpatient care management based on data from the Ribeirão Preto

  15. Effect of down-regulation of voltage-gated sodium channel Nav1.7 on activation of astrocytes and microglia in DRG in rats with cancer pain

    Jun; Pan; Xiang-Jin; Lin; Zhi-Heng; Ling; You-Zhi; Cai

    2015-01-01

    Objective:To evaluate the effect of down-regulation of Nav1.7 on the activation of astrocytes and microglia in DRG of rats with cancer pain,and explore the transmission of the nociceptive information.Methods:Lentiviral vector harboring RNAi sequence targeting the Navl.7 gene was constructed,and Walker 256 breast cancer cell and morphine was injected to build the bone cancer pain model and morphine tolerance model in rats.Lentiviral vector was injected.Rats in each model were divided into 4 groups:model group,PBS group,vehicle group and LV-Nav1.7 group.The expression levels of GFAP and OX42 in dorsal root ganglia(DRG) were measured.Results:After the animal model was built,the level of Navl.7,GFAP and OX42 was improved obviously with the time prolonged,which was statistically significant(P<0.05).The expression level of GFAP and OX42 in the DRG in the LV-Navl.7 group declined obviously compared to the model group,PBS group and vehicle group(P<0.05).Conclusions:Intrathecal injection of Navl.7 shRNA lentiviral vector can reduce the expression of Nav1.7and inhibit the activation of astrocytes and microglia in DRG.The effort is also effective in morphine tolerance bone cancer pain model rats.

  16. Increased Nerve Growth Factor Signaling in Sensory Neurons of Early Diabetic Rats Is Corrected by Electroacupuncture

    Stefania Lucia Nori

    2013-01-01

    Full Text Available Diabetic polyneuropathy (DPN, characterized by early hyperalgesia and increased nerve growth factor (NGF, evolves in late irreversible neuropathic symptoms with reduced NGF support to sensory neurons. Electroacupuncture (EA modulates NGF in the peripheral nervous system, being effective for the treatment of DPN symptoms. We hypothesize that NGF plays an important pathogenic role in DPN development, while EA could be useful in the therapy of DPN by modulating NGF expression/activity. Diabetes was induced in rats by streptozotocin (STZ injection. One week after STZ, EA was started and continued for three weeks. NGF system and hyperalgesia-related mediators were analyzed in the dorsal root ganglia (DRG and in their spinal cord and skin innervation territories. Our results show that four weeks long diabetes increased NGF and NGF receptors and deregulated intracellular signaling mediators of DRG neurons hypersensitization; EA in diabetic rats decreased NGF and NGF receptors, normalized c-Jun N-terminal and p38 kinases activation, decreased transient receptor potential vanilloid-1 ion channel, and possibly activated the nuclear factor kappa-light-chain-enhancer of activated B cells (Nf-κB. In conclusion, NGF signaling deregulation might play an important role in the development of DPN. EA represents a supportive tool to control DPN development by modulating NGF signaling in diabetes-targeted neurons.

  17. Gabapentin inhibits high-threshold calcium channel currents in cultured rat dorsal root ganglion neurones

    Sutton, K G; Martin, D. J.; Pinnock, R. D.; Lee, K.; Scott, R. H.

    2002-01-01

    This study examined the action of gabapentin (gabapentin,1-(aminomethyl) cyclohexane acetic acid (Neurontin®)) on voltage-gated calcium (Ca2+) channel influx recorded in cultured rat dorsal root ganglion (DRG) neurones.Voltage-gated Ca2+ influx was monitored using both fura-2 based fluorescence Ca2+ imaging and the whole-cell patch clamp technique.Imaging of intracellular Ca2+ transients revealed that gabapentin inhibited KCl (30 mM)-evoked voltage-dependent Ca2+ influx. Both the duration for...

  18. Co-culture of osteocytes and neurons on a unique patterned surface

    Boggs, Mary E.; Thompson, William R.; Farach-Carson, Mary C.; Duncan, Randall L.; Beebe, Thomas P.

    2011-01-01

    Neural and skeletal communication is essential for the maintenance of bone mass and transmission of pain, yet the mechanism(s) of signal transduction between these tissues is unknown. The authors established a novel system to co-culture murine long bone osteocyte-like cells (MLO-Y4) and primary murine dorsal root ganglia (DRG) neurons. Assessment of morphology and maturation marker expression on perlecan domain IV peptide (PlnDIV) and collagen type-1 (Col1) demonstrated that PlnDIV was an opt...

  19. Predictors of High Profit and High Deficit Outliers under SwissDRG of a Tertiary Care Center.

    Tarun Mehra

    Full Text Available Case weights of Diagnosis Related Groups (DRGs are determined by the average cost of cases from a previous billing period. However, a significant amount of cases are largely over- or underfunded. We therefore decided to analyze earning outliers of our hospital as to search for predictors enabling a better grouping under SwissDRG.28,893 inpatient cases without additional private insurance discharged from our hospital in 2012 were included in our analysis. Outliers were defined by the interquartile range method. Predictors for deficit and profit outliers were determined with logistic regressions. Predictors were shortlisted with the LASSO regularized logistic regression method and compared to results of Random forest analysis. 10 of these parameters were selected for quantile regression analysis as to quantify their impact on earnings.Psychiatric diagnosis and admission as an emergency case were significant predictors for higher deficit with negative regression coefficients for all analyzed quantiles (p<0.001. Admission from an external health care provider was a significant predictor for a higher deficit in all but the 90% quantile (p<0.001 for Q10, Q20, Q50, Q80 and p = 0.0017 for Q90. Burns predicted higher earnings for cases which were favorably remunerated (p<0.001 for the 90% quantile. Osteoporosis predicted a higher deficit in the most underfunded cases, but did not predict differences in earnings for balanced or profitable cases (Q10 and Q20: p<0.00, Q50: p = 0.10, Q80: p = 0.88 and Q90: p = 0.52. ICU stay, mechanical and patient clinical complexity level score (PCCL predicted higher losses at the 10% quantile but also higher profits at the 90% quantile (p<0.001.We suggest considering psychiatric diagnosis, admission as an emergency case and admission from an external health care provider as DRG split criteria as they predict large, consistent and significant losses.

  20. The effects of huwentoxin-I on the voltage-gated sodium channels of rat hippocampal and cockroach dorsal unpaired median neurons.

    Wang, Meichi; Rong, Mingqiang; Xiao, Yucheng; Liang, Songping

    2012-03-01

    Huwentoxin-I (HWTX-I) is a 33-residue peptide isolated from the venom of Ornithoctonus huwena and could inhibit TTX-sensitive voltage-gated sodium channels and N-type calcium channels in mammalian dorsal root ganglion (DRG) neurons. However, the effects of HWTX-I on mammalian central neuronal and insect sodium channel subtypes remain unknown. In this study, we found that HWTX-I potently inhibited sodium channels in rat hippocampal and cockroach dorsal unpaired median (DUM) neurons with the IC(50) values of 66.1±5.2 and 4.80±0.58nM, respectively. Taken together with our previous work on DRG neurons (IC(50)≈55nM), the order of sodium channel sensitivity to HWTX-I inhibition was insect central DUM≫mammalian peripheral>mammalian central neurons. HWTX-I exhibited no effect on the steady-state activation and inactivation of sodium channels in rat hippocampal and cockroach DUM neurons. PMID:22094230

  1. Association between tetrodotoxin resistant channels and lipid rafts regulates sensory neuron excitability.

    Alessandro Pristerà

    Full Text Available Voltage-gated sodium channels (VGSCs play a key role in the initiation and propagation of action potentials in neurons. Na(V1.8 is a tetrodotoxin (TTX resistant VGSC expressed in nociceptors, peripheral small-diameter neurons able to detect noxious stimuli. Na(V1.8 underlies the vast majority of sodium currents during action potentials. Many studies have highlighted a key role for Na(V1.8 in inflammatory and chronic pain models. Lipid rafts are microdomains of the plasma membrane highly enriched in cholesterol and sphingolipids. Lipid rafts tune the spatial and temporal organisation of proteins and lipids on the plasma membrane. They are thought to act as platforms on the membrane where proteins and lipids can be trafficked, compartmentalised and functionally clustered. In the present study we investigated Na(V1.8 sub-cellular localisation and explored the idea that it is associated with lipid rafts in nociceptors. We found that Na(V1.8 is distributed in clusters along the axons of DRG neurons in vitro and ex vivo. We also demonstrated, by biochemical and imaging studies, that Na(V1.8 is associated with lipid rafts along the sciatic nerve ex vivo and in DRG neurons in vitro. Moreover, treatments with methyl-β-cyclodextrin (MβCD and 7-ketocholesterol (7KC led to the dissociation between rafts and Na(V1.8. By calcium imaging we demonstrated that the lack of association between rafts and Na(V1.8 correlated with impaired neuronal excitability, highlighted by a reduction in the number of neurons able to conduct mechanically- and chemically-evoked depolarisations. These findings reveal the sub-cellular localisation of Na(V1.8 in nociceptors and highlight the importance of the association between Na(V1.8 and lipid rafts in the control of nociceptor excitability.

  2. Effect of knockout of α2δ-1 on action potentials in mouse sensory neurons.

    Margas, Wojciech; Ferron, Laurent; Nieto-Rostro, Manuela; Schwartz, Arnold; Dolphin, Annette C

    2016-08-01

    Gene deletion of the voltage-gated calcium channel auxiliary subunit α2δ-1 has been shown previously to have a cardiovascular phenotype, and a reduction in mechano- and cold sensitivity, coupled with delayed development of neuropathic allodynia. We have also previously shown that dorsal root ganglion (DRG) neuron calcium channel currents were significantly reduced in α2δ-1 knockout mice. To extend our findings in these sensory neurons, we have examined here the properties of action potentials (APs) in DRG neurons from α2δ-1 knockout mice in comparison to their wild-type (WT) littermates, in order to dissect how the calcium channels that are affected by α2δ-1 knockout are involved in setting the duration of individual APs and their firing frequency. Our main findings are that there is reduced Ca(2+) entry on single AP stimulation, particularly in the axon proximal segment, reduced AP duration and reduced firing frequency to a 400 ms stimulation in α2δ-1 knockout neurons, consistent with the expected role of voltage-gated calcium channels in these events. Furthermore, lower intracellular Ca(2+) buffering also resulted in reduced AP duration, and a lower frequency of AP firing in WT neurons, mimicking the effect of α2δ-1 knockout. By contrast, we did not obtain any consistent evidence for the involvement of Ca(2+)-activation of large conductance calcium-activated potassium (BK) and small conductance calcium-activated potassium (SK) channels in these events. In conclusion, the reduced Ca(2+) elevation as a result of single AP stimulation is likely to result from the reduced duration of the AP in α2δ-1 knockout sensory neurons.This article is part of the themed issue 'Evolution brings Ca(2+) and ATP together to control life and death'. PMID:27377724

  3. Long-Term Activation of Group I Metabotropic Glutamate Receptors Increases Functional TRPV1-Expressing Neurons in Mouse Dorsal Root Ganglia

    Masuoka, Takayoshi; Kudo, Makiko; Yoshida, Junko; Ishibashi, Takaharu; Muramatsu, Ikunobu; Kato, Nobuo; Imaizumi, Noriko; Nishio, Matomo

    2016-01-01

    Damaged tissues release glutamate and other chemical mediators for several hours. These chemical mediators contribute to modulation of pruritus and pain. Herein, we investigated the effects of long-term activation of excitatory glutamate receptors on functional expression of transient receptor potential vaniloid type 1 (TRPV1) in dorsal root ganglion (DRG) neurons and then on thermal pain behavior. In order to detect the TRPV1-mediated responses in cultured DRG neurons, we monitored intracellular calcium responses to capsaicin, a TRPV1 agonist, with Fura-2. Long-term (4 h) treatment with glutamate receptor agonists (glutamate, quisqualate or DHPG) increased the proportion of neurons responding to capsaicin through activation of metabotropic glutamate receptor mGluR1, and only partially through the activation of mGluR5; engagement of these receptors was evident in neurons responding to allylisothiocyanate (AITC), a transient receptor potential ankyrin type 1 (TRPA1) agonist. Increase in the proportion was suppressed by phospholipase C (PLC), protein kinase C, mitogen/extracellular signal-regulated kinase, p38 mitogen-activated protein kinase or transcription inhibitors. Whole-cell recording was performed to record TRPV1-mediated membrane current; TRPV1 current density significantly increased in the AITC-sensitive neurons after the quisqualate treatment. To elucidate the physiological significance of this phenomenon, a hot plate test was performed. Intraplantar injection of quisqualate or DHPG induced heat hyperalgesia that lasted for 4 h post injection. This chronic hyperalgesia was attenuated by treatment with either mGluR1 or mGluR5 antagonists. These results suggest that long-term activation of mGluR1/5 by peripherally released glutamate may increase the number of neurons expressing functional TRPV1 in DRG, which may be strongly associated with chronic hyperalgesia. PMID:27064319

  4. Long-Term Activation of Group I Metabotropic Glutamate Receptors Increases Functional TRPV1-Expressing Neurons in Mouse Dorsal Root Ganglia.

    Masuoka, Takayoshi; Kudo, Makiko; Yoshida, Junko; Ishibashi, Takaharu; Muramatsu, Ikunobu; Kato, Nobuo; Imaizumi, Noriko; Nishio, Matomo

    2016-01-01

    Damaged tissues release glutamate and other chemical mediators for several hours. These chemical mediators contribute to modulation of pruritus and pain. Herein, we investigated the effects of long-term activation of excitatory glutamate receptors on functional expression of transient receptor potential vaniloid type 1 (TRPV1) in dorsal root ganglion (DRG) neurons and then on thermal pain behavior. In order to detect the TRPV1-mediated responses in cultured DRG neurons, we monitored intracellular calcium responses to capsaicin, a TRPV1 agonist, with Fura-2. Long-term (4 h) treatment with glutamate receptor agonists (glutamate, quisqualate or DHPG) increased the proportion of neurons responding to capsaicin through activation of metabotropic glutamate receptor mGluR1, and only partially through the activation of mGluR5; engagement of these receptors was evident in neurons responding to allylisothiocyanate (AITC), a transient receptor potential ankyrin type 1 (TRPA1) agonist. Increase in the proportion was suppressed by phospholipase C (PLC), protein kinase C, mitogen/extracellular signal-regulated kinase, p38 mitogen-activated protein kinase or transcription inhibitors. Whole-cell recording was performed to record TRPV1-mediated membrane current; TRPV1 current density significantly increased in the AITC-sensitive neurons after the quisqualate treatment. To elucidate the physiological significance of this phenomenon, a hot plate test was performed. Intraplantar injection of quisqualate or DHPG induced heat hyperalgesia that lasted for 4 h post injection. This chronic hyperalgesia was attenuated by treatment with either mGluR1 or mGluR5 antagonists. These results suggest that long-term activation of mGluR1/5 by peripherally released glutamate may increase the number of neurons expressing functional TRPV1 in DRG, which may be strongly associated with chronic hyperalgesia. PMID:27064319

  5. Shaping the System – The DRG Evaluation Project of the German Society for Gynaecology and Obstetrics (Deutsche Gesellschaft für Gynäkologie und Geburtshilfe, DGGG)

    Fiori, W.; Renner, S. P.; Siam, K.; Babapirali, J.; Roeder, N.; Dausch, E.; Hildebrandt, T.; Hillemanns, P.; Nehmzow, M.; Zygmunt, M.; Piroth, D.; Schem, C.; Schwenzer, T.; Friese, K.; Wallwiener, D.; Beckmann, M. W.

    2013-01-01

    Introduction: The German DRG system is annually adapted to the changing services provided. For the further development, the self-governing body and its DRG Institute (InEK) depend on participation of the users. Methods: For one of the DRG evaluation projects initiated by DGGG, cost and performance data for the year 2011 from 16 hospitals were available. After plausibility checks and corrections, analyses for service and cost homogeneity were performed. In cases of inadequate DRG-representation attributes were sought that would make an appropriate reimbursement possible. Conspicuities and potential solutions were checked for clinical plausibility. Results: 44 concrete modification proposals for further development of the G-DRG system were formulated and submitted in due time to the InEK. In addition, 3 modification proposals were addressed to the German Institute for Medical Documentation and Information (Deutsches Institut für Medizinische Dokumentation und Information, DIMDI) for further development of the diagnosis classification ICD-10-GM. For all modification proposals care was taken to minimise misdirected incentives and to reduce the potential for disputes with the cost bearers and their auditors services in settlements. Discussion: The publication of the G-DRG system 2014 shows which modification proposals have been realised. Essentially, an appropriate redistribution of the resources among the gynaecological and obstetrics departments is to be expected. The financial pressure that is caused by the generally inadequate financing of hospitals will not be reduced by a further development of the G-DRG system. PMID:24771931

  6. Clenbuterol, a beta(2)-agonist, retards atrophy in denervated muscles

    Zeman, Richard J.; Ludemann, Robert; Etlinger, Joseph D.

    1987-01-01

    The effects of a beta(2) agonist, clenbuterol, on the protein content as well as on the contractile strength and the muscle fiber cross-sectional area of various denervated muscles from rats were investigated. It was found that denervated soleus, anterior tibialis, and gastrocnemius muscles, but not the extensor digitorum longus, of rats treated for 2-3 weeks with clenbuterol contained 95-110 percent more protein than denervated controls. The twofold difference in the protein content of denervated solei was paralleled by similar changes in contractile strength and muscle fiber cross-sectional area.

  7. A simple, highly efficient method for heterologous expression in mammalian primary neurons using cationic lipid-mediated mRNA transfection

    Damian J Williams

    2010-11-01

    Full Text Available Expression of heterologous proteins in adult mammalian neurons is a valuable technique for the study of neuronal function. The postmitotic nature of mature neurons prevents effective DNA transfection using simple, cationic lipid-based methods. Adequate heterologous protein expression is often only achievable using complex techniques that, in many cases, are associated with substantial toxicity. Here, a simple method for high efficiency transfection of mammalian primary neurons using in vitro-transcribed mRNA and the cationic lipid transfection reagent Lipofectamine 2000 is described. Optimal transfection conditions were established in adult mouse dissociated dorsal root ganglion (DRG neurons using a 96-well based luciferase activity assay. Using these conditions, a transfection efficiency of 25% was achieved in DRG neurons transfected with EGFP mRNA. High transfection efficiencies were also obtained in dissociated rat superior cervical ganglion (SCG neurons and mouse cortical and hippocampal cultures. Endogenous Ca2+ currents in EGFP mRNA-transfected SCG neurons were not significantly different from untransfected neurons, which suggested that this technique is well suited for heterologous expression in patch clamp recording experiments. Functional expression of a cannabinoid receptor (CB1R, a G protein inwardly-rectifying K+ channel (GIRK4 and a dominant-negative G protein α-subunit mutant (GoA G203T indicate that the levels of heterologous protein expression attainable using mRNA transfection are suitable for most functional protein studies. This study demonstrates that mRNA transfection is a straightforward and effective method for heterologous expression in neurons and is likely to have many applications in neuroscience research.

  8. Ligustrazine inhibits high voltage-gated Ca2+ and TTX-resistant Na+ channels of primary sensory neuron and thermal nociception in the rat:a study on peripheral mechanism

    2006-01-01

    Objective Ligustrazine, also named as tetramethylpyrazine, is a compound purified from Ligusticum chuanxiong hort and has ever been testified to be a calcium antagonist. The present investigation was to determine the antinociceptive effect of ligustrazine and, if any, the peripheral ionic mechanism involved. Methods Paw withdrawal Latency(PWL) to noxious heating was measured in vivo and whole-cell patch recording was performed on small dorsal root ganglion (DRG) neurons. Results Intraplantar injection of ligustrazine (0.5 mg in 25 μl) significantly prolonged the withdrawal latency of ipsilateral hindpaw to noxious heating in the rat. Ligustrazine not only reversibly inhibited high-voltage gated calcium current of dorsal root ganglion (DRG) neuron in dose-dependent manner with IC50 of 1.89 mmol/L, but also decreased tetrodotoxin (TTX) -resistant sodium current in relatively selective and dose-dependent manner with IC50 of2.49 mmol/L. Conclusion The results suggested that ligustrazine could elevate the threshold of thermal nociception through inhibiting the high-voltage gated calcium current and TTX-resistant sodium current of DRG neuron .in the rat.

  9. An Approach to Enhance Alignment and Myelination of Dorsal Root Ganglion Neurons.

    Liu, Chun; Chan, Christina

    2016-01-01

    Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to bridge the lesion site, properly organized axonal alignment is required. Since demyelination after nerve injury strongly impairs the conductive capacity of surviving axons, remyelination is critical for successful functioning of regenerated nerves. Previously, we demonstrated that mesenchymal stem cells (MSCs) aligned on a pre-stretch induced anisotropic surface because the cells can sense a larger effective stiffness in the stretched direction than in the perpendicular direction. We also showed that an anisotropic surface arising from a mechanical pre-stretched surface similarly affects alignment, as well as growth and myelination of axons. Here, we provide a detailed protocol for preparing a pre-stretched anisotropic surface, the isolation and culture of dorsal root ganglion (DRG) neurons on a pre-stretched surface, and show the myelination behavior of a co-culture of DRG neurons with Schwann cells (SCs) on a pre-stretched surface. PMID:27585118

  10. CPEB3 Deficiency Elevates TRPV1 Expression in Dorsal Root Ganglia Neurons to Potentiate Thermosensation.

    Fong, Sitt Wai; Lin, Hsiu-Chen; Wu, Meng-Fang; Chen, Chih-Cheng; Huang, Yi-Shuian

    2016-01-01

    Cytoplasmic polyadenylation element binding protein 3 (CPEB3) is a sequence-specific RNA-binding protein that downregulates translation of multiple plasticity-related proteins (PRPs) at the glutamatergic synapses. Activity-induced synthesis of PRPs maintains long-lasting synaptic changes that are critical for memory consolidation and chronic pain manifestation. CPEB3-knockout (KO) mice show aberrant hippocampus-related plasticity and memory, so we investigated whether CPEB3 might have a role in nociception-associated plasticity. CPEB3 is widely expressed in the brain and peripheral afferent sensory neurons. CPEB3-KO mice with normal mechanosensation showed hypersensitivity to noxious heat. In the complete Freund's adjuvant (CFA)-induced inflammatory pain model, CPEB3-KO animals showed normal thermal hyperalgesia and transiently enhanced mechanical hyperalgesia. Translation of transient receptor potential vanilloid 1 (TRPV1) RNA was suppressed by CPEB3 in dorsal root ganglia (DRG), whereas CFA-induced inflammation reversed this inhibition. Moreover, CPEB3/TRPV1 double-KO mice behaved like TRPV1-KO mice, with severely impaired thermosensation and thermal hyperalgesia. An enhanced thermal response was recapitulated in non-inflamed but not inflamed conditional-KO mice, with cpeb3 gene ablated mostly but not completely, in small-diameter nociceptive DRG neurons. CPEB3-regulated translation of TRPV1 RNA may play a role in fine-tuning thermal sensitivity of nociceptors. PMID:26915043

  11. Auto-oxidation products of epigallocatechin gallate activate TRPA1 and TRPV1 in sensory neurons.

    Kurogi, Mako; Kawai, Yasushi; Nagatomo, Katsuhiro; Tateyama, Michihiro; Kubo, Yoshihiro; Saitoh, Osamu

    2015-01-01

    The sensation of astringency is elicited by catechins and their polymers in wine and tea. It has been considered that catechins in green tea are unstable and auto-oxidized to induce more astringent taste. Here, we examined how mammalian transient receptor potential V1 (TRPV1) and TRPA1, which are nociceptive sensors, are activated by green tea catechins during the auto-oxidation process. Neither TRPV1 nor TRPA1 could be activated by any of the freshly prepared catechin. When one of the major catechin, epigallocatechin gallate (EGCG), was preincubated for 3h in Hank's balanced salt solution, it significantly activated both TRP channels expressed in HEK293 cells. Even after incubation, other catechins showed much less effects. Results suggest that only oxidative products of EGCG activate both TRPV1 and TRPA1. Dorsal root ganglion (DRG) sensory neurons were also activated by the incubated EGCG through TRPV1 and TRPA1 channels. Liquid chromatography-mass spectrometry revealed that theasinensins A and D are formed during incubation of EGCG. We found that purified theasinensin A activates both TRPV1 and TRPA1, and that it stimulates DRG neurons through TRPV1 and TRPA1 channels. Results suggested a possibility that TRPV1 and TRPA1 channels are involved in the sense of astringent taste of green tea. PMID:25422365

  12. Motor Neuron Diseases

    ... Awards Enhancing Diversity Find People About NINDS Motor Neuron Diseases Fact Sheet See a list of all ... can I get more information? What are motor neuron diseases? The motor neuron diseases (MNDs) are a ...

  13. Neuronal Migration Disorders

    ... Enhancing Diversity Find People About NINDS NINDS Neuronal Migration Disorders Information Page Table of Contents (click to ... being done? Clinical Trials Organizations What are Neuronal Migration Disorders? Neuronal migration disorders (NMDs) are a group ...

  14. Calibration on Pegase of a selective D.R.G. installation for short life and long life fission gas

    Pegase irradiation loops are equipped with a detection installation which measures the global activity of short-life and long-life fission gases which are released in CO2, but the reduced size of circuits in the loop results in an accumulation of long life fission gases, and therefore in problems in the interpretation of measured signals. Thus, the authors propose an additional detection installation which allows long-life fission gases to be separately measured. The principle is to ensure a partial decay of the sampled gas by imposing an additional transit time in order to get rid of short-life fission gases which have a radioactive period of some tenths of a second. A second detector is then used to measure the residual activity of long-life fission gases. The author describes the installation (the normal circuit and the modified circuit), reports the performed tests and the calibration, presents and discusses the obtained results and the installation sensitivity (for short-life and long-life fission gases), and reports their application to the relationship between DRG (sheath failure detection) signals obtained on Pegase and on EDF and EL4 reactors

  15. Micronuclei formation in non-neuronal cells of foetal rat dorsal root ganglia (DRG) induced in vitro by cisplatin and carboplatin

    Jirsová, Kateřina; Mandys, Václav

    Zemplínská Teplica : Oganizátoři symposia, 1995. s. 34. [Bilateral Symposium of Workers with Cell Culotures, Slovak and Czech Republic /1./. 24.05.1995-26.05.1995, Zemplínská Teplica] R&D Projects: GA ČR GA304/95/0255

  16. DNA polymerase-beta is expressed early in neurons of Alzheimer's disease brain and is loaded into DNA replication forks in neurons challenged with beta-amyloid

    A. Copani; J.J.M. Hoozemans; F. Caraci; M. Calafiore; E.S. van Haastert; R. Veerhuis; A.J.M. Rozemuller; E. Aronica; M.A. Sortino; F. Nicoletti

    2006-01-01

    Cultured neurons exposed to synthetic beta-amyloid (A beta) fragments reenter the cell cycle and initiate a pathway of DNA replication that involves the repair enzyme DNA polymerase-beta (DNA pol-beta) before undergoing apoptotic death. In this study, by performing coimmunoprecipitation experiments

  17. Role of T-type calcium current in identified D-hair mechanoreceptor neurons studied in vitro

    Dubreuil, A.S.; Boukhaddaoui, H.; Desmadryl, G; Martinez-Salgado, C.; Moshourab, R.; Lewin, G. R.; Carroll, P.; Valmier, J; Scamps, F.

    2004-01-01

    Different subsets of dorsal root ganglion (DRG) mechanoreceptors transduce low- and high-intensity mechanical stimuli. It was shown recently that, in vivo, neurotrophin-4 (NT-4)-dependent D-hair mechanoreceptors specifically express a voltage-activated T-type calcium channel (Ca(v)3.2) that may be required for their mechanoreceptive function. Here we show that D-hair mechanoreceptors can be identified in vitro by a rosette-like morphology in the presence of NT-4 and that these rosette neurons...

  18. Chromosome aberration analysis based on a beta-binomial distribution

    Analyses carried out here generalized on earlier studies of chromosomal aberrations in the populations of Hiroshima and Nagasaki, by allowing extra-binomial variation in aberrant cell counts corresponding to within-subject correlations in cell aberrations. Strong within-subject correlations were detected with corresponding standard errors for the average number of aberrant cells that were often substantially larger than was previously assumed. The extra-binomial variation is accomodated in the analysis in the present report, as described in the section on dose-response models, by using a beta-binomial (B-B) variance structure. It is emphasized that we have generally satisfactory agreement between the observed and the B-B fitted frequencies by city-dose category. The chromosomal aberration data considered here are not extensive enough to allow a precise discrimination between competing dose-response models. A quadratic gamma ray and linear neutron model, however, most closely fits the chromosome data. (author)

  19. Roles of mitochondria and temperature in the control of intracellular calcium in adult rat sensory neurons

    Kang, S. H.; Carl, A; McHugh, J.M.; Goff, H.R.; Kenyon, J L

    2007-01-01

    We recorded Ca2+ current and intracellular Ca2+ ([Ca2+]i) in isolated adult rat dorsal root ganglion (DRG) neurons at 20 and 30 °C. In neurons bathed in tetraethylammonium and dialyzed with cesium, warming reduced resting average [Ca2+]i from 87 to 49 nM and the time constant of the decay of [Ca2+]i transients (τr) from 1.3 s to 0.99 s (Q10 = 1.4). The Buffer Index, the ratio between Ca2+ influx and Δ[Ca2+]i (∫ICa·dt/Δ[Ca2+]i), increased 2- to 3-fold with warming. Neither inhibition of the pl...

  20. A Method to Target and Isolate Airway-innervating Sensory Neurons in Mice.

    Kaelberer, Melanie Maya; Jordt, Sven-Eric

    2016-01-01

    Somatosensory nerves transduce thermal, mechanical, chemical, and noxious stimuli caused by both endogenous and environmental agents. The cell bodies of these afferent neurons are located within the sensory ganglia. Sensory ganglia innervate a specific organ or portion of the body. For instance, the dorsal root ganglia (DRG) are located in the vertebral column and extend processes throughout the body and limbs. The trigeminal ganglia are located in the skull and innervate the face, and upper airways. Vagal afferents of the nodose ganglia extend throughout the gut, heart, and lungs. The nodose neurons control a diverse array of functions such as: respiratory rate, airway irritation, and cough reflexes. Thus, to understand and manipulate their function, it is critical to identify and isolate airway specific neuronal sub-populations. In the mouse, the airways are exposed to a fluorescent tracer dye, Fast Blue, for retrograde tracing of airway-specific nodose neurons. The nodose ganglia are dissociated and fluorescence activated cell (FAC) sorting is used to collect dye positive cells. Next, high quality ribonucleic acid (RNA) is extracted from dye positive cells for next generation sequencing. Using this method airway specific neuronal gene expression is determined. PMID:27168016

  1. NMDA receptor subunit expression and PAR2 receptor activation in colospinal afferent neurons (CANs during inflammation induced visceral hypersensitivity

    Caudle Robert M

    2009-09-01

    Full Text Available Abstract Background Visceral hypersensitivity is a clinical observation made when diagnosing patients with functional bowel disorders. The cause of visceral hypersensitivity is unknown but is thought to be attributed to inflammation. Previously we demonstrated that a unique set of enteric neurons, colospinal afferent neurons (CANs, co-localize with the NR1 and NR2D subunits of the NMDA receptor as well as with the PAR2 receptor. The aim of this study was to determine if NMDA and PAR2 receptors expressed on CANs contribute to visceral hypersensitivity following inflammation. Recently, work has suggested that dorsal root ganglion (DRG neurons expressing the transient receptor potential vanilloid-1 (TRPV1 receptor mediate inflammation induced visceral hypersensitivity. Therefore, in order to study CAN involvement in visceral hypersensitivity, DRG neurons expressing the TRPV1 receptor were lesioned with resiniferatoxin (RTX prior to inflammation and behavioural testing. Results CANs do not express the TRPV1 receptor; therefore, they survive following RTX injection. RTX treatment resulted in a significant decrease in TRPV1 expressing neurons in the colon and immunohistochemical analysis revealed no change in peptide or receptor expression in CANs following RTX lesioning as compared to control data. Behavioral studies determined that both inflamed non-RTX and RTX animals showed a decrease in balloon pressure threshold as compared to controls. Immunohistochemical analysis demonstrated that the NR1 cassettes, N1 and C1, of the NMDA receptor on CANs were up-regulated following inflammation. Furthermore, inflammation resulted in the activation of the PAR2 receptors expressed on CANs. Conclusion Our data show that inflammation causes an up-regulation of the NMDA receptor and the activation of the PAR2 receptor expressed on CANs. These changes are associated with a decrease in balloon pressure in response to colorectal distension in non-RTX and RTX lesioned

  2. The RNA binding and transport proteins staufen and fragile X mental retardation protein are expressed by rat primary afferent neurons and localize to peripheral and central axons.

    Price, T J; Flores, C M; Cervero, F; Hargreaves, K M

    2006-09-15

    Neuronal proteins have been traditionally viewed as being derived solely from the soma; however, accumulating evidence indicates that dendritic and axonal sites are capable of a more autonomous role in terms of new protein synthesis. Such extra-somal translation allows for more rapid, on-demand regulation of neuronal structure and function than would otherwise be possible. While mechanisms of dendritic RNA transport have been elucidated, it remains unclear how RNA is trafficked into the axon for this purpose. Primary afferent neurons of the dorsal root (DRG) and trigeminal (TG) ganglia have among the longest axons in the neuraxis and such axonal protein synthesis would be advantageous, given the greater time involved for protein trafficking to occur via axonal transport. Therefore, we hypothesized that these primary sensory neurons might express proteins involved in RNA transport. Rat DRG and TG neurons expressed staufen (stau) 1 and 2 (detected at the mRNA level) and stau2 and fragile x mental retardation protein (FMRP; detected at the protein level). Stau2 mRNA was also detected in human TG neurons. Stau2 and FMRP protein were localized to the sciatic nerve and dorsal roots by immunohistochemistry and to dorsal roots by Western blot. Stau2 and FMRP immunoreactivities colocalized with transient receptor potential channel type 1 immunoreactivity in sensory axons of the sciatic nerve and dorsal root, suggesting that these proteins are being transported into the peripheral and central terminals of nociceptive sensory axons. Based on these findings, we propose that stau2 and FMRP proteins are attractive candidates to subserve RNA transport in sensory neurons, linking somal transcriptional events to axonal translation. PMID:16809002

  3. Structures of A[beta]-Related Peptide−Monoclonal Antibody Complexes

    Gardberg, Anna; Dice, Lezlee; Pridgen, Kathleen; Ko, Jan; Patterson, Paul; Ou, Susan; Wetzel, Ronald; Dealwis, Chris; (Case Western); (CIT); (Tennessee-K)

    2009-06-15

    Passive immunotherapy (PI) is being explored as a potential therapeutic against Alzheimer's disease. The most promising antibodies (Abs) used in PI target the EFRH motif of the A{beta} N-terminus. The monoclonal anti-A{beta} Ab PFA1 recognizes the EFRH epitope of A{beta}. PFA1 has a high affinity for A{beta} fibrils and protofibrils (0.1 nM), as well as good affinity for A{beta} monomers (20 nM). However, PFA1 binds the toxic N-terminally modified pyroglutamate peptide pyro-Glu3-A{beta} with a 77-fold loss in affinity compared to the WT A{beta}(1-8). Furthermore, our earlier work illustrated PFA1's potential for cross-reactivity. The receptor tyrosine kinase Ror2, which plays a role in skeletal and bone formation, possesses the EFRH sequence. PFA1 Fab binds the Ror2(518-525) peptide sequence REEFRHEA with a 3-fold enhancement over WT A{beta}(1-8). In this work, the crystal structures of the hybridoma-derived PFA1 Fab in complex with pyro-Glu3-A{beta} peptide and with a cross-reacting peptide from Ror2 have been determined at resolutions of 1.95 and 2.7 {angstrom}, respectively. As with wild-type A{beta}, these peptides bind to the Fab via a combination of charge- and shape-complementarity, hydrogen-bonding, and hydrophobic interactions. Comparison of the structures of the four peptides A{beta}(1-8), Grip1, pyro-Glu3-A{beta}(3-8), and Ror2 in complex with PFA1 shows that the greatest conformational flexibility occurs at residues 2 to 3 and 8 of the peptide. These structures provide a molecular basis of the specificity tolerance of PFA1 and its ability to recognize A{beta} N-terminal heterogeneity. The structures provide clues to improving mAb specificity and affinity for pyroglutamate A{beta}.

  4. Autophagy inhibition in endogenous and nutrient-deprived conditions reduces dorsal root ganglia neuron survival and neurite growth in vitro.

    Clarke, Joseph-Patrick; Mearow, Karen

    2016-07-01

    Peripheral neuropathies can result in cytoskeletal changes in axons, ultimately leading to Wallerian degeneration and cell death. Recently, autophagy has been studied as a potential target for improving axonal survival and growth during peripheral nerve damage. This study investigates the influence of autophagy on adult dorsal root ganglia (DRG) neuron survival and axonal growth under control and nutrient deprivation conditions. Constitutive autophagy was modulated with pharmacological activators (rapamycin; Rapa) and inhibitors (3-methyladenine, bafilomycin A1) in conjunction with either a nutrient-stable environment (standard culture medium) or a nutrient-deprived environment (Hank's balanced salt solution + Ca(2+) /Mg(2+) ). The results demonstrated that autophagy inhibition decreased cell viability and reduced neurite growth and branching complexity. Although autophagy was upregulated with nutrient deprivation compared with the control, it was not further activated by rapamycin, suggesting a threshold level of autophagy. Overall, both cellular and biochemical approaches combined to show the influence of autophagy on adult DRG neuron survival and growth. © 2016 Wiley Periodicals, Inc. PMID:27018986

  5. Juvenil neuronal ceroid lipofuscinosis

    Ostergaard, J R; Hertz, Jens Michael

    1998-01-01

    Neuronal ceroid-lipofuscinosis is a group of neurodegenerative diseases which are characterized by an abnormal accumulation of lipopigment in neuronal and extraneuronal cells. The diseases can be differentiated into several subgroups according to age of onset, the clinical picture...

  6. NEURON and Python

    Michael Hines; Davison, Andrew P.; Eilif Muller

    2009-01-01

    The NEURON simulation program now allows Python to be used, alone or in combination with NEURON's traditional Hoc interpreter. Adding Python to NEURON has the immediate benefit of making available a very extensive suite of analysis tools written for engineering and science. It also catalyzes NEURON software development by offering users a modern programming tool that is recognized for its flexibility and power to create and maintain complex programs. At the same time, nothing is lost because ...

  7. Refractory Neuron Circuits

    Sarpeshkar, Rahul; Watts, Lloyd; Mead, Carver

    1992-01-01

    Neural networks typically use an abstraction of the behaviour of a biological neuron, in which the continuously varying mean firing rate of the neuron is presumed to carry information about the neuron's time-varying state of excitation. However, the detailed timing of action potentials is known to be important in many biological systems. To build electronic models of such systems, one must have well-characterized neuron circuits that capture the essential behaviour of real neur...

  8. Motor Neurons that Multitask

    Goulding, Martyn

    2012-01-01

    Animals use a form of sensory feedback termed proprioception to monitor their body position and modify the motor programs that control movement. In this issue of Neuron, Wen et al. (2012) provide evidence that a subset of motor neurons function as proprioceptors in C. elegans, where B-type motor neurons sense body curvature to control the bending movements that drive forward locomotion.

  9. Valutazione dell'appropriatezza dei ricoveri in un Policlinico Universitario: analisi mediante l'uso comparativo dei sistemi di classificazione isogravitá APR-DRG e Disease Staging e del PRUO

    M. Volpe

    2003-05-01

    Full Text Available

    Obiettivi: valutare l’appropriatezza organizzativa dei ricoveri effettuati in un Policlinico Universitario attraverso la comparazione di due metodi, dei quali uno basato sui dati della scheda di dimissione ospedaliera ed utilizza, rispettivamente, i sistemi di classificazione iso-gravità APR-DRG e Disease Staging e l’altro sulla revisione delle cartelle cliniche mediante il PRUO.

    Metodi: oggetto di analisi sono i ricoveri ordinari effettuati nell’anno 2001 ed afferenti ai DRG inclusi nella delibera della Giunta Regionale del Lazio 864/2002 che recepisce il D.P.C.M. 29/11/2001 sui livelli essenziali di assistenza.

    Risultati: i risultati evidenziano che le due varianti del
    metodo basato sulla SDO (metodo APPRO mostrano quote di ricoveri inappropriati sovrapponibili rispetto al complesso dei ricoveri oggetto di analisi, ma con differenze anche rilevanti tra APR-DRG e Disease Staging in relazione ai singoli DRG considerati, riconducibili ai diversi algoritmi di attribuzione del livello di severità utilizzati dai due sistemi. L’analisi campionaria effettuata con il metodo PRUO su casi afferenti ai DRG della DGR 864/2002 caratterizzati da livelli di severità minimi evidenzia una proporzione di ricoveri inappropriati superiore a quella determinata tramite i metodi basati sulla SDO. Tale differenza è verosimilmente dovuta al ruolo del valore delle soglie percentuali di accettabilità individuate dalla Regione Lazio per ciascun DRG: le quote di ricoveri che eccedono tali soglie sono considerate inappropriate.

    Conclusioni: sulla base dei risultati ottenuti gli autori
    descrivono gli interventi organizzativi adottati per ottimizzare il contesto di erogazione delle prestazioni
    oggetto di analisi, discutono vantaggi e limiti dei metodi SDO-based e del metodo analitico PRUObased e ne propongono l

  10. Changes in skin levels of two neutotrophins (glial cell line derived neurotrohic factor and neurotrophin-3) cause alterations in cutaneous neuron responses to mechanical stimuli

    Jeffrey Lawson; Sabrina L. Mcllwrath; H. Richard Koerber

    2008-01-01

    Neurotrophins are important for the development and maintenance of both high and low threshold mechanoreceptors (HTMRs and LTMRs). In this series of studies, the effects of constitutive overexpression of two different neurotrophins, neurotrophin-3 (NT-3) and glial cell line derived neurotrohic factor (GDNF), were examined. Previous studies indicated that both of them may be implicated in the normal development of mouse dorsal root ganglion (DRG) neurons. Neurons from mice transgenically altered to overexpress NT-3 or GDNF (NT-3-OE or GDNF-OE mice) in the skin were examined using several physiological, immunohistochemi-cal and molecular techniques. Ex vivo skin/nerve/DRG/spinal cord and skin/nerve preparations were used to determine the response characteristics of the cutaneous neurons; immunohistochemistry was used to examine the biochemical phenotype of DRG cells and the skin; RT-PCR was used to examine the levels of candidate ion channels in skin and DRG that may correlate with changes in physiologi-cal responses. In GDNF-OE mice, I-isolectin B4 (IB4)-immunopositive C-HTMRs (nociceptors), a large percentage of which are sensitive to GDNF, had significantly lower mechanical thresholds than wildtype (WT) neurons. Heat thresholds for the same cells were not different. Mechanical sensitivity changes in GDNF-OE mice were correlated with significant increases in acid sensing ion channels 2a (ASIC2a) and 2b (ASIC2b) and transient receptor potential channel AI (TRPAI), all of which are putative mechanosensitive ion channels. Overexpression of NT-3 affected the responses of A-LTMRs and A-HTMRs, hut had no effect on C-HTMRs. Slowly adapting type 1 (SA1) LTMRs and A-HTMRs had increased mechanical sensitivity compared to WT. Mechanical sensitivity was correlated with significant increases in acid-sensing ion channels ASIC1 and ASIC3. This data indicates that both neurotrophins play roles in determining mechanical thresholds of cutaneous HTMRs and LTMRs and that sensitivity

  11. Involvement of IKAP in peripheral target innervation and in specific JNK and NGF signaling in developing PNS neurons.

    Anastasia Abashidze

    Full Text Available A splicing mutation in the ikbkap gene causes Familial Dysautonomia (FD, affecting the IKAP protein expression levels and proper development and function of the peripheral nervous system (PNS. Here we attempted to elucidate the role of IKAP in PNS development in the chick embryo and found that IKAP is required for proper axonal outgrowth, branching, and peripheral target innervation. Moreover, we demonstrate that IKAP colocalizes with activated JNK (pJNK, dynein, and β-tubulin at the axon terminals of dorsal root ganglia (DRG neurons, and may be involved in transport of specific target derived signals required for transcription of JNK and NGF responsive genes in the nucleus. These results suggest the novel role of IKAP in neuronal transport and specific signaling mediated transcription, and provide, for the first time, the basis for a molecular mechanism behind the FD phenotype.

  12. Underlying mechanism of regulatory actions of diclofenac, a nonsteroidal anti-inflammatory agent, on neuronal potassium channels and firing: an experimental and theoretical study.

    Huang, C W; Hung, T Y; Liao, Y K; Hsu, M C; Wu, S N

    2013-06-01

    Diclofenac (DIC), a nonsteroidal anti-inflammatory drug, is known to exert anti-nociceptive and anti-convulsant actions; however, its effects on ion currents, in neurons remain debatable. We aimed to investigate (1) potential effects of diclofenac on membrane potential and potassium currents in differentiated NSC-34 neuronal cells and dorsal root ganglion (DRG) neurons with whole-cell patch-clamp technology, and (2) firing of action potentials (APs), using a simulation model from hippocampal CA1 pyramidal neurons based on diclofenac's effects on potassium currents. In the NSC-34 cells, diclofenac exerted an inhibitory effect on delayed-rectifier K⁺ current (I(KDR)) with an IC₅₀ value of 73 μM. Diclofenac not merely inhibited the I(KDR) amplitude in response to membrane depolarization, but also accelerated the process of current inactivation. The inhibition by diclofenac of IK(DR) was not reversed by subsequent application of either naloxone. Importantly, diclofenac (300 μM) increased the amplitude of M-type K⁺ current (I)(KM)), while flupirtine (10 μM) or meclofenamic acid (10 μM) enhanced it effectively. Consistently, diclofenac (100 μM) increased the amplitude of I(KM) and diminished the I(KDR) amplitude, with a shortening of inactivation time constant in DRG neurons. Furthermore, by using the simulation modeling, we demonstrated the potential electrophysiological mechanisms underlying changes in AP firing caused by diclofenac. During the exposure to diclofenac, the actions on both I(KM) and I(KDR) could be potential mechanism through which it influences the excitability of fast-spiking neurons. Caution needs to be made in attributing the effects of diclofenac primarily to those produced by the activation of I(KM). PMID:23959723

  13. Endosome-mediated retrograde axonal transport of P2X3 receptor signals in primary sensory neurons

    Xu-Qiao Chen; BinWang; Chengbiao Wu; Jin Pan; Bo Yuan; Yuan-Yuan Su; Xing-Yu Jiang; Xu Zhang; Lan Bao

    2012-01-01

    Neurotrophins and their receptors adopt signaling endosomes to transmit retrograde signals.However,the mechanisms of retrograde signaling for other ligand/receptor systems are poorly understood.Here,we report that the signals of the purinergic (P)2X3 receptor,an ATP-gated ion channel are retrogradely transported in dorsal root ganglion (DRG) neuron axons.We found that Rab5,a small GTPase,controls the early sorting of P2X3 receptors into endosomes,while Rab7 mediates the fast retrograde transport of P2X3 receptors.Intraplantar injection and axonal application into the microfluidic chamber of α,β-methylene-ATP (α,β-MeATP),a P2X selective agonist,enhanced the endocytosis and retrograde transport of P2X3 receptors.The α,β-MeATP-induced Ca2+ influx activated a pathway comprised of protein kinase C,rat sarcoma viral oncogene and extracellular signal-regulated protein kinase (ERK),which associated with endocytic P2X3 receptors to form signaling endosomes.Disruption of the lipid rafts abolished the α,β-MeATP-induced ERK phosphorylation,endocytosis and retrograde transport of P2X3 receptors.Furthermore,treatment of peripheral axons with α,β-MeATP increased the activation level of ERK and cAMP response element-binding protein in the cell bodies of DRG neurons and enhanced neuronal excitability.Impairment of either microtubule-based axonal transport in vivo or dynein function in vitro blocked α,β-MeATP-induced retrograde signals.These results indicate that P2X3 receptor-activated signals are transmitted via retrogradely transported endosomes in primary sensory neurons and provide a novel signaling mechanism for ligand-gated channels.

  14. Hsp27 and axonal growth in adult sensory neurons in vitro

    Mearow Karen M

    2005-04-01

    Full Text Available Abstract Background Neurite growth can be elicited by growth factors and interactions with extracellular matrix molecules like laminin. Among the targets of the signalling pathways activated by these stimuli are cytoskeletal elements, such as actin, tubulin and neurofilaments. The cytoskeleton can also be modulated by other proteins, such as the small heat shock protein Hsp27. Hsp27 interacts with actin and tubulin in non-neuronal cells and while it has been suggested to play a role in the response of some neurons to injury, there have been no direct studies of its contribution to axonal regeneration. Results We have investigated neurite initiation and process extension using cultures of adult dorsal root ganglion (DRG sensory neurons and a laminin stimulation paradigm. Employing confocal microscopy and biochemical analyses we have examined localization of Hsp27 at early and later stages of neurite growth. Our results show that Hsp27 is colocalized with actin and tubulin in lamellopodia, filopodia, focal contacts and mature neurites and growth cones. Disruption of the actin cytoskeleton with cytochalasin D results in aberrant neurite initiation and extension, effects which may be attributable to alterations in actin polymerization states. Inhibition of Hsp27 phosphorylation in our cultures results in an atypical growth pattern that may be attributable to an effect of pHsp27 on the stability of the actin cytoskeleton. Conclusion We observed colocalization of the phosphorylated and non-phosphorylated forms of Hsp27 with actin and tubulin in both very early and later stages of neurite growth from cultured adult DRG neurons. The colocalization of Hsp27 and pHsp27 with actin in lamellopodia and focal contacts at early stages of neurite growth, and in processes, branch points and growth cones at later stages, suggests that Hsp27 may play a role in neuritogenesis and subsequent neurite extension, and potentially in the patterning of this growth. Hsp27

  15. Deletion of ENTPD3 does not impair nucleotide hydrolysis in primary somatosensory neurons or spinal cord [v1; ref status: indexed, http://f1000r.es/3rm

    Eric McCoy

    2014-07-01

    Full Text Available Ectonucleotidases are membrane-bound or secreted proteins that hydrolyze extracellular nucleotides.  Recently, we identified three ectonucleotidases that hydrolyze extracellular adenosine 5’-monophosphate (AMP to adenosine in primary somatosensory neurons.  Currently, it is unclear which ectonucleotidases hydrolyze ATP and ADP in these neurons.  Ectonucleoside triphosphate diphosphohydrolases (ENTPDs comprise a class of enzymes that dephosphorylate extracellular ATP and ADP.  Here, we found that ENTPD3 (also known as NTPDase3 or CD39L3 was located in nociceptive and non-nociceptive neurons of the dorsal root ganglion (DRG, in the dorsal horn of the spinal cord, and in free nerve endings in the skin.  To determine if ENTPD3 contributes directly to ATP and ADP hydrolysis in these tissues, we generated and characterized an Entpd3 knockout mouse.  This mouse lacks ENTPD3 protein in all tissues examined, including the DRG, spinal cord, skin, and bladder.  However, DRG and spinal cord tissues from Entpd3-/- mice showed no reduction in histochemical staining when ATP, ADP, AMP, or UTP were used as substrates.  Additionally, using fast-scan cyclic voltammetry (FSCV, adenosine production was not impaired in the dorsal spinal cord of Entpd3-/- mice when the substrate ADP was applied.  Further, Entpd3-/- mice did not differ in nociceptive behaviors when compared to wild-type mice, although Entpd3-/- mice showed a modest reduction in β-alanine-mediated itch.  Taken together, our data indicate that deletion of Entpd3 does not impair ATP or ADP hydrolysis in primary somatosensory neurons or in dorsal spinal cord.  Moreover, our data suggest there could be multiple ectonucleotidases that act redundantly to hydrolyze nucleotides in these regions of the nervous system.

  16. Deletion of ENTPD3 does not impair nucleotide hydrolysis in primary somatosensory neurons or spinal cord [v2; ref status: indexed, http://f1000r.es/4dl

    Eric McCoy

    2014-09-01

    Full Text Available Ectonucleotidases are membrane-bound or secreted proteins that hydrolyze extracellular nucleotides.  Recently, we identified three ectonucleotidases that hydrolyze extracellular adenosine 5’-monophosphate (AMP to adenosine in primary somatosensory neurons.  Currently, it is unclear which ectonucleotidases hydrolyze ATP and ADP in these neurons.  Ectonucleoside triphosphate diphosphohydrolases (ENTPDs comprise a class of enzymes that dephosphorylate extracellular ATP and ADP.  Here, we found that ENTPD3 (also known as NTPDase3 or CD39L3 was located in nociceptive and non-nociceptive neurons of the dorsal root ganglion (DRG, in the dorsal horn of the spinal cord, and in free nerve endings in the skin.  To determine if ENTPD3 contributes directly to ATP and ADP hydrolysis in these tissues, we generated and characterized an Entpd3 knockout mouse.  This mouse lacks ENTPD3 protein in all tissues examined, including the DRG, spinal cord, skin, and bladder.  However, DRG and spinal cord tissues from Entpd3-/- mice showed no reduction in histochemical staining when ATP, ADP, AMP, or UTP were used as substrates.  Additionally, using fast-scan cyclic voltammetry (FSCV, adenosine production was not impaired in the dorsal spinal cord of Entpd3-/- mice when the substrate ADP was applied.  Further, Entpd3-/- mice did not differ in nociceptive behaviors when compared to wild-type mice, although Entpd3-/- mice showed a modest reduction in β-alanine-mediated itch.  Taken together, our data indicate that deletion of Entpd3 does not impair ATP or ADP hydrolysis in primary somatosensory neurons or in dorsal spinal cord.  Moreover, our data suggest there could be multiple ectonucleotidases that act redundantly to hydrolyze nucleotides in these regions of the nervous system.

  17. Treating High Blood Pressure: Is a Beta-Blocker Drug Right for You?

    ... High Blood Pressure: Is a Beta-blocker Drug Right for You? What are beta-blockers? Beta-blockers ... talk with your doctor about which drugs are right for you. If your blood pressure is slightly ...

  18. Study of a New Neuron

    Adler, S. L.; Bhanot, G. V.; Weckel, J. D.

    1994-01-01

    We study a modular neuron alternative to the McCulloch-Pitts neuron that arises naturally in analog devices in which the neuron inputs are represented as coherent oscillatory wave signals. Although the modular neuron can compute $XOR$ at the one neuron level, it is still characterized by the same Vapnik-Chervonenkis dimension as the standard neuron. We give the formulas needed for constructing networks using the new neuron and training them using back-propagation. A numerical study of the mod...

  19. Impact of the DRG System in Arizona. Hearing before the Subcommittee on Health and Long-Term Care of the Select Committee on Aging. House of Representatives, Ninety-Ninth Congress, First Session (September 14, 1985, Tucson, AZ).

    Congress of the U.S., Washington, DC. House Select Committee on Aging.

    Text of a Congressional hearing held in Tucson, Arizona, to examine health care and the diagnostic related group (DRG) system is presented in this document. Opening statements are delivered by Representatives Kolbe and McCain. Witnesses testifying include: (1) two older Arizona residents who had experienced problems related to diagnostic related…

  20. Kalman Filter Neuron Training

    Murase, Haruhiko; KOYAMA, Shuhei; HONAMI, Nobuo; Kuwabara, Takao

    1991-01-01

    An attempt of implementing Kalman filter algorithm in the procedure for training the neural network was made and evaluated. The Kalman filter neuron training program (KNT) was coded. The performance of Kalman filter in KNT was compared to commonly used neuron training algorithm. The study revealed that KNT requires much less calculation time to accomplish neuron training than commonly used other algorithms do. KNT also gave much smaller final error than any other algorithms tested in this study.

  1. Mesmerising mirror neurons.

    Heyes, Cecilia

    2010-06-01

    Mirror neurons have been hailed as the key to understanding social cognition. I argue that three currents of thought-relating to evolution, atomism and telepathy-have magnified the perceived importance of mirror neurons. When they are understood to be a product of associative learning, rather than an adaptation for social cognition, mirror neurons are no longer mesmerising, but they continue to raise important questions about both the psychology of science and the neural bases of social cognition. PMID:20167276

  2. P2X7 receptors in satellite glial cells mediate high functional expression of P2X3 receptors in immature dorsal root ganglion neurons

    Chen Yong

    2012-02-01

    Full Text Available Abstract Background The purinergic P2X3 receptor (P2X3R expressed in the dorsal root ganglion (DRG sensory neuron and the P2X7 receptor (P2X7R expressed in the surrounding satellite glial cell (SGC are two major receptors participating in neuron-SGC communication in adult DRGs. Activation of P2X7Rs was found to tonically reduce the expression of P2X3Rs in DRGs, thus inhibiting the abnormal pain behaviors in adult rats. P2X receptors are also actively involved in sensory signaling in developing rodents. However, very little is known about the developmental change of P2X7Rs in DRGs and the interaction between P2X7Rs and P2X3Rs in those animals. We therefore examined the expression of P2X3Rs and P2X7Rs in postnatal rats and determined if P2X7R-P2X3R control exists in developing rats. Findings We immunostained DRGs of immature rats and found that P2X3Rs were expressed only in neurons and P2X7Rs were expressed only in SGCs. Western blot analyses indicated that P2X3R expression decreased while P2X7R expression increased with the age of rats. Electrophysiological studies showed that the number of DRG neurons responding to the stimulation of the P2XR agonist, α,β-meATP, was higher and the amplitudes of α,β-meATP-induced depolarizations were larger in immature DRG neurons. As a result, P2X3R-mediated flinching responses were much more pronounced in immature rats than those found in adult rats. When we reduced P2X7R expression with P2X7R-siRNA in postnatal and adult rats, P2X3R-mediated flinch responses were greatly enhanced in both rat populations. Conclusions These results show that the P2X7R expression increases as rats age. In addition, P2X7Rs in SGCs exert inhibitory control on the P2X3R expression and function in sensory neurons of immature rats, just as observed in adult rats. Regulation of P2X7R expression is likely an effective way to control P2X3R activity and manage pain relief in infants.

  3. Axotomy of tributaries of the pelvic and pudendal nerves induces changes in the neurochemistry of mouse dorsal root ganglion neurons and the spinal cord.

    McCarthy, Carly J; Tomasella, Eugenia; Malet, Mariana; Seroogy, Kim B; Hökfelt, Tomas; Villar, Marcelo J; Gebhart, G F; Brumovsky, Pablo R

    2016-05-01

    Using immunohistochemical techniques, we characterized changes in the expression of several neurochemical markers in lumbar 4-sacral 2 (L4-S2) dorsal root ganglion (DRG) neuron profiles (NPs) and the spinal cord of BALB/c mice after axotomy of the L6 and S1 spinal nerves, major tributaries of the pelvic (targeting pelvic visceral organs) and pudendal (targeting perineum and genitalia) nerves. Sham animals were included. Expression of cyclic AMP-dependent transcription factor 3 (ATF3), calcitonin gene-related peptide (CGRP), transient receptor potential cation channel subfamily V, member 1 (TRPV1), tyrosine hydroxylase (TH) and vesicular glutamate transporters (VGLUT) types 1 and -2 was analysed seven days after injury. L6-S1 axotomy induced dramatic de novo expression of ATF3 in many L6-S1 DRG NPs, and parallel significant downregulations in the percentage of CGRP-, TRPV1-, TH- and VGLUT2-immunoreactive (IR) DRG NPs, as compared to their expression in uninjured DRGs (contralateral L6-S1-AXO; sham mice); VGLUT1 expression remained unaltered. Sham L6-S1 DRGs only showed a small ipsilateral increase in ATF3-IR NPs (other markers were unchanged). L6-S1-AXO induced de novo expression of ATF3 in several lumbosacral spinal cord motoneurons and parasympathetic preganglionic neurons; in sham mice the effect was limited to a few motoneurons. Finally, a moderate decrease in CGRP- and TRPV1-like-immunoreactivities was observed in the ipsilateral superficial dorsal horn neuropil. In conclusion, injury of a mixed visceral/non-visceral nerve leads to considerable neurochemical alterations in DRGs matched, to some extent, in the spinal cord. Changes in these and potentially other nociception-related molecules could contribute to pain due to injury of nerves in the abdominopelvic cavity. PMID:25749859

  4. Structural characterization and expression analysis of a beta-thymosin homologue (Tβ) in disk abalone, Haliotis discus discus.

    Kasthuri, Saranya Revathy; Premachandra, H K A; Umasuthan, Navaneethaiyer; Whang, Ilson; Lee, Jehee

    2013-09-15

    Repertoires of proteins and small peptides play numerous physiological roles as hormones, antimicrobial peptides, and cellular signaling factors. The beta-thymosins are a group of small acidic peptides involved in processes such as actin sequestration, neuronal development, wound healing, tissue repair, and angiogenesis. Recent characterization of the beta thymosins as immunological regulators in invertebrates led to our identification and characterization of a beta-thymosin homologue (Tβ) from Haliotis discus discus. The cDNA possessed an ORF of 132 bp encoding a protein of 44 amino acids with a molecular mass of 4977 Da. The amino acid sequence shows high identity with another molluskan beta-thymosin and has a characteristic actin binding motif (LKKTET) and glutamyl donors. Phylogenetic analysis showed a close relationship with molluskan homologues, as well as its distinct identity and common ancestral origin. Genomic analysis revealed a 3 exon-2 intron structure similar to the other homologues. In silico promoter analysis also revealed significant transcription factor binding sites, providing evidence for the expression of this gene under different cellular conditions, including stress or pathogenic attack. Tissue distribution profiling revealed a ubiquitous presence in all the examined tissues, but with the highest expression in mantle and hemocyte. Immune challenge with lipopolysaccharide, poly I:C and Vibrio parahemolyticus induced beta-thymosin expression in gill and hemocytes, affirming an immune-related role in invertebrates. PMID:23680646

  5. The Expenditures for Academic Inpatient Care of Inflammatory Bowel Disease Patients Are Almost Double Compared with Average Academic Gastroenterology and Hepatology Cases and Not Fully Recovered by Diagnosis-Related Group (DRG Proceeds.

    Daniel C Baumgart

    Full Text Available Crohn's disease (CD and ulcerative colitis (UC challenge economies worldwide. Detailed health economic data of DRG based academic inpatient care for inflammatory bowel disease (IBD patients in Europe is unavailable.IBD was identified through ICD-10 K50 and K51 code groups. We took an actual costing approach, compared expenditures to G-DRG and non-DRG proceeds and performed detailed cost center and type accounting to identify coverage determinants.Of all 3093 hospitalized cases at our department, 164 were CD and 157 UC inpatients in 2012. On average, they were 44.1 (CD 44.9 UC 43.3 all 58 years old, stayed 10.1 (CD 11.8 UC 8.4 vs. all 8 days, carried 5.8 (CD 6.4 UC 5.2 vs. all 6.8 secondary diagnoses, received 7.4 (CD 7.7 UC 7 vs. all 6.2 procedures, had a higher cost weight (CD 2.8 UC 2.4 vs. all 1.6 and required more intense nursing. Their care was more costly (means: total cost IBD 8477€ CD 9051€ UC 7903€ vs. all 5078€. However, expenditures were not fully recovered by DRG proceeds (means: IBD 7413€, CD 8441€, UC 6384€ vs all 4758€. We discovered substantial disease specific mismatches in cost centers and types and identified the medical ward personnel and materials budgets to be most imbalanced. Non-DRG proceeds were almost double (IBD 16.1% vs. all 8.2%, but did not balance deficits at total coverage analysis, that found medications (antimicrobials, biologics and blood products, medical materials (mostly endoscopy items to contribute most to the deficit.DRGs challenge sophisticated IBD care.

  6. Electrophysiological evidence for the existence of a rare population of C-fiber low threshold mechanoreceptive (C-LTM) neurons in glabrous skin of the rat hindpaw.

    Djouhri, Laiche

    2016-02-01

    The mammalian skin in innervated by distinct classes of low-threshold mechanoreceptive (LTM) primary afferent neurons that are classified as Aβ-, Aδ- or C-LTMs according to their axonal conduction velocities (CVs). C-LTMs are thought to signal pleasant and erotic touch sensations in humans, and to exist only in the hairy skin of primates and other species. Using intracellular recordings from rat L4/L5 dorsal root ganglion (DRG) neurons that were classified in vivo as C-nociceptors or C-LTMs, according to their dorsal root CVs and their responses to mechanical and thermal stimuli, the present study provides the first electrophysiological evidence that C-LTMs exist in the glabrous skin of the rat's hindpaw. Indeed 6.4% (5/78) of the total sample of lumbar C-fiber DRG neurons with receptive fields in the glabrous skin of the rat hindpaw were C-LTMs. The electrophysiological properties of this rare subpopulation of C-fiber neurons (mean CV=0.48±0.06m/s) are distinct from those of C-fiber high threshold mechanoreceptors (HTMs). Indeed, their mean mechanical (1.7±1.1mN) and electrical (4.0±0.4V) thresholds was significantly different from that of C-HTMs. They also exhibited faster action potential and afterhyperpolarization kinetics than C-HTMs. The present study lends support to previous studies that have provided indirect evidence for the presence of C-LTMs in glabrous skin. If C-LTMs are present in human glabrous skin, they may, in this type of skin, represent a novel peripheral neuronal substrate for the pleasant/social touch sensation, and account for or contribute to touch hypersensitivity after injury. PMID:26752785

  7. Regulation of voltage-gated Ca(2+) currents by Ca(2+)/calmodulin-dependent protein kinase II in resting sensory neurons.

    Kostic, Sandra; Pan, Bin; Guo, Yuan; Yu, Hongwei; Sapunar, Damir; Kwok, Wai-Meng; Hudmon, Andy; Wu, Hsiang-En; Hogan, Quinn H

    2014-09-01

    Calcium/calmodulin-dependent protein kinase II (CaMKII) is recognized as a key element in encoding depolarization activity of excitable cells into facilitated voltage-gated Ca(2+) channel (VGCC) function. Less is known about the participation of CaMKII in regulating VGCCs in resting cells. We examined constitutive CaMKII control of Ca(2+) currents in peripheral sensory neurons acutely isolated from dorsal root ganglia (DRGs) of adult rats. The small molecule CaMKII inhibitor KN-93 (1.0μM) reduced depolarization-induced ICa by 16-30% in excess of the effects produced by the inactive homolog KN-92. The specificity of CaMKII inhibition on VGCC function was shown by the efficacy of the selective CaMKII blocking peptide autocamtide-2-related inhibitory peptide in a membrane-permeable myristoylated form, which also reduced VGCC current in resting neurons. Loss of VGCC currents is primarily due to reduced N-type current, as application of mAIP selectively reduced N-type current by approximately 30%, and prior N-type current inhibition eliminated the effect of mAIP on VGCCs, while prior block of L-type channels did not reduce the effect of mAIP on total ICa. T-type currents were not affected by mAIP in resting DRG neurons. Transduction of sensory neurons in vivo by DRG injection of an adeno-associated virus expressing AIP also resulted in a loss of N-type currents. Together, these findings reveal a novel molecular adaptation whereby sensory neurons retain CaMKII support of VGCCs despite remaining quiescent. PMID:25064143

  8. Regulation of Voltage-Gated Ca2+ Currents by Ca2+/Calmodulin-dependent Protein Kinase II in Resting Sensory Neurons

    Kostic, Sandra; Pan, Bin; Guo, Yuan; Yu, Hongwei; Sapunar, Damir; Kwok, Wai-Meng; Hudmon, Andy; Wu, Hsiang-En; Hogan, Quinn H.

    2014-01-01

    Calcium/calmodulin-dependent protein kinase II (CaMKII) is recognized as a key element in encoding depolarization activity of excitable cells into facilitated voltage-gated Ca2+ channel (VGCC) function. Less is known about the participation of CaMKII in regulating VGCCs in resting cells. We examined constitutive CaMKII control of Ca2+ currents in peripheral sensory neurons acutely isolated from dorsal root ganglia (DRGs) of adult rats. The small molecule CaMKII inhibitor KN-93 (1.0μM) reduced depolarization-induced ICa by 16 – 30% in excess of the effects produced by the inactive homolog KN-92. The specificity of CaMKII inhibition on VGCC function was shown by efficacy of the selective CaMKII blocking peptide autocamtide-2-related inhibitory peptide in a membrane-permeable myristoylated form, which also reduced VGCC current in resting neurons. Loss of VGCC currents is primarily due to reduced N-type current, as application of mAIP selectively reduced N-type current by approximately 30%, and prior N-type current inhibition eliminated the effect of mAIP on VGCCs, while prior block of L-type channels did not reduce the effect of mAIP on total ICa. T-type currents were not affected by mAIP in resting DRG neurons. Transduction of sensory neurons in vivo by DRG injection of an adeno-associated virus expressing AIP also resulted in a loss of N-type currents. Together, these findings reveal a novel molecular adaptation whereby sensory neurons retain CaMKII support of VGCCs despite remaining quiescent. PMID:25064143

  9. Kappe neurons, a novel population of olfactory sensory neurons

    Ahuja, Gaurav; Nia, Shahrzad Bozorg; Zapilko, Veronika; Shiriagin, Vladimir; Kowatschew, Daniel; Oka, Yuichiro; Korsching, Sigrun I.

    2014-01-01

    Perception of olfactory stimuli is mediated by distinct populations of olfactory sensory neurons, each with a characteristic set of morphological as well as functional parameters. Beyond two large populations of ciliated and microvillous neurons, a third population, crypt neurons, has been identified in teleost and cartilaginous fishes. We report here a novel, fourth olfactory sensory neuron population in zebrafish, which we named kappe neurons for their characteristic shape. Kappe neurons ar...

  10. Neuronal Response Clamp

    Avner Wallach

    2011-04-01

    Full Text Available Responses of individual neurons to ongoing input are highly variable, reflecting complex threshold dynamics. Experimental access to this threshold dynamics is required in order to fully characterize neuronal input-output relationships. The challenge is practically intractable using present day experimental paradigms due to the cumulative, nonlinear interactions involved. Here we introduce the Neuronal Response Clamp, a closed-loop technique enabling control over the instantaneous response probability of the neuron. The potential of the technique is demonstrated by showing direct access to threshold dynamics of cortical neuron in-vitro using extracellular recording and stimulation, over timescales ranging from seconds to many hours. Moreover, the method allowed us to expose the sensitivity of threshold dynamics to spontaneous input from the network in which the neuron is embedded. The Response Clamp technique follows the rationale of the voltage-clamp and dynamic-clamp approaches, extending it to the neuron's spiking behavior. The general framework offered here is applicable in the study of other neural systems, beyond the single neuron level.