Sample records for chick embryo formation

  1. Neurotoxic effect of the dithiocarbamate tecoram on the chick embryo

    Steenis, G. van; Logten, M.J. van


    Tecoram, when administered at doses of 0.01, 0.1, 1 or 10 mg per egg in propylene glycol or in saline to chick embryos caused paralysis, shortening of the extremities, muscular atrophy, dwarfing and death. Microscopically there were signs of peripheral neuropathy, mainly confined to the distal parts

  2. The effect of insecticide Deltamethrin on development of chick embryos

    This study was conducted to evaluate the cyto and the embryo toxicity of Deltamethrin and its commercial formulation DECIS 50 EC in chick embryo during its critical embryonic development period before and in the organogenesis. The embryos were incubated in well closed plastic caps containing the complete egg composition at 38 o. the Deltamethrin and DECIS were found to cause histological and morphological malformations, specially in the brain, also they reduced the majority of the synthetic activities of the DNA, RNA, and proteins in the embryonic and the vascular areas. The flow cytometric analysis showed alterations in frequency of cells in both embryonic and vascular areas in the treated embryo during the cell cycle phases. Our study also showed that the DECIS had greater cyto and embryo toxicity than the Seltamethrin for analysis (author). 149 refs., 36 figs., 16 tabs

  3. [Effects of chronic paralysis of chick embryo by flaxedil on the development of the neuromuscular junction].

    Bourgeois, J P; Betz, H; Changuex, J P


    Chronic paralysis of Chick embryos by the cholinergic antagonist flaxedil blocks the subsynaptic accumulation of acetylcholinesterase but not the formation of acetylcholine receptor cluster. Flaxedil paralysis also causes an increase of the total muscle content of acetylcholine receptor without altering the half-life of the receptor protein. The spontaneous activity of the embryon therefore "shuts off" the synthesis of extrasynpatic acetylcholine receptor. PMID:417864

  4. Chick embryo fibroblasts produce two forms of hyaluronidase

    Orkin, RW; Toole, BP


    Cultured chick embryo fibroblasts derived from skin and skeletal muscle exhibit hyaluronidase activity both associated with the cell layer and secreted into the medium. Although both forms of the enzyme have a number of similar characteristics (R.W. Orkin and B.P. Toole, 1980, J. Biol. CHem. 255), they differ in thermal stability at neutral pH and in behavior on ion-exchange chromatography. Both forms of the enzyme are equally stable at acidic pH for long intervals, but the cell-associated hy...


    Chandrapragasam Vani


    Full Text Available Silica is one of the most effective stored seedand food grain protecting pesticide agent, usedfor ages Silica nanoparticles are hard and strong andresistant to brittle fracture under an imposed stress. The aim of our study is to synthesize and evaluate the toxicity of silica nanoparticles at various concentrations using chick embryo, as it serves as a bridging model between in vivo and ex vivo studies. Silica nanoparticles of 70 nm sizewith concentrations of 10, 20, 40 and 80 PPM were injected in vivo and incubated for 19 days as per IACUC guidelines. The study was carried out on dissected chick embryo after the 19th day of incubation. Vital tissues such as liver and heart tissues were subjected to toxicity assays. Biochemical assay of antioxidant enzymes like superoxide dismutase and glutathione peroxide were assessed. The results showed a decreased level of MDA (Malondialdehyde, an end product of lipid Peroxidation, whereas increased activity of superoxide dismutase and glutathione peroxide were observed. Therefore silica nanoparticles may be used as a pesticide and also as a biomaterial for therapeutic application in the field of medicine.

  6. BMP is an important regulator of proepicardial identity in the chick embryo.

    Schlueter, Jan; Männer, Jörg; Brand, Thomas


    The proepicardium (PE) is a transient structure formed by pericardial coelomic mesothelium at the venous pole of the embryonic heart and gives rise to several cell types of the mature heart. In order to study PE development in chick embryos, we have analyzed the expression pattern of the marker genes Tbx18, Wt1, and Cfc. During PE induction, the three marker genes displayed a left-right asymmetric expression pattern. In each case, expression on the right side was stronger than on the left side. The left-right asymmetric gene expression observed here is in accord with the asymmetric formation of the proepicardium in the chick embryo. While initially the marker genes were expressed in the primitive sinus horn, subsequently, expression became confined to the PE mesothelium. In order to search for signaling factors involved in PE development, we studied Bmp2 and Bmp4 expression. Bmp2 was bilaterally expressed in the sinus venosus. In contrast, Bmp4 expression was initially expressed unilaterally in the right sinus horn and subsequently in the PE. In order to assess its functional role, BMP signaling was experimentally modulated by supplying exogenous BMP2 and by inhibiting endogenous BMP signaling through the addition of Noggin. Both supplying BMP and blocking BMP signaling resulted in a loss of PE marker gene expression. Surprisingly, both experimental situations lead to cardiac myocyte formation in the PE cultures. Careful titration experiments with exogenously added BMP2 or Noggin revealed that PE-specific marker gene expression depends on a low level of BMP signaling. Implantation of BMP2-secreting cells or beads filled with Noggin protein into the right sinus horn of HH stage 11 embryos resulted in downregulation of Tbx18 expression, corresponding to the results of the explant assay. Thus, a distinct level of BMP signaling is required for PE formation in the chick embryo. PMID:16677627

  7. Bilateral neuro-retinitis following chick embryo cell anti-rabies vaccination – a case report

    Rai Harminder


    Full Text Available Abstract Background The Optic nerve is rarely involved after sheep brain anti-rabies vaccination in the form of retrobulbar neuritis or papillitis. Bilateral neuroretinitis after chick embryo cell antirabies vaccination has not been reported. Case presentation We report the case of a 56 year old male who developed bilateral neuro-retinitis following three injections of antirabies vaccine prepared from the chick embryo. Conclusion The chick embryo cell antirabies vaccine can cause bilateral neuroretinits which has not been reported previously.

  8. Method of Electroporation for the Early Chick Embryo

    Hatakeyama, Jun; Shimamura, Kenji

    Chick embryos have long been one of the favored model systems in the field of embryology and developmental biology. Recent advances in the gene manipulation technologies (Muramatsu et al., 1997; Nakamura et al., 2004) make this model system even more attractive for the developmental biologists (see review by Stern, 2005). Thanks to its two dimensional geometry, easiness in accessibility and observation, and well-established fate maps (e.g. Couly and Le Douarin, 1988; Garcia-Martinez et al., 1993; Hatada and Stern, 1994; Psychoyos and Stern, 1996; Sawada and Aoyama, 1999; Cobos et al., 2001; Lopez-Sanchez et al., 2001; Redkar et al., 2001; Fernandez-Garre et al., 2002; Kimura et al., 2006; Matsushita et al., 2008), it has great advantages especially for studies at the early embryonic stages, such as the processes of gastrulation, neural induction, left-right patterning, etc. For such purposes, a whole embryo culture system, originally invented by Dennis A. T. New (New, 1955), and its derivatives (Flamme, 1987; Sundin and Eichele, 1992; Stern, 1993; Chapman et al., 2001) have been widely used.

  9. Development of teeth in chick embryos after mouse neural crest transplantations

    Mitsiadis, Thimios A.; Chéraud, Yvonnick; Sharpe, Paul; Fontaine-Pérus, Josiane


    Teeth were lost in birds 70–80 million years ago. Current thinking holds that it is the avian cranial neural crest-derived mesenchyme that has lost odontogenic capacity, whereas the oral epithelium retains the signaling properties required to induce odontogenesis. To investigate the odontogenic capacity of ectomesenchyme, we have used neural tube transplantations from mice to chick embryos to replace the chick neural crest cell populations with mouse neural crest cells. The mouse/chick ...

  10. Gc protein-derived macrophage-activating factor (GcMAF) stimulates cAMP formation in human mononuclear cells and inhibits angiogenesis in chick embryo chorionallantoic membrane assay.

    Pacini, Stefania; Morucci, Gabriele; Punzi, Tiziana; Gulisano, Massimo; Ruggiero, Marco


    The effects of Gc protein-derived macrophage-activating factor (GcMAF) have been studied in cancer and other conditions where angiogenesis is deregulated. In this study, we demonstrate for the first time that the mitogenic response of human peripheral blood mononuclear cells (PBMCs) to GcMAF was associated with 3'-5'-cyclic adenosine monophosphate (cAMP) formation. The effect was dose dependent, and maximal stimulation was achieved using 0.1 ng/ml. Heparin inhibited the stimulatory effect of GcMAF on PBMCs. In addition, we demonstrate that GcMAF (1 ng/ml) inhibited prostaglandin E(1)- and human breast cancer cell-stimulated angiogenesis in chick embryo chorionallantoic membrane (CAM) assay. Finally, we tested different GcMAF preparations on CAM, and the assay proved to be a reliable, reproducible and inexpensive method to determine the relative potencies of different preparations and their stability; we observed that storage at room temperature for 15 days decreased GcMAF potency by about 50%. These data could prove useful for upcoming clinical trials on GcMAF. PMID:21170647

  11. Effect of Low Dose Radiation Upon Antioxidant Parameters in Skeletal Muscle of Chick Embryo

    In this paper an attempt was made to determine the effect of irradiation of eggs with low dose ionizing radiation upon lipid peroxide (TBARS) level, glutathione (GSH) level, activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) in skeletal muscle of chick embryo and newly hatched chicks. The eggs of a heavy breeding chickens were irradiated with a dose of 0.3 Gy gamma radiation (60Co source) on the 19th day of incubation. Along with the irradiated chick embryos, there was a control group of non-irradiated chick embryos. The antioxidant parameters were measured in breast muscle (m. pectoralis superficialis) and thigh muscle (m. biceps femoris) of chick embryos on 1, 3, 6, 24 and 72 h after egg irradiation. All parameters were determined spectrophotometrically. Lipid peroxidation, GSH level and CAT activity decreased in the breast and thigh muscle of chick embryos on the first hour after irradiation, while the activity of GSH-Px increased in the thigh muscle on the 1st hour after irradiation. CAT activity decreased in the breast muscle of chick embryos on the hour 24 after irradiation. The GSH level increased in the breast and thigh muscle of chick embryos on the hour 72 after irradiation while the activity of GSH-Px increased in the breast muscle. At the same time CAT activity decreased in breast muscle while lipid peroxidation decreased in thigh muscle. The obtained results showed that acute irradiation of chicken eggs on the 19th day of incubation with the dose of 0.3 Gy gamma radiation could be an oxidative stress in both types of muscles immediately after irradiation. However, at the one-day old chicks (72 hours after irradiation) this dose could have a stimulating effect upon GSH level in both breast and thigh muscle.(author)

  12. A shell-less chick embryo culturing technique, reproduced successfully under local circumstances

    The goal of this project was to demonstrate shell-less chick embryo culturing as a potential experimental model in the field of developmental anatomy. Freshly laid, fertilized chicken eggs of Egyptian Fayoumi breed were obtained from Poultry Research Institute Punjab, Rawalpindi. The fertilized chicken eggs were preincubated for 33 hours under standard conditions of 37.5 degree C and 65-75% humidity, to bring them to stage 9 (29-33 hours embryo, 7 somites) of Hamburger and Hamilton staging system. After this period, the eggs were taken out of the incubator, placed horizontally, wiped with 70% ethanol and permitted to air-dry for 10 minutes to reduce contamination from the egg surface and also to ensure that the embryo was properly positioned. The eggs contents were then transferred into the culture containers by cracking the undersides against an edge. The formation and growth of the embryonic membranes, the central nervous system - beginning from the vesicle stage, the circulatory system - including the heart, the eyes, beak, limbs, skin, feathers, wings and folding of the body were directly observed. Repeated successful culturing was attempted, tracing the developmental process of the embryo upto the 15th day of embryonic life at least after which the survivability period varied in different embryo cultures. The most advanced age reached in this project was day 19 of the embryonic life, which in researchers understanding is the latest developmental stage in shellless environment described as yet. The normal hatching time of this breed is 21-22 days. The size of these embryos was smaller as compared to the embryos of the same age that carried out their development inside their shells. (author)

  13. Cysteic acid and taurine synthesis from sulphate in the chick embryo

    The formation of taurine from sulphate was studied in the chick embryo using the radioisotopes of: sulphur, carbon and hydrogen. The following reactions occur: 1) reduction of sulphate to sulphite; 2) fixation of the sulphite on a carbon chain with an amino group, resulting from desulphydration of L-cysteine, which leads to the formation of L-cysteic acid; 3) decarboxylation of L-cysteic acid. Reaction (1) takes place only in the endo-dermal cells of the yolk sac; reaction (2) in these same cells and in the yolk; reaction (3) is general, localized in the yolk sac, in the yolk as well as in the tissues of the embryo itself. The enzyme which catalyses reaction (2) has been purified; the coenzyme is pyridoxal phosphate. The desulphydration of cysteine by this enzyme is a reversible reaction. In non-physiological conditions of concentration and temperature, pyridoxal phosphate catalyses in the presence of metallic ions, the desulphydration of cysteine and the formation of cysteic acid from sulphite. (author)

  14. 3-Dimensional modelling of chick embryo eye development and growth using high resolution magnetic resonance imaging.

    Goodall, Nicola; Kisiswa, Lilian; Prashar, Ankush; Faulkner, Stuart; Tokarczuk, Paweł; Singh, Krish; Erichsen, Jonathan T; Guggenheim, Jez; Halfter, Willi; Wride, Michael A


    Magnetic resonance imaging (MRI) is a powerful tool for generating 3-dimensional structural and functional image data. MRI has already proven valuable in creating atlases of mouse and quail development. Here, we have exploited high resolution MRI to determine the parameters necessary to acquire images of the chick embryo eye. Using a 9.4 Tesla (400 MHz) high field ultra-shielded and refrigerated magnet (Bruker), MRI was carried out on paraformaldehyde-fixed chick embryos or heads at E4, E6, E8, and E10. Image data were processed using established and custom packages (MRICro, ImageJ, ParaVision, Bruker and mri3dX). Voxel dimensions ranged from 62.5 microm to 117.2 microm. We subsequently used the images obtained from the MRI data in order to make precise measurements of chick embryo eye surface area, volume and axial length from E4 to E10. MRI was validated for accurate sizing of ocular tissue features by direct comparison with previously published literature. Furthermore, we demonstrate the utility of high resolution MRI for making accurate measurements of morphological changes due to experimental manipulation of chick eye development, thereby facilitating a better understanding of the effects on chick embryo eye development and growth of such manipulations. Chondroitin sulphate or heparin were microinjected into the vitreous cavity of the right eyes of each of 3 embryos at E5. At E10, embryos were fixed and various eye parameters (volume, surface area, axial length and equatorial diameter) were determined using MRI and normalised with respect to the un-injected left eyes. Statistically significant alterations in eye volume (p < 0.05; increases with chondroitin sulphate and decreases with heparin) and changes in vitreous homogeneity were observed in embryos following microinjection of glycosaminoglycans. Furthermore, in the heparin-injected eyes, significant disturbances at the vitreo-retinal boundary were observed as well as retinal folding and detachment

  15. Hypoxia adaptation and hemoglobin mutation in Tibetan chick embryo

    GOU; Xiao; LI; Ning; LIAN; Linsheng; YAN; Dawei; ZHANG; Hao


    Tibetan chick lives at high altitudes between 2600 and 4200 m with a high hatchability and low land breeds survive rarely with a hatchability of 3.0% under hypoxia of simulated 4200 m. Under hypoxia of whole 21 d, the hatchability of Tibetan chick and Recessive White Feather broiler differed with a greatest disparity from day 4 to 11 and also significantly in other stages except from day 1 to 3. Hypoxia in each stage did not reduce significantly survival rate of this stage except hatchability. These two results indicated that the hypoxia in the early stage had an adverse effect on the later stage. All exons encoding chick hemoglobins were sequenced to analyze gene polymorphism. The functional mutation Met-32(B13)-Leu, related with hypoxia, was found in αD globin chain and the mutation frequency increased with increased altitude. In addition, under hypoxic conditions, the population with higher mutation frequency had a higher hatchability. The automated homology model building was carried out using crystal structure coordinates of chick HbD. The results indicated that the substitution Met-32(B13)-Leu provides a more hydrophobic environment which leads to higher stability of heme and oxygen affinity of hemoglobin. The occurrence of the mutation Met-32(B13)-Leu is related to the origin of Tibetan chick.

  16. Effects of white light on the pineal gland of the chick embryo

    Aige-Gil, Vicente; Murillo-Ferrol, Narciso


    Chick embryos were directly exposed to a source of white light during incubation and sacrificed before hatching. The light caused a number of teratological effects such as high mortality, delay in development, celosomy, hepatomegaly, auricular dilation and micrognatia. The pineal gland of the illuminated embryos showed an increase in number and size of the intracytoplasmic lipid droplets of the follicular pinealocytes. These findings suggest that the pineal...

  17. Investigation of the chick embryo as a potential alternative to the mouse for evaluation of radiopharmaceuticals

    Introduction: The chick embryo is an emerging in vivo model in several areas of pre-clinical research including radiopharmaceutical sciences. Herein, it was evaluated as a potential test system for assessing the biodistribution and in vivo stability of radiopharmaceuticals. For this purpose, a number of radiopharmaceuticals labeled with 18F, 125I, 99mTc, and 177Lu were investigated in the chick embryo and compared with the data obtained in mice. Methods: Chick embryos were cultivated ex ovo for 17–19 days before application of the radiopharmaceutical directly into the peritoneum or intravenously using a vein of the chorioallantoic membrane (CAM). At a defined time point after application of radioactivity, the embryos were euthanized by shock-freezing using liquid nitrogen. Afterwards they were separated from residual egg components for post mortem imaging purposes using positron emission tomography (PET) or single photon emission computed tomography (SPECT). Results: SPECT images revealed uptake of [99mTc]pertechnetate and [125I]iodide in the thyroid of chick embryos and mice, whereas [177Lu]lutetium, [18F]fluoride and [99mTc]-methylene diphosphonate ([99mTc]-MDP) were accumulated in the bones. [99mTc]-dimercaptosuccinic acid (99mTc-DMSA) and the somatostatin analog [177Lu]-DOTATOC, as well as the folic acid derivative [177Lu]-DOTA-folate showed accumulation in the renal tissue whereas [99mTc]-mebrofenin accumulated in the gall bladder and intestine of both species. In vivo dehalogenation of [18F]fallypride and of the folic acid derivative [125I]iodo-tyrosine-folate was observed in both species. In contrast, the 3′-aza-2′-[18F]fluorofolic acid ([18F]-AzaFol) was stable in the chick embryo as well as in the mouse. Conclusions: Our results revealed the same tissue distribution profile and in vivo stability of radiopharmaceuticals in the chick embryo and the mouse. This observation is promising with regard to a potential use of the chick embryo as an inexpensive


    Shweta P. Alai et al.


    Full Text Available There are various drugs used in daily life. Some of them show hazardous effect on various organs and some of them shows even lethal effect. Such drugs are required to banned. One of them is Etizolam (marketed under the brand name Sedekopan, Pasaden or Depas is a drug which is a thiebenzothiodiazepines analog. The aim of this work is done to compare the effect of administration of the etizolam drug at (Zero-day before incubation and at the (5-day first week of incubation, to get an idea about the extent of the teratogenic effect of this drug on the development of chick embryo. So that morphological and biochemical studies were done on the chick embryo of 8- and 16-days old chick embryos treated with (0.5mg/ml egg of etizolam on the (zero and 5-th days of incubation. Etizolam caused abnormality in concentration dependent manner. Enlarged remains of primitive streak was seen at 0.1mg/ml and o.3mg/ml concentration of Etizolam embryo dies. Etizolam inhibit development of somites of the development of 5-day at of incubation also it does not allow the development of notochord. The toxic effects of the drug was seen also more at 16-days treated chick embryos and the drug induced death of chick embryo at both 8- and 16-days treated with 0.1mg/ml and o.3mg/ml. The conclusion from this study was the etizolam drug has dangerous effects on tissues and with powerful toxicity on the cells, because the drug absorbed rapidly with peak plasma levels achieved between 30 minutes and 2 hours and has a mean elimination half life of about 3 and a half hours.

  19. The Protective Effects of Different Sources of Maternal Selenium on Oxidative Stressed Chick Embryo Liver.

    Xiao, Xue; Yuan, Dong; Wang, Yong-Xia; Zhan, Xiu-An


    The experiment was conducted to investigate the protective effects of different sources of maternal selenium (Se) on oxidative stressed chick embryo. A total of 270 Lingnan Yellow broiler breeders were randomly allocated into three treatments with five replicates for 18 birds each. Breeders were fed with basal diet (BD) including 0.04 mg/kg Se or BD supplemented with sodium selenite (SS) or selenomethionine (SM) at a level of 0.15 mg Se/kg. The rearing experiment lasted for 8 weeks after an 8-week pre-test. Twenty eggs were collected from each replicate during the last 10-day, then incubated in a commercial incubator. On embryonic 17th, fertile eggs were transferred into 39.5 °C temperature stimulation for 6 h. Afterward, five eggs were randomly selected from each replicate for collecting chick embryo sample. The results showed that Se supplementation in the diet of breeders resulted in lower reactive oxygen species (ROS), heat shock protein 70 (HSP70), malondialdehyde (MDA), carbonyl and 8-hydroxydeoxyguanosine (8-OHdG) concentrations and higher glutathione peroxidase (GPx), total superoxide dismutase (T-SOD), and catalase (CAT) activities in heat stress treated chick embryo (P reduce ROS concentration and oxidative damage by upregulating the expression of antioxidative selenoprotein, and maternal SM is superior to SS in heat stress treated chick embryo. PMID:26554950

  20. Development of a green fluorescent protein metastatic-cancer chick-embryo drug-screen model

    Bobek, V.; Plachý, Jiří; Pintérová, D.; Kološtová, K.; Boubelík, Michael; Jiang, P.; Yang, M.; Hoffman, R. M.


    Roč. 21, č. 4 (2004), s. 347-352. ISSN 0262-0898 Institutional research plan: CEZ:AV0Z5052915 Keywords : chick embryo * Lewis lung carcinoma * metastasis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.048, year: 2004

  1. Development of remnant kidney in the chick embryos with unilateral renal agenesis

    Zemanová, Zdeňka; Jirsová, Z.; Murphy, M. J.

    Berlin : Institute of biology, 2005. s. 61-61. [Combined workshop of fundamental physiology of the european working group of physiology and perinatal development poultry /2./. 23.09.2005-25.09.2005, Berlin] R&D Projects: GA ČR GA304/04/0972 Keywords : chick embryo * renal agenesis * mesonephros Subject RIV: EA - Cell Biology

  2. Bendiocarb effect on liver and central nervous system in the chick embryo

    Petrovová, E.; Sedmera, David; Lešník, František; Luptáková, L.


    Roč. 44, č. 4 (2009), s. 383-388. ISSN 0360-1234 Institutional research plan: CEZ:AV0Z50450515 Keywords : Bendiocarb * Chick embryo * Toxicity Subject RIV: EA - Cell Biology Impact factor: 1.097, year: 2009

  3. Morphogenetic Activities of Bendiocarb as Cholinesterase Inhibitor on Development of the Chick Embryo

    Petrovová, E.; Luptáková, L.; Mazenský, D.; Danko, J.; Sedmera, David

    Rijeka: InTech, 2011 - (Stoytcheva, M.), s. 469-494 ISBN 978-953-307-457-3 Institutional research plan: CEZ:AV0Z50110509 Keywords : bendiocarb * chick embryo * toxicity * development * pesticides * central nervous system * cell death Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine

  4. Adaptation of infectious bronchitis virus in primary cells of the chick embryo chorioallantoic membrane

    M. H. Mohammed


    Full Text Available The susceptibility of the primary chick embryo chorioallontoic membrane cells to infectious bronchitis virus was evaluated after twenty consecutive passages in chick embryo chorioallontoic membrane cells. Virus replication was monitored by cytopathic observation, indirect immunoperoxidase, and reverse transcription polymerase chain reaction (RT-PCR. At 72 hours post-infection (p.i. in third passage, the cytopathic effect was characterized by rounding up of cells, monolayer detachment, intracytoplasmic brownish colouration was readily observed by immunoperoxidase from 24 hours p.i in third passage, and at all times the extracted viral RNA from IBV-infected monolayers was demonstrated by RT-PCR. Tissue culture ineffective dose50 (TCID50 was used to measure virus titration performed on primary chick embryo chorioallontoic membrane cells and the titre in twenty passage was 108.6 TCID50/ml. The results obtained in this study suggested that the primary chick embryo chorioallontoic membrane cells can be used for adaptation infectious bronchitis virus (IBV and may be considered a step forward for the use of these cells in the future for IBV vaccine production

  5. A method for detecting MFO induction by Japanese pulp mill effluents with chick embryo

    Tatarazako, Norihisa; Kamaya, Yasushi [Japan Pulp and Paper Research Inst. Inc., Tokodai, Tsukuba (Japan)


    As a biomarker of physiological responses in fish exposed to pulp and paper mill effluents, mixed function oxygenase (MFO) induction has been investigated by many researchers. The induction and/or inhibition of MFOs is generally affected by various factors of fish such as species, maturity, sex, size and spawning status, and by other environmental variables. Therefore, the method demands technical skills to get a constant result. In addition, the test is costly and also time consuming. In this paper, the authors propose a MFOs induction method using chick embryo, instead of fish, for the assessment of pulp mill effluent. The merits of this method are as follows; inexpensive and commercially available test organisms, easy maintenance of the organisms, no feeding, high uniformity of the developing stage, sensitive responses to xenobiotics, low sample volume requirements, easy testing of various samples at one time and minimal training. P450 1A1 has been found in the microsome of chick embryo. Metabolic function of the P450 1A1 can be regarded basically the same as that of fish. Small amount of xenobiotics, about 100 {micro}l, were injected into the air chamber of 16-day-old chick embryos. Liver microsomes were isolated 48 h after administration. Ethoxyresorufin-O-deethylase (EROD) activity was determined by the direct fluorimetric method. Using the chick embryo method, the authors analyzed various chloro-organics, model bleached kraft pulp mill effluents, model black liquor and several total effluents of Japanese pulp mills. Methylcholanthrene and 2,3,7,8-TCDD were used as positive controls. In this paper, the authors will report the details of the chick embryo method and also some results of the assessment using the method.

  6. Chick embryo partial ischemia model: a new approach to study ischemia ex vivo.

    Syamantak Majumder

    Full Text Available BACKGROUND: Ischemia is a pathophysiological condition due to blockade in blood supply to a specific tissue thus damaging the physiological activity of the tissue. Different in vivo models are presently available to study ischemia in heart and other tissues. However, no ex vivo ischemia model has been available to date for routine ischemia research and for faster screening of anti-ischemia drugs. In the present study, we took the opportunity to develop an ex vivo model of partial ischemia using the vascular bed of 4(th day incubated chick embryo. METHODOLOGY/PRINCIPAL FINDINGS: Ischemia was created in chick embryo by ligating the right vitelline artery using sterile surgical suture. Hypoxia inducible factor- 1 alpha (HIF-1alpha, creatine phospho kinase-MB and reactive oxygen species in animal tissues and cells were measured to confirm ischemia in chick embryo. Additionally, ranolazine, N-acetyl cysteine and trimetazidine were administered as an anti-ischemic drug to validate the present model. Results from the present study depicted that blocking blood flow elevates HIF-1alpha, lipid peroxidation, peroxynitrite level in ischemic vessels while ranolazine administration partially attenuates ischemia driven HIF-1alpha expression. Endothelial cell incubated on ischemic blood vessels elucidated a higher level of HIF-1alpha expression with time while ranolazine treatment reduced HIF-1alpha in ischemic cells. Incubation of caprine heart strip on chick embryo ischemia model depicted an elevated creatine phospho kinase-MB activity under ischemic condition while histology of the treated heart sections evoked edema and disruption of myofibril structures. CONCLUSIONS/SIGNIFICANCE: The present study concluded that chick embryo partial ischemia model can be used as a novel ex vivo model of ischemia. Therefore, the present model can be used parallel with the known in vivo ischemia models in understanding the mechanistic insight of ischemia development and in

  7. Influence of red laser irradiation on the embryo development and condition of Japanese quails' chicks

    We researched the influence of low-level laser irradiation of Japanese Quail hatching eggs on the embryo development, mortality, hatch ability, and physiological state of one-day old chicks and showed the stimulation of embryo development, a statistically reliable decrease in early embryo mortality, and an increase in hatch ability of quail as compared with control. It is detected that one-day old quails from irradiated eggs have higher indices of red blood and a reliable lower level of lipid peroxidation in liver compare with the control group

  8. Endocrine cells in atresic chick embryo intestine: histochemical and immunohistochemical study

    T. Renda


    Full Text Available Intestinal motility disorders are an important problem in the postoperative management of patients with intestinal atresia. Intestinal motility could be initiated by luminal factors that activate intrinsic and extrinsic primary afferent nerves involved in the peristaltic reflex. Endocrine cells act as a key point, because they transfer information regarding the intestinal contents and intraluminal pressure to nerve fibers lying in close proximity to the basolateral surface of the epithelium. In chick embryo, experimental intestinal atresia is associated with disorders in the development of the enteric nervous system, related to the severity of intestinal dilation. Our aim was to investigate the distribution pattern of endocrine cells in the developing endocrine system of chick embryo small intestine with experimentally-induced atresia on day 12 and on day 16. Changes in enteroendocrine population were examined in gut specimens (excised proximal and distal to the atresia from experimental embryos 19 days old and in control sham-operated chick embryos at the same age. Sections from proximal and distal bowel and control bowel were stained with Grimelius silver stain, a valuable histochemical method for detecting the argyrophil and argentophilic cells, and with an immunohistochemical procedure for detecting serotonin and neurotensin immunoreactive cells. In chick embryo proximal bowel, intestinal dilation differed in the various embryos. We found significantly higher enteroendocrine cell counts in proximal bowel than in distal and control bowel. The differences depended on the precociousness of surgery and the severity of dilation. Considering the major contribution of enteroendocrine cells to the peristaltic reflex, our data may help to explain the pathogenesis of motility disorders related to intestinal atresia.

  9. Effects of ionizing radiation on free amino acid content of chick embryo blood plasma. [. gamma. radiation

    Todorov, B.; Dryanovski, P.; Simeonovska, M.


    Using thin-layer chromatography on cellulose film, it was demonstrated that there is a decrease in blood plasma free amino acids after exposing chick embryos to radiation (1000 rad). Only lysine and histidine content decreases on the 1st day. On the 2d, 3d, and 10th days, there is a decrease in lysine, histidine, arginine, glycine, glutamine, serine, aspartic and glutamic acids, threonine, and alanine. There was no change in amounts of leucine, isoleucine, phenylalanine, methionine, valine, and tyrosine throughout the observation period. It was demonstrated that there is a change in rate of migration of nonessential amino acids from egg albumin to embryo blood after exposure to radiation.

  10. Fusion mechanism of the myoblasts in the myotome of the chick embryo

    González Santander, R.; Toledo Lobo, M.V.; Martínez Alonso, F.J.; Martínez Cuadrado, G.


    We have studied the fusion process of rnyoblasts in the mytome corresponding to H.H. stages 22, 23 and 24 from calcitonin-treated chick embryos and their untreated controls. The micrograph images of this process were studied to detect the ultrastructural changes in myoblast morphology that could be associated with the known hormonal and biochen~ical changes that take place in preparation of fusion. Once actin and myosin myofilament differentiation and sarcomero...

  11. Biochemical and cytochemical evidence indicates that coated vesicles in chick embryo myotubes contain newly synthesized acetylcholinesterase


    We have isolated highly purified coated vesicles from 17-d-old chick embryo skeletal muscle. These isolated coated vesicles contain acetylcholinesterase (AChE) in a latent, membrane-protected form as demonstrated enzymatically and morphologically using the Karnovsky and Roots histochemical procedure (J. Histochem. Cytochem., 1964, 12:219- 221). By the use of appropriate inhibitors the cholinesterase activity can be shown to be specific for acetylcholine. It also can be concluded that most of ...

  12. Effect of ionizing radiation on the free amino acid content of blood plasma of chick embryos

    As has been shown by thin-layer chromatography irradiation (1000 rad) of chick embryos decreases the free amino acid content of blood plasma. The quantity of lysine and histidine was only decreased 24 hours after exposure. The contents of lysine, histidine, arginine, glycine, glutamine, serine, aspartic and glutamic acids, threonine and alanine reduced 2, 3 and 10 days after irradiation. The amount of leucine, isoleucine, phenylalanine, methionine, valine and tyrosine remained invariable throughout the entire perjod of observation

  13. Systematic assessment of blood circulation time of functionalized upconversion nanoparticles in the chick embryo

    Nadort, Annemarie; Liang, Liuen; Grebenik, Ekaterina; Guller, Anna; Lu, Yiqing; Qian, Yi; Goldys, Ewa; Zvyagin, Andrei


    Nanoparticle-based delivery of drugs and contrast agents holds great promise in cancer research, because of the increased delivery efficiency compared to `free' drugs and dyes. A versatile platform to investigate nanotechnology is the chick embryo chorioallantoic membrane tumour model, due to its availability (easy, cheap) and accessibility (interventions, imaging). In our group, we developed this model using several tumour cell lines (e.g. breast cancer, colon cancer). In addition, we have synthesized in-house silica coated photoluminescent upconversion nanoparticles with several functional groups (COOH, NH2, PEG). In this work we will present the systematic assessment of their in vivo blood circulation times. To this end, we injected chick embryos grown ex ovo with the functionalized UCNPs and obtained a small amount of blood at several time points after injection to create blood smears The UCNP signal from the blood smears was quantified using a modified inverted microscope imaging set-up. The results of this systematic study are valuable to optimize biochemistry protocols and guide nanomedicine advancement in the versatile chick embryo tumour model.

  14. Proanthocyanidins Prevent High Glucose-Induced Eye Malformation by Restoring Pax6 Expression in Chick Embryo

    Rui-Rong Tan


    Full Text Available Gestational diabetes mellitus (GDM is one of the leading causes of offspring malformations, in which eye malformation is an important disease. It has raised demand for therapy to improve fetal outcomes. In this study, we used chick embryo to establish a GDM model to study the protective effects of proanthocyanidins on eye development. Chick embryos were exposed to high glucose (0.2 mmol/egg on embryo development day (EDD 1. Proanthocyanidins (1 and 10 nmol/egg were injected into the air sac on EDD 0. Results showed that both dosages of proanthocyanidins could prevent the eye malformation and rescue the high glucose-induced oxidative stress significantly, which the similar effects were showed in edaravone. However, proanthocyanidins could not decrease the glucose concentration of embryo eye. Moreover, the key genes regulating eye development, Pax6, was down-regulated by high glucose. Proanthocyanidins could restore the suppressed expression of Pax6. These results indicated proanthocyanidins might be a promising natural agent to prevent high glucose-induced eye malformation by restoring Pax6 expression.

  15. Morphometric and autoradiographic analysis of frontonasal development in the chick embryo

    Dimensional changes in the nasal processes were measured in chick embryos from Hamburger and Hamilton (1951) stages 20 through 27.5. Transverse measurements in the frontonasal region of freshly fixed embryos were compared to frontal sections of the nasal region of comparably staged embryos. These observations were correlated with autoradiographic studies of cell movement employing an implant labeling technique. Morphometric analysis indicated that between stages 20 and 25 the separation of the nasal pit orifices increased coincidentally with rapid forebrain enlargement. Since the separation of the nasal pit fundi increased more rapidly, the orientation of the nasal pits changed. Autoradiographic studies indicated that lateral movement of medial nasal process mesenchyme into the base of the nasal groove and medial area at the base of the lateral nasal process had occurred. After stage 25, the separation of the nasal orifices declined dramatically, coincidental with rapid orbital enlargement. In contrast, the separation of the nasal pit fundi was maintained. It is proposed that nasal development of the chick embryo may be governed initially by forebrain enlargement and associated lateral movements of mesenchyme in the medial nasal processes, resulting in reorientation of the invaginating nasal placodes; subsequently, orbital enlargement and an associated medial redirection of growth of the lateral nasal processes assumes greater significance to the continued development of the frontonasal region

  16. A new gestational diabetes mellitus model: hyperglycemia-induced eye malformation via inhibition of Pax6 in the chick embryo

    Zhang, Shi-Jie; Li, Yi-Fang; Tan, Rui-Rong; Tsoi, Bun; Huang, Wen-Shan; Huang, Yi-Hua; Tang, Xiao-Long; Hu, Dan; Yao, Nan; Yang, Xuesong; Kurihara, Hiroshi; Wang, Qi; He, Rong-Rong


    ABSTRACT Gestational diabetes mellitus (GDM) is one of the leading causes of fetal malformations. However, few models have been developed to study the underlying mechanisms of GDM-induced fetal eye malformation. In this study, a high concentration of glucose (0.2 mmol per egg) was injected into the air sac of chick embryos on embryo development day (EDD) 1 to develop a hyperglycemia model. Results showed that 47.3% of embryonic eye malformation happened on EDD 5. In this model, the key genes regulating eye development, Pax6, Six3 and Otx2, were downregulated by hyperglycemia. Among these genes, the expression of Pax6 was the most vulnerable to hyperglycemia, being suppressed by 70%. A reduction in Pax6 gene expression induced eye malformation in chick embryos. However, increased expression of Pax6 in chick embryos could rescue hyperglycemia-induced eye malformation. Hyperglycemia stimulated O-linked N-acetylglucosaminylation, which caused oxidative stress in chick embryos. Pax6 was found to be vulnerable to free radicals, but the antioxidant edaravone could restore Pax6 expression and reverse eye malformation. These results illustrated a successful establishment of a new chick embryo model to study the molecular mechanism of hyperglycemia-induced eye malformation. The suppression of the Pax6 gene is probably mediated by oxidative stress and could be a crucial target for the therapy of GDM-induced embryonic eye malformation. PMID:26744353

  17. Chick embryos have the same pattern of hypoxic lower-brain activation as fetal mammals.

    Landry, Jeremy P; Hawkins, Connor; Lee, Aaron; Coté, Alexandra; Balaban, Evan; Pompeiano, Maria


    cFos expression (indicating a particular kind of neuronal activation) was examined in embryonic day (E) 18 chick embryos after exposure to 4 h of either normoxia (21% O2), modest hypoxia (15% O2), or medium hypoxia (10% O2). Eight regions of the brainstem and hypothalamus were surveyed, including seven previously shown to respond to hypoxia in late-gestation mammalian fetuses (Breen et al., 1997; Nitsos and Walker, 1999b). Hypoxia-related changes in chick embryo brain activation mirrored those found in fetal mammals with the exception of the medullary Raphe, which showed decreased hypoxic activation, compared with no change in mammals. This difference may be explained by the greater anapyrexic responses of chick embryos relative to mammalian fetuses. Activation in the A1/C1 region was examined in more detail to ascertain whether an O2-sensitive subpopulation of these cells containing heme oxygenase 2 (HMOX2) may drive hypoxic brain responses before the maturation of peripheral O2-sensing. HMOX2-positive and -negative catecholaminergic cells and interdigitating noncatecholaminergic HMOX2-positive cells all showed significant changes in cFos expression to hypoxia, with larger population responses seen in the catecholaminergic cells. Hypoxia-induced activation of lower-brain regions studied here was significantly better correlated with activation of the nucleus of the solitary tract (NTS) than with that of HMOX2-containing A1/C1 neurons. Together, these observations suggest that (1) the functional circuitry controlling prenatal brain responses to hypoxia is strongly conserved between birds and mammals, and (2) NTS neurons are a more dominant driving force for prenatal hypoxic cFos brain responses than O2-sensing A1/C1 neurons. PMID:25964066

  18. Apical accumulation of MARCKS in neural plate cells during neurulation in the chick embryo

    Arruti Cristina


    Full Text Available Abstract Background The neural tube is formed by morphogenetic movements largely dependent on cytoskeletal dynamics. Actin and many of its associated proteins have been proposed as important mediators of neurulation. For instance, mice deficient in MARCKS, an actin cross-linking membrane-associated protein that is regulated by PKC and other kinases, present severe developmental defects, including failure of cranial neural tube closure. Results To determine the distribution of MARCKS, and its possible relationships with actin during neurulation, chick embryos were transversely sectioned and double labeled with an anti-MARCKS polyclonal antibody and phalloidin. In the neural plate, MARCKS was found ubiquitously distributed at the periphery of the cells, being conspicuously accumulated in the apical cell region, in close proximity to the apical actin meshwork. This asymmetric distribution was particularly noticeable during the bending process. After the closure of the neural tube, the apically accumulated MARCKS disappeared, and this cell region became analogous to the other peripheral cell zones in its MARCKS content. Actin did not display analogous variations, remaining highly concentrated at the cell subapical territory. The transient apical accumulation of MARCKS was found throughout the neural tube axis. The analysis of another epithelial bending movement, during the formation of the lens vesicle, revealed an identical phenomenon. Conclusions MARCKS is transiently accumulated at the apical region of neural plate and lens placode cells during processes of bending. This asymmetric subcellular distribution of MARCKS starts before the onset of neural plate bending. These results suggest possible upstream regulatory actions of MARCKS on some functions of the actin subapical meshwork.

  19. Free energy of multicomponent embryo formation

    Kurasov, Victor


    An expression for the free energy of embryo formation is constructed and analyzed. The Gibbs dividing surfaces method is used to attain a coincidence between Laplace formula for critical embryo and integral definitions of concenterations inside the embryo. A global structure of free energy is analyzed and some useful properties are extracted.

  20. Investigation of the Effects of Pre-Incubation Formaldehyde Fumigation on the Tracheal Epithelium of Chicken Embryos and Chicks

    HAYRETDAĞ, Sibel; KOLANKAYA, Dürdane


    This study aimed to evaluate the effects of pre-incubation formaldehyde fumigation on the tracheal epithelium of chicken embryos and chicks. Throughout the study pre-incubation formaldehyde fumigation was applied to 18-day-old embryos and 1-day-old chicks only once, at 1 of 2 different concentrations (3x, 42 ml of formalin and 21 g of potassium permanganate per m3 and 4x, 56 ml of formalin and 28 g of potassium permanganate per m3) for 1 of 2 different durations (20 min and 40 min). Tracheal ...

  1. Effect of a single sublethal dose of permethrin on the development of heart in chick embryo

    Present study was carried out to evaluate the toxic effects of single sublethal doses (0.05 ml) of four different concentrations of permethrin (25, 50, 100 and 200 ppm) on the development of heart in 16/sup th/ day old chick embryo. Saline homogenate/extract of heart tissue was analyzed for the estimation of a few enzyme activities and some of the biochemical components. Enzymes included the amylase, alkaline phosphatase (AkP), acid phosphatase (AcP), Glutamate Oxaloacetate Transaminase (AST), Glutamate Pyruvate Transaminase (ALT) and Lactate dehydrogenase (LDH) and the biochemical components included the glucose, glycpen, total proteins, soluble proteins, free amino acids, total lipids, cholesterol, urea, uric acid, DNA and RNA contents. Activities of some enzymes were decreased whereas others increased. Significant decrease was observed, in alkaline phosphatase (AkP) activity, glutamate pyruvate transmainase (ALT) activity and in glutamate oxaloacetate transaminase (AST) activity at different dose levels. Significant increase was observed in the activities of amylase and acid phosphatase. However, the activity of lactate dehydrogenase (LDH) remained unaffected. Some of the biochemical components were decreased whereas others increased. Significant increases were noted in glucose, Glycogen, soluble protein, free amino acid, cholesterol and DNA contents. Significant decreases were observed in total protein, urea, uric acid, total lipids and RNA contents. Results of this experiment indicates that permethrin seriously affects the function of the heart of the developing chick embryo through damaging its muscles. (author)

  2. The developmental gross morphology of pancreas in chick embryo after prenatal administration of valproic acid

    To determine the effects of prenatal administration of valproic acid on the developmental gross morphology of pancreas in chick embryo. Study Design: Experimental study. Place and Duration of Study: Anatomy Department, Regional Centre, College of Physicians and Surgeons, Islamabad, from February 2010 to February 2011. Methodology: An experimental group-A and control group-B, comprised of 30 eggs each. Freshly laid fertilized chicken eggs of experimental group were injected with valproic acid, incubated and hatched. Eggs of control group underwent sham treatment using normal saline. The chicks were sacrificed on hatching or day 22 of incubation, whichever was earlier. The pancreata of only alive chicks of both groups were dissected out, and evaluated for gross morphology in terms of length and weight by statistically comparing with control ones. Then pancreata were stained with aldehyde fuchsin and orange-G stain to study other obvious histological effects, if any. Results: Chicken embryos exposed to valproic acid in ovo, showed significant decrease in length and weight of pancreata. The mean of length (cm) of pancreata in group-A was 2.208 +- 0.166, and group-B was 2.300 +- 0.102 (p=0.008). The mean of weight (g) of pancreata in group-A was 0.032 +- 0.009, and group-B was 0.048 +- 0.005 (p=0.001). Conclusion: Valproic acid exposure showed retarding effect on the gross development of pancreas as depicted by decrease in the length and weight of pancreata. (author)

  3. Regulation of the glycosylations of collagen hydroxylysine in chick embryo tendon and cartilage cells.

    Anttinen, H; Hulkko, A


    The regulation of the glycosylations of hydroxylysine was studied in isolated chick-embryo cells by labelling with a [14C]lysine pulse. The course of the procollagen lysyl modifications was compared in tendon and cartilage cells, and the effect on the gycosylations of the degree of lysyl hydroxylation and the concentration of Mn2+ and Fe2+ were also studied, in tendon cells. Procollagen triple helix formation was inhibited in most experiments in order to eliminate the effect of this process on the continuation of the reactions. Both in the tendon and cartilage cells the intracellular lysyl modifications proceeded in a biphasic fashion. After an initial sharp linear increase, the reactions did not cease but were protracted at a slower but constant rate. Lysyl hydroxylation was followed by rapid galactosylation in both cell types and this was followed almost immediately by rapid glucosylation, suggesting a close association of the corresponding enzymes. The data further suggest that other factors must also exist, in addition to the differences in the timing of triple helix formation and the actual hydroxylysine content, which are responsible for the different amounts of galactose in the collagens synthesized by these cell types. The amount of glucosylgalactosylhydroxylysine nevertheless seemed to be determined by the available acceptor sites, i.e., the amount of galactosylhydroxylysine. In further experiments with tendon cells the oxygen participating in lysyl hydroxylation was displaced by nitrogen at various points in time. When the degree of lysyl hydroxylation was reduced to less than one-third of the original, the total amounts of glycosylated residues decreased correspondingly, but their proportion relative to total hydroxylysine remained unchanged. Extra Mn2+ increased the proportion of galactosylated hydroxylysine, suggesting that the activity of hydrosylysyl galactosyltransferase is not saturating in respect of the catalyzed reaction. Experiments on the

  4. Chick embryo xenograft model reveals a novel perineural niche for human adipose-derived stromal cells

    Ingrid R. Cordeiro


    Full Text Available Human adipose-derived stromal cells (hADSC are a heterogeneous cell population that contains adult multipotent stem cells. Although it is well established that hADSC have skeletal potential in vivo in adult organisms, in vitro assays suggest further differentiation capacity, such as into glia. Thus, we propose that grafting hADSC into the embryo can provide them with a much more instructive microenvironment, allowing the human cells to adopt diverse fates or niches. Here, hADSC spheroids were grafted into either the presumptive presomitic mesoderm or the first branchial arch (BA1 regions of chick embryos. Cells were identified without previous manipulations via human-specific Alu probes, which allows efficient long-term tracing of heterogeneous primary cultures. When grafted into the trunk, in contrast to previous studies, hADSC were not found in chondrogenic or osteogenic territories up to E8. Surprisingly, 82.5% of the hADSC were associated with HNK1+ tissues, such as peripheral nerves. Human skin fibroblasts showed a smaller tropism for nerves. In line with other studies, hADSC also adopted perivascular locations. When grafted into the presumptive BA1, 74.6% of the cells were in the outflow tract, the final goal of cardiac neural crest cells, and were also associated with peripheral nerves. This is the first study showing that hADSC could adopt a perineural niche in vivo and were able to recognize cues for neural crest cell migration of the host. Therefore, we propose that xenografts of human cells into chick embryos can reveal novel behaviors of heterogeneous cell populations, such as response to migration cues.

  5. Aluminum effects on blood chemistry and long bone development in the chick embryo.

    Firling, C E; Severson, A R; Hill, T A


    Body growth, blood chemistry, and long bone development of 10- to 16-day chick embryos (Gallus gallus) treated with aluminum (Al) citrate, sodium (Na) citrate, or sodium chloride (NaCl) were investigated. Two administration protocols were used. Acutely-treated embryos received 6.0 mumol Al citrate or Na citrate on day 8 of incubation. Chronically-treated embryos received a daily dose of 1.5 mumol Al citrate or Na citrate beginning on day 8 of incubation. For both protocols, Al citrate and Na citrate had no significant influence on viability or body weight. Al citrate-treated embryos had: (a) significantly shorter mean tibia lengths by day 16 of incubation, (b) a consistently lower ratio of tibia length: body weight on all days investigated, and (c) a persistent mid-diaphyseal malformation (angulation) of the femur and tibia. Spatially correlated with the malformation was a calcification defect detected by alizarin red S staining of intact tibias and the accumulation of aluminum as demonstrated by acid solochrome azurine staining of histological sections. Aluminum was localized at the mineralization front of the osteogenic collar surrounding the cartilage core of the tibia. Aluminum citrate or Na citrate had no significant effect on serum total calcium, inorganic phosphorus, total alkaline phosphatase activity, or creatinine, except for a transitory hypercalcemia (day 10) and phosphatemia (days 10 and 12) in Al citrate-treated embryos. The concomitant localization of Al and the early calcification defect in the region of tibial malformation implicate aluminum in the pathogenesis of the skeletal abnormality. PMID:7998819

  6. Effect of gamma irradiation on the immune system of the chick embryo and the induction of tolerance

    The effect of low doses of gamma ray (1, 2.5 and 5 gray) irradiation on the lymphoid organs (bursa of fabricius - BF-, the thymus- Thy - and the bone marrow) of the developing chick were investigated.The embryos were irradiated at stage 34 (about 8 days of incubation)

  7. Anatomical and functional recovery following spinal cord transection in the chick embryo.

    Shimizu, I; Oppenheim, R W; O'Brien, M; Shneiderman, A


    Following complete transection of the thoracic spinal cord at various times during embryonic development, chick embryos and posthatched animals exhibited various degrees of anatomical and functional recovery depending upon the age of injury. Transection on embryonic day 2 (E2), when neurogenesis is still occurring and before descending or ascending fiber tracts have formed, produced no noticeable behavioral or anatomical deficits. Embryos hatched on their own and were behaviorally indistinguishable from control hatchlings. Similar results were found following transection on E5, an age when neurogenesis is complete and when ascending and descending fiber tracts have begun to project through the thoracic region. Within 48 h following injury on E5, large numbers of nerve fibers were observed growing across the site of transection. By E8, injections of horse-radish peroxidase (HRP) administered caudal to the lesion, retrogradely labelled rostral spinal and brainstem neurons. Embryos transected on E5 were able to hatch and could stand and locomote posthatching in a manner that was indistinguishable from controls. Following spinal cord transections on E10, anatomical recovery of the spinal cord at the site of injury was not quite as complete as after E5 transection. Nonetheless, anatomical continuity was restored at the site of injury, axons projected across this region, and rostral spinal and brainstem neurons could be retrogradely labelled following HRP injections administered caudal to the lesion. At least part of this anatomical recovery may be mediated by the regeneration or regrowth of lesioned axons. Although none of the embryos transected on E10 that survived to hatching were able to hatch on their own, because several sham-operated embryos were also unable to hatch, we do not attribute this deficit to the spinal transection. When E10-transected embryos were aided in escaping from the shell, they were able to support their own weight, could stand, and locomote

  8. Toxicity and EROD-inducing potency of 24 polycyclic aromatic hydrocarbons (PAHs) in chick embryos

    Brunstroem, B. (Uppsala Univ. (Sweden). Dept. of Zoophysiology); Broman, D. (Stockholm Univ. (Sweden). Dept. of Zoology); Naef, C. (Stockholm Univ. (Sweden). Dept. of Analytical Chemistry)


    The toxicities (embryolethality) of 24 polycyclic aromatic hydrocarbons (PAHs) were determined in chick embryos using a 72-h test. The substances, dissolved in peanut oil, were injected into the air sacs of eggs preincubated for 7 days. LD{sub 50} values were determined for the four most toxic of the 24 compounds. Benzo(k)fluoranthene was the most potent, with an LD{sub 50} of 14 {mu}g (56 nmol)/kg egg. Dibenz(a,h)anthracene, benz(a)lanthracene and benzol(b)naphtho(2,3-d)thiophene were a few times less toxic (LD{sub 50} = 39 {mu}g (140 nmol)/kg, 79 {mu}g (349 nmol)/kg and 82 {mu}g (350 nmol)/kg, respectively). The LD{sub 50} of benzo(k)fluoranthene was only about 5 times higher than that previously found for the most potent coplanar polychlorinated biphenyl (PCB), 3,3',4,4',5-pentachlorobiphenyl (LD{sub 50} = 3.1 {mu}g (9.4 nmol)/kg), in the same kind of test. The toxicities of 18 of the PAHs in this study have also been evaluated previously using a 2-week test in chick embryos. Dibenz(a,h)anthracene, which had not been studied earlier in the 2-week test, proved to be almost as toxic as previously found for benzo(k)fluoranthene in that test. Several of the PAHs studied induced EROD activity in chick embryos, and, in general, the most toxic PAHs were also the most potent inducers of EROD. The highest enzyme activities were found after treatment with indeno(1,2,3-c,d)pyrene (12 times the control value) and dibenz(a,h)anthracene (8 times the control value). However, due to the high toxicity of dibenz(a,h)anthracene, the dose used was 7 times lower than that of indeno(1,2,3-c,d)pyrene. Following injection of PAHs on day 7, the EROD activities on day 10 were considerably lower than those obtained after a corresponding treatment with coplanar PCBs in an earlier study. Of the PAHs studied, some exhibited very high embryotoxicity. The most toxic PAHs induced EROD activity, suggesting that their toxicity was at least partly mediated via binding to the Ah receptor

  9. Formation of Youth Identity in Indonesian Islamic Chick Lit

    Novita Dewi


    This paper is to argue that literature studies may help reveal the formation of young Indonesian female Muslim identity by looking at the books they read and write. It will particularly discuss two popular Islamic Chick Lit Santri Semelekete [Funky Islamic Boarding School Girl] (2005) by Ma’rifatun Baroroh and Jilbab Britney Spears: Catatan Harian Sabrina [Britney Spears’ Headscarf: Sabrina’s Diary] (2004) by Herlinatiens. The first part of the discussion will examine some external aspects su...

  10. Early expression of hypocretin/orexin in the chick embryo brain.

    Kyle E Godden

    Full Text Available Hypocretin/Orexin (H/O neuropeptides are released by a discrete group of neurons in the vertebrate hypothalamus which play a pivotal role in the maintenance of waking behavior and brain state control. Previous studies have indicated that the H/O neuronal development differs between mammals and fish; H/O peptide-expressing cells are detectable during the earliest stages of brain morphogenesis in fish, but only towards the end of brain morphogenesis (by ∼ 85% of embryonic development in rats. The developmental emergence of H/O neurons has never been previously described in birds. With the goal of determining whether the chick developmental pattern was more similar to that of mammals or of fish, we investigated the emergence of H/O-expressing cells in the brain of chick embryos of different ages using immunohistochemistry. Post-natal chick brains were included in order to compare the spatial distribution of H/O cells with that of other vertebrates. We found that H/O-expressing cells appear to originate from two separate places in the region of the diencephalic proliferative zone. These developing cells express the H/O neuropeptide at a comparatively early age relative to rodents (already visible at 14% of the way through fetal development, thus bearing a closer resemblance to fish. The H/O-expressing cell population proliferates to a large number of cells by a relatively early embryonic age. As previously suggested, the distribution of H/O neurons is intermediate between that of mammalian and non-mammalian vertebrates. This work suggests that, in addition to its roles in developed brains, the H/O peptide may play an important role in the early embryonic development of non-mammalian vertebrates.

  11. Analysis of cerebro-spinal fluid protein composition in early developmental stages in chick embryos.

    Gato, A; Martín, P; Alonso, M I; Martín, C; Pulgar, M A; Moro, J A


    Foetal cerebro-spinal fluid (CSF) has a very high protein concentration when compared to adult CSF, and in many species five major protein fractions have been described. However, the protein concentration and composition in CSF during early developmental stages remains largely unknown. Our results show that in the earliest stages (18 to 30 H.H.) of chick development there is a progressive increase in CSF protein concentration until foetal values are attained. In addition, by performing electrophoretic separation and high-sensitivity silver staining, we were able to identify a total of 21 different protein fractions in the chick embryo CSF. In accordance with the developmental pattern of their concentration, these can be classified as follows: A: high-concentration fractions which corresponded with the ones described in foetal CSF by other authors; B: low-concentration fractions which remained stable throughout the period studied; C: low-concentration fractions which show changes during this period. The evolution and molecular weight of the latter group suggest the possibility of an important biological role. Our data demonstrate that all the CSF protein fractions are present in embryonic serum; this could mean that the specific transport mechanisms in neuroepithelial cells described in the foetal period evolve in very early stages of development. In conclusion, this paper offers an accurate study of the protein composition of chick embryonic CSF, which will help the understanding of the influences on neuroepithelial stem cells during development and, as a result, the appropriate conditions for the in vitro study of embryonic/foetal nervous tissue cells. PMID:15039986

  12. Embryonic cerebrospinal fluid regulates neuroepithelial survival, proliferation, and neurogenesis in chick embryos.

    Gato, Angel; Moro, J A; Alonso, M I; Bueno, D; De La Mano, A; Martín, C


    Early in development, the behavior of neuroepithelial cells is controlled by several factors, which act in a developmentally regulated manner. Diffusible factors are secreted locally by the neuroepithelium itself, although other nearby structures may also be involved. Evidence suggests a physiological role for the cerebrospinal fluid in the development of the brain. Here, using organotypic cultures of chick embryo neuroepithelial explants from the mesencephalon, we show that the neuroepithelium in vitro is not able to self-induce cell survival, replication, and neurogenesis. We also show that the embryonic cerebrospinal fluid (E-CSF) promotes neuroepithelial stem cell survival and induces proliferation and neurogenesis in mesencephalic explants. These data strongly suggest that E-CSF is involved in the regulation of neuroepithelial cells behavior, supporting the hypothesis that this fluid plays a key role during the early development of the central nervous system. PMID:15803475

  13. Ethanol- and/or Taurine-Induced Oxidative Stress in Chick Embryos

    Berning, Emily J.; Noah Bernhardson; Kelly Coleman; Farhat, Dina A.; Courtney M. Gushrowski; Alison Lanctot; Maddock, Benjamin H.; Michels, Kathryn G.; Luke A. Mugge; Nass, Catherine M.; Yearsley, Sarah M.; Miller, Robert R.


    Because taurine alleviates ethanol- (EtOH-) induced lipid peroxidation and liver damage in rats, we asked whether exogenous taurine could alleviate EtOH-induced oxidative stress in chick embryos. Exogenous EtOH (1.5 mmol/Kg egg or 3 mmol/Kg egg), taurine (4 μmol/Kg egg), or EtOH and taurine (1.5 mmol EtOH and 4 μmol taurine/Kg egg or 3 mmol EtOH and 4 μmol taurine/Kg egg) were injected into fertile chicken eggs during the first three days of embryonic development (E0–2). At 11 days of develop...

  14. NMR Based Cerebrum Metabonomic Analysis Reveals Simultaneous Interconnected Changes during Chick Embryo Incubation.

    Yue Feng

    Full Text Available To find out if content changes of the major functional cerebrum metabolites are interconnected and formed a network during the brain development, we obtained high-resolution magic-angle-spinning (HR-MAS 1H NMR spectra of cerebrum tissues of chick embryo aged from incubation day 10 to 20, and postnatal day 1, and analyzed the data with principal component analysis (PCA. Within the examined time window, 26 biological important molecules were identified and 12 of them changed their relative concentration significantly in a time-dependent manner. These metabolites are generally belonged to three categories, neurotransmitters, nutrition sources, and neuronal or glial markers. The relative concentration changes of the metabolites were interconnected among/between the categories, and, more interestingly, associated with the number and size of Nissl-positive neurons. These results provided valuable biochemical and neurochemical information to understand the development of the embryonic brain.

  15. High glucose environment inhibits cranial neural crest survival by activating excessive autophagy in the chick embryo

    Wang, Xiao-Yu; Li, Shuai; Wang, Guang; Ma, Zheng-Lai; Chuai, Manli; Cao, Liu; Yang, Xuesong


    High glucose levels induced by maternal diabetes could lead to defects in neural crest development during embryogenesis, but the cellular mechanism is still not understood. In this study, we observed a defect in chick cranial skeleton, especially parietal bone development in the presence of high glucose levels, which is derived from cranial neural crest cells (CNCC). In early chick embryo, we found that inducing high glucose levels could inhibit the development of CNCC, however, cell proliferation was not significantly involved. Nevertheless, apoptotic CNCC increased in the presence of high levels of glucose. In addition, the expression of apoptosis and autophagy relevant genes were elevated by high glucose treatment. Next, the application of beads soaked in either an autophagy stimulator (Tunicamycin) or inhibitor (Hydroxychloroquine) functionally proved that autophagy was involved in regulating the production of CNCC in the presence of high glucose levels. Our observations suggest that the ERK pathway, rather than the mTOR pathway, most likely participates in mediating the autophagy induced by high glucose. Taken together, our observations indicated that exposure to high levels of glucose could inhibit the survival of CNCC by affecting cell apoptosis, which might result from the dysregulation of the autophagic process. PMID:26671447

  16. Optical coherence tomography. A new high-resolution imaging technology to study cardiac development in chick embryos

    Yelbuz, T.M.; Choma, M.A.; Thrane, L.;


    Background-Optical coherence tomography (OCT) is a depth-resolved, noninvasive, non-destructive imaging modality, the use of which has yet to be fully realized in developmental biology. Methods and Results-We visualized embryonic chick hearts at looping stages using an OCT system with a 22 mum...... axial and 27 mum lateral resolution and an acquisition rate of 4000 A-scans per second. Normal chick embryos from stages 14 to 22 and sham-operated and cardiac neural crest-ablated embryos from stages 15 and 18 were scanned by OCT. Three-dimensional data sets were acquired and processed to create...... volumetric reconstructions and short video clips. The OCT-scanned embryos (2 in each group) were photographed after histological sectioning in comparable planes to those visualized by OCT. The optical and histological results showing cardiovascular microstructures such as myocardium, the cardiac jelly, and...

  17. Investigation of Interaction between Deferoxamine and Low Frequency Electromagnetic Field on Angiogenesis in Chick Embryo

    Atena Dashtizadeh


    Full Text Available Background: Deferoxamine (DFO is an iron chelator. In the present research, the synergic effects of deferoxamine and electromagnetic field (with 50 H frequency and 100 Gauss intensity on angiogenesis of chick chorioallantoic membrane were investigated. Materials and Methods: In this experimental study 80 fertilized egg used and randomly divided 8 group: control group, laboratory control groups of 1 and 2, experimental group 1 (treatment with electromagnetic field, 2 and 3 (treatment with deferoxamine 10, 100 µmol, respectively, 4 and 5 (treatment both deferoxamine 10 and 100 µmol respectively and electromagnetic field. On 8th day of incubation, 2 and 4 groups were incubated with 10 µL deferoxamine and for 3 and 5 groups were incubated with 10 µL deferoxamine 100 µmol. On 10th day, 1, 4 and 5 groups were put in electromagnetic field. On 12th day, the number and length of vessels in all samples was measured by Image J software. Data were analyzed by SPSS-19, ANOVA and t-test. Results: The mean number and length of vessels in the control and experimental cases did not show any significant differences. Comparison between mean number of vessels in the control and group 2, 3, 4, 5 showed a significant decrease (p<0.05 and groups 2 and 4 was showed a significant decrease in the mean length of vessels compared with the controls (p<0.05. Conclusion: Using deferoxamine with low frequency electromagnetic field (50 Hz and 100 G cause inhibition of angiogenesis in chick embryo chorioallantoic membrane.

  18. Characterisation of genes induced during memory formation in the chick

    Full text: Memory formation can be divided into short-term and long-term. Short-term memory involves electro-chemical activity in the neurons whereas long-term memory requires a permanent change that includes protein synthesis. One of the problems involved with identifying late memory related genes is determining an optimal system in which to study gene expression. We have used a discriminated passive avoidance task in chicks to identify genes that are differentially regulated during memory formation. A mRNA subtraction method was previously used to specifically identify several genes that are expressed in the chick intermediate medial hyperstriatum ventrale (IMHV) within two hours of training. Eight bands ranging in size from 400bp to 1100bp were obtained in the initially screen. We are currently cloning these PCR products into suitable vectors for further analysis. Two of these clones have been sequenced and analysed using both the blastn and blastx programs in ANGIS. The first clone was found to correspond to cytochrome c oxidase subunit 2. Cytochrome C oxidase (COX) is a transmembrane protein localized in the inner mitochondrial membrane and forms part of the mitochondrial respiratory chain complex. The second clone codes for the ferritin heavy chain. Ferritin is a ubiquitous protein that is involved in iron homeostasis. At present it is unclear what role these two proteins play in memory formation but further studies are being undertaken to determine the expression profiles of these genes following memory induction. Copyright (2002) Australian Neuroscience Society

  19. Measuring the electric activity of chick embryos heart through 16 bit audio card monitored by the Goldwavetm software

    Silva, Dilson; Cortez, Celia Martins


    In the present work we used a high-resolution, low-cost apparatus capable of detecting waves fit inside the sound bandwidth, and the software package GoldwaveTM for graphical display, processing and monitoring the signals, to study aspects of the electric heart activity of early avian embryos, specifically at the 18th Hamburger & Hamilton stage of the embryo development. The species used was the domestic chick (Gallus gallus), and we carried out 23 experiments in which cardiographic spectra of QRS complex waves representing the propagation of depolarization waves through ventricles was recorded using microprobes and reference electrodes directly on the embryos. The results show that technique using 16 bit audio card monitored by the GoldwaveTM software was efficient to study signal aspects of heart electric activity of early avian embryos.

  20. A set of stage-specific gene transcripts identified in EK stage X and HH stage 3 chick embryos

    Cho Seoae; Moon Sunjin; Park Tae; Kim Heebal; Lee Bo; Park Taesung; Lim Jeong; Han Jae


    Abstract Background The embryonic developmental process in avian species is quite different from that in mammals. The first cleavage begins 4 h after fertilization, but the first differentiation does not occur until laying of the egg (Eyal-Giladi and Kochav (EK) stage X). After 12 to 13 h of incubation (Hamburger and Hamilton (HH) stage 3), the three germ layers form and germ cell segregation in the early chick embryo are completed. Thus, to identify genes associated with early embryonic deve...

  1. Fractal analysis of extra-embryonic vessels of chick embryos under the effect of glucosamine and chondroitin sulfates.

    de Souza Lins Borba, Fernanda Katharine; Felix, Giovanni Loos Queiroz; Costa, Edbhergue Ventura Lola; Silva, Lisie; Dias, Paulo Fernando; de Albuquerque Nogueira, Romildo


    Like heparan sulfate proteoglycans, some monosaccharides and glycosaminoglycans, such as sulfated glucosamine (GS) and chondroitin (CS), integrate the vascular extracellular matrix and may influence vascular endothelial cell growth. To assess the effects of these substances on blood vessel formation, we used the chick yolk sac membrane (YSM) model and fractal geometry quantification, which provided an objective in vivo method for testing potential agents that promote vasculogenesis and angiogenesis. An image processing method was developed to evaluate YSM capillary vessels after they were implanted in a methylcellulose disk of GS or CS at a concentration between 0.001-0.1mg/disk (performed on 2-day old embryos). This method resulted in a binary image of the microvascular network (white vessels on a black background). Fractal box-counting (DBC) and information (DINF) dimensions were used to quantify the activity of GS and CS in vasculogenesis and angiogenesis. YSM treated with GS (0.001-0.1mg) and CS (0.03-0.1mg) showed an increase in fractal dimensions that corresponded to vitelline vessel growth compared to the control group (vehicle), with GS displaying higher fractal dimension values. PMID:26873109

  2. The Bioaccumulation and Toxicity of Platinum Group Metals in Developing Chick Embryos

    Pavel, Ioana; Monahan, Jennifer; Markopoulos, Marjorie; Gagnon, Zofia; Nejame, Britney; Cawley, Jacob; Reens, David


    Recent studies showed that platinum group metals (PGMs) such as Pt, Pd, and Rh from automobile catalytic converters, can accumulate in the soft tissues of a variety of living organisms. However, the effects of PGMs on bone and organs development of animals are not clearly understood. To examine these aspects, developing chick embryos were injected with 0.1, 1.0, 5, or 10 ppm solutions of Pt, Rh, Pd, or with a PGMs mixture. 1) Pathological Changes: were observed for all PGM treatments above 1 ppm. Bone Cells Assesment: Chondrocyte cells in thibiotarsus showed decreased diameter and length. 2) PGMs Accumulation in Tissues: was quantified by GFAAS spectrometry on finely ground tissue powder. 3) Bone Demineralization: was detected by micro-Raman spectroscopy imaging on paraffin embedded bone sections. 4) DNA Damage in Cells: was determined by using a Comet assay and fluorescence spectroscopy. Oxidative Damage in Tissues: was analyzed using a glutathione peroxidase assay. The overall results indicated that PGMs presence in our environment raises concerns about their long-term health effects on all organisms.

  3. Study on the kinetics of photorepair of dimers in the DNA of chick embryo fibroblasts

    We studied quantitative aspects of photoreactivation of UV irradiated chick embryo fibroblast cells (C.E.F.), by monitoring the disappearance of U.V. endonuclease sensitive sites (E.S.S.) from their DNA. We used sucrose gradient sedimentation to measure the molecular weight of the DNA treated with the UV endonuclease and the molecular weight of the same DNA not treated with the endonuclease. Comparing the two gives us the number of remaining E.S.S. A dose of 10 J/m2 (260 nm) yields about 2.6 E.S.S. per 107 dalton. It was found that C.E.F. are proficient in dark repair, this repair taking place at a rate of about 0.1 E.S.S. per 107 dalton per hour. Under influence of blacklight (365 nm) a much greater repair rate was found. We estimate the sensitivity, obtained with our methods and techniques thus far, to be better than 0.1 E.S.S./107 dalton, but we discuss several ways in which we can improve this number. From our results, we estimate that the average number of effective enzyme molecules is less than 0.7/107 dalton, resulting in an upper estimate of 105 photorepair enzyme molecules per C.E.F. cell

  4. Mechanisms of reoxygenation-induced calcium overload in cultured chick embryo heart cells

    We examined mechanisms by which Ca enters cultured myocardial cells during posthypoxic reoxygenation. Monolayer cultures of chick embryo ventricular cells were prepared from hearts 10 days in ovo. Cells were exposed to hypoxic conditions (PO2 less than 1.5 Torr), and 45Ca uptake during subsequent reoxygenation was then examined in the absence and presence of modulators of Ca channel-dependent Ca entry and Na-Ca exchange. Modulation of Ca entry by free radical-scavenging enzymes was also examined. Hypoxia for 120 min followed by reoxygenation increased Ca content from 1.9 to 6.1 nmol/mg protein (P less than 0.05) at 30 min. Verapamil (10(-5) M) added before reoxygenation reduced Ca overload to 3.1 +/- 0.2 nmol/mg protein (P less than 0.05), but both verapamil and BAY K 8644 were without effect on modulating Ca entry if added 30 min after reoxygenation. 24Na content of cells increased from 70 nmol/mg protein in control cells to 157 nmol/mg protein (P less than 0.05) after hypoxia and reoxygenation, favoring Ca entry via Na-Ca exchange. Dichlorobenzamil significantly ameliorated reoxygenation-induced Ca overload, as did catalase and superoxide dismutase. We conclude that reoxygenation-induced Ca overload is unlikely to occur via the Ca channel. It occurs in part via Na-Ca exchange and is substantially ameliorated by enzymatic O2 free radical scavengers

  5. Formation of Youth Identity in Indonesian Islamic Chick Lit

    Novita Dewi


    Full Text Available This paper is to argue that literature studies may help reveal the formation of young Indonesian female Muslim identity by looking at the books they read and write. It will particularly discuss two popular Islamic Chick Lit Santri Semelekete [Funky Islamic Boarding School Girl] (2005 by Ma’rifatun Baroroh and Jilbab Britney Spears: Catatan Harian Sabrina [Britney Spears’ Headscarf: Sabrina’s Diary] (2004 by Herlinatiens. The first part of the discussion will examine some external aspects such as physical presentation, biographical details of the authors, and publication-related matters. Then, using such side-line information, the next part will discuss the novels’ contents to see in what way they offer some cultural analysis of youth identity in contemporary Indonesian society.

  6. Gc protein-derived macrophage-activating factor (GcMAF) stimulates cAMP formation in human mononuclear cells and inhibits angiogenesis in chick embryo chorionallantoic membrane assay

    S. Pacini; G.Morucci; T.Punzi; Gulisano, M; Ruggiero, M


    Abstract: The effects of Gc protein-derived macrophage-activating factor (GcMAF) have been studied in cancer and other conditions where angiogenesis is deregulated. In this study, we demonstrate for the first time that the mitogenic response of human peripheral blood mononuclear cells (PBMCs) to GcMAF was associated with 3'-5'-cyclic adenosine monophosphate (cAMP) formation. The effect was dose dependent, and maximal stimulation was achieved using 0.1 ng/ml. Heparin inhibited the stimulatory ...

  7. Coregulation of calcium channels and beta-adrenergic receptors in cultured chick embryo ventricular cells

    To examine mechanisms whereby the abundance of functional Ca channels may be regulated in excitable tissue, Ca channel number was estimated by binding of the dihydropyridine (DHP) antagonist 3H (+)PN200-110 to monolayers of intact myocytes from chick embryo ventricle. Beta adrenergic receptor properties were studied in cultured myocytes using [3H]CGP12177, an antagonist ligand. Physiological correlates for alterations in DHP binding site number included 45Ca uptake and contractile response to (+)BAYk 8644, a specific L-type Ca channel activator. All binding and physiological determinations were performed in similar intact cell preparations under identical conditions. 4-h exposure to 1 microM isoproterenol reduced cell surface beta-adrenergic receptor number from 44 +/- 3 to 17 +/- 2 fmol/mg (P less than 0.05); DHP binding sites declined in number from 113 +/- 25 to 73 +/- 30 fmol/mg (P less than 0.03). When protein kinase A was activated by a non-receptor-dependent mechanism, DHP binding declined similarly to 68% of control. Exposure to diltiazem, a Ca channel antagonist, for 18-24 h had no effect on number of DHP binding sites. After 4-h isoproterenol exposure, 45Ca uptake stimulated by BAYk 8644 declined from 3.3 +/- 0.2 nmol/mg to 2.9 +/- 0.3 nmol/mg (P less than 0.01) and BAYk 8644-stimulated increase in amplitude of contraction declined from 168 +/- 7 to 134 +/- 11% (P = 0.02). Thus, elevation of [cAMP] in myocytes is associated with a time-dependent decline in Ca channel abundance as estimated by DHP binding and a decline in physiological responses that are in part dependent on abundance of Ca channels. Binding of a directly acting Ca channel antagonist for 18-24 h does not modulate the number of DHP binding sites

  8. Studies on novel drug development using developing chick embryo and its future aspect

    Described is the use of developing chick embryo (DCE), an advantageous alternative experimental animal, in studies on development of novel drugs like radio-sensitizer and on toxicological evaluation. Authors have established DCE transplanted with mouse mammary gland tumor EMT6/KU cells and have found that etanidazole, a radio-sensitizing nitroimidazole derivative, exerts the significant tumor shrinking activity in this tumor-bearing DCE when irradiated by 8 Gy X-ray. They are to test the sensitizing activity of their synthetic nitroimidazole derivatives and other structure-related sensitizers like clinically used (in Denmark) nimorazole. In addition, as the antioxidant activity in vivo can be hardly tested, authors are trying to make DCE system for it since they have studied the activity of derivatives of artepillin C, an active principle of propolis. They are also using DCE chorioallantoic membrane (CAM) for testing the anti-angiogenic activity aiming to develop an antitumor agent from compounds related to fingolimod (FTY-720). For the test of anti-tumor activity in various cell types, their observation is that some of tumor cell types cannot take in DCE. Recently DCE has been used in toxicology for prediction of cardiovascular system like bradycardia and QT elongation, and for lethality test. Hen's egg test-CAM (HET-CAM) and cultured cell system to test irritation are reported to be more valid than other standard methods. DCE system is simple, inexpensive, and unnecessary for particular equipment and facility and desirably becomes an alternative experimental animal next to those like zebra-fish, rat and mouse. (author)

  9. Ethanol- and/or Taurine-Induced Oxidative Stress in Chick Embryos

    Emily J. Berning


    Full Text Available Because taurine alleviates ethanol- (EtOH- induced lipid peroxidation and liver damage in rats, we asked whether exogenous taurine could alleviate EtOH-induced oxidative stress in chick embryos. Exogenous EtOH (1.5 mmol/Kg egg or 3 mmol/Kg egg, taurine (4 μmol/Kg egg, or EtOH and taurine (1.5 mmol EtOH and 4 μmol taurine/Kg egg or 3 mmol EtOH and 4 μmol taurine/Kg egg were injected into fertile chicken eggs during the first three days of embryonic development (E0–2. At 11 days of development (midembryogenesis, serum taurine levels and brain caspase-3 activities, homocysteine (HoCys levels, reduced glutathione (GSH levels, membrane fatty acid composition, and lipid hydroperoxide (LPO levels were measured. Early embryonic EtOH exposure caused increased brain apoptosis rates (caspase-3 activities; increased brain HoCys levels; increased oxidative-stress, as measured by decreased brain GSH levels; decreased brain long-chain polyunsaturated levels; and increased brain LPO levels. Although taurine is reported to be an antioxidant, exogenous taurine was embryopathic and caused increased apoptosis rates (caspase-3 activities; increased brain HoCys levels; increased oxidative-stress (decreased brain GSH levels; decreased brain long-chain polyunsaturated levels; and increased brain LPO levels. Combined EtOH and taurine treatments also caused increased apoptosis rates and oxidative stress.

  10. A freeze-fracture study on the developing satellite cells of spinal ganglia in the chick embryo

    Claudio A. Ferraz de Carvalho


    Full Text Available A freeze-fracture analysis of the satellite cells of spinal ganglia of the chick embryo was performed in 8 successive stages of development, from the 5th incubation day to hatching. The characteristic laminar disposition of the cells were first observed on the 7th day. Tight junctions were found at the 20th incubation day. Small groups or irregular aggregates of particles, but not gap junctions, were described on the 7th and 8th days. Pinocytotic vesicles were pointed out in the different stages considered.

  11. Angiogenic Potential of Human Neonatal Foreskin Stromal Cells in the Chick Embryo Chorioallantoic Membrane Model

    Radhakrishnan Vishnubalaji


    Full Text Available Several studies have demonstrated the multipotentiality of human neonatal foreskin stromal cells (hNSSCs as being able to differentiate into adipocytes and osteoblasts and potentially other cell types. Recently, we demonstrated that hNSSCs play a role during in vitro angiogenesis and appear to possess a capacity to differentiate into endothelial-like cells; however, their angiogenic potential within an ex vivo environment remains unclear. Current study shows hNSSCs to display significant migration potential in the undifferentiated state and high responsiveness in the in vitro wound healing scratch assay. When hNSSCs were seeded onto the top of the CAM, human von Willebrand factor (hVWF, CD31, smooth muscle actin (SMA, and factor XIIIa positive cells were observed in the chick endothelium. CAMs transplanted with endothelial-differentiated hNSSCs displayed a higher number of blood vessels containing hNSSCs compared to CAMs transplanted with undifferentiated hNSSCs. Interestingly, undifferentiated hNSSCs showed a propensity to differentiate towards ectoderm with indication of epidermal formation with cells positive for CD1a, CK5/6, CK19, FXIIIa, and S-100 cells, which warrant further investigation. Our findings imply a potential angiogenic role for hNSSCs ex vivo in the differentiated and undifferentiated state, with potential contribution to blood vessel formation and potential application in tissue regeneration and vascularization.

  12. The effects of thermal manipulations during embryogenesis of broiler chicks on growth of embryo and skeletal traits

    Aygün, Ali; Narinç, Doǧan


    Incubation temperature is one of the important environmental factors that can induce epigenetic thermal adaptation of different physiological control systems. Thus, post hatch thermo tolerance ability of birds may be gained using these manipulations during different incubation periods. The current study was carried out to reveal the effects of temperature manipulations during early and late embryogenesis on weight of embryo and size of skeletal bilateral traits (face, wings, metatarsus, tibia, and femur) in broiler chicken embryos. One thousand commercial broiler eggs from 46 week old breeder flock were used in study. Treatments consisted of eggs incubated at 37.8°C and 55% relative humidity throughout (control; DG1), heated to 36.9°C and supplied 60% relative humidity for 6 hours daily from day 0 to 8 (DG2), heated to 36.9°C and supplied 60% relative humidity for 6 hours daily from day 10 to 18 (DG3), heated to 41°C and supplied 65% relative humidity for 3 hours daily from day 8 to 10 (DG4), and heated to 41°C and supplied 65% relative humidity for 3 hours daily from day 16 to 18 (DG5). Measurements of embryo weight and bilateral traits were obtained at 20 day of incubation and at hatch (at day 21). It was determined that the live weights of embryo and chick were affected significantly by treatment; DG3 group has shown higher mean values than the other treatment groups (P<0.05). There were differences in lengths of femur, tibia and metatarsus among treatment groups at hatch. Particularly, the high incubator temperatures at the second half of incubation accelerated growth of body and bone in embryos. These consequences of the treatments performed at different temperatures and times indicate that the different metabolic shifts realized by the embryos.

  13. Dual labeling of neural crest cells and blood vessels within chicken embryos using Chick(GFP) neural tube grafting and carbocyanine dye DiI injection.

    Delalande, Jean-Marie; Thapar, Nikhil; Burns, Alan J


    All developing organs need to be connected to both the nervous system (for sensory and motor control) as well as the vascular system (for gas exchange, fluid and nutrient supply). Consequently both the nervous and vascular systems develop alongside each other and share striking similarities in their branching architecture. Here we report embryonic manipulations that allow us to study the simultaneous development of neural crest-derived nervous tissue (in this case the enteric nervous system), and the vascular system. This is achieved by generating chicken chimeras via transplantation of discrete segments of the neural tube, and associated neural crest, combined with vascular DiI injection in the same embryo. Our method uses transgenic chick(GFP) embryos for intraspecies grafting, making the transplant technique more powerful than the classical quail-chick interspecies grafting protocol used with great effect since the 1970s. Chick(GFP)-chick intraspecies grafting facilitates imaging of transplanted cells and their projections in intact tissues, and eliminates any potential bias in cell development linked to species differences. This method takes full advantage of the ease of access of the avian embryo (compared with other vertebrate embryos) to study the co-development of the enteric nervous system and the vascular system. PMID:26065540

  14. Evidence against a direct role for oxidative stress in cadmium-induced axial malformation in the chick embryo

    Cadmium (Cd) is a powerful inducer of oxidative stress. It also causes ventral body wall defects in chick embryos treated at Hamburger-Hamilton stages 16-17. By measuring malondialdehyde levels (TBARS method) and cotreating with antioxidants (tempol, ascorbate, and N-acetylcysteine), we sought to determine if oxidative stress were directly related to teratogenesis. We also investigated the expression of mRNAs for antioxidant enzymes superoxide dismutase (SOD) -1 and -2, catalase (CAT), and glutathione peroxidase (GPx). RT-PCR showed reductions in SOD-1, SOD-2, and CAT 1 hour after treatment with Cd. MDA levels increased 4 hours after Cd, and remained elevated 24 hours after treatment. Of the antioxidants, only N-acetylcysteine reduced MDA levels to control values. Nonetheless, no antioxidant could reduce embryo lethality or malformation rates. Furthermore, MDA levels 24 hours after treatment were identical in malformed and normal embryos exposed to Cd. Hence, we conclude that oxidative stress may not have a direct role in Cd teratogenesis.

  15. Planetary Embryo Bow Shocks as a Mechanism for Chondrule Formation

    Mann, Christopher R.; Boley, Aaron C.; Morris, Melissa M.


    We use radiation hydrodynamics with direct particle integration to explore the feasibility of chondrule formation in planetary embryo bow shocks. The calculations presented here are used to explore the consequences of a Mars-size planetary embryo traveling on a moderately excited orbit through the dusty, early environment of the solar system. The embryo's eccentric orbit produces a range of supersonic relative velocities between the embryo and the circularly orbiting gas and dust, prompting t...

  16. Activity and isoenzyme spectrum of alkaline phosphatase in liver and blood plasma of gamma-irradiated hen and chick embryos

    Twelve day and 20 day-old embryos and one-day old chicks were gamma-irradiated with a single dose of 100 rad. On the 1st, 2nd and 72nd hours after treatment the alkaline phosphatase (AP) activity and isozyme spectrum in liver homogenates and blood plasma were determined. AP in the liver was demonstrated histochemically as well. The results show that the initial damage of the liver parenchyma by irradiation treatment is characterized by a slight decrease of liver AP activity and a marked increase in total plasma enzyme activity. The isozyme spectrum changes (increase of AP1 in the early period and increase of AP1 and AP5 in the later period) show that the initial liver parenchymal damage is followed by bile duct damage as well. Comparison of the results of biochemical and histochemical studies indicate the presence of direct correlation between serum and liver alkaline phosphatase activities. (A.B.)

  17. Relation of cell division to the acquisition of responsiveness to cortisol in the neural retina of the chick embryo

    Ben-Or, S.; Eshel, M. (Hebrew Univ., Jerusalem (Israel). Hadassah Medical School)


    Responsiveness of the neural retina to cortisol, as measured by cortisol-induced glutamine synthetase activity, is acquired in the chick embryo during the second week of embryogenesis. The magnitude of the response is inversely related to the growth rate of the neural retina. This developmental event is also acquired by the 8-d-old neural retina under organ culture conditions. The acquisition of competence to respond to the hormonal stimulation can be reversibly abolished by inhibition of DNA synthesis with 0.01 mM cytosine arabinoside; the magnitude of response that resumes after withdrawal of the drug, is characterized by the stage of growth of the neural retina. Responsiveness to cortisol in the embryonic neural retina is apparently coupled to the number of Muller cells (the targets for cortisol action) that have withdrawn from the cell cycle.

  18. Relation of cell division to the acquisition of responsiveness to cortisol in the neural retina of the chick embryo

    Responsiveness of the neural retina to cortisol, as measured by cortisol-induced glutamine synthetase activity, is acquired in the chick embryo during the second week of embryogenesis. The magnitude of the response is inversely related to the growth rate of the neural retina. This developmental event is also acquired by the 8-d-old neural retina under organ culture conditions. The acquisition of competence to respond to the hormonal stimulation can be reversibly abolished by inhibition of DNA synthesis with 0.01 mM cytosine arabinoside; the magnitude of response that resumes after withdrawal of the drug, is characterized by the stage of growth of the neural retina. Responsiveness to cortisol in the embryonic neural retina is apparently coupled to the number of Muller cells (the targets for cortisol action) that have withdrawn from the cell cycle. (author)

  19. Efficacy of the Frame and Hu mathematical model for the quantitative analysis of agents influencing growth of chick embryo fibroblasts

    The experiments on the effect of various sera and substratum surface area upon growth of chick embryo fibroblasts-like in secondary cultures are described and discussed on the grounds of a mathematical model for growth of anchorage-dependent cells proposed by Frame and Hu. The model and presented results demonstrate the mutual independence of the effects of agent influencing of rate of cell proliferation (i.e. accelerating or retarding growth) and the agents that modify the limitation of cell proliferation (i.e. maximum cell density at confluence). The model proposed by Frame and Hu due to its relative simplicity offers and easy mode of description and quantitative evaluation of experiments concerning cell growth regulation. It is shown that various sera added at constant concentration significantly modify the rate of cell proliferation with little effect upon the maximum cell density attainable. The cells grew much more slowly in the presence of calf serum than in the presence of chick serum and the addition of iron and zinc complexes to calf serum significantly accelerated cell growth. An increase in the substratum surface area by the addition of glass wool to culture vessels significantly increased cell density per constant volume of medium even when retardation of growth was observed. The results presented point to the need of direct cell counting for estimation of cell growth curves and discussion of effects of agents influencing parameters characterizing cell proliferation. (author). 34 refs, 5 figs, 2 tabs

  20. FGF2 plays a key role in embryonic cerebrospinal fluid trophic properties over chick embryo neuroepithelial stem cells.

    Martín, C; Bueno, D; Alonso, M I; Moro, J A; Callejo, S; Parada, C; Martín, P; Carnicero, E; Gato, A


    During early stages of brain development, neuroepithelial stem cells undergo intense proliferation as neurogenesis begins. Fibroblast growth factor 2 (FGF2) has been involved in the regulation of these processes, and although it has been suggested that they work in an autocrine-paracrine mode, there is no general agreement on this because the behavior of neuroepithelial cells is not self-sufficient in explants cultured in vitro. In this work, we show that during early stages of development in chick embryos there is another source of FGF2, besides that of the neuroepithelium, which affects the brain primordium, since the cerebrospinal fluid (E-CSF) contains several isoforms of this factor. We also demonstrate, both in vitro and in vivo, that the FGF2 from the E-CSF has an effect on the regulation of neuroepithelial cell behavior, including cell proliferation and neurogenesis. In order to clarify putative sources of FGF2 in embryonic tissues, we detected by in situ hybridization high levels of mRNA expression in notochord, mesonephros and hepatic primordia, and low levels in brain neuroectoderm, corroborated by semiquantitative PCR analysis. Furthermore, we show that the notochord segregates several FGF2 isoforms which modify the behavior of the neuroepithelial cells in vitro. In addition, we show that the FGF2 ligand is present in the embryonic serum; and, by means of labeled FGF2, we prove that this factor passes via the neuroepithelium from the embryonic serum to the E-CSF in vivo. Considering all these results, we propose that, in chick embryos, the behavior of brain neuroepithelial stem cells at the earliest stages of development is influenced by the action of the FGF2 contained within the E-CSF which could have an extraneural origin, thus suggesting a new and complementary way of regulating brain development. PMID:16916506

  1. Triploidy and haploid-triploid mosaicism among chick embryos (Gallus domesticus).

    Zartman, D L; Smith, A L


    Homomorphic, chromosomally abnormal roosters were mated to normal hens. The 23 hens produced 67 embryos, including two triploids and a haploid-triploid mosaic at about 26 hours of incubation. Both of the triploid embryos were conceived within a 5-day period. The presence of a single genome of paternal origin with marker chromosomes in each triploid led to the conclusion that these embryos were derived from diploid, ZW-type ova fertilized by haploid, Z-type spermatozoa. The inheritance pattern of the mosaic embryo was clearly due to a spermatozoal origin for the haploid cell line; and one genome of the three in the triploid cell line was paternal. The sec chromosomes were Z/ZZZ, with one Z of each cell line being a translocation product of paternal derivation. PMID:1192842

  2. The chick embryo as a model for the effects of prenatal exposure to alcohol on craniofacial development.

    Kiecker, Clemens


    Prenatal exposure to ethanol results in fetal alcohol spectrum disorder (FASD), a syndrome characterised by a broad range of clinical manifestations including craniofacial dysmorphologies and neurological defects. The characterisation of the mechanisms by which ethanol exerts its teratogenic effects is difficult due to the pleiotropic nature of its actions. Different experimental model systems have been employed to investigate the aetiology of FASD. Here, I will review studies using these different model organisms that have helped to elucidate how ethanol causes the craniofacial abnormalities characteristic of FASD. In these studies, ethanol was found to impair the prechordal plate-an important embryonic signalling centre-during gastrulation and to negatively affect the induction, migration and survival of the neural crest, a cell population that generates the cartilage and most of the bones of the skull. At the cellular level, ethanol appears to inhibit Sonic hedgehog signalling, alter levels of retionoic acid activity, trigger a Ca(2+)-CamKII-dependent pathway that antagonises WNT signalling, affect cytoskeletal dynamics and increase oxidative stress. Embryos of the domestic chick Gallus gallus domesticus have played a central role in developing a working model for the effects of ethanol on craniofacial development because they are easily accessible and because key steps in craniofacial development are particularly well established in the avian embryo. I will finish this review by highlighting some potential future avenues of fetal alcohol research. PMID:26777098

  3. Immunocharacteristics of oestrogen and androgen target cells in the anterior pituitary gland of the chick as embryo demonstrated by a combined method of autoradiography and immunohistochemistry

    The distribution of oestrogen and androgen target cells in the anterior pituitary gland of the chick embryo on days 10, 12 and 15 of incubation was studied 1 h after the injection of tritium-labelled steroid hormone using the thaw-mount autoradiographic technique. Oestradiol target cells were localized in the caudal zone that corresponds to the so-called 'caudal lobe', while androgen target cells were found throughout the rostral and caudal lobes of the anterior gland. With a combined autoradiography and immunohistochemistry technique, most of the oestrogen target cells showed immunoreactivity to turkey LH antiserum but not to adrenocorticotrophin (1-24) and β-thyrotrophin antisera. In contrast, androgen target cells did not show positive immunoreactivity to the three antisera used. The results suggested a direct and early involvement of oestrogens but not of androgens in the feedback regulation of pituitary gonadotrophin secretion in the chick embryo. (U.K.)

  4. Derivation of feline vaccine-associated fibrosarcoma cell line and its growth on chick embryo chorioallantoic membrane – a new in vivo model for veterinary oncological studies

    Zabielska, K.; Lechowski, R.; M. Król; Pawłowski, K. M.; Motyl, T.; Dolka, I.; Żbikowski, A.


    Feline vaccine associated fibrosarcomas are the second most common skin tumor in cats. Methods of treatment are: surgery, chemotherapy and radiotherapy. Nevertheless, the usage of cytostatics in feline vaccine associated sarcoma therapy is limited due to their adverse side effects, high toxicity and low biodistribution after i.v. injection. Therefore, much research on new therapeutic drugs is being conducted. In human medicine, the chick embryo chorioallantoic membrane (CAM) model is used as ...

  5. Antibody response of patients after postexposure rabies vaccination with small intradermal doses of purified chick embryo cell vaccine or purified Vero cell rabies vaccine

    Briggs, D. J.; A. Banzhoff; U. Nicolay; Sirikwin, S.; B. Dumavibhat; S. Tongswas; C. Wasi


    Although the introduction of tissue culture vaccines for rabies has dramatically improved the immunogenicity and safety of rabies vaccines, they are often prohibitively expensive for developing countries. To examine whether smaller doses of these vaccines could be used, we tested the safety and immunogenicity of purified chick embryo cell vaccine (PCECV) on 211 patients in Thailand with World Health Organization (WHO) category II and III exposures to rabies. The patients presented at two Thai...

  6. Dynamics of skeletal pattern formation in developing chick limb.

    Newman, S A; Frisch, H L


    During development of the embryonic chick limb the skeletal pattern is laid out as cartilaginous primordia, which emerge in a proximodistal sequence over a period of 4 days. The differentiation of cartilage is preceded by changes in cellular contacts at specific locations in the precartilage mesenchyme. Under realistic assumptions, the biosynthesis and diffusion through the extracellular matrix of a cell surface protein, such as fibronectin, will lead to spatial patterns of this molecule that could be the basis of the emergent primordia. As cellular differentiation proceeds, the size of the mesenchymal diffusion chamber is reduced in descrete steps, leading to sequential reorganizations of the morphogen pattern. The successive patterns correspond to observed rows of skeletal elements, whose emergence, in theory and in practice, depends on the maintenance of a unique boundary condition at the limb bud apex. PMID:462174

  7. The role of auxin signaling in early embryo pattern formation

    Smit, Margot E.; Weijers, Dolf


    Pattern formation of the early Arabidopsis embryo generates precursors to all major cell types, and is profoundly controlled by the signaling molecule auxin. Here we discuss recent milestones in our understanding of auxin-dependent embryo patterning. Auxin biosynthesis, transport and response mec

  8. Lectin binding in the ependymal cells of the cephalic portion of the nervous system in the chick embryo.

    Gheri, Gherardo; Vichi, Debora; Sgambati, Eleonora


    The content, distribution and changes of the glycoconjugates oligosaccharides in the ependymal cells of the cephalic portion of the nervous system, in the chick embryo from 5 days of incubation till hatching and in the 3 days old chicken, were investigated. For this purpose a battery of six HRP-conjugated lectins were used (WGA, SBA, UEA I, LTA, PNA, ConA). Enzyme and chemical treatments were performed on some sections prior to staining with HRP-lectins. Our findings showed a large amount of all the investigated sugar residues at the apical portion of the ependymal cells, for the whole considered period of incubation and in the 3 days old chicken. This could indicate that also the immature ependymal cells (spongiobasts) begin to play a tipical role of the mature cells. The presence of cytoplasmic sopranuclear granules, containing D-glucosamine, D-galactose-(beta --> 3)-N-acetil-D-galactosamine and sialic acid in the early stages of incubation, might represent a secretion by the ependymal cells to integrate a not yet fully functioning secretion by the choroid plexuses. At the ciglia a large amount of oligosaccharides were detected in the second part of the period of incubation and in 3 days old chicken. These oligosaccharides could be involved in determining and mantaining the movement of the ciglia to facilitate the flow of the CSF. PMID:16981397

  9. Effects of detergent on the sulphation of chondroitin by cell-free preparations from chick-embryo epiphyseal cartilage.

    Sugumaran, G; Silbert, J E


    The effects of the non-ionic detergent Triton X-100 on 6-sulphation of two species of endogenous nascent proteochondroitin by a chick-embryo cartilage microsomal system was examined. Sulphation of the larger (Type I) species with adenosine 3'-phosphate 5'-phosphosulphate was slightly diminished when Triton X-100 was present, whereas sulphation of the smaller (Type II) species was slightly enhanced. An ordered rather than random pattern of sulphation was obtained for the smaller proteoglycan, but with a considerably lower degree of sulphation than that of the larger proteochondroitin. These differences were consistent with other differences between these two species as described previously. Sulphation of exogenous [14C]chondroitin and exogenous proteo[3H]chondroitin by the microsomal system with Triton X-100 present produced ordered rather than random sulphation patterns. When a 100,000 g supernatant fraction was utilized for sulphation of [14C]chondroitin or proteo[3H]chondroitin, Triton X-100 was not needed, and ordered sulphation was still obtained. When hexasaccharide was used, sulphation of multiple N-acetylgalactosamine residues of the individual hexasaccharides resulted. This was relatively independent of Triton X-100 or the concentration of the hexasaccharide acceptors. With soluble enzyme, sulphation of multiple N-acetylgalactosamine residues on the individual hexasaccharide molecules was even greater, so that tri-sulphated products were found. This suggests that ordered rather than random sulphation of chondroitin with these enzyme preparations is due to enzyme-substrate interaction rather than to membrane organization. PMID:1637348

  10. Planetary Embryo Bow Shocks as a Mechanism for Chondrule Formation

    Mann, Christopher R; Morris, Melissa M


    We use radiation hydrodynamics with direct particle integration to explore the feasibility of chondrule formation in planetary embryo bow shocks. The calculations presented here are used to explore the consequences of a Mars-size planetary embryo traveling on a moderately excited orbit through the dusty, early environment of the solar system. The embryo's eccentric orbit produces a range of supersonic relative velocities between the embryo and the circularly orbiting gas and dust, prompting the formation of bow shocks. Temporary atmospheres around these embryos, which can be created via volatile outgassing and gas capture from the surrounding nebula, can non-trivially affect thermal profiles of solids entering the shock. We explore the thermal environment of solids that traverse the bow shock at different impact radii, the effects that planetoid atmospheres have on shock morphologies, and the stripping efficiency of planetoidal atmospheres in the presence of high relative winds. Simulations are run using adia...

  11. Tunicamycin inhibits biosynthesis of acid mucopolysaccharides in cultures of chick embryo fibroblasts

    The time course of the incorporation of 14C-glucosamine into extracellular mucopolysaccharides (MPS) and that of H235SO4 into intracellular and extracellular MPS were experimentally studied. The monolayer cultures of chick embrio fibroblasts in test tubes were prepared with modified Eagle's minimal essential medium supplemented with 5% calf serum. After confluence, the medium was replaced with the prewarmed fresh one containing 0.4 μCi/ml glucosamine-1-14C or 2 μCi/ml H235SO4. Tunicamycin (TM) was added at the same time as the radioactive compounds. At the appropriate time intervals of incubation at 370C, cultured cells were sampled, and the medium was withdrawn. The cell sheets were rinsed twice with 1 ml each of could distilled water. Acid MPS were precipitated with cetylpyridinium chloride. The precipitate was collected by centrifugation. Insoluble matter was collected on glass fiber filter. After drying, the radioactivity retained on the filter was counted with a scintillation counter. This fraction is of extracellular MPS. In the case of H235SO4 incorporation, the step of NaOH treatment was omitted. This fraction is of intracellular MPS. As a result, slight difference was observed in the degree of inhibition by 1.0 μg/ml TM between the two fractions. TM also inhibited the incorporation of H235SO4. The degree of inhibition of H235SO4 incorporation into intracellular MPS was similar to that of 14C-glucosamine. The inhibition of MPS biosynthesis by TM suggests the possibility of participation of lipid-linked intermediate in the biosynthesis of MPS. (Iwakiri, K.)

  12. Effect of Mobile Phone Radiation on Cardiovascular Development of Chick Embryo.

    Ye, W; Wang, F; Zhang, W; Fang, N; Zhao, W; Wang, J


    The biological effects on cardiovascular development of chicken embryos were examined after radiation exposure using mobile phone (900 MHz; specific absorption rate˜1.07 W/kg) intermittently 3 h per day during incubation. Samples were selected by morphological and histological methods. The results showed the rate of embryonic mortality and cardiac deformity increased significantly in exposed group (P Comet assay data showed the haemocyte mean tail in the exposed group was significantly larger than that of the control (P < 0.01). The arterial vascular wall of exposed group was thicker (P < 0.05) than that of the control on D13, which was reversed to normal in later stages. Our findings suggest that long-term exposure of MPR may induce myocardium pathological changes, DNA damage and increased mortality; however, there was little effect on vascular development. PMID:26171674

  13. Direct embryo formation in leaves of Camillia japonica L.

    Pedroso, M C; Pais, M S


    The culture conditions for direct embryo formation in leaves of Camellia japonica L. were established. An auxin treatment followed by incubation during 11 days in darkness on diluted Murashige and Skoog modified basal medium induced direct morphogenesis. The number of subcultures, subculture interval and leaf age affected in vitro leaf response. The results showed that the cells from a cultured leaf respond differently to the same culture conditions by forming embryos, roots, and non-morphogenic as well as organogenic callus. Direct embryo formation occurred only in the marginal leaf regions. Direct root formation only occurred in a well-defined region of the midrib whereas callus was preferentially formed on the leaf basis. The results suggest the existence of differences in morphogenic competence according to leaf regions. Plantlet regeneration was successfully achieved from somatic embryos and from leaf basisderived callus, via shoot bud induction. PMID:24201879

  14. Focal adhesion kinase as a mechanotransducer during rapid brain growth of the chick embryo.

    Desmond, Mary E; Knepper, Janice E; DiBenedetto, Angela J; Malaugh, Elizabeth; Callejo, Sagrario; Carretero, Raquel; Alonso, Maria-Isabel; Gato, Angel


    Expansion of the hollow fluid-filled embryonic brain occurs by an increase in intraluminal pressure created by accumulation of cerebrospinal fluid (CSF). Experiments have shown a direct correlation between cavity pressure and cell proliferation within the neuroepithelium. These findings lead us to ask how mechanistically this might come about. Are there perhaps molecules on the luminal surface of the embryonic neuroepithelium, such as focal adhesion kinases (FAKs) known to respond to tension in other epithelial cells? Immunodetection using antibodies to total FAK and p-FAK was performed with subsequent confocal analysis of the pattern of their activation under normal intraluminal pressure and induced chronic pressure. Western analysis was also done to look at the amount of FAK expression, as well as its activation under these same conditions. Using immunolocalization, we have shown that FAK is present and activated on both apical and basolateral surfaces and within the cytoplasm of the neuroepithelial cells. This pattern changed profoundly when the neuroepithelium was under pressure. By Western blot, we have shown that FAK was upregulated and activated in the neuroepithelium of the embryos just after the neural tube becomes a closed pressurized system, with phosphorylation detected on the luminal instead of the basal surface, along with an increase in cell proliferation. Chronic hyper-pressure does not induce an increase in phosphorylation of FAK. In conclusion, here we show that neuroepithelial cells respond to intraluminal pressure via FAK phosphorylation on the luminal surface. PMID:24860993

  15. Regionalization and intersegmental coordination of rhythm-generating networks in the spinal cord of the chick embryo.

    Ho, S; O'Donovan, M J


    We have examined the regionalization and coordination of rhythm-generating networks in the isolated spinal cord of the chick embryo between embryonic days 9 and 13, by recording the pattern of rhythmic activity recorded from muscle nerves and ventral roots following a variety of lesions. We found that the capacity for rhythmic activity is distributed along the rostrocaudal axis of the cord but can be expressed in a single, isolated segment. Specializations within the lumbosacral cord were investigated by isolating particular regions and recording their motor output. The rostral part of the lumbosacral cord generates more cycles than the caudal part, and this difference becomes more pronounced with development. In the unlesioned cord, motoneuron activity is synchronized along the rostrocaudal axis. Lesion experiments revealed that the synchronization of motoneuron activity and the synaptic drive to caudal motoneurons is mediated in part by propriospinal pathways traveling in the ventrolateral white matter tracts and by synaptic interactions within the gray matter. The dorsal fiber tracts may also be involved but their effects appear to be weak. Lesions in dorsal-ventral and mediolateral planes were used to localize regions critical for rhythmogenesis and for the alternation of flexor and extensor motoneurons. Rhythmic activity with alternation persisted in spinal cords in which the dorsal and medial half had been removed. Severe medial or dorsal lesions, resulting in a thin strip of lateral or ventral gray matter, altered the phasing of motoneuron activity from alternating to synchronous without effects on cycle timing. These results suggest that the critical neural components for alternation are located close to and dorsomedial to the lateral motor column, and that the capacity for rhythmogenesis is distributed widely throughout the ventral gray matter and is not localized to specific nuclei. PMID:8463824

  16. Binding characteristics of brain-derived neurotrophic factor to its receptors on neurons from the chick embryo

    Brain-derived neurotrophic factor (BDNF), a protein known to support the survival of embryonic sensory neurons and retinal ganglion cells, was derivatized with 125I-Bolton-Hunter reagent and obtained in a biologically active, radioactive form (125I-BDNF). Using dorsal root ganglion neurons from chick embryos at 9 d of development, the basic physicochemical parameters of the binding of 125I-BDNF with its receptors were established. Two different classes of receptors were found, with dissociation constants of 1.7 x 10(-11) M (high-affinity receptors) and 1.3 x 10(-9) M (low-affinity receptors). Unlabeled BDNF competed with 125I-BDNF for binding to the high-affinity receptors with an inhibition constant essentially identical to the dissociation constant of the labeled protein: 1.2 x 10(-11) M. The association and dissociation rates from both types of receptors were also determined, and the dissociation constants calculated from these kinetic experiments were found to correspond to the results obtained from steady-state binding. The number of high-affinity receptors (a few hundred per cell soma) was 15 times lower than that of low-affinity receptors. No high-affinity receptors were found on sympathetic neurons, known not to respond to BDNF, although specific binding of 125I-BDNF to these cells was detected at a high concentration of the radioligand. These results are discussed and compared with those obtained with nerve growth factor on the same neuronal populations

  17. Developmental regulation of the 3-methylcholanthrene- and dioxin-inducible CYP1A5 gene in chick embryo liver in vivo.

    Bentivegna, C S; Ihnat, M A; Baptiste, N S; Hamilton, J W


    The cDNA sequences for two dioxin-inducible cytochrome P450s in chicken, CYP1A4 and CYP1A5, have recently been reported which correspond to two dioxin-inducible forms of P450 previously designated as TCDDAHH and TCDDAA, respectively. The developmental expression of CYP1A4-associated aryl hydrocarbon (benzo[a]pyrene) hydroxylase (AHH) activity and its association with expression of the Ah receptor had previously been characterized in chick embryo liver. The purpose of this study was to examine the developmental regulation of the second dioxin-inducible P450 gene, CYP1A5, in chick embryo liver. A partial gene sequence for CYP1A5 indicated that the intron/exon organization of this gene was identical to that of the CYP1A1 and CYP1A2 mammalian genes and was present in a single copy in the genome. CYP1A5 mRNA was expressed basally in chick embryo liver and was highly inducible by the Ah receptor ligands, 3-methylcholanthrene, beta-naphthoflavone, and 3,4,3', 4'-tetrachlorobiphenyl (TCB), but not by the phenobarbital analog, glutethimide. CYP1A5 mRNA levels were increased 40- to 50-fold within 5 h after a single TCB treatment, corresponding to a 30- to 40-fold increase in the transcription rate of the CYP1A5 gene at this time point. In contrast to a previous report that CYP1A5 mRNA expression was inducible by estradiol, we observed no effects of estradiol or dexamethasone on CYP1A5 mRNA expression, either alone or in combination with TCB. Basal and TCB-inducible CYP1A5 mRNA expression was maximal in liver at 8 days of development and remained high throughout the remainder of embryonic development. Thus, CYP1A5 appears to be regulated in a very similar manner to CYP1A4 in chick embryo liver. PMID:9705900

  18. The effect of parathion on the synthesis of DNA, RNA and protein in the primary erythrocytes, and on the nervous system differentiation in chick embryo

    In the experiments, chick embryos between the primitive streak stage and the 6-somites stage were used. These embryos were cultured in vitro, with the presence of Parathion for 12, 24, 36 hours. The result showed that Parathion has a teratogenic effect on the early stages of embryogenesis. Morphological and histological changes in brain, embryonic axis, heart mesenchyme and erythrocytes were found. Parathion's effect at molecular level quantitatively and qualitatively was studied, using autoradiography and radioactivity measurements after incorporation of 3H thymidine, 3H uridine and 3H delta amino levulinic acid. An inhibitory effect of Parathion on the synthesis of DNA, RNA and hemoglobin was shown. The inhibitory ratio increases with the increase of exposure period to Parathion. These results suggested that Parathion affects other mechanisms which have no relation with acetylcholinesterase activity. This activity was inhibited in a more advanced stages by Parathion. (author)

  19. Direct binding of recombinant plasminogen kringle 1-3 to angiogenin inhibits angiogenin-induced angiogenesis in the chick embryo CAM.

    Youn, Mi-Ran; Park, Mee-Hee; Choi, Chang-Ki; Ahn, Byung-Cheol; Kim, Hak Yong; Kang, Sang Sun; Hong, Yong-Kil; Joe, Young Ae; Kim, Jong-Soo; You, Weon-Kyoo; Lee, Hyo-Sil; Chung, Soo-Il; Chang, Soo-Ik


    Angiogenin is one of the most potent angiogenesis-inducing proteins. Angiostatin is one of the most potent angiogenesis inhibitors, and it contains the first four kringle domains of plasminogen (K1-4). Recombinant human plasminogen kringle 1-3 (rK1-3) was expressed in Escherichia coli and purified to homogeneity. The binding of t-4-aminomethylcyclohexanecarboxylic acid with the purified kringle 1-3 was determined by changes in intrinsic fluorescence. rK1-3 exhibits comparable ligand-binding properties as native human plasminogen kringle 1-3. The purified rK1-3 inhibits neovascularization in the chick embryo chorioallantoic membrane (CAM) assay. Interaction of angiogenin with rK1-3 was examined by immunological binding assay and surface plasmon resonance kinetic analysis, and the equilibrium dissociation constants for the complex, Kd, are 0.89 and 0.18 microM, respectively. rK1-3 inhibits angiogenin-induced angiogenesis in the chick embryo CAM in a concentration-dependent manner. These results indicate that rK1-3 directly binds to angiogenin and thus rK1-3 inhibits the angiogenic activity of angiogenin. PMID:16564503

  20. Trichloroethylene Exposure during Cardiac Valvuloseptal Morphogenesis Alters Cushion Formation and Cardiac Hemodynamics in the Avian Embryo

    Drake, Victoria J.; Koprowski, Stacy L.; Lough, John; Hu, Norman; Susan M. Smith


    It is controversial whether trichloroethylene (TCE) is a cardiac teratogen. We exposed chick embryos to 0, 0.4, 8, or 400 ppb TCE/egg during the period of cardiac valvuloseptal morphogenesis (2–3.3 days’ incubation). Embryo survival, valvuloseptal cellularity, and cardiac hemodynamics were evaluated at times thereafter. TCE at 8 and 400 ppb/egg reduced embryo survival to day 6.25 incubation by 40–50%. At day 4.25, increased proliferation and hypercellularity were observed within the atriovent...

  1. Planetary Embryo Bow Shocks as a Mechanism for Chondrule Formation

    Mann, Christopher R.; Boley, Aaron C.; Morris, Melissa A.


    We use radiation hydrodynamics with direct particle integration to explore the feasibility of chondrule formation in planetary embryo bow shocks. The calculations presented here are used to explore the consequences of a Mars-size planetary embryo traveling on a moderately excited orbit through the dusty, early environment of the solar system. The embryo’s eccentric orbit produces a range of supersonic relative velocities between the embryo and the circularly orbiting gas and dust, prompting the formation of bow shocks. Temporary atmospheres around these embryos, which can be created via volatile outgassing and gas capture from the surrounding nebula, can non-trivially affect thermal profiles of solids entering the shock. We explore the thermal environment of solids that traverse the bow shock at different impact radii, the effects that planetoid atmospheres have on shock morphologies, and the stripping efficiency of planetoidal atmospheres in the presence of high relative winds. Simulations are run using adiabatic and radiative conditions, with multiple treatments for the local opacities. Shock speeds of 5, 6, and 7 km s-1 are explored. We find that a high-mass atmosphere and inefficient radiative conditions can produce peak temperatures and cooling rates that are consistent with the constraints set by chondrule furnace studies. For most conditions, the derived cooling rates are potentially too high to be consistent with chondrule formation.

  2. Effectiveness of a Prudhoe Bay crude oil and its aliphatic, aromatic and heterocyclic fractions in inducing mortality and aryl hydrocarbon hydroxylase in chick embryo in ovo

    Walters, P.; Khan, S.; O' Brien, P.J.O.; Rahimtula, A.T.; Payne, J.F.


    Prudhoe Bay crude oil (PBCO) and its aliphatic, aromatic and heterocyclic fractions were tested on the developing chick embryo for (i) embryotoxicity (ii) their ability to induce hepatic and renal cytochrome P450 levels as well as hepatic, renal and pulmonary aryl hydrocarbon hydroxylase activities. On the basis of its concentration in PBCO, the aromatic fraction was responsible for most of the embryotoxicity as well as for the enzyme inducing ability. The NOS fraction constituted less than 7% (w/v) of PbCO but, on a weight equivalent basis, was roughly as potent as the aromatic fraction in causing embryotoxicity and in inducing cytochrome P450 levels and aryl hydrocarbon hydroxylase. The aliphatic fraction was found to be essentially inactive. The results are consistent with the concept that elevation of aryl hydrocarbon hydroxylase levels by certain components of PBCO may lead to increased embroyotoxicity.

  3. Metabolic and anatomic development of the chick embryo as studied by phosphorus-31 magnetic resonance spectroscopy and proton MRI

    Thirty fertilized chick eggs were studied sequentially over the 22-day maturation period of the egg by phosphorous-31 (P-31) magnetic resonance spectroscopy (MRS) and proton (H-1) magnetic resonance imaging (MRI) at 2.0 Tesla. The total MR observable P-31 volume in the egg decreased by nearly 80% from day 4 through day 22, suggesting P-31 uptake into tissues in thich the P-31 signal is not visible by MR, such as phospholipid bilayers, membranes, cellular stuctures, and bone mineral phosphates. Of the remaining visible phosphates, the relative phospholipid peak area decreased and the relative peak areas of the phosphocreatine and nucleoside di- and triphosphate metabolites increased. Inorganic phosphate also showed an increase in peak area during maturation. The metabolic development correlated with the anatomic development of visible structures such as brain, eyes, spine and organs in the chick as seen by MRI. In-vivo MR techniques offers the potential to follow changes in prenatal development and maturation. (orig.)

  4. Hydrocortisone and Vitamin D3 stimulation of 32Psub(i)-phosphate accumulation by organ-cultured chick embryo duodenum

    Either vitamin D3 (or 1 α,25-(OH)2-D3) or hydrocortisone (HC) stimulated phosphate accumulation by organ-cultured embryonic chick duodenum. In combination, these two steroids stimulated phosphate uptake synergistically. Phosphate accumulation appeared to be independent of other vitamin D3-stimulated processes: CaBP concentration, cAMP concentration, or alkaline phosphataseactivity. L-phenylalanine, a reported alkaline phosphate inhibitor, when added to the culture medium progressively inhibited either D3- or HC-stimulated phosphate uptake subsequent to culture, but did not inhibit the synergistic action under these conditions L-phenylalanine had no consistent effect on alkaline phosphotase activity but unexpectedly, greatly inhibited vitamin D3 - stimulated CaBP concentration, but only in the absence of HC. Some limited suggestion of an intestinal phosphoprotein sensitve to either vitamin D3 or HC was observed. (orig.)

  5. Cysteic acid and taurine synthesis from sulphate in the chick embryo; Synthese de l'acide cysteique et de la taurine a partir du sulfate dans l'oeuf embryonne de poule

    Chapeville, F. [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires


    The formation of taurine from sulphate was studied in the chick embryo using the radioisotopes of: sulphur, carbon and hydrogen. The following reactions occur: 1) reduction of sulphate to sulphite; 2) fixation of the sulphite on a carbon chain with an amino group, resulting from desulphydration of L-cysteine, which leads to the formation of L-cysteic acid; 3) decarboxylation of L-cysteic acid. Reaction (1) takes place only in the endo-dermal cells of the yolk sac; reaction (2) in these same cells and in the yolk; reaction (3) is general, localized in the yolk sac, in the yolk as well as in the tissues of the embryo itself. The enzyme which catalyses reaction (2) has been purified; the coenzyme is pyridoxal phosphate. The desulphydration of cysteine by this enzyme is a reversible reaction. In non-physiological conditions of concentration and temperature, pyridoxal phosphate catalyses in the presence of metallic ions, the desulphydration of cysteine and the formation of cysteic acid from sulphite. (author) [French] On a montre que la formation de taurine a partir de sulfate dans I'oeuf embryonne de poule, etudiee a l'aide des radioisotopes, du soufre, du carbone et de l'hydrogene, correspond aux reactions suivantes: 1) reduction du sulfate en sulfite; 2) fixation du sulfite sur une chaine tricarbonee et aminee provenant de la desulfhydration de la L-cysteine, fixation conduisant a la formation d'acide L-cysteique; 3) decarboxylation de l'acide L-cysteique. La reaction (1) a lieu uniquement dans les cellules de l'endoderme du sac vitellin; la reaction (2) dans les memes cellules et dans le vitellus; la reaction (3) est plus generale, elle est localisee dans le sac vitellin, dans le vitellus et dans les tissus de l'embryon. L'enzyme qui catalyse la reaction (2) a ete purifie; il possede le phosphate de pyridoxal comme coenzyme. La desulfhydration de la cysteine par cet enzyme est une reaction reversible. Dans les conditions non

  6. Effects of nitric oxide modulating activities on development of enteric nervous system mediated gut motility in chick embryo model

    Hossein-Ali Arab; Samad Muhammadnejad; Seyed-Muhammad Faghihi; Hossein Hassanpour; Ahad Muhammadnejad


    The enteric nervous system (ENS) arises from the enteric neural crest-derived cells (ENCCs), and many molecules and biochemical processes may be involved in its development. This study examined the effects of modulating embryonic nitric oxide (NO) activity on the intestinal motility induced by ENS. One-hundred-and-twenty fertilized chicken eggs were assigned to three main groups and incubated at 37°C and 60% humidity. The eggs were treated with -nitro-L-arginine methyl ester (L-NAME), sodium nitroprusside (SNP), L-arginine (L-Arg) or vehicle from days 3 (1st group), 7 (2nd group) and 10 (3rd group) of incubation and continued up to day 18. On day 19, the embryos were sacrificed, the jejunal and colorectal segments were taken and the intestinal motility was assessed using isolated organ system. The intestinal motility was recorded normally and following cholinergic, adrenergic and non-adrenergic non-cholinergic (NANC) stimulations. The ENS structure was assessed by immunohistochemistry (IHC) using glial fibrillary acidic protein (GFAP). Rhythmic intestinal contractions were seen in all treatment groups, but inhibition of NO in the L-NAME-treated embryos caused significant decrease ( < 0.01) in the frequency and amplitude of the contraction. The responsiveness to adrenergic, cholinergic and NANC stimulations was also significantly decreased ( <0.05). The GFAP expression was significantly ( < 0.05) reduced in the L-NAME-treated embryos. This study showed that the inhibition of NO caused a deficient development of the ENS, leading to a decrease in the frequency and amplitude of the intestinal contractions and reduced the responsiveness to adrenergic, cholinergic and NANC signalling.

  7. In ovo monitoring of smooth muscle fiber development in the chick embryo: diffusion tensor imaging with histologic correlation.

    Jianrong Xu

    Full Text Available BACKGROUND: Magnetic resonance imaging is a noninvasive method of evaluating embryonic development. Magnetic resonance diffusion tensor imaging, which is based on the measuring the directional diffusivity of water molecules, is an established method of evaluating tissue structure. Prolonged imaging times have precluded the use of embryonic diffusion tensor imaging due to motion artifact. Using temperature-based motion suppression, we aimed to investigate whether diffusion tensor imaging can be used to monitor embryonic smooth muscle development in ovo, and to determine the correlation between histologically-derived muscle fiber fraction, day of incubation and diffusion tensor imaging fractional anisotropy values and length of tracked fibers. METHODOLOGY/PRINCIPAL FINDINGS: From a set of 82 normally developing fertile chicken eggs, 5 eggs were randomly chosen each day from incubation days 5 to 18 and cooled using a dual-cooling technique prior to and during magnetic resonance imaging at 3.0 Tesla. Smooth muscle fibers of the gizzard were tracked using region of interests placed over the gizzard. Following imaging, the egg was cracked and the embryo was fixated and sectioned, and a micrograph most closely corresponding to the acquired magnetic resonance image was made. Smooth muscle fiber fraction was determined using an automated computer algorithm. CONCLUSIONS/SIGNIFICANCE: We show that diffusion tensor images of smooth muscle within the embryonic gizzard can be acquired in ovo from incubation day 11 through hatching. Length of tracked fibers and day of incubation were found to have statistical significance (p<0.05 by multiple linear regression correlation with histologic specimens of sacrificed embryos from day 11 of incubation through hatching. The morphologic pattern of development in our histologic specimens corresponds to the development of embryonic gizzard as reported in the literature. These results suggest that diffusion tensor imaging

  8. Pattern formation during gastrulation in the sea urchin embryo.

    McClay, D R; Armstrong, N A; Hardin, J


    The sea urchin embryo follows a relatively simple cell behavioral sequence in its gastrulation movements. To form the mesoderm, primary mesenchyme cells ingress from the vegetal plate and then migrate along the basal lamina lining the blastocoel. The presumptive secondary mesenchyme and endoderm then invaginate from the vegetal pole of the embryo. The archenteron elongates and extends across the blastocoel until the tip of the archenteron touches and attaches to the opposite side of the blastocoel. Secondary mesenchyme cells, originally at the tip of the archenteron, differentiate to form a variety of structures including coelomic pouches, esophageal muscles, pigment cells and other cell types. After migration of the secondary mesenchyme cells from their original position at the tip of the archenteron, the endoderm fuses with an invagination of the ventral ectoderm (the stomodaem), to form the mouth and complete the process of gastrulation. A larval skeleton is made by primary mesenchyme cells during the time of archenteron and mouth formation. A number of experiments have established that these morphogenetic movements involve a number of cell autonomous behaviors plus a series of cell interactions that provide spatial, temporal and scalar information to cells of the mesoderm and endoderm. The cell autonomous behaviors can be demonstrated by the ability of micromeres or endoderm to perform their morphogenetic functions if either is isolated and grown in culture. The requirement for cell interactions has been demonstrated by manipulative experiments where it has been shown that axial information, temporal information, spatial information and scalar information is obtained by mesoderm and endoderm from other embryonic cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1299366

  9. Antibody response of patients after postexposure rabies vaccination with small intradermal doses of purified chick embryo cell vaccine or purified Vero cell rabies vaccine

    D.J. Briggs


    Full Text Available Although the introduction of tissue culture vaccines for rabies has dramatically improved the immunogenicity and safety of rabies vaccines, they are often prohibitively expensive for developing countries. To examine whether smaller doses of these vaccines could be used, we tested the safety and immunogenicity of purified chick embryo cell vaccine (PCECV on 211 patients in Thailand with World Health Organization (WHO category II and III exposures to rabies. The patients presented at two Thai hospitals and were randomized into three groups. Patients in Group 1 received 0.1 ml PCECV intradermally at two sites on days 0, 3, 7, and at one site on days 30 and 90. Group 2 was treated similarly, except that purified Vero cell rabies vaccine (PVRV was used instead of PCECV. Group 3 received 1.0 ml PCECV intramuscularly on days 0, 3, 7, 14, 30 and 90. After 0, 3, 7, 14, 30 and 90 days serum was collected from the subjects and the geometric mean titres (GMTs of rabies virus neutralizing antibody determined. After 14 days the GMT of 59 patients vaccinated intradermally with PCECV was equivalent to that of patients who received PVRV. Adverse reactions were more frequent in patients who received vaccines intradermally, indicating the reactions were associated with the route of injection, rather than the vaccine per se. We conclude that PCECV is a safe and highly immunogenic vaccine for postexposure rabies vaccination when administered intradermally in 0.1-ml doses using the two-site method (‘‘2,2,2,0,1,1’’ recommended by WHO.

  10. Involvement of cell surface phosphatidylinositol-anchored glycoproteins in cell-cell adhesion of chick embryo myoblasts


    During myogenesis myoblasts fuse to form multinucleate cells that express muscle-specific proteins. A specific cell-cell adhesion process precedes lipid bilayer union during myoblast fusion (Knudsen, K. A., and A. F. Horwitz. 1977. Dev. Biol. 58:328-338) and is mediated by cell surface glycoproteins (Knudsen, K. A., 1985. J. Cell Biol. 101:891- 897). In this paper we show that myoblast adhesion and myotube formation are inhibited by treating fusion-competent myoblasts with phosphatidylinosito...

  11. Safety and immunogenicity of the Intradermal thai red cross (2-2-2-0-1-1) post exposure vaccination regimen in the Indian population using purified chick embryo cell rabies vaccine

    Chhabra Mala; Ichhpujani R; Bhardwaj M; Tiwari K; Panda R; Lal S


    PURPOSE: To test the immunogenicity of the WHO recommended "2-2-2-0-1-1" post-exposure rabies vaccination regimen in Indian subjects to determine the feasibility of replacing crude sheep brain nerve tissue rabies vaccine with modern tissue culture rabies vaccine at major anti-rabies treatment centers throughout India. METHODS: Purified chick embryo cell vaccine (PCECV) was administered in the dosage of 0.1mL per site to 53 Indian subjects. RESULTS: All subjects produced rabies antibodies abov...

  12. Differential effects of protein kinase inhibitors and activators on memory formation in the 2-day-old chick.

    Serrano, P A; Beniston, D S; Oxonian, M G; Rodriguez, W A; Rosenzweig, M R; Bennett, E L


    Thirteen protein kinase inhibitors (PKIs) were investigated in chicks for their in vitro effects on PKC activity and for their in vivo effects on memory formation for a peak-avoidance task. Amnesia occurred by 15-30 min post-training when agents that inhibit primarily Ca2+/calmodulin were injected into brain. Amnesia occurred by 60 min post-training when agents that inhibit PKC-, PKA-, and/or PKG-dependent protein kinases, but not Ca2+/calmodulin, were injected. Enhancement of memory formation was accomplished by injecting bradykinin, but not forskolin. Both of these agents, however, attenuated the amnesia produced by H-7. These results are discussed as relevant neural processes involved in memory and synaptic plasticity. PMID:8129687

  13. Harvesting clues from genome wide transcriptome analysis for exploring thalidomide mediated anomalies in eye development of chick embryo: Nitric oxide rectifies the thalidomide mediated anomalies by swinging back the system to normal transcriptome pattern.

    Kumar, Pavitra; Kasiviswanathan, Dharanibalan; Sundaresan, Lakshmikirupa; Kathirvel, Priyadarshan; Veeriah, Vimal; Dutta, Priya; Sankaranarayanan, Kavitha; Gupta, Ravi; Chatterjee, Suvro


    Thalidomide, the notorious teratogen is known to cause various developmental abnormalities, among which a range of eye deformations are very common. From the clinical point of view, it is necessary to pinpoint the mechanisms of teratogens that tune the gene expression. However, to our knowledge, the molecular basis of eye deformities under thalidomide treatmenthas not been reported so far. Present study focuses on the possible mechanism by which thalidomide affects eye development and the role of Nitric Oxide in recovering thalidomide-mediated anomalies of eye development using chick embryo and zebrafish models with transcriptome analysis. Transcriptome analysis showed that 403 genes were up-regulated and 223 genes were down-regulated significantly in thalidomide pre-treated embryos. 8% of the significantly modulated genes have been implicated in eye development including Pax6, OTX2, Dkk1 and Shh. A wide range of biological process and molecular function was affected by thalidomide exposure. Biological Processes including structural constituent of eye lens and Molecular functions such as visual perception and retinal metabolic process formed strong annotation clustersindicating the adverse effects of thalidomide on eye development and function. Here, we have discussed the whole embryo transcriptome with the expression of PAX6, SOX2, and CRYAAgenes from developing eyes. Our experimental data showing structural and functional aspects includingeye size, lens transparency and optic nerve activity and bioinformatics analyses of transcriptome suggest that NO could partially protect thalidomide treated embryos from its devastating effects on eye development and function. PMID:26717904

  14. WOX2 and polar auxin transport during spruce embryo pattern formation

    Palovaara, Joakim; Hakman, Inger


    The WOX family of transcription factors and polar auxin transport (PAT) are both essential for embryonic patterning and thus normal embryo development in angiosperms. Recent analysis by us of WOX-related genes in Picea and Pinus suggests that they play fundamental roles during embryo development also in conifers.1 It has been proposed that there is a connection between the spatial separation of WOX2 and WOX8, and PAT in the formation of the apical-basal axis in Arabidopsis embryos and that bo...

  15. Effects of gravity on spicule formation in cultured micromeres of sea urchin embryo

    Izumi-Kurotani, A.; Kiyomoto, M.; Imai, M.; Eguchi, H.


    To investigate the effects of gravity on morphogenesis at the cellular level, we have proposed a new experimental system with micromeres from sea urchin embryos [Izumi-Kurotani, A., Kiyomoto, M. Morphogenesis and gravity in a whole Amphibian Embryo and in isolated blastomeres of sea urchins, in: Marthy, H. -J. (Ed.), Developmental Biology Research in Space. Adv. Space Biol. Med. vol. 9, Elsevier, Amsterdam, pp. 83 99, 2003]. We studied spicule formation in cultured micromeres of sea urchin embryo under various conditions of gravity: hypergravity by a centrifuge and simulated microgravity in a vertical clinostat. Spicule elongation was suppressed under both experimental conditions.

  16. Klf5 regulates lineage formation in the pre-implantation mouse embryo

    Lin, Suh-Chin J.; Wani, Maqsood A.; Whitsett, Jeffrey A.; Wells, James M.


    Kruppel-like transcription factors (Klfs) are essential for the induction and maintenance of pluripotency of embryonic stem cells (ESCs), yet little is known about their roles in establishing the three lineages of the pre-implantation embryo. Here, we show that Klf5 is required for the formation of the trophectoderm (TE) and the inner cell mass (ICM), and for repressing primitive endoderm (PE) development. Although cell polarity appeared normal, Klf5 mutant embryos arrested at the blastocyst ...

  17. Formation of sulphite, cysteic acid and taurine from sulphate by the egg embryo

    It is shown that the formation of taurine from sulphate by the chicken embryo involves the reduction of sulphate to sulphite (I), the synthesis of cysteic acid (II) and its decarboxylation (Ill). The reaction (I) takes place in the vitellin sac. The reaction (II) results from the condensation of the sulphite with a-amino-acrylic acid and is carried out by the yolk. The enzymes responsible for the decarboxylation (III) are distributed both in the embryo and in its appendages. (author)

  18. The chick somitogenesis oscillator is arrested before all paraxial mesoderm is segmented into somites

    McGrew Michael J


    Full Text Available Abstract Background Somitogenesis is the earliest sign of segmentation in the developing vertebrate embryo. This process starts very early, soon after gastrulation has initiated and proceeds in an anterior-to-posterior direction during body axis elongation. It is widely accepted that somitogenesis is controlled by a molecular oscillator with the same periodicity as somite formation. This periodic mechanism is repeated a specific number of times until the embryo acquires a defined specie-specific final number of somites at the end of the process of axis elongation. This final number of somites varies widely between vertebrate species. How termination of the process of somitogenesis is determined is still unknown. Results Here we show that during development there is an imbalance between the speed of somite formation and growth of the presomitic mesoderm (PSM/tail bud. This decrease in the PSM size of the chick embryo is not due to an acceleration of the speed of somite formation because it remains constant until the last stages of somitogenesis, when it slows down. When the chick embryo reaches its final number of somites at stage HH 24-25 there is still some remaining unsegmented PSM in which expression of components of the somitogenesis oscillator is no longer dynamic. Finally, we identify a change in expression of retinoic acid regulating factors in the tail bud at late stages of somitogenesis, such that in the chick embryo there is a pronounced onset of Raldh2 expression while in the mouse embryo the expression of the RA inhibitor Cyp26A1 is downregulated. Conclusions Our results show that the chick somitogenesis oscillator is arrested before all paraxial mesoderm is segmented into somites. In addition, endogenous retinoic acid is probably also involved in the termination of the process of segmentation, and in tail growth in general.

  19. Transferin concentration and location during formation of chick retina: developmental correlates

    The amount of transferrin in chick retina was measured during development and compared to transferrin location seen immunocytochemically. Between embryonic day 6 (E6), and 5 days post hatching, two periods occur in which transferrin concentrations rise sharply and decline. During the first, transferrin concentration rises 5-fold between E6 and 10, then rapidly declines by E14. A second increase begins on E17 and peaks by E19-20. Immunocytochemical findings demonstrate that during the first rise in concentration, transferrin is located primarily in neuritic layers. Later in development, when levels again increase, newly forming photoreceptor outer segments are strongly transferrin positive. These findings are discussed in light of developmental events occurring during retinal maturation (author)

  20. Wnt/β-catenin-signaling and the formation of Müller glia-derived progenitors in the chick retina.

    Gallina, Donika; Palazzo, Isabella; Steffenson, Lillia; Todd, Levi; Fischer, Andy J


    Müller glia can be stimulated to de-differentiate, proliferate, and form Müller glia-derived progenitor cells (MGPCs) that are capable of producing retinal neurons. The signaling pathways that influence the de-differentiation of mature Müller glia and proliferation of MGPCs may include the Wnt-pathway. The purpose of this study was to investigate how Wnt-signaling influences the formation of MGPCs in the chick retina in vivo. In NMDA-damaged retinas where MGPCs are known to form, we find dynamic changes in retinal levels of potential readouts of Wnt-signaling, including dkk1, dkk3, axin2, c-myc, tcf-1, and cd44. We find accumulations of nuclear β-catenin in MGPCs that peaks at 3 days and rapidly declines by 5 days after NMDA-treatment. Inhibition of Wnt-signaling with XAV939 in damaged retinas suppressed the formation of MGPCs, increased expression of ascl1a and decreased hes5, but had no effect upon the differentiation of progeny produced by MGPCs. Activation of Wnt-signaling, with GSK3β-inhibitors, in the absence of retinal damage, failed to stimulate the formation of MGPCs, whereas activation of Wnt-signaling in damaged retinas stimulated the formation of MGPCs. In the absence of retinal damage, FGF2/MAPK-signaling stimulated the formation of MGPCs by activating a signaling network that includes Wnt/β-catenin. In FGF2-treated retinas, inhibition of Wnt-signaling reduced numbers of proliferating MGPCs, whereas activation of Wnt-signaling failed to influence the formation of proliferating MGPCs. Our findings indicate that Wnt-signaling is part of a network initiated by FGF2/MAPK or retinal damage, and activation of canonical Wnt-signaling is required for the formation of proliferating MGPCs. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 983-1002, 2016. PMID:26663639

  1. Maternal Mga is required for Wnt signaling and organizer formation in the early Xenopus embryo

    Fei Gu; Huijuan Shi; Li Gao; Haiyan Zhang; Qinghua Tao


    Maternal Wnt11 is both necessary and sufficient for the formation of Spemann organizer in Xenopus embryo.Xnr3 and Siamois have been identified as the direct target genes of maternal Wnt11/β-catenin during organizer induction.The depletion of maternal XTcf3 resulted in the ectopic expression of Xnr3 and Siamois,suggesting the activity of β-catenin/XTcf3 is strictly regulated in the early Xenopus embryos.Here,we show that Xenopus mga (Xmga) is a maternal gene required for dorsal axis formation.Overexpression experiments indicate that mouse Mga potentiates the activity of β-catenin in the induction of organizer-specific genes.Depletion of maternal Xmga results in the dramatic decrease of the expression of organizer genes and ventralization phenotype,indicating that Xmga is required for β-catenin function and organizer formation.Depletion of XTcf3 cannot rescue organizer gene expression and axis formation in Xmga-depleted embryos,suggesting Xmga is downstream of XTcf3 during organizer induction.We conclude that maternal Xmga is critical for the function of β-catenin during organizer formation and dorsal development of Xenopus embryo.To our knowledge,this is a report for the first time to implicate Mga in regulating Wnt signaling.

  2. Maternal Mga is required for Wnt signaling and organizer formation in the early Xenopus embryo.

    Gu, Fei; Shi, Huijuan; Gao, Li; Zhang, Haiyan; Tao, Qinghua


    Maternal Wnt11 is both necessary and sufficient for the formation of Spemann organizer in Xenopus embryo. Xnr3 and Siamois have been identified as the direct target genes of maternal Wnt11/β-catenin during organizer induction. The depletion of maternal XTcf3 resulted in the ectopic expression of Xnr3 and Siamois, suggesting the activity of β-catenin/XTcf3 is strictly regulated in the early Xenopus embryos. Here, we show that Xenopus mga (Xmga) is a maternal gene required for dorsal axis formation. Overexpression experiments indicate that mouse Mga potentiates the activity of β-catenin in the induction of organizer-specific genes. Depletion of maternal Xmga results in the dramatic decrease of the expression of organizer genes and ventralization phenotype, indicating that Xmga is required for β-catenin function and organizer formation. Depletion of XTcf3 cannot rescue organizer gene expression and axis formation in Xmga-depleted embryos, suggesting Xmga is downstream of XTcf3 during organizer induction. We conclude that maternal Xmga is critical for the function of β-catenin during organizer formation and dorsal development of Xenopus embryo. To our knowledge, this is a report for the first time to implicate Mga in regulating Wnt signaling. PMID:23070227

  3. Formation and growth of embryos of the Earth-Moon system

    Ipatov, Sergei I.


    Galimov and Krivtsov [1] made computer simulations of the formation of the embryos of the Earth and the Moon as a result of contraction of a rarefied condensation. The angular momentum needed for such contraction could not be acquired during formation of the condensation from a protoplanetary disk. Using the formulas presented in [2], we obtained that the angular momentum of the present Earth-Moon system could be acquired at a collision of two rarefied condensations with a total mass not smaller than 0.1M_{e}, where M_{e} is the Earth mass. In principle, the angular momentum of the condensation needed for formation of the Earth-Moon system could be acquired by accumulation only of small objects, but for such model, the parental condensations of Venus and Mars could also get the angular momentum that was enough for formation of large satellites. Probably, the condensations that contracted and formed the embryos of the terrestrial planets other than the Earth did not collide with massive condensations, and therefore they did not get a large enough angular momentum needed to form massive satellites. The embryos formed as a result of contraction of the condensation grew by accumulation of solid planetesimals. The mass of the rarefied condensation that was a parent for the embryos of the Earth and the Moon could be relatively small (0.02M_{e} or even less), if we take into account the growth of the angular momentum of the embryos at the time when they accumulated planetesimals. There could be also the second main collision of the parental rarefied condensation with another condensation, at which the radius of the Earth's embryo condensation was smaller than the semi-major axis of the orbit of the Moon's embryo. The second main collision (or a series of similar collisions) could change the tilt of the Earth to its present value. For large enough eccentricities of planetesimals, the effective radii of proto-Earth and proto-Moon were proportional to r (where r is the

  4. Serotonin mediation of early memory formation via 5HT2B receptor-induced glycogenolysis in the day-old chick

    Marie Elizabeth Gibbs; Leif eHertz


    Investigation of the effects of serotonin on memory formation in the chick revealed an action on at least two 5-HT receptors. Serotonin injected intracerebrally produced a biphasic effect on memory consolidation with enhancement at low doses and inhibition at higher doses. The non-selective 5-HT receptor antagonist methiothepin and the selective 5-HT2B/C receptor antagonist SB221284 both inhibited memory, suggesting actions of serotonin on at least two different receptor subtypes. The 5-HT2B/...

  5. Development of otoconia in the embryonic chick (Gallus domesticus)

    Fermin, C. D.; Igarashi, M.


    In the chick (Gallus domesticus) embryo, otoconium formation started first over the macula sacculi around the 4th day of incubation, and a day later over the macula utriculi. It was determined that each otoconium formed as a result of the segmentation of the immature otolithic membrane, and that the calcium responsible for otoconium calcification was incorporated into the organic matrix of each otoconium in the form of small electron-dense granules (20-150 nm in. diameter). The presence of calcium in these granules was confirmed by histochemical staining with osmic-potassium pyroantimonate, by EDTA chelation, and by X-ray micronanalysis under the electron microscope.

  6. Thermal Evolution And Core Formation In Planetesimals And Planetary Embryos

    Sramek, O.; Labrosse, S.; Ricard, Y. R.; milelli, L.


    Recent dating of iron meteorites shows that they were formed almost as early as the oldest known objects of the solar system, the CAIs. Moreover, several meteorites show a magnetization that is thought to originate from the action of a dynamo at the early stages of the planetesimals evolution. Core formation requires melting of the metal which then can percolate toward the center, providing the solid matrix deforms and compacts. The energy source for melting of the metal comes from a combination of short lived radionuclides, mostly 26Al, and accretion energy for bodies larger than about 1000 km. We considered a suite of numerical calculations solving for the coupled problem of thermal evolution, melt percolation and matrix compaction, systematically exploring the different accretion histories, final body size and initial concentration in 26Al. Our model handles simultaneously metal and silicates in both solid and liquid states. Depending on the accretion rate, melting occurs from the center outward, in a shallow outer shell progressing inward, or in the two locations. Segregation of the protocore decreases the efficiency of radiogenic heating by confining the 26Al in the outer silicate shell. Various types of planetesimals partly differentiated and sometimes differentiated in multiple metal-silicate layers can be obtained. We discuss the thermal profiles of the accreted bodies in relation to possible early dynamo action as evidenced by remanent magnetization observed on some meteorite samples.

  7. Formation and reparation of the AP-sites into DNA from the gamma-irradiated embryo of bombyx mori

    Full text: It is well known that radiosensitivity of an organism is in dependence on the DNA reparation systems functioning into cells. Sharp difference in the radioresistance of silkworm embryo at different stage of growth showed by us earlier (Agaev F.A. et al, 1991) can provide to suggest that DNA reparation system into cells of 3-daily embryo (more radiosensitive stage) and 7-daily embryo (more radioresistance stage) may be functioning with various efficiency. It was shown that quantity of the AP-sites (i.e. apurine and apirimidine sites) registered into DNA of 3-daily embryo is 1,2 - 1,4 time more, that into DNA of 7-daily embryo g-irradiated at the same dozes. The increasing of the difference between the registered AP-sites into DNA of 3- and 7-daily embryo has been observed also at the increasing of the radiation doze. At the postradiation incubation of the 3- and 7- daily embryo the lowering of AP-sites quantity into DNA was observed. This fact allowed to testify that the reparation system of damages, such as DNA-apurinization and apirimidinization are functioned into these embryo cells. At the same time the rate of the AP-sites reparation into embryo cells is varied. For example, the remanent quantity of AP-sites into DNA of 7-daily embryo after 45 min of postradiation period consists of 30% those registered immediately after embryo irradiation. The remanent quantity of AP-sites into DNA of 3-daily embryo is lowered on 50%. The difference in the rate of cells reparation is keeping at the constant level irrespective of g-irradiation doze. The binding reaction between the /14 centigrade/-methoxyamine and AP-sites in DNA in vitro has been showed that the reparation time of the 50% AP-sites for 3-daily embryo is 45 min and for 7-daily embryo is 30 min in respective to registered value at once after 100 Gy irradiation doze. In spite of essential difference in the both AP-sites formation into DNA of 3- and 7-daily embryo at once after irradiation and the

  8. Safety and immunogenicity of the Intradermal thai red cross (2-2-2-0-1-1 post exposure vaccination regimen in the Indian population using purified chick embryo cell rabies vaccine

    Chhabra Mala


    Full Text Available PURPOSE: To test the immunogenicity of the WHO recommended "2-2-2-0-1-1" post-exposure rabies vaccination regimen in Indian subjects to determine the feasibility of replacing crude sheep brain nerve tissue rabies vaccine with modern tissue culture rabies vaccine at major anti-rabies treatment centers throughout India. METHODS: Purified chick embryo cell vaccine (PCECV was administered in the dosage of 0.1mL per site to 53 Indian subjects. RESULTS: All subjects produced rabies antibodies above 0.5IU/mL by day 14 post-vaccination. Only minor adverse reactions including swelling (6.6%, erythema (5.4% and pain (1.4% were observed for which no treatment was required. CONCLUSIONS: This study demonstrated that PCECV is safe and highly immunogenic in Indian subjects when administered intradermally as 0.1mL/site using the "2-2-2-0-1-1" post-exposure regimen.

  9. Hyperglycemia impairs left-right axis formation and thereby disturbs heart morphogenesis in mouse embryos.

    Hachisuga, Masahiro; Oki, Shinya; Kitajima, Keiko; Ikuta, Satomi; Sumi, Tomoyuki; Kato, Kiyoko; Wake, Norio; Meno, Chikara


    Congenital heart defects with heterotaxia are associated with pregestational diabetes mellitus. To provide insight into the mechanisms underlying such diabetes-related heart defects, we examined the effects of high-glucose concentrations on formation of the left-right axis in mouse embryos. Expression of Pitx2, which plays a key role in left-right asymmetric morphogenesis and cardiac development, was lost in the left lateral plate mesoderm of embryos of diabetic dams. Embryos exposed to high-glucose concentrations in culture also failed to express Nodal and Pitx2 in the left lateral plate mesoderm. The distribution of phosphorylated Smad2 revealed that Nodal activity in the node was attenuated, accounting for the failure of left-right axis formation. Consistent with this notion, Notch signal-dependent expression of Nodal-related genes in the node was also down-regulated in association with a reduced level of Notch signaling, suggesting that high-glucose concentrations impede Notch signaling and thereby hinder establishment of the left-right axis required for heart morphogenesis. PMID:26351675

  10. Hyperglycemia impairs left–right axis formation and thereby disturbs heart morphogenesis in mouse embryos

    Hachisuga, Masahiro; Oki, Shinya; Kitajima, Keiko; Ikuta, Satomi; Sumi, Tomoyuki; Kato, Kiyoko; Wake, Norio; Meno, Chikara


    Congenital heart defects with heterotaxia are associated with pregestational diabetes mellitus. To provide insight into the mechanisms underlying such diabetes-related heart defects, we examined the effects of high-glucose concentrations on formation of the left–right axis in mouse embryos. Expression of Pitx2, which plays a key role in left–right asymmetric morphogenesis and cardiac development, was lost in the left lateral plate mesoderm of embryos of diabetic dams. Embryos exposed to high-glucose concentrations in culture also failed to express Nodal and Pitx2 in the left lateral plate mesoderm. The distribution of phosphorylated Smad2 revealed that Nodal activity in the node was attenuated, accounting for the failure of left–right axis formation. Consistent with this notion, Notch signal-dependent expression of Nodal-related genes in the node was also down-regulated in association with a reduced level of Notch signaling, suggesting that high-glucose concentrations impede Notch signaling and thereby hinder establishment of the left–right axis required for heart morphogenesis. PMID:26351675

  11. Effect of irradiation and colchicine on callus and somatic embryo formation in cassava (Manihot esculenta Crantz)

    A study was conducted to assess the mutagenic effect of gamma radiation on sprouting and height in four local cassava accessions. The four cassava accessions were assessed for their callus induction and somatic embryo formation ability from leaf lobes from gamma irradiated stakes as well as colchicine treated leaf lobes on different concentrations of plant growth regulators, incorporated into Murashige and skoog, (1962) (MS) basal medium. The cassava accessions were irradiated at 0, 32, 35, 45 and 50 Gy and planted in pots filled with loamy soil. The height of the shoots was measured with rule after sprouting. The leaf lobes were collected from the shoots and cultured on MS medium supplemented with 8 mg/l 2, 4-D and 16 mg/l Picloram. Another set of leaf lobes were treated with 0.0, 0.05, 0.1, 0.2, 0.25 g/l colchicine for one hour and thereafter culture on MS medium supplemented with 8 mg/l 2,4-D and 16 mg/l Picloram as described above. Callus induction from leaf lobes in 45 and 50 Gy were significantly (p≤0.05) affected by the irradiation. On the other hand, Callus induction from leaf lobes in 0.1-0.25 g/l colchicine were significantly (p≤0.05) affected by the mutagenic treatment whereas callus induced from leaf lobes in 0.05 g/l colchicine was not significantly (p≤0.05) affected. Callus induced on 8 mg/l 2, 4-D and 16 mg/l picloram gave the best response in Ankrah and all control tested while Tomfa recorded the least. Colchicine at a concentration of 0.05 g/l and radiation dose of 32 Gy treatments gave the best response of callusing. Callus induction decreased with increasing colchicine concentration and gamma irradiation. Callus derived from irradiated and colchicine leaf lobes appeared soft but friable and tiny, compact, respectively, predominately with creamy to brown colouration. Calli obtained were sub-cultures on embryo regeneration medium consisting of MS supplemented with 0.01mg/1 NAA and o.1 mg/1 BAP. There was no plantlet regeneration. Instead

  12. Retinoic Acid Signaling Is Essential for Valvulogenesis by Affecting Endocardial Cushions Formation in Zebrafish Embryos.

    Li, Junbo; Yue, Yunyun; Zhao, Qingshun


    Retinoic acid (RA) plays important roles in many stages of heart morphogenesis. Zebrafish embryos treated with exogenous RA display defective atrio-ventricular canal (AVC) specification. However, whether endogenous RA signaling takes part in cardiac valve formation remains unknown. Herein, we investigated the role of RA signaling in cardiac valve development by knocking down aldh1a2, the gene encoding an enzyme that is mainly responsible for RA synthesis during early development, in zebrafish embryos. The results showed that partially knocking down aldh1a2 caused defective formation of primitive cardiac valve leaflets at 108 hpf (hour post-fertilization). Inhibiting endogenous RA signaling by 4-diethylaminobenzal-dehyde revealed that 16-26 hpf was a key time window when RA signaling affects the valvulogenesis. The aldh1a2 morphants had defective formation of endocardial cushion (EC) at 76 hpf though they had almost normal hemodynamics and cardiac chamber specification at early development. Examining the expression patterns of AVC marker genes including bmp4, bmp2b, nppa, notch1b, and has2, we found the morphants displayed abnormal development of endocardial AVC but almost normal development of myocardial AVC at 50 hpf. Being consistent with the reduced expression of notch1b in endocardial AVC, the VE-cadherin gene cdh5, the downstream gene of Notch signaling, was ectopically expressed in AVC of aldh1a2 morphants at 50 hpf, and overexpression of cdh5 greatly affected the formation of EC in the embryos at 76 hpf. Taken together, our results suggest that RA signaling plays essential roles in zebrafish cardiac valvulogenesis. PMID:26671342

  13. Chick development and high dose of bendiocarb

    Petrovová, E.; Sedmera, David; Luptáková, L.; Mazenský, D.; Danko, J.


    Roč. 47, č. 9 (2012), s. 1312-1318. ISSN 1093-4529 Institutional research plan: CEZ:AV0Z50110509 Keywords : Bendiocarb * central nervous system * chick embryo * liver * toxicity Subject RIV: EA - Cell Biology Impact factor: 1.252, year: 2012

  14. Precambrian phosphatized embryos and larvae from the Doushantuo Formation and their affinities, Guizhou (SW China)

    CHEN Junyuan; CHI Huimei


    Weng,an phosphates of the Precambrian Doushantuo Formation, Guizhou (southwestern China) preserve a large number of exquisite biological structures, which are mostly micro-spherical and represent seaweeds, acritarchs and developing eggs related to various groups of metazoans. Here is a report of a variety of developing eggs and larvae, which are most probably of Cnidarian affinity. The eggs examined in the study are composed of early cleavage embryos and two-layered gastrulae. The early cleavage embryos are radial and total cleavage with equal-size blastomeres. The gastrulae mostly bear a large archenteron, which is filled with yolk-degrading organic matter. Ovoid to fusiform planula-like larvae identified in thin sections under light microscope are mostly mouthless and their gastrovascular cavity is filled with possible yolk-degrading organic matter. They are likely representatives of non-feeding larva. The uncommon planula-like structures are hollow, with each having a mouth-like structure on its narrow end. We interpret them as feeding larva. Study of these embryos with possible Cnidarian affinities shed new insight on the origin of metazoans.

  15. A biomechanical analysis of ventral furrow formation in the Drosophila melanogaster embryo.

    Vito Conte

    Full Text Available The article provides a biomechanical analysis of ventral furrow formation in the Drosophila melanogaster embryo. Ventral furrow formation is the first large-scale morphogenetic movement in the fly embryo. It involves deformation of a uniform cellular monolayer formed following cellularisation, and has therefore long been used as a simple system in which to explore the role of mechanics in force generation. Here we use a quantitative framework to carry out a systematic perturbation analysis to determine the role of each of the active forces observed. The analysis confirms that ventral furrow invagination arises from a combination of apical constriction and apical-basal shortening forces in the mesoderm, together with a combination of ectodermal forces. We show that the mesodermal forces are crucial for invagination: the loss of apical constriction leads to a loss of the furrow, while the mesodermal radial shortening forces are the primary cause of the internalisation of the future mesoderm as the furrow rises. Ectodermal forces play a minor but significant role in furrow formation: without ectodermal forces the furrow is slower to form, does not close properly and has an aberrant morphology. Nevertheless, despite changes in the active mesodermal and ectodermal forces lead to changes in the timing and extent of furrow, invagination is eventually achieved in most cases, implying that the system is robust to perturbation and therefore over-determined.

  16. The Early Stages of Heart Development: Insights from Chicken Embryos

    Johannes G. Wittig


    Full Text Available The heart is the first functioning organ in the developing embryo and a detailed understanding of the molecular and cellular mechanisms involved in its formation provides insights into congenital malformations affecting its function and therefore the survival of the organism. Because many developmental mechanisms are highly conserved, it is possible to extrapolate from observations made in invertebrate and vertebrate model organisms to humans. This review will highlight the contributions made through studying heart development in avian embryos, particularly the chicken. The major advantage of chick embryos is their accessibility for surgical manipulation and functional interference approaches, both gain- and loss-of-function. In addition to experiments performed in ovo, the dissection of tissues for ex vivo culture, genomic, or biochemical approaches is straightforward. Furthermore, embryos can be cultured for time-lapse imaging, which enables tracking of fluorescently labeled cells and detailed analysis of tissue morphogenesis. Owing to these features, investigations in chick embryos have led to important discoveries, often complementing genetic studies in mice and zebrafish. As well as including some historical aspects, we cover here some of the crucial advances made in understanding early heart development using the chicken model.

  17. Effects of arctigenin on angiogenesis of chick embryo chorioallantoic membrane%牛蒡子苷元对鸡胚绒毛尿囊膜血管新生的影响

    刘奋; 薛芳喜; 关永霞; 姚景春; 闫莹


    目的:观察牛蒡子苷元对鸡胚绒毛尿囊膜( CAM)血管新生的影响。方法采用乙醇结晶法制备牛蒡子苷元,开窗法制备CAM模型。将CAM模型中的40只鸡胚随机分为阴性对照组、低剂量组、中剂量组、高剂量组,每组各10只。分别将含有DMSO、2.5、5.0、10.0 g/L牛蒡子苷元的药物载体置于阴性对照组、低剂量组、中剂量组、高剂量组CAM中央血管稀少区,孵育48 h后,剪下CAM,在解剖显微镜下观察药物载体覆盖区及CAM周围的新生血管数量、形态,观察各组新生血管等级,记录各组药物载体边缘5 mm内新生血管的数目,计算血管抑制率( IR)。结果低、中、高剂量组CAM新生血管发育均受抑制。中、高剂量组新生血管等级3+、4+的数量多于阴性对照组(P<0.05),阴性对照组、低剂量组、中剂量组、高剂量组新生血管数目分别为(78.26±5.62)、(59.22±4.16)、(37.43±6.51)、(24.17±3.23)条,低剂量组新生血管数低于阴性对照组(P<0.05),中、高剂量组新生血管数均低于阴性对照组(P<0.01),且中、高剂量组新生血管数均低于低剂量组(P<0.05)。低剂量组、中剂量组、高剂量组新生血管IR分别为24.33%、52.17%及69.12%。结论牛蒡子苷元具有抑制鸡胚CAM血管新生的作用。%Objective To observe the influence of arctigenin on the angiogenesis of chick embryo chorioallantoic membrane ( CAM) .Methods Arctigenin was prepared by the procedure of ethanol crystallization from Fructus Arctii and we used the windowing method to prepare the CAM model .Forty chick embryos of CAM models were randomly divided into the negative control group, low (2.5 g/L), medium (5.0 g/L), and high (10.0 g/L) doses of arctigenin groups, 10 em-bryos in each group .Then the negative control solution containing DMSO and each dose of liquid containing

  18. Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer.

    Tomioka, Ikuo; Mizutani, Eiji; Yoshida, Tomoyuki; Sugawara, Atsushi; Inai, Kentaro; Sasada, Hiroshi; Sato, Eimei


    The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization. PMID:17446658

  19. The impact of food intake and social habits on embryo quality and the likelihood of blastocyst formation.

    Braga, Daniela Paes Almeida Ferreira; Halpern, Gabriela; Setti, Amanda S; Figueira, Rita Cássia S; Iaconelli, Assumpto; Borges, Edson


    The aim of this study was to evaluate the influence of patients' lifestyle factors and eating habits on embryo development. A total of 2659 embryos recovered from 269 patients undergoing intracytoplasmic sperm injection cycles were included. The frequency of intake of food items and social habits were registered and its influences on embryo development evaluated. The consumption of cereals, vegetables and fruits positively influenced the embryo quality at the cleavage stage. The quality of the embryo at the cleavage stage was also negatively correlated with the consumption of alcoholic drinks and smoking habits. The consumption of fruits influenced the likelihood of blastocyst formation, which was also positively affected by the consumption of fish. Being on a weight-loss diet and consumption of red meat had a negative influence on the likelihood of blastocyst formation. The likelihood of blastocyst formation was also negatively influenced by the consumption of alcoholic drinks and by smoking habits. The consumption of red meat and body mass index had a negative effect on the implantation rate and the likelihood of pregnancy. In addition, being on a weight-loss diet had a negative influence on implantation rate. Our evidence suggests a possible relationship between environmental factors and ovary biology. PMID:25982093

  20. Specification of germ layer identity in the chick gastrula

    Cai Qin; Matsumoto Kiyoshi; Chapman Susan C; Schoenwolf Gary C


    Abstract Background Chick definitive endoderm is an important source of signals that pattern the early embryo forming a central structure around which the body plan is constructed. Although the origin of definitive endoderm has been mapped in the chick, arising principally from rostral streak at elongating streak stages, it is not known when this layer first becomes fully committed to its germ layer fate, an important issue to resolve in light of its critical role in subsequent patterning of ...

  1. Altered callose deposition during embryo sac formation of multi-pistil mutant (mp1) in Medicago sativa.

    Zhou, H C; Jin, L; Li, J; Wang, X J


    Whether callose deposition is the cause or result of ovule sterility in Medicago sativa remains controversial, because it is unclear when and where changes in callose deposition and dissolution occur during fertile and sterile embryo sac formation. Here, alfalfa spontaneous multi-pistil mutant (mp1) and wild-type plants were used to compare the dynamics of callose deposition during embryo sac formation using microscopy. The results showed that both mutant and wild-type plants experienced megasporogenesis and megagametogenesis, and there was no significant difference during megasporogenesis. In contrast to the wild-type plants, in which the mature embryo sac was observed after three continuous cycles of mitosis, functional megaspores of mutant plants developed abnormally after the second round of mitosis, leading to degeneration of synergid, central, and antipodal cells. Callose deposition in both mutant and wild-type plants was first observed in the walls of megasporocytes, and then in the megaspore tetrad walls. After meiosis, the callose wall began to degrade as the functional megaspore underwent mitosis, and almost no callose was observed in the mature embryo sac in wild-type plants. However, callose deposition was observed in mp1 plants around the synergid, and increased with the development of the embryo sac, and was mainly deposited at the micropylar end. Our results indicate that synergid, central, and antipodal cells, which are surrounded by callose, may degrade owing to lack of nutrition. Callose accumulation around the synergid and at the micropylar end may hinder signals required for the pollen tube to enter the embryo sac, leading to abortion. PMID:27323128

  2. Bendiocarbamate toxicity in the chick embryo

    Petrovová, E.; Sedmera, David; Míšek, Ivan; Lešník, František; Luptáková, L.


    Roč. 55, č. 1 (2009), s. 61-65. ISSN 0015-5500 Grant ostatní: VEGA(SK) 1/4430/07 Institutional research plan: CEZ:AV0Z50450515 Keywords : carbamate * pesticides * embryotoxicity Subject RIV: EA - Cell Biology Impact factor: 0.924, year: 2009

  3. Retroviral Vector-Mediated Gene Transfer into the Chick Optic Vesicle by In Ovo Electroporation

    Sakuta, Hiraki; Suzuki, Ryoko; Noda, Masaharu

    The chick embryo offers many advantages for developmental studies over other vertebrate embryos as it allows easy access for in ovo surgical manipulations, such as tissue transplantation and the implantation of cultured cells or chemically treated beads for the local release of humoral factors. In particular, owing to its external position in the embryo, the chick eye is a popular model for studying the patterning mechanism of the central nervous system (CNS). This patterning has a crucial role in shaping functional organization because it is the basis of the specific wiring in the CNS. Genetic analysis is not easy in the chick, as compared with the mouse for which transgene introduction or gene targeting techniques have been well established. However, because methods for the expression of exogenous genes and for gene silencing in the chick embryo have been recently developed, the functional analysis of genes has become possible in combination with classical techniques of developmental biology and neurobiology.


    Euclidez Braga Malheiros


    Full Text Available

    The influence of sex and egg weight on small intestine villi height and crypt depth was analyzed at 18 d of incubation, at hatching, and 7 days post-hatch. Amounts of duodenum. jejunum and ileum were removed and fixed in Bouin solution for 24 hours. After rinse in 5% ethanol, the amounts were dehydrated in a graded alcohol series up to absolute ethanol (70%, 80%, 90% and 100%, diafanized in absolute xylene and embedded in paraplast. Sections were stained with hematoxylin and eosin. Dta were submitted to analysis of variance. And significant treatment means were separated by Tukey´s test (p<0.05. In all three analyzed ages, birds from heavy eggs presented the heaviest body weights. At 7 days of age, female chicks were heavier than males. In the jejunum, villi were higher in male embryos as compared to female embryos, whereas in the ileum, villi were higher in females as compared to males. In the three intestinal segments, crypts were deeper in males than in females. Duodenum, jejunum, and ileum villi height and jejunum crypt depth were higher in embryos derived from heavy eggs. At hatching, duodenal crypts were deeper in male than in female chicks, while jejunum and ileum crypts were deeper in female chicks. Newly-hatched chicks from heavy eggs presented the highest villi. In the three intestinal segments, crypt was shallower in chicks hatched from light as compared to heavy eggs. At 7 days of age, jejunum villi were higher in male than in female chicks, whereas crypts were deeper in females in all intestinal segments. Chicks hatched from heavy eggs presented the highest jejunum and ileum villi, and the deepest ileum depth. The results of this study show that body weight and the intestinal mucosa are influenced by sex and egg weight, and that females from heavy eggs were the heaviest chicks, and their jejunum and ileum mucosa presented the highest growth.

    KEY WORDS: Broiler embryos, chicks

  5. Kid depletion in mouse oocytes associated with multinucleated blastomere formation and inferior embryo development.

    Egashira, Akiyoshi; Yamauchi, Nobuhiko; Islam, Md Rashedul; Yamagami, Kazuki; Tanaka, Asami; Suyama, Hikaru; El-Sayed, El-Sharawy Mohamed; Tabata, Shoji; Kuramoto, Takashi


    This study investigated the knockdown (KD) of Kid on maturation developmental competence and multinucleation of mouse germinal vesicle (GV) oocytes after parthenogenetic activation. Data revealed that Kid messenger RNA (mRNA) was expressed in GV and MII stage oocyte and 1- and 2-cell embryos. Additionally, Kid mRNA expression in the Kid KD group decreased by nearly 46% compared to the control small interfering RNA (siRNA) groups. The rate of multinucleated embryos in the Kid KD group (52.4%) was significantly higher (P  4-cell stage (28.6% vs. 53.5%) and the blastocyst stage (2.4% vs. 23.3%) compared to the control siRNA groups. Suppression of Kid using siRNA caused multinucleation in early embryos with high frequency and it may increase 2- to 4-cell arrested embryos and reduce the developmental competence to blastocyst. PMID:26890962

  6. Serotonin mediation of early memory formation via 5HT2B receptor-induced glycogenolysis in the day-old chick

    Marie Elizabeth Gibbs


    Full Text Available Investigation of the effects of serotonin on memory formation in the chick revealed an action on at least two 5HT receptors. Serotonin injected intracerebrally produced a biphasic effect on memory consolidation with enhancement at low doses and inhibition at higher doses. The non-selective 5HT receptor antagonist methiothepin and the selective 5HT2B/C receptor antagonist SB221284 both inhibited memory, suggesting actions of serotonin on at least 2 different receptor subtypes. The 5HT2B/C and astrocyte-specific 5-HT receptor agonists, fluoxetine and paroxetine, enhanced memory and the effect was attributed to glycogenolysis. Inhibition of glycogenolysis with a low dose of DAB prevented both serotonin and fluoxetine from enhancing memory during short-term memory but not during intermediate memory. The role of serotonin on the 5HT2B/C receptor appears to involve glycogen breakdown in astrocytes during short-term memory, whereas other published evidence attributes the second period of glycogenolysis to noradrenaline.

  7. 鸡胚低分子提取物对D-半乳糖拟衰老小鼠线粒体DNA缺失突变的影响%Effect of low molecular weight substance extracted from chick embryo on mitochondrial DNA deletion in aging mice induced by D-galactose

    陈姝; 何蕴韶; 程刚; 高劲松


    Objective To investigate the effect of low molecular weight substance extracted from chick embryo on mitochondrial DNA(mtDNA)deletion in senile mice induced bv D-galactose.Methods Senile mice induced by D-galactose were treated with low molecular weight substance extracted from chick embryo.The deleted fragment of mtDNA was examined by using polymerase chain reaction technique and agarose gel electrophoresis.A relative quantitation of band densities was performed by using densitometry scanning techniques.The deletion was identified by using direct sequencing analysis. Results The 4239 bD mtDNA deletion were present in liver,cerebral cortex and hippocampus tissues in all of the mice.and it was significantly higher in senile model mice than in control mice(all P<0.01).Low molecular weight substance extracted from chick embryo reduced the mtDNA deletion in various tissues in senile model mice(all P<0.05).Deletion was more abundant in liver than in cerebral cortex and hippoeampus. Conclusions The 4239 bp mtDNA deletion is a common deletion,and low molecular weight substance extracted from chick embryo could decrease the incidence of 4239 bD mtDNA deletion.%目的 探讨鸡胚低分子提取物对D-半乳糖诱导的衰老小鼠线粒体DNA(mtDNA)缺失突变的影响.方法 采用D-半乳糖诱导制备衰老小鼠模型并用鸡胚低分子提取物处理.用聚合酶链反应技术和琼脂糖凝胶电泳检测mtDNA缺失片段,密度扫描技术对扩增片段进行相对定量.缺失片段构建质粒后,直接测序进行鉴定.结果 全部小鼠的肝、大脑皮质与海马组织内均存在4239 bp的mtDNA缺失片段.衰老模型鼠不同组织mtDNA缺失的相对百分含量均明显高于正常对照组(均为P%0.01),而鸡胚低分子提取物可明显降低模型鼠肝、大脑皮质与海马组织内mtDNA缺失的比例(均为P<0.05).肝组织中mtDNA缺失的比例较大脑皮质和海马组织更高.结论 4239 bp的mtDNA缺失片段普遍存在

  8. Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos

    Jun Dai, Xiaojuan Cui, Zuoyan Zhu, Wei Hu


    Full Text Available This study focused on concatemer formation and integration pattern of transgenes in zebrafish embryos. A reporter plasmid based on enhanced green fluorescent protein (eGFP driven by Cytomegalovirus (CMV promoter, pCMV-pax6in-eGFP, was constructed to reflect transgene behavior in the host environment. After removal of the insertion fragment by double digestion with various combinations of restriction enzymes, linearized pCMV-pax6in-eGFP vectors were generated with different combinations of 5′-protruding, 3′-protruding, and blunt ends that were microinjected into zebrafish embryos. Repair of double-strand breaks (DSBs was monitored by GFP expression following religation of the reporter gene. One-hundred-and-ninety-seven DNA fragments were amplified from GFP-positive embryos and sequenced to analyze the repair characteristics of different DSB end combinations. DSBs involving blunt and asymmetric protruding ends were repaired efficiently by direct ligation of blunt ends, ligation after blunting and fill-in, or removed by cutting. Repair of DSBs with symmetric 3′-3′ protrusions was less efficient and utilized template-directed repair. The results suggest that non-homologous end joining (NHEJ was the principal mechanism of exogenous gene concatemer formation and integration of transgenes into the genome of transgenic zebrafish.

  9. Supraadditive formation of micronuclei in preimplantation mouse embryos in vitro after combined treatment with X-rays and caffeine

    The influence of caffeine (0.1 or 2 mM), X-rays (0.24 Gy or 0.94 Gy, or of a combination of both on the formation of micronuclei in early stages of preimplantation mouse embryos in vitro was studied. X-rays as well as caffeine induced micronuclei. The dose-effect curve after irradiation was linear for the dose range measured. Caffeine did not induce micronuclei if the concentration was 1 mM or less; between 1 mM and 7 mM, however, there was a linear increase in the number of micronuclei. A considerable enhancement of the number of radiation-induced micronuclei was observed when irradiation of the embryos was followed by a treatment with caffeine. Not only was the sum of the single effects exceeded by the combination effects, but the combination results even lay in the range of supraadditivity of the envelope of additivity

  10. Indicators of functional differentiation of the chick embryonic kidney

    Zemanová, Zdeňka; Ujec, Evžen; Jirsová, Z.; Maňáková, E.


    Roč. 131, č. 4 (2002), s. 847-860. ISSN 1095-6433 R&D Projects: GA ČR GA304/98/0604 Institutional research plan: CEZ:AV0Z5011922 Keywords : Chick embryo * Mesonephros * Glomerular filtration Subject RIV: ED - Physiology Impact factor: 1.274, year: 2002

  11. Zebrafish embryo screen for mycobacterial genes involved in the initiation of granuloma formation reveals a newly identified ESX-1 component

    Esther J. M. Stoop


    The hallmark of tuberculosis (TB is the formation of granulomas, which are clusters of infected macrophages surrounded by additional macrophages, neutrophils and lymphocytes. Although it has long been thought that granulomas are beneficial for the host, there is evidence that mycobacteria also promote the formation of these structures. In this study, we aimed to identify new mycobacterial factors involved in the initial stages of granuloma formation. We exploited the zebrafish embryo Mycobacterium marinum infection model to study initiation of granuloma formation and developed an in vivo screen to select for random M. marinum mutants that were unable to induce granuloma formation efficiently. Upon screening 200 mutants, three mutants repeatedly initiated reduced granuloma formation. One of the mutants was found to be defective in the espL gene, which is located in the ESX-1 cluster. The ESX-1 cluster is disrupted in the Mycobacterium bovis BCG vaccine strain and encodes a specialized secretion system known to be important for granuloma formation and virulence. Although espL has not been implicated in protein secretion before, we observed a strong effect on the secretion of the ESX-1 substrates ESAT-6 and EspE. We conclude that our zebrafish embryo M. marinum screen is a useful tool to identify mycobacterial genes involved in the initial stages of granuloma formation and that we have identified a new component of the ESX-1 secretion system. We are confident that our approach will contribute to the knowledge of mycobacterial virulence and could be helpful for the development of new TB vaccines.

  12. Inactivation of the Huntington's disease gene (Hdh impairs anterior streak formation and early patterning of the mouse embryo

    Conlon Ronald A


    Full Text Available Abstract Background Huntingtin, the HD gene encoded protein mutated by polyglutamine expansion in Huntington's disease, is required in extraembryonic tissues for proper gastrulation, implicating its activities in nutrition or patterning of the developing embryo. To test these possibilities, we have used whole mount in situ hybridization to examine embryonic patterning and morphogenesis in homozygous Hdhex4/5 huntingtin deficient embryos. Results In the absence of huntingtin, expression of nutritive genes appears normal but E7.0–7.5 embryos exhibit a unique combination of patterning defects. Notable are a shortened primitive streak, absence of a proper node and diminished production of anterior streak derivatives. Reduced Wnt3a, Tbx6 and Dll1 expression signify decreased paraxial mesoderm and reduced Otx2 expression and lack of headfolds denote a failure of head development. In addition, genes initially broadly expressed are not properly restricted to the posterior, as evidenced by the ectopic expression of Nodal, Fgf8 and Gsc in the epiblast and T (Brachyury and Evx1 in proximal mesoderm derivatives. Despite impaired posterior restriction and anterior streak deficits, overall anterior/posterior polarity is established. A single primitive streak forms and marker expression shows that the anterior epiblast and anterior visceral endoderm (AVE are specified. Conclusion Huntingtin is essential in the early patterning of the embryo for formation of the anterior region of the primitive streak, and for down-regulation of a subset of dynamic growth and transcription factor genes. These findings provide fundamental starting points for identifying the novel cellular and molecular activities of huntingtin in the extraembryonic tissues that govern normal anterior streak development. This knowledge may prove to be important for understanding the mechanism by which the dominant polyglutamine expansion in huntingtin determines the loss of neurons in

  13. Angiogenesis is repressed by ethanol exposure during chick embryonic development.

    Wang, Guang; Zhong, Shan; Zhang, Shi-Yao; Ma, Zheng-Lai; Chen, Jian-Long; Lu, Wen-Hui; Cheng, Xin; Chuai, Manli; Lee, Kenneth Ka Ho; Lu, Da-Xiang; Yang, Xuesong


    It is now known that excess alcohol consumption during pregnancy can cause fetal alcohol syndrome to develop. However, it is not known whether excess ethanol exposure could directly affect angiogenesis in the embryo or angiogenesis being indirectly affected because of ethanol-induced fetal alcohol syndrome. Using the chick yolk sac membrane (YSM) model, we demonstrated that ethanol exposure dramatically inhibited angiogenesis in the YSM of 9-day-old chick embryos, in a dose-dependent manner. Likewise, the anti-angiogenesis effect of ethanol could be seen in the developing vessel plexus (at the same extra-embryonic regions) during earlier stages of embryo development. The anti-angiogenic effect of ethanol was found associated with excess reactive oxygen species (ROS) production; as glutathione peroxidase activity increased while superoxide dismutase 1 and 2 activities decreased in the YSMs. We further validated this observation by exposing chick embryos to 2,2'-azobis-amidinopropane dihydrochloride (a ROS inducer) and obtained a similar anti-angiogenesis effect as ethanol treatment. Semiquantitative reverse transcription-polymerase chain reaction analysis of the experimental YSMs revealed that expression of angiogenesis-related genes, vascular endothelial growth factor and its receptor, fibroblast growth factor 2 and hypoxia-inducible factor, were all repressed following ethanol and 2,2'-azobis-amidinopropane dihydrochloride treatment. In summary, our results suggest that excess ethanol exposure inhibits embryonic angiogenesis through promoting superfluous ROS production during embryo development. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26177723

  14. The gametophytic expression of the gene for sensor histidine kinase CKI1 is essential for embryo sac formation in Arabidopsis

    Hejátko, Jan; Pernisová, M.; Eneva, T.; Palme, K.; Brzobohatý, Břetislav

    Brno : Masarykova univerzita v Brně, 2003 - (Dubová, J.). s. 67 ISBN 80-210-3194-8. [International Conference on Plant Embryology /9./. 01.09.2003-03.09.2003, Brno] R&D Projects: GA MŠk VS96096; GA MŠk LN00A081 Grant ostatní: QLRT(XE) 2000-0020; QLRT-CT(XE) 2001-01871 Institutional research plan: CEZ:AV0Z5004920 Keywords : embryo sac formation * CKI1 * histidine kinase Subject RIV: BO - Biophysics

  15. Direct Somatic Embryo Formation from Roots of In Vitro-Seedlings of Oil Palm (Elaeis Guineesis Jacq.

    Stapaporn KERDSUWAN


    Full Text Available The induction of somatic embryos (SEs directly from the roots of oil palm plantlets raised in vitro-seedling and origin of somatic embryos. SEs were directly induced from the roots of oil palm plantlets regenerated from immature oil palm fruit (3 months after pollination: 3MAP and provided by Golden Tenera Company. The plants were cultured on OPCM medium supplemented with 3 % sucrose, 200 mg/l ascorbic acid and 0.5 mg/l NAA. The culture was maintained under light conditions at an intensity of 15 µmol/m2/s, 14 h photoperiod, 28±1 °C for 2 months. Histological studies revealed that SEs were initiated from epidermal cells and a few parenchymatous cells. Parenchyma cells had a large nucleus, dense cytoplasm with a broad basal area in contact with maternal tissue. Those cells underwent rapid cell division to form proembryonic mass (PEM and finally developed into SEs through somatic embryogenesis (SE. The highest number of SEs was 1.2 SEs/root and the percentage of SE formation was recorded to be 80. The present study confirms direct SE formation from roots and describes the origin of SE.

  16. Low dose trichloroethylene alters cytochrome P450 - 2C subfamily expression in the developing chick heart

    Makwana, Om; Ahles, Lauren; Lencinas, Alejandro; Selmin, Ornella I.; Runyan, Raymond B.


    Trichloroethylene (TCE) is an organic solvent and common environmental contaminant. TCE exposure is associated with heart defects in humans and animal models. Primary metabolism of TCE in adult rodent models is by specific hepatic cytochrome P450 enzymes (Lash et al., 2000). As association of TCE exposure with cardiac defects is in exposed embryos prior to normal liver development, we investigated metabolism of TCE in the early embryo. Developing chick embryos were dosed in ovo with environme...

  17. Production of Interspecific Germline Chimeras via Embryo Replacement.

    Choi, Hee Jung; Lee, Hyung Chul; Kang, Kyung Soo; Lee, Hyo Gun; Ono, Tamao; Nagai, Hiroki; Sheng, Guojun; Han, Jae Yong


    In avian species, primordial germ cells (PGCs) use the vascular system to reach their destination, the genital ridge. Because of this unique migratory route of avian germ cells, germline chimera production can be achieved via germ cell transfer into a blood vessel. This study was performed to establish an alternative germ cell-transfer system for producing germline chimeras by replacing an original host embryo with a donor embryo, while retaining the host extraembryonic tissue and yolk, before circulation. First, to test the migratory capacity of PGCs after embryo replacement, Korean Oge (KO) chick embryos were used to replace GFP transgenic chick embryos. Four days after replacement, GFP-positive cells were detected in the replaced KO embryonic gonads, and genomic DNA PCR analysis with the embryonic gonads demonstrated the presence of the GFP transgene. To produce an interspecific germline chimera, the original chick embryo proper was replaced with a quail embryo onto the chick yolk. To detect the gonadal PGCs in the 5.5-day-old embryonic gonads, immunohistochemistry was performed with monoclonal antibodies specific to either quail or chick PGCs, i.e., QCR1 and anti-stage-specific embryonic antigen-1 (SSEA-1), respectively. Both the QCR1-positive and SSEA-1-positive cells were detected in the gonads of replaced quail embryos. Forty percent of the PGC population in the quail embryos was occupied by chick extraembryonically derived PGCs. In conclusion, replacement of an embryo onto the host yolk before circulation can be applied to produce interspecies germline chimeras, and this germ cell-transfer technology is potentially applicable for reproduction of wild or endangered bird species. PMID:26063873

  18. Daily supplementation with ghrelin improves in vitro bovine blastocysts formation rate and alters gene expression related to embryo quality.

    Dovolou, Eleni; Periquesta, Eva; Messinis, Ioannis E; Tsiligianni, Theodora; Dafopoulos, Konstantinos; Gutierrez-Adan, Alfonso; Amiridis, Georgios S


    Ghrelin is a gastric peptide having regulatory role in the reproductive system functionality, acting mainly at central level. Because the expression of ghrelin system (ghrelin and its receptor) has been detected in the bovine ovary, the objectives of the present study were to investigate whether ghrelin can affect the developmental potential of in vitro-produced embryos, and to test their quality in terms of relative abundance of various genes related to metabolism, apoptosis and oxidation. In the first experiment, in vitro-produced zygotes were cultured in the absence (control [C]) and in the presence of three concentrations of acylated ghrelin (200 pg/mL [Ghr200], 800 pg/mL [Ghr800]; and 2000 pg/mL [Ghr2000]); blastocyst formation rates were examined on Days 7, 8, and 9. In the second experiment, only the 800 pg/mL dose of ghrelin was used. Zygotes were produced as in experiment 1 and 24 hours post insemination they were divided into 4 groups; in two groups (C; without ghrelin; Ghr800 with ghrelin), embryos were cultured without medium replacement; in the remaining two groups (Control N and GhrN), the culture medium was daily renewed. A pool of Day-7 blastocysts were snap frozen for relative mRNA abundance of various genes related to metabolism, oxidation, implantation, and apoptosis. In experiment 3, embryos were produced as in experiment 2, but in the absence of serum (semi-defined culture medium). In experiment 1, no differences were detected between C, Ghr200, and Ghr2000, although fewer blastocysts were produced in group Ghr800 compared with C. In experiment 2, the lowest blastocysts yield was found in Ghr800, whereas daily renewal of ghrelin (Ghr800N) resulted to increased blastocysts formation rate, which on Day 7 was the highest among groups (P quality than controls. Our results imply a specific role of ghrelin in early embryonic development; however, the specific mode of its action needs further investigation. PMID:24332928

  19. The maternal-effect gene cellular island encodes aurora B kinase and is essential for furrow formation in the early zebrafish embryo.

    Taijiro Yabe


    Full Text Available Females homozygous for a mutation in cellular island (cei produce embryos with defects in cytokinesis during early development. Analysis of the cytoskeletal events associated with furrow formation reveal that these defects include a general delay in furrow initiation as well as a complete failure to form furrow-associated structures in distal regions of the blastodisc. A linkage mapping-based candidate gene approach, including transgenic rescue, shows that cei encodes the zebrafish Aurora B kinase homologue. Genetic complementation analysis between the cei mutation and aurB zygotic lethal mutations corroborate gene assignment and reveal a complex nature of the maternal-effect cei allele, which appears to preferentially affect a function important for cytokinesis in the early blastomeres. Surprisingly, in cei mutant embryos a short yet otherwise normal furrow forms in the center of the blastodisc. Furrow formation is absent throughout the width of the blastodisc in cei mutant embryos additionally mutant for futile cycle, which lack a spindle apparatus, showing that the residual furrow signal present in cei mutants is derived from the mitotic spindle. Our analysis suggests that partially redundant signals derived from the spindle and astral apparatus mediate furrow formation in medial and distal regions of the early embryonic blastomeres, respectively, possibly as a spatial specialization to achieve furrow formation in these large cells. In addition, our data also suggest a role for Cei/AurB function in the reorganization of the furrow-associated microtubules in both early cleavage- and somite-stage embryos. In accordance with the requirement for cei/aurB in furrow induction in the early cleavage embryo, germ plasm recruitment to the forming furrow is also affected in embryos lacking normal cei/aurB function.

  20. Yolk hormones influence in ovo chemosensory learning, growth, and feeding behavior in domestic chicks.

    Bertin, Aline; Meurisse, Maryse; Arnould, Cécile; Leterrier, Christine; Constantin, Paul; Cornilleau, Fabien; Vaudin, Pascal; Burlot, Thierry; Delaveau, Joel; Rat, Christophe; Calandreau, Ludovic


    In this study, we assessed whether prenatal exposure to elevated yolk steroid hormones can influence in ovo chemosensory learning and the behavior of domestic chicks. We simulated a maternal environmental challenge by experimentally enhancing yolk progesterone, testosterone, and estradiol concentrations in hen eggs prior to incubation. The embryos from these hormones-treated eggs (HO) as well as sham embryos (O) that had received the vehicle-only were exposed to the odor of fish oil (menhaden) between embryonic Days 11 and 20. An additional group of control embryos (C) was not exposed to the odor. All chicks were tested following hatching for their feeding preferences between foods that were or were not odorized with the menhaden odor. In the 3-min choice tests, the behavior of O chicks differed significantly according to the type of food whereas C and HO chicks showed no preference between odorized and non-odorized food. Our result suggests weaker response in HO chicks. In addition, HO chicks showed impaired growth and reduced intake of an unfamiliar food on the 24-h time scale compared to controls. Our data suggest that embryonic exposure to increased yolk hormone levels can alter growth, chemosensory learning, and the development of feeding behaviors. PMID:26419601

  1. Microarray analysis of normal and abnormal chick ventricular myocardial development

    Krejčí, Eliška; Peševski, Živorad; Dealmeida, A.C.; Mrug, M.; Fresco, V.M.; Argraves, W.S.; Barth, J.L.; Cui, X.; Sedmera, David


    Roč. 61, Suppl.1 (2012), S137-S144. ISSN 0862-8408 R&D Projects: GA ČR(CZ) GAP302/11/1308; GA ČR(CZ) GD204/09/H084 Institutional research plan: CEZ:AV0Z50110509 Institutional support: RVO:67985823 Keywords : chick embryo * gene chip array * hypoplastic left heart syndrome * hemodynamics * gene expression Subject RIV: EA - Cell Biology Impact factor: 1.531, year: 2012

  2. Conserved and divergent expression patterns of markers of axial development in reptilian embryos: Chinese soft-shell turtle and Madagascar ground gecko.

    Yoshida, Michio; Kajikawa, Eriko; Kurokawa, Daisuke; Noro, Miyuki; Iwai, Tatsuhiro; Yonemura, Shigenobu; Kobayashi, Kensaku; Kiyonari, Hiroshi; Aizawa, Shinichi


    The processes of development leading up to gastrulation have been markedly altered during the evolution of amniotes, and it is uncertain how the mechanisms of axis formation are conserved and diverged between mouse and chick embryos. To assess the conservation and divergence of these mechanisms, this study examined gene expression patterns during the axis formation process in Chinese soft-shell turtle and Madagascar ground gecko preovipositional embryos. The data suggest that NODAL signaling, similarly to avian embryos but in contrast to eutherian embryos, does not have a role in epiblast and hypoblast development in reptilian embryos. The posterior marginal epiblast (PME) is the initial molecular landmark of axis formation in reptilian embryos prior to primitive plate development. Ontogenetically, PME may be the precursor of the primitive plate, and phylogenetically, Koller's sickle and posterior marginal zone in avian development may have been derived from the PME. Most of the genes expressed in the mouse anterior visceral endoderm (AVE genes), especially signaling antagonist genes, are not expressed in the hypoblast of turtle and gecko embryos, though they are expressed in the avian hypoblast. This study proposes that AVE gene expression in the hypoblast and the visceral endoderm could have been independently established in avian and eutherian lineages, similar to the primitive streak that has been independently acquired in these lineages. PMID:27174471

  3. Development of space-fertilized eggs and formation of primordial germ cells in the embryos of medaka fish

    Ijiri, K.

    In the second International Microgravity Laboratory (IML-2) mission in 1994, four small Japanese killifish (Medaka, Oryzias latipes) made a space travel of 15 days aboard a space shuttle. These four adult Medaka fish successfully mated in space for the first time among vertebrate animals. Moreover, the eggs they laid developed normally, at least in their external appearance, hatching as fry (baby fish) in space. Fish mated and laid eggs every day during the first week. Near the end of the mission most of the eggs had a well-developed body with two pigmented eyes. In total, 43 eggs were laid (detected), out of which 8 fry hatched in space, as truly `space-originated' babies. A further 30 fry hatched within 3 days after landing. This is the normal hatching rate, compared with the ground-based data. Among the 8 space-originated fry, four were killed for histological sections, and germ cells at the gonadal region were counted for each fry. Their numbers were in the range of the germ cells of the normal control fry (ground-kept samples). Thus, as embryos developed normally in their external appearance, inside the embryos the formation of primordial germ cells took place normally in space, and their migration to the genital ridges was not hindered by microgravity. The two of the remaining space-originated fry have grown up and been creating their offspring in the laboratory. This proved that the primordial germ cells formed in space were also normal from a functional point of view. The four space-travelled adult fish re-started mating and laying eggs on the 7th day after landing and continued to do so every day afterward. Fertilization rate and hatchability of these eggs were as high as the eggs laid by the laboratory-kept fish. This fact implies that in gametogenesis of adult fish, there are no specific stages of germ cells extremely susceptible to microgravity.

  4. Ni2+ treatment causes cement gland formation in ectoderm explants of Xenopus laevis embryo



    We found T-type calcium channel blocker Ni2+ can efficiently induce the formation of cement gland in Xenopus laevis animal cap explants.Nother T-typer specific calcium channel blocker Amiloride can also induce the formation of cement gland,while L-type specific calcium channel blocker Nifedipine as no inductive effect.These results may offer us an new approach to study the differentiation of cement gland through the change of intracelluar calcium concentration.

  5. Biological Effects Produced by Intense Pulsed Light (Xe-Cl on the Cartilage of the Tongue Chick Embryo Using Various Filters Efectos Biológicos Producidos por la Luz Pulsada Intensa (Xe-Cl sobre el Cartílago Lingual del Embrión de Pollo Usando Diversos Filtros

    R. E Avila


    Full Text Available The Laser used correctly in the medical practice offers clear advantages compared with traditional therapies. The improvement and even the elimination of many significant skin lesions can be achieved with reduced risks to patients. However, it is important to keep security measures and understand the possible effects on an experimental model. The chick embryo is a good model to evaluate the direct effects of non-ionizing radiation for its easy handling and availability. The purpose of this communication is to show our histological findings in organs of the chick embryo with and without protective barrier to be subjected to radiation excimer. We used the following issuers: intense pulsed light (excimer Xe-Cl laser of 308 nm wavelength. It was irradiated embryos through an open window on eggshells. Aseptically the eggs were kept for 24 hours in an incubator. The protective barriers were used with and without colored glass, latex, cellophane, paper, polycarbonate of different colors and thicknesses. The most outstanding results, with no barrier and barriers with transparent and green were intense marked congestion in capillaries, edema and focus the necrosis. We concluded that the tissue changes observed are consistent with possible side effects of these radiations fototérmicos we warned about possible side effects when they are applied indiscriminately. We believe it is important to explore different means to safeguard the safety of operators and patients.El láser utilizado correctamente en la práctica médica ofrece claras ventajas cuando se compara con las terapias tradicionales. La mejoría e incluso la eliminación significativa de muchas lesiones cutáneas se pueden lograr con riesgos reducidos para los pacientes. Sin embargo, es importante guardar medidas de seguridad y conocer los posibles efectos en un modelo experimental. El embrión de pollo es un buen modelo para evaluar los efectos directos de radiaciones no ionizantes por su f

  6. The chick chorioallantoic membrane as an in vivo xenograft model for Burkitt lymphoma

    Burkitt lymphoma (BL) is an aggressive malignancy that arises from B-cells and belongs to the group of Non-Hodgkin lymphomas (NHL). Due to the lack of appropriate in vivo models NHL research is mainly performed in vitro. Here, we studied the use of the chick chorioallantoic membrane (CAM) for the generation of human BL xenograft tumors, which we compared with known characteristics of the human disease. In order to generate experimental BL tumors, we inoculated human BL2B95 and BL2-GFP cells on the CAM. BL2B95 xenograft-tumors were grown for seven days and subsequently analyzed with transmission electron and immunofluorescence microscopy, as well as histological staining approaches. BL2-GFP cells were studied at regular intervals up to seven days, and their metastatic behavior was visualized with intravital immunofluorescence techniques. Xenografted BL2B95 cells formed solid tumors in the CAM model with a Ki67-index greater than 90%, preservation of typical tumor markers (CD10, CD19, CD20), a ‘starry sky’ morphology, production of agyrophilic fibers in the stroma, formation of blood and lymphatic vessels and lymphogenic dissemination of BL2B95 to distant sites. We identified macrophages, lymphocytes and heterophilic granulocytes (chick homolog of neutrophils) as the most abundant immune cells in the experimental tumors. BL2-GFP cells could be traced in real-time during their distribution in the CAM, and the first signs for their dissemination were visible after 2-3 days. We show that xenografted BL2B95 cells generate tumors in the CAM with a high degree of cellular, molecular and proliferative concord with the human disease, supporting the application of the CAM model for NHL research with a focus on tumor-stroma interactions. Additionally we report that BL2-GFP cells, grafted on the CAM of ex ovo cultured chick embryos, provide a powerful tool to study lymphogenic dissemination in real-time

  7. Interaction of human and chick DNA repair functions in UV-irradiated xeroderma pigmentosum-chick erythrocyte heterokaryons

    Fusion of chick erythrocytes with human primary fibroblasts results in the formation of heterokaryons in which the inactive chick nuclei become reactivated. The expression of chick DNA repair functions was investigated by the analysis of the DNA repair capacity after exposure to ultraviolet (UV) irradiation of such heterokaryons obtained after fusion of chick erythrocytes with normal human or xeroderma pigmentosum (XP) cells of complementation groups A, B, C and D. Unscheduled DNA synthesis (UDS) in normal human nuclei in these heterokaryons is suppressed during the first 2-4 days after fusion. The extent and duration of this suppression is positively correlated with the number of chick nuclei in the heterokaryons. Suppression is absent in heterokaryons obtained after fusion of chicken embryonic fibroblasts with XP cells (complementation group A and C). Restoration of DNA repair synthesis is found after fusion in XP nuclei of all complementation groups studied. It occurs rapidly in XP group A nuclei, starting one day after fusion and reaching near normal human levels after 5-8 days. In nuceli of the B, C and D group increased levels of UDS are found 5 days after fusion. At 8 days after fusion the UDS level is about 50% of that found in normal human nuclei. The pattern of UDS observed in the chick nuclei parallels that of the human counterpart in the fusion. In heterokaryons obtained after fusion of chick fibroblasts with XP group C cells UDS remains at the level of chick cells. These suggest that reactivation of chick erythrocyte nuclei results in expression of repair functions which are able to complement the defects in the XP complementation groups A, B, C and D

  8. High-resolution Magnetic Resonance Imaging analysis of breast cancer xenograft on the chick chorioallantoic membrane

    ZUO, ZHI


    In this thesis an age-adapted cooling regime for immobilization of the chick embryo is proposed. Reliable immobilization completely avoided motion artifacts, enabling high resolution magnetic resonance imaging (MRI) of the chicken embryo and also tumor xenograft on the chorioallantoic membrane (CAM). Tumor growth monitoring was firstly evaluated after xenotransplantation of MDA-MB-231 cells on the CAM. Tumor volumes were monitored from day 4 to day 9 after grafting applying a T2-weighted R...

  9. Absorption of the trypan blue in the proximal tubules of the chick mesonephros

    Jirsová, Z.; Zemanová, Zdeňka; Martínek, J.

    Brno : Masarykova univerzita, 2005. s. 56-56. ISBN 80-210-3793-8. [Congress of the Czech anatomical society /43./, Lojda symposium /42./. 04.09.2005-07.09.2005, Brno] R&D Projects: GA ČR(CZ) GA304/04/0972 Keywords : chick embryo * mesonephros * proximal tubule * trypan blue Subject RIV: EA - Cell Biology

  10. Changes in activation sequence of embryonic chick atria correlate with developing myocardial architecture

    Sedmera, David; Wessels, A.; Trusk, T. C.; Thompson, R. P.; Hewett, K. W.; Gourdie, R. G.


    Roč. 291, 4 (2006), H1646-H1652. ISSN 0363-6135 Grant ostatní: NIH RR-16434; NIH HL-50582; NIH HL-52813; NIH HD-39446 Institutional research plan: CEZ:AV0Z50450515 Keywords : chick embryo * optical mapping * conduction system Subject RIV: ED - Physiology Impact factor: 3.724, year: 2006

  11. Increased ventricular preload is compensated by myocyte proliferation in normal and hypoplastic fetal chick left ventricle

    Dealmeida, A.; McQuinn, T. C.; Sedmera, David


    Roč. 100, - (2007), s. 1363-1370. ISSN 0009-7330 Institutional research plan: CEZ:AV0Z50450515 Keywords : chick embryo * hemodynamics * fetal surgery * hypoplastic left heart syndrome Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery Impact factor: 9.721, year: 2007

  12. Brain asymmetry modulates perception of biological motion in newborn chicks (Gallus gallus).

    Rugani, Rosa; Rosa Salva, Orsola; Regolin, Lucia; Vallortigara, Giorgio


    Few light-points on the joints of a moving animal give the impression of biological motion (BM). Day-old chicks prefer BM to non-BM, suggesting a conserved predisposition to attend to moving animals. In humans and other mammals a network of regions, primarily in the right hemisphere, provides the neural substrate for BM perception. However, this has not been investigated in avians. In birds the information from each eye is mainly feeding to the contralateral hemisphere. To study brain asymmetry, we recorded the eye spontaneously used by chicks to inspect a BM stimulus. We also investigated the effect of lateralization following light exposure of the embryos. In Experiment 1, highly lateralized chicks aligned with the apparent direction of motion only when they were exposed to a BM-stimulus moving rightward first, monitoring it with the left-eye-system. In Experiment 2 weakly lateralized chicks did not show any behavioral asymmetry. Moreover, they counter aligned with the apparent direction of motion. Brain lateralization affects chicks behavior while processing and approaching a BM stimulus. Highly lateralized chicks aligned their body with the apparent direction of the BM, a behavior akin to a following response, monitoring the stimulus preferentially with their left eye. This suggests a right hemisphere dominance in BM processing. Weakly lateralized chicks counter-aligned with the apparent direction of the BM, facing it during interaction, and monitored it equally with both eyes. Environmental factors (light stimulation) seem to affect the development of lateralization, and consequently social behavior. PMID:25930217

  13. Proteome analysis of chick embryonic cerebrospinal fluid.

    Parada, Carolina; Gato, Angel; Aparicio, Mariano; Bueno, David


    During early stages of embryo development, the brain cavity is filled with embryonic cerebrospinal fluid (E-CSF), a complex fluid containing different protein fractions that contributes to the regulation of the survival, proliferation and neurogenesis of the neuroectodermal stem cells. Using 2-DE, protein sequencing and database searches, we identified and analyzed the proteome of the E-CSF from chick embryos (Gallus gallus). We identified 26 different gene products, including proteins related to the extracellular matrix, proteins associated with the regulation of osmotic pressure and metal transport, proteins related to cell survival, MAP kinase activators, proteins involved in the transport of retinol and vitamin D, antioxidant and antimicrobial proteins, intracellular proteins and some unknown proteins. Most of these gene products are involved in the regulation of developmental processes during embryogenesis in systems other than E-CSF. Interestingly, 14 of them are also present in adult human CSF proteome, and it has been reported that they are altered in the CSF of patients suffering neurodegenerative diseases and/or neurological disorders. Understanding these molecules and the mechanisms they control during embryonic neurogenesis is a key contribution to the general understanding of CNS development, and may also contribute to greater knowledge of these human diseases. PMID:16287170

  14. An ultraviolet-sensitive maternal mRNA encoding a cytoskeletal protein may be involved in axis formation in the ascidian embryo

    Ultraviolet (uv) irradiation of the vegetal hemisphere of fertilized eggs during ooplasmic segregation inhibits subsequent gastrulation and axis formation in ascidian embryos. The molecular basis of this phenomenon was investigated in by comparing in vivo protein synthesis and in vitro mRNA translation in normal and uv-irradiated embryos of the ascidian Styela clava. Analysis of protein synthesis by [35S]methionine incorporation, two-dimensional (2D) gel electrophoresis, and autoradiography showed that only 21 of 433 labeled polypeptides were missing or decreased in labeling intensity in uv-irradiated embryos. The most prominent of these was a 30,000 molecular weight (pI 6.0) polypeptide (p30). Extraction of gastrulae with the nonionic detergent Triton X-100 showed that p30 is retained in the detergent insoluble residue, suggesting that it is associated with the cytoskeleton. Several lines of evidence suggest that p30 may be involved in axis formation. First, p30 labeling peaks during gastrulation, when the embryonic axis is being established. Second, axis formation and p30 labeling are abolished by the same threshold uv dose, which is distinct from that required to inactivate muscle cell development. Third, the uv sensitivity period for abolishing p30 labeling and axis formation are both restricted to ooplasmic segregation. In vitro translation of egg RNA followed by 2D gel electrophoresis and autoradiography of the protein products showed that p30 is encoded by a maternal mRNA. The translation of p30 mRNA was abolished by uv irradiation of fertilized eggs during ooplasmic segregation suggesting that this message is a uv-sensitive target. The results are consistent with the hypothesis that uv irradiation blocks gastrulation and axis formation by inhibiting the translation of maternal mRNA localized in the vegetal hemisphere of the fertilized egg

  15. Phosphorylation of the zebrafish M6Ab at serine 263 contributes to filopodium formation in PC12 cells and neurite outgrowth in zebrafish embryos.

    Kai-Yun Huang

    Full Text Available BACKGROUND: Mammalian M6A, a member of the proteolipid protein (PLP/DM20 family expressed in neurons, was first isolated by expression cloning with a monoclonal antibody. Overexpression of M6A was shown to induce filopodium formation in neuronal cells; however, the underlying mechanism of is largely unknown. Possibly due to gene duplication, there are two M6A paralogs, M6Aa and M6Ab, in the zebrafish genome. In the present study, we used the zebrafish as a model system to investigate the role of zebrafish M6Ab in filopodium formation in PC12 cells and neurite outgrowth in zebrafish embryos. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrated that zebrafish M6Ab promoted extensive filopodium formation in NGF-treated PC12 cells, which is similar to the function of mammalian M6A. Phosphorylation at serine 263 of zebrafish M6Ab contributed to this induction. Transfection of the S263A mutant protein greatly reduced filopodium formation in PC12 cells. In zebrafish embryos, only S263D could induce neurite outgrowth. CONCLUSIONS/SIGNIFICANCE: Our results reveal that the phosphorylation status of zebrafish M6Ab at serine 263 is critical for its role in regulating filopodium formation and neurite outgrowth.

  16. Inhibition of MEK and GSK3 supports ES cell-like domed colony formation from avian and reptile embryos.

    Nakanoh, Shota; Okazaki, Kenji; Agata, Kiyokazu


    As amniotes diversified, mammals may have modified mechanisms of cellular pluripotency along with the acquisition of a placenta. What then defined pluripotent states in the ancestral amniotes? To study the evolutionary background of pluripotency in amniotes, we tested the effects of extracellular effectors on primary culture cells from avian and reptile embryos in serum-free medium. When treated with a combination of a MEK inhibitor and a GSK3 inhibitor (2i condition), chicken early embryos formed domed colonies (DCs), which were morphologically indistinguishable from the colonies formed by mouse and rat naïve embryonic stem cells. However, no DCs formed when cells from further-developed embryos were cultured in the 2i condition, indicating that there is a clear boundary of DC-forming ability at around the stage of primitive streak elongation. Quail embryos at the blastoderm and cleavage stages also formed DCs in the 2i condition, which is consistent with the notion that the appearance of DCs corresponds with the presence of pluripotent cells in embryos. Gecko blastoderms also formed DCs in the 2i condition, but gastrulas did not. ERK activation by bFGF caused an effect opposite to that of the 2i condition, namely, it dispersed colonies of cells even from early embryos in all species examined. These results suggest that the regulation of pluripotency by FGF/ERK signaling may date back at least to the common ancestor of mammals, birds, and reptiles. However, gene expression analysis indicated the possibility that mammalian pluripotency transcription factors function differently in non-mammalian amniotes. PMID:23829214

  17. Direct Somatic Embryo Formation from Roots of In Vitro-Seedlings of Oil Palm (Elaeis Guineesis Jacq.)

    Stapaporn KERDSUWAN; Sompong TE-CHATO


    The induction of somatic embryos (SEs) directly from the roots of oil palm plantlets raised in vitro-seedling and origin of somatic embryos. SEs were directly induced from the roots of oil palm plantlets regenerated from immature oil palm fruit (3 months after pollination: 3MAP) and provided by Golden Tenera Company. The plants were cultured on OPCM medium supplemented with 3 % sucrose, 200 mg/l ascorbic acid and 0.5 mg/l NAA. The culture was maintained under light conditions at an intensi...

  18. Amiodarone Induces Overexpression of Similar to Versican b to Repress the EGFR/Gsk3b/Snail Signaling Axis during Cardiac Valve Formation of Zebrafish Embryos

    Lee, Hung-Chieh; Lo, Hao-Chan; Lo, Dao-Ming; Su, Mai-Yan; Hu, Jia-Rung; Wu, Chin-Chieh; Chang, Sheng-Nan; Dai, Ming-Shen; Tsai, Chia‐Ti; Tsai, Huai-Jen


    Although Amiodarone, a class III antiarrhythmic drug, inhibits zebrafish cardiac valve formation, the detailed molecular pathway is still unclear. Here, we proved that Amiodarone acts as an upstream regulator, stimulating similar to versican b (s-vcanb) overexpression at zebrafish embryonic heart and promoting cdh-5 overexpression by inhibiting snail1b at atrioventricular canal (AVC), thus blocking invagination of endocardial cells and, as a result, preventing the formation of cardiac valves. A closer investigation showed that an intricate set of signaling events ultimately caused the up-regulation of cdh5. In particular, we investigated the role of EGFR signaling and the activity of Gsk3b. It was found that knockdown of EGFR signaling resulted in phenotypes similar to those of Amiodarone-treated embryos. Since the reduced phosphorylation of EGFR was rescued by knockdown of s-vcanb, it was concluded that the inhibition of EGFR activity by Amiodarone is s-vcanb-dependent. Moreover, the activity of Gsk3b, a downstream effector of EGFR, was greatly increased in both Amiodarone-treated embryos and EGFR-inhibited embryos. Therefore, it was concluded that reduced EGFR signaling induced by Amiodarone treatment results in the inhibition of Snail functions through increased Gsk3b activity, which, in turn, reduces snail1b expression, leading to the up-regulation the cdh5 at the AVC, finally resulting in defective formation of valves. This signaling cascade implicates the EGFR/Gsk3b/Snail axis as the molecular basis for the inhibition of cardiac valve formation by Amiodarone. PMID:26650936

  19. In Ovo Electroporations of HH Stage 10 Chicken Embryos

    Blank, Marissa C.; Chizhikov, Victor; Millen, Kathleen J.


    Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology. With the advent of molecular biological techniques, the chick has become a useful system in which to study gene regulation and function. By electroporating DNA or RNA constructs into the developing chicken embryo, genes can be expressed or knocked down in order to analyze in vivo gene function. Similarly, reporter constructs can be used for fate mapping or to e...

  20. 不同鼻咽癌细胞株对鸡胚尿囊膜模型血管生成影响的比较研究%Angiogenic effect of different nasopharyngeal carcinoma cell lines on the chick embryo chorioallantoic membrane

    林少雄; 李湘平; 鲁娟; 梁碧君; 刘雄


    Objective The purpose of this study was to compare the angiogenic effect of different nasopharyngeal carcinoma (NPC) cell lines on the chick embryo chorioallantoic membrane (CAM). Methods Six days old fertilized eggs were used in this study. A square window on the shell was cut, and the the NPC cell lines 5-8F, 6-10B and CNE-2 were inocluated into the CAM (n=10) with PBS as blank control. The eggs were then incubated for 6 days before the enumeration of the vessels number (VN). The ratios between the area covered by the vessel and the CAM (VA/ CAM) were analyzed. Results All the 3 NPC cell lines grow on the CAMs (100%) with a cell number of 2×l06/CAM. The VN was significantly decreased (P<0.05). The VA/CAM of 5-8F was significantly higher than the other two cell lines (P<0.05). The VA/CAM from the three NPC cell lines inoculated embryos were significantly higher than the control (P<0.05). Conclusion The results indicated that the angiogenic effects of these NPC cell line are different with each other, therefore the selection of these NPC cell lines may depend on the different CAM experimental conditions.%目的 比较不同鼻咽癌细胞株在鸡胚尿囊膜模型上促进血管生成能力的差异.方法 选用6日龄鸡胚,开窗后分别种植5-8F、6-10B及CNE-2三种鼻咽癌细胞株,三种细胞株细胞数为2×106/鸡胚者各一组,细胞数为5×105/鸡胚者各一组,另一组为对照组(PBS),每组10个鸡胚.孵育6日后,统计分析新生血管数及血管面积/鸡胚面积比.结果 种植细胞数为5×105/鸡胚时,5-8F、6-10B细胞部分鸡胚有移植瘤形成,CNE-2细胞鸡胚均未见成瘤.种植细胞数为2× 106/鸡胚时,三种鼻咽癌细胞株均可100%成瘤,其新生血管数依次递减,分别为(38.7±2.50)、(33.5±4.43)、(29.7±2.71),差异有统计学意义(P<0.05);血管面积/鸡胚面积比分别为(22.2±2.18)%、(18.7±2.45)%、(16.9±2.62)%,均高于对照组的(9.5±1.86)

  1. Neurofibrillar differentiation in 8- to 14- somites chick embryos

    Eros Abrantes Erhart


    Full Text Available Com o intuito de estudar o início da diferenciação neurofibrilar de neuro-blastos foram preparados cortes seriados de embriões de galinha impregnados pela prata, modificação de De Castro do método de Cajal. A diferenciação neurofibrilar inicia-se em fases mais jovens que as descritas na literatura. Em embriões de galinha de 8 somitos, os primeiros neuroblastos são encontrados na região rombencefálica do tubo neural, ainda aberto, na altura da invaginação ótica. Aos 12 somitos, os neuroblastos começam a aparecer, também, na junção prosencéfalo-mesencefálica, e aos 14 somitos, bem mais diferenciados, apresentam evidente, aumento numérico. Os neuroblastos localizados na junção prosencéfalo-mesencefálica enviam seus axônios caudalmente, constituindo o que parece ser o primórdio do fascículo longitudinal medial. Os neuroblastos rombencefálicos enviam seus axônios ventral, cranial e caudalmente, na espessura das paredes neurais, constituindo, provàvelmente, os primórdios dos sistemas cordonais e de associação; por apresentarem-se mais ou menos grupados nos níveis do trigêmio, facial e vago, talvez induzam, também, a pré-organização dos primórdios dos núcleos motores dos nervos V, VII e do complexo vagal.

  2. Trichloroethylene: a cardiac teratogen in developing chick embryos.

    Loeber, C P; Hendrix, M J; Diez De Pinos, S; Goldberg, S J


    Prior studies have evaluated the teratogenicity of TCE, a contaminant of drinking water. However, none specifically examined effects of TCE on cardiogenesis. The purpose of this study was to determine if TCE is a cardiac teratogen in early embryogenesis in an avian model. Fertile White Leghorn chicken eggs were incubated under standard conditions. At stage 6, 12, 18, or 23, TCE, in concentrations of 5 to 25 microM (2 to 28 micrograms/g body weight) was injected into the air space of the egg (vol = 0.03 ml). Mineral oil and saline served as control solutions. For this double-blinded study, solutions were coded and remained so until all observations were made and recorded. Embryonic hearts (n = 1055) were examined at stage 29, 34, or 44. Gross examination was performed, followed by microdissection. Cardiac malformations were found in 7.3% of TCE-treated hearts, compared to 2.3% of saline controls (p less than 0.01), and 1.5% of mineral oil controls (p less than 0.001). No significant difference in incidence of malformations was found when comparing saline and mineral oil controls. Cardiac defects include septal defects, cor biloculare, conotruncal abnormalities, atrioventricular canal defects, and abnormal cardiac muscle. These data demonstrate that TCE is a cardiac teratogen in an avian model. PMID:3205631

  3. Effect of Cyclophosphamide on Neural Tube Development in Chick Embryos



    Full Text Available Cyclophosphamide is a nitrogen mustard alkylating agent. CP has potent immunosuppressive properties and issued clinically in a number of autoimmune disorders like Wegener’s granulomatosis, rheumatoid arthritis, nephritic syndrome, systemic lupus erythematous and has also been used to prevent organ rejection after transplantation. In the present study fertilized eggs were administered with cyclophosphamide and the development of neural tube was studied after 21 days. The histological and gross features of neural tube were identified. Cyclophosphamide cytotoxicity results in depression of proliferation of cell activity associated with malformations and embryonic death. Injection of the drug causes depression of mitotic activity by day 2 which produces malformations.

  4. IGF-1/IGFBP-1 increases blastocyst formation and total blastocyst cell number in mouse embryo culture and facilitates the establishment of a stem-cell line

    Hsu Teng-Tsao


    Full Text Available Abstract Background Apoptosis occurs frequently for blastocysts cultured in vitro, where conditions are suboptimal to those found in the natural environment. Insulin-like growth factor-1 (IGF-1 plays an important role in preventing apoptosis in the early development of the embryo, as well as in the progressive regulation of organ development. We hypothesize that IGF-1 and its dephosphorylated binding protein (IGFBP-1 may be able to improve embryo culture with an associated reduced cell death, and that the resultant increase in the total cell number of the embryo could increase the chances of establishing an embryonic stem-cell line. Results In vivo fertilized zygotes were cultured in medium containing supplementary IGF-1, or IGFBP-1/IGF-1. The stages of the resultant embryos were evaluated at noon on day five post-hCG injection. The extent of apoptosis and necrosis was evaluated using Annexin V and propidium iodine staining under fluorescent microscopy. The establishment of embryonic stem-cell lines was performed using the hatching blastocysts that were cultured in the presence of IGF-1 or IGFBP-1/IGF-1. The results show that the rate of blastocyst formation in a tissue-culture system in the presence of IGF-1 was 88.7% and IGFBP-1/IGF-1 it was 94.6%, respectively, and that it was significantly greater than the figure for the control group (81.9%. IGFBP-1/IGF-1 also resulted in a higher hatching rate than was the case for the control group (68.8% vs. 48.6% respectively. IGF-1 also increased the number of Annexin V-free and propidium iodine-free blastocysts in culture (86.8% vs. 75.9% respectively. Total cell number of blastocyst in culture was increased by 18.9% for those examples cultured with dephosphorylated IGFBP-1/IGF-1. For subsequent stem-cell culture, the chances of the successful establishment of a stem-cell line was increased for the IGF-1 and IGFBP-1/IGF-1 groups (IGF-1 vs. IGFBP-1/IGF-1 vs. control: 45.8% vs. 59.6% vs. 27

  5. Avian maternal response to chick distress

    Edgar, J. L.; Lowe, J. C.; Paul, E. S.; C. J. Nicol


    The extent to which an animal is affected by the pain or distress of a conspecific will depend on its capacity for empathy. Empathy most probably evolved to facilitate parental care, so the current study assessed whether birds responded to an aversive stimulus directed at their chicks. Domestic hens were exposed to two replicates of the following conditions in a counterbalanced order: control (C; hen and chicks undisturbed), air puff to chicks (APC; air puff directed at chicks at 30 s interva...

  6. Effect of species-specific sound stimulation on the development and hatching of broiler chicks.

    Tong, Q; McGonnell, I M; Romanini, C E B; Bergoug, H; Roulston, N; Exadaktylos, V; Berckmans, D; Bahr, C; Guinebretière, M; Eterradossi, N; Garain, P; Demmers, T


    1. Previous research has reported that chicken embryos develop a functionary auditory system during incubation and that prenatal sound may play an important role in embryo development and alter the hatch time. In this study the effects of prenatal auditory stimulation on hatch process, hatch performance, the development of embryo and blood parameters were investigated. 2. Four batches of Ross 308 broiler breeder eggs were incubated either in control or in sound-stimulated groups. The sound-stimulated embryos were exposed to a discontinuous sound of species-specific calls by means of a speaker at 72 dB for 16 h a day: maternal calls from d 10 to d 19 of incubation time and embryo/chick calls from d 19 until hatching. The species-specific sound was excluded from the control group. 3. The onset of hatch was delayed in the sound-stimulated group compared to the controls. This was also supported by comparison of the exact hatching time of individual focal chicks within the two groups. However, the sound-stimulated embryos had a lower hatchability than the control group, mainly due to significantly increased numbers of late deaths. 4. The embryos exhibited a similar growth pattern between the sound-stimulated group and the control group. Although sound exposure decreased body weight at d 16, no consistent effect of sound on body weight at incubation stage was observed. Species-specific sound stimulation also had no impact on chick quality, blood values and plasma corticosterone concentrations during hatch. PMID:25559058

  7. Multiple embryos in the Lepidocyclina pustulosa group as possible indicators of palaeoenvironmental conditions: The case of the Late Eocene Toluviejo Formation (Sinú Domain, Caribbean, Colombia

    Baumgartner-Mora, Claudia; Baumgartner, Peter O.; Rincón Martínez, Daniel; Salazar Ortiz, Edward A.


    The Middle-Late Eocene Toluviejo Formation of the Sinú-San Jacinto folded belt (Caribbean, Colombia) contains 15-75 m thick, grey, massive limestone sequences that are interbedded with terrigenous nearshore to offshore mudstones to quartzose sandstones and conglomerates. The formation accumulated in a transitional continent-ocean setting, probably on oceanic crust. We studied over 80 polished thin sections under light microscopy and cathodoluminescence (CL) to analyse carbonate microfacies and Larger Benthic Foraminifera (LBF). The overall facies distribution is 3-fold: (1) Facies close to the shoreline are dominated by low diversity operculinid assemblages, rich in detrital elements. (2) Shallow offshore facies are characterized by round lepidocyclinids, associated with, and often encrusted by, corallinacean algae, Sporolithon and structureless microbial crusts. The matrix is rich in carbonate/detrital mud and sand-sized detritals. (3) Distal offshore facies on structural highs show abundance of flat, current-sorted lepidoclinids without a noticeable detrital component. Facies 1 and 2 are clearly under the influence of suspension- and dissolved-nutrient input, probably carried offshore by freshwater lids of river plumes. Facies (2) contains abundant specimens of the Lepidocyclina pustulosa group of which the macrospheric forms show complicated embryonic apparatuses, which suppose double or multiple embryos. Detailed observation under CL allows to count up to 6-8 embryos, often seen aligned in the equatorial plane. The embryonic apparatus occupies often more than half of the diameter of specimens. The tendency towards large, very flat embryonic apparatuses (diameter measured in axial cuts up to 2,5 mm for a thickness of 0.2-0.3 mm) is closely correlated with abundant coralgal and microbial encrustations, oxide-stained carbonate/detrital mud and probably some preserved organic matter. The presence of multiple embryos has been reported by several authors and has

  8. The avian embryo responding to microgravity of space flight

    Hullinger, Ronald L.


    Of all the many potential and real microenvironmental influences, only gravity would appear to have remained relatively constant and ubiquitous for developing organisms. Histo- and organogenesis as well as differential growth of the embryo and fetus may have evolved with a constant environmental factor of gravity. Chick embryos of 2-day and 9-day stages of incubation were flown in an incubator on the Space Shuttle during a 9-day mission. Significant differences in embryo response to this microgravity environment were observed. This paper offers an analysis and suggests mechanisms which may contribute to these results.

  9. Pattern of trypan blue storage in the chick embryonic kidney reflects development of absorptive capacity of the proximal tubules

    Zemanová, Zdeňka


    Roč. 15, č. 3-4 (2004), s. 160-165. ISSN 1470-2061 R&D Projects: GA ČR GA304/04/0972 Institutional research plan: CEZ:AV0Z5011922 Keywords : chick embryo * mesonephric nephrons * trypan blue Subject RIV: ED - Physiology Impact factor: 0.724, year: 2004

  10. Fibroblast Growth Factor-2 regulates proliferation of cardiac myocytes in normal and hypoplastic left ventricles in the developing chick

    Dealmeida, A.; Sedmera, David


    Roč. 19, č. 2 (2009), s. 159-169. ISSN 1047-9511 R&D Projects: GA ČR GA304/08/0615 Institutional research plan: CEZ:AV0Z50450515; CEZ:AV0Z50110509 Keywords : Chick embryo * Hypoplastic left heart syndrome * Adenovirus Subject RIV: EA - Cell Biology Impact factor: 1.183, year: 2009

  11. Bambi and Sp8 Expression Mark Digit Tips and Their Absence Shows That Chick Wing Digits 2 and 3 Are Truncated

    Jesús C Casanova; Claudio Badia-Careaga; Verónica Uribe; Juan José Sanz-Ezquerro


    An often overlooked aspect of digit development is the special nature of the terminal phalanx, a specialized structure with characteristics distinct from other phalanges, for example the presence of ectodermal derivatives such as nails and claws. Here, we describe the unique ossification pattern of distal phalanges and characteristic gene expression in the digit tips of chick and duck embryos. Our results show that the distal phalanx of chick wing digit 1 is a genuine tip with a characteristi...

  12. Toward a Deterministic Model of Planetary Formation VI: Dynamical Interaction and Coagulation of Multiple Rocky Embryos and Super-Earth Systems around Solar Type Stars

    Ida, S


    Radial velocity and transit surveys indicate that solar-type stars bear super-Earths, with mass and period up to ~ 20 M_E and a few months, are more common than those with Jupiter-mass gas giants. In many cases, these super-Earths are members of multiple-planet systems in which their mutual dynamical interaction has influenced their formation and evolution. In this paper, we modify an existing numerical population synthesis scheme to take into account protoplanetary embryos' interaction with their evolving natal gaseous disk, as well as their close scatterings and resonant interaction with each other. We show that it is possible for a group of compact embryos to emerge interior to the ice line, grow, migrate, and congregate into closely-packed convoys which stall in the proximity of their host stars. After the disk-gas depletion, they undergo orbit crossing, close scattering, and giant impacts to form multiple rocky Earths or super-Earths in non-resonant orbits around ~ 0.1AU with moderate eccentricities of ~...


    Radial velocity and transit surveys indicate that solar-type stars bear super-Earths, with masses up to ∼20 M + and periods up to a few months, that are more common than those with Jupiter-mass gas giants. In many cases, these super-Earths are members of multiple-planet systems in which their mutual dynamical interaction has influenced their formation and evolution. In this paper, we modify an existing numerical population synthesis scheme to take into account protoplanetary embryos' interaction with their evolving natal gaseous disks, as well as their close scatterings and resonant interaction with each other. We show that it is possible for a group of compact embryos to emerge interior to the ice line, grow, migrate, and congregate into closely packed convoys which stall in the proximity of their host stars. After the disk-gas depletion, they undergo orbit crossing, close scattering, and giant impacts to form multiple rocky Earths or super-Earths in non-resonant orbits around ∼0.1 AU with moderate eccentricities of ∼0.01-0.1. We suggest that most refractory super-Earths with periods in the range of a few days to weeks may have formed through this process. These super-Earths differ from Neptune-like ice giants by their compact sizes and lack of a substantial gaseous envelope.

  14. In ovo electroporations of HH stage 10 chicken embryos.

    Blank, Marissa C; Chizhikov, Victor; Millen, Kathleen J


    Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology. With the advent of molecular biological techniques, the chick has become a useful system in which to study gene regulation and function. By electroporating DNA or RNA constructs into the developing chicken embryo, genes can be expressed or knocked down in order to analyze in vivo gene function. Similarly, reporter constructs can be used for fate mapping or to examine putative gene regulatory elements. Compared to similar experiments in mouse, chick electroporation has the advantages of being quick, easy and inexpensive. This video demonstrates first how to make a window in the eggshell to manipulate the embryo. Next, the embryo is visualized with a dilute solution of India ink injected below the embryo. A glass needle and pipette are used to inject DNA and Fast Green dye into the developing neural tube, then platinum electrodes are placed parallel to the embryo and short electrical pulses are administered with a pulse generator. Finally, the egg is sealed with tape and placed back into an incubator for further development. Additionally, the video shows proper egg storage and handling and discusses possible causes of embryo loss following electroporation. PMID:18989448

  15. Partial purification and biological activities and properties of chick growth factors.



    Full Text Available Cellular stimulating factors on cell proliferation in the supernatants of chick embryo carcases and adult muscles were studied. There were plural stimulating factors in embryonic and adult muscular supernatants that promoted cell proliferation without any supplement of sera and other materials. Salting-out methods with ammonium sulfate, ethanol fractionation, and isoelectric precipitation were used to isolate the stimulating factors, and these three methods proved the presence of plural stimulants on cell proliferation in the supernatants of chick embryo and adult muscles. The stimulants had altered physico-chemical properties and biological activities due to embryological development. The embryonic stimulants enhanced the synthesis of DNA and protein remarkably, and RNA synthesis in whole cell systems slightly. The muscular stimulants enhanced protein synthesis without any stimulation of DNA and RNA synthesis. Partial purification of the stimulants from the ethanol fractions was performed by DEAE-cellulose chromatography and Sephadex gel chromatography.

  16. Altered Cardiac Function and Ventricular Septal Defect in Avian Embryos Exposed to Low-Dose Trichloroethylene

    Echoleah S Rufer; Hacker, Timothy A.; Flentke, George R.; Drake, Victoria J.; Brody, Matthew J.; Lough, John; Susan M. Smith


    Trichloroethylene (TCE) is the most frequently reported organic groundwater contaminant in the United States. It is controversial whether gestational TCE exposure causes congenital heart defects. The basis for TCE’s proposed cardiac teratogenicity is not well understood. We previously showed that chick embryos exposed to 8 ppb TCE during cardiac morphogenesis have reduced cardiac output and increased mortality. To further investigate TCE’s cardioteratogenic potential, we exposed in ovo chick ...

  17. Neurochemical phenotype and birthdating of specific cell populations in the chick retina

    Karin da Costa calaza; Patricia F. Gardino


    The chick embryo is one of the most traditional models in developing neuroscience and its visual system has been one of the most exhaustively studied. The retina has been used as a model for studying the development of the nervous system. Here, we describe the morphological features that characterize each stage of the retina development and studies of the neurogenesis period of some specific neurochemical subpopulations of retinal cells by using a combination of immunohistochemistry and autor...

  18. Effects of mechanical loading on early conduction system differentiation in the chick

    Šaňková, Barbora; Machálek, J.; Sedmera, David


    Roč. 298, č. 5 (2010), H1571-H1576. ISSN 0363-6135 R&D Projects: GA ČR(CZ) GA304/08/0615 Institutional research plan: CEZ:AV0Z50110509; CEZ:AV0Z50450515 Keywords : chick embryo * optical mapping * cardiac conduction system Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery Impact factor: 3.880, year: 2010

  19. Zinc bioavailability in the chick

    Methods for assessing zinc bioavailability were evaluated in the chick. A low-zinc chick diet was developed using rehydrated, spray-dried egg white autoclaved at 121 C for 30 min as the primary protein source. The relative bioavailability of zinc from soy flour and beef was determined by whole-body retention of extrinsic 65Zn, and in slope ratio assays for growth rate and tissue zinc. Compared to zinc carbonate added to an egg white-based diet, all methods gave similar estimates of approximately 100% zinc bioavailability for beef but estimates for soy flour varied widely. The slope ratio assay for growth rate gave the best estimate of zinc bioavailability for soy flour. True absorption, as measured by percent isotope retention from extrinsically labeled soy flour, was 47%

  20. Effect of cultivars and culture medium on callus formation and plant regeneration from mature embryos of wheat triticum aestivum L.)

    Experiments were conducted to investigate callus induction, plant regeneration and somaclonal variations in 9 locally developed wheat cultivars viz., Ghaznavi-98, Fakhr-e-Sarhad, Inqilab-91, Tatara, Takbeer, Margalla, Pirsabak-85, SARC-3 and Khattakwal and one line from ICARDA (ICP-3) on hormones free and various concentrations of 2,4-dichorophenoxyaceitc acid (2, 4-D) in LS/MS medium. Cultivars responded differently to medium and 2, 4-D concentration for callus induction. Maximum calli induction (67.5%) was noted in ICP-3 followed by SARC-3 (65.5%) on MS. In our studies MS medium was more effective for all the wheat cultivars compared with LS medium for callus induction. When the effect of different concentration of 2, 4-D on callus induction was studied, MS medium containing 2 mg l/sup -1/ 2, 4-D produced the greatest number of calli. Morphology of these cultivars were further studied on both LS/MS medium containing only 2 mg l/sup -1/ 2, 4-D. Embroyogenic and non-embryogenic calli were observed in all the cultivars studied. Embryogenic calli were generally compact and rapidly growing whereas non-embryogenic calli were loose and slow growing. For regeneration, LS/MS medium was supplemented with various combinations of IAA and BAP. Significant differences were detected in plant regeneration, culture efficiency and regeneration capacity when mature embryos of 9 locally developed cultivars and one line from ICARDA (ICP-3) were compared. Six cultivars responded efficiently to LS medium while four showed better performance on MS medium for plant regeneration. Majority of the somaclones regenerated in this study were found to be inferior for agronomic traits except plant height, days to heading and maturity when compared with their respective controls. Our results showed that callus derived from this wheat cultivar is amendable and could be used for genetic transformation studies. (author)

  1. FoxA4 Favours Notochord Formation by Inhibiting Contiguous Mesodermal Fates and Restricts Anterior Neural Development in Xenopus Embryos

    Murgan, Sabrina; Castro Colabianchi, Aitana Manuela; Monti, Renato José; Boyadjián López, Laura Elena; Aguirre, Cecilia E.; Stivala, Ernesto González; López, Silvia L.


    In vertebrates, the embryonic dorsal midline is a crucial signalling centre that patterns the surrounding tissues during development. Members of the FoxA subfamily of transcription factors are expressed in the structures that compose this centre. Foxa2 is essential for dorsal midline development in mammals, since knock-out mouse embryos lack a definitive node, notochord and floor plate. The related gene foxA4 is only present in amphibians. Expression begins in the blastula –chordin and –noggin expressing centre (BCNE) and is later restricted to the dorsal midline derivatives of the Spemann's organiser. It was suggested that the early functions of mammalian foxa2 are carried out by foxA4 in frogs, but functional experiments were needed to test this hypothesis. Here, we show that some important dorsal midline functions of mammalian foxa2 are exerted by foxA4 in Xenopus. We provide new evidence that the latter prevents the respecification of dorsal midline precursors towards contiguous fates, inhibiting prechordal and paraxial mesoderm development in favour of the notochord. In addition, we show that foxA4 is required for the correct regionalisation and maintenance of the central nervous system. FoxA4 participates in constraining the prospective rostral forebrain territory during neural specification and is necessary for the correct segregation of the most anterior ectodermal derivatives, such as the cement gland and the pituitary anlagen. Moreover, the early expression of foxA4 in the BCNE (which contains precursors of the whole forebrain and most of the midbrain and hindbrain) is directly required to restrict anterior neural development. PMID:25343614

  2. Significance of chick quality score in broiler production

    Ven, van de, J.; Wagenberg, van, A.V.; Uitdehaag, K.A.; Groot Koerkamp, P.W.G.; Kemp, B; Brand, van den, MGJ Mark


    The quality of day old chicks is crucial for profitable broiler production, but a difficult trait to define. In research, both qualitative and quantitative measures are used with variable predictive value for subsequent performance. In hatchery practice, chick quality is judged on a binomial scale, as chicks are divided into first grade (Q1-saleable) and second grade (Q2) chicks right after hatch. Incidences and reasons for classifying chicks as Q2, and potential of these chicks for survival ...

  3. Development of basal and induced testosterone hydroxylase activity in the chicken embryo in ovo

    Paolini, Moreno; Pozzetti, Laura; Sapone, Andrea; Luigi Biagi, Gian; Cantelli-Forti, Giorgio


    The sensitivity of the developing embryo to xenobiotics is highly dependent on the expression of metabolizing enzymes including cytochromes P450 (CYP). In the present study, therefore, the ontogeny of the CYP-dependent system in the chick was investigated with testosterone hydroxylase activity as a marker of CYP expression.Chicken embryo livers were assayed for basal and phenobarbitone (PB)-induced regio- and stereo-selective testosterone hydroxylase activity, from the first appearance of the...

  4. Specification of germ layer identity in the chick gastrula

    Cai Qin


    Full Text Available Abstract Background Chick definitive endoderm is an important source of signals that pattern the early embryo forming a central structure around which the body plan is constructed. Although the origin of definitive endoderm has been mapped in the chick, arising principally from rostral streak at elongating streak stages, it is not known when this layer first becomes fully committed to its germ layer fate, an important issue to resolve in light of its critical role in subsequent patterning of the early embryo. Results Through gene expression screening of chick gastrula, we identified molecular markers of definitive endoderm restricted to rostral (Sox17 and caudal (Gata5/6 regions, suggesting that at least two subpopulations of definitive endodermal cells exist during ingression. We show (1 that presumptive mesoderm cells migrate to the middle layer and remain mesenchymal when transplanted to rostral primitive streak, and prospective endoderm cells enter the lower layer and become epithelial when transplanted to caudal primitive streak; and (2 that presumptive endoderm cells and mesoderm cells lose normal gene expression (Sox17 and Wnt8c, respectively when transplanted outside of their normal position of origin. Moreover, when rostral or caudal primitive streak segments are transplanted into rostral blastoderm isolates (RBIs, both types of transplants express Sox17 4–6 hours later–consistent with their new position, regardless of their presumptive germ layer origin–and prospective mesoderm transplants, which normally express Wnt8c, turn off expression, suggesting that signals within the rostral blastoderm induce endoderm gene expression, and repress mesoderm gene expression, during gastrulation. Conclusion Our results demonstrate that germ layer identity is fixed at the time populations of endoderm and mesoderm cells ingress through the primitive streak, whereas their gene expression patterns remain labile. In addition, our results show that

  5. Quantitative analysis of embryonic kidney impairment by confocal microscopy and stereology: effect of 1,2-dibromoethane in the chick mesonephros

    Holejšovská, Iva; Radochová, Barbora; Novotná, Božena; Janáček, Jiří; Jirkovská, M.; Kubínová, Lucie


    Roč. 46, č. 6 (2005), s. 661-667. ISSN 0007-1668 R&D Projects: GA AV ČR(CZ) KJB5039302 Institutional research plan: CEZ:AV0Z50390512; CEZ:AV0Z50110509 Keywords : chick embryo * confocal microscopy * stereology Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 0.813, year: 2005

  6. The effect of gamma irradiation and ethyl methan sulfonate on somatic embryo formation of soybean (Glycine max L.)

    Soybean is a source of protein and vegetable oil. Global climate change affect the productivity of soybean, so that new cultivars that have superior characteristic can be produced. In vitro techniques through somaclonal variation and mutation is one alternative for obtaining new varieties when genetic material as the material selection is not available. Mutation induction can be performed on embryogenic cell populations using gamma irradiation or chemical compounds, such as Ethyl Methane Sulfonate (EMS). Both of these methods have been widely used to increase the genetic diversity of plants and have produced new clones with superior characteristic. The main component that must be controlled in the implementation of these technologies is somatic cells regeneration after mutation treatment in order to get in vitro shoots. Regeneration methods which are successfully applied to certain varieties, often is not successfully for other varieties of the same species. Some factors that influence it, are such as explants source, genotype, medium composition, genotype, medium composition, etc. Somaclonal variation and mutation treatment can cause cell damage that is sometimes necessary need modifications of the regeneration method that has been produced before. The aim of the experiment was to get cell population and planlet mutation with gamma irradiation and Ethyl Methan Sulfonate (EMS). Young embryozygotic was used as explant came from young pod that was harvested at 12-20 days after fertilization of Willis, Burangrang and Baluran varieties and accession No B 3592. Embryogenic callus induction was done by using MS media with vitamin B5 added with 20 mg/l of 2,4-D and 3% sucrose. The callus were irradiated by gamma rays 400 rad or dilute in EMS solution with 0.1%, 0.3% and 0.5% concentration for 1, 2, and 3 hours. After mutation treatment, the callus were sub culture for seed somatic induction. The results showed that callus formation was influenced by plant genotype. All

  7. Efficacy of using radio transmitters to monitor least tern chicks

    Whittier, Joanna B.; Leslie, David M., Jr.


    Little is known about Least Tern (Sterna antillarum) chicks from the time they leave the nest until fledging because they are highly mobile and cryptically colored. We evaluated the efficacy of using radiotelemetry to monitor Interior Least Tern (S. a. athalassos) chicks at Salt Plains National Wildlife Refuge, Oklahoma. In 1999, we attached radio transmitters to 26 Least Tern chicks and tracked them for 2-17 days. No adults abandoned their chicks after transmitters were attached. Transmitters did not appear to alter growth rates of transmittered chicks (P = 0.36) or prevent feather growth, although dermal irritation was observed on one chick. However, without frequent reattachment, transmitters generally did not remain on chicks transmitter removal, presumably by adult terns. Although the presence of transmitters did not adversely affect Least Tern chicks, future assessments should investigate nonintrusive methods to improve retention of transmitters on young chicks and reduce the number of times that chicks need to be handled.

  8. Temperature and CO2 during the hatching phase. II. Effects on chicken embryo physiology

    Maatjens, C.M.; Reijrink, I.A.M.; Anker, van den I.; Molenaar, R.; Pol, van der C.W.; Kemp, B.; Brand, van den H.


    The objective of this study was to investigate the effect of eggshell temperature (EST) and carbon dioxide concentration during only the hatching phase on physiological characteristics of embryos and chicks. Three groups of eggs were incubated at an EST of 37.8°C until d 19 of incubation (E19). From

  9. Developmental competence of porcine chimeric embryos produced by aggregation

    Li, Juan; Jakobsen, Jannik E.; Xiong, Qiang;


    either by parthenogenetic activation (PA) or handmade cloning (HMC). Results showed that the developmental competence of chimeric embryos, evaluated based on their blastocyst rate and total cell number per blastocyst, was increased when two whole 2-cell stage embryos (PA or HMC) were aggregated. In...... comparison, when two blastomeres were aggregated, the developmental competence of the chimeric embryos decreased if the blastomeres were either from PA or from HMC embryos, but not if they were from different sources, i.e. one PA and one HMC blastomere. To evaluate the cell contribution in embryo formation......The purpose of our study was to compare the developmental competence and blastomere allocation of porcine chimeric embryos formed by micro-well aggregation. Chimeras were created by aggregating either two blastomeres originating from 2-cell embryos or two whole embryos, where embryos were produced...

  10. Compensatory adaptations of the remnant kidney to the overload in the chick model of the unilateral renal agenesis

    Zemanová, Zdeňka; Jirsová, Z.; Hubičková, L.; Klusoňová, Petra; Tůma, P.

    Berlin : Freie Universität, 2007. s. 47-47. [Combined workshop on fundamental physiology and perinatal development in poultry /3./. 05.10.2007-07.10.2007, Berlin] R&D Projects: GA ČR(CZ) GA304/04/0972 Institutional research plan: CEZ:AV0Z50110509 Keywords : spr2 * chick embryo * unilateral renal agenesis * compensatory renal agenesis Subject RIV: EA - Cell Biology

  11. The de novo formation of a vascular network, in warm-blooded embryos, occurs via a self-assembly process that spans multiple length and time scales

    Little, Charles D.


    Taking advantage of wide-field, time-lapse microscopy we examined the assembly of vascular polygonal networks in whole bird embryos and in explanted embryonic mouse tissue (allantois). Primary vasculogenesis assembly steps range from cellular (1-10 μm) to tissue (100μm-1mm) level events: Individual vascular endothelial cells extend protrusions and move with respect to the extracellular matrix/surrounding tissue. Consequently, long-range, tissue-level, deformations directly influence the vascular pattern. Experimental perturbation of endothelial-specific cell-cell adhesions (VE-cadherin), during mouse vasculogenesis, permitted dissection of the cellular motion required for sprout formation. In particular, cells are shown to move actively onto vascular cords that are subject to strain via tissue deformations. Based on the empirical data we propose a simple model of preferential migration along stretched cells. Numerical simulations reveal that the model evolves into a quasi-stationary pattern containing linear segments, which interconnect above a critical volume fraction. In the quasi-stationary state the generation of new branches offsets the coarsening driven by surface tension. In agreement with empirical data, the characteristic size of the resulting polygonal pattern is density-independent within a wide range of volume fractions. These data underscore the potential of combining physical studies with experimental embryology as a means of studying complex morphogenetic systems. In collaboration with Brenda J. Rongish^1, Andr'as Czir'ok^1,2, Erica D. Perryn^1, Cheng Cui^1, and Evan A. Zamir^1 ^1Department of Anatomy and Cell Biology, the University of Kansas Medical Center, Kansas City, KS ^2Department of Biological Physics, E"otv"os Lor'and University, Budapest, Hungary.

  12. HNK-1 immunoreactivity during early morphogenesis of the head region in a nonmodel vertebrate, crocodile embryo

    Kundrát, Martin


    The present study examines HNK-1 immunoidentification of a population of the neural crest (NC) during early head morphogenesis in the nonmodel vertebrate, the crocodile ( Crocodylus niloticus) embryos. Although HNK-1 is not an exclusive NC marker among vertebrates, temporospatial immunoreactive patterns found in the crocodile are almost consistent with NC patterns derived from gene expression studies known in birds (the closest living relatives of crocodiles) and mammals. In contrast to birds, the HNK-1 epitope is immunoreactive in NC cells at the neural fold level in crocodile embryos and therefore provides sufficient base to assess early migratory events of the cephalic NC. I found that crocodile NC forms three classic migratory pathways in the head: mandibular, hyoid, and branchial. Further, I demonstrate that, besides this classic phenotype, there is also a forebrain-derived migratory population, which consolidates into a premandibular stream in the crocodile. In contrast to the closely related chick model, crocodilian premandibular and mandibular NC cells arise from the open neural tube suggesting that species-specific heterochronic behavior of NC may be involved in the formation of different vertebrate facial phenotypes.

  13. Neural crest induction by paraxial mesoderm in Xenopus embryos requires FGF signals.

    Monsoro-Burq, Anne-Hélène; Fletcher, Russell B; Harland, Richard M


    At the border of the neural plate, the induction of the neural crest can be achieved by interactions with the epidermis, or with the underlying mesoderm. Wnt signals are required for the inducing activity of the epidermis in chick and amphibian embryos. Here, we analyze the molecular mechanisms of neural crest induction by the mesoderm in Xenopus embryos. Using a recombination assay, we show that prospective paraxial mesoderm induces a panel of neural crest markers (Slug, FoxD3, Zic5 and Sox9), whereas the future axial mesoderm only induces a subset of these genes. This induction is blocked by a dominant negative (dn) form of FGFR1. However, neither dnFGFR4a nor inhibition of Wnt signaling prevents neural crest induction in this system. Among the FGFs, FGF8 is strongly expressed by the paraxial mesoderm. FGF8 is sufficient to induce the neural crest markers FoxD3, Sox9 and Zic5 transiently in the animal cap assay. In vivo, FGF8 injections also expand the Slug expression domain. This suggests that FGF8 can initiate neural crest formation and cooperates with other DLMZ-derived factors to maintain and complete neural crest induction. In contrast to Wnts, eFGF or bFGF, FGF8 elicits neural crest induction in the absence of mesoderm induction and without a requirement for BMP antagonists. In vivo, it is difficult to dissociate the roles of FGF and WNT factors in mesoderm induction and neural patterning. We show that, in most cases, effects on neural crest formation were parallel to altered mesoderm or neural development. However, neural and neural crest patterning can be dissociated experimentally using different dominant-negative manipulations: while Nfz8 blocks both posterior neural plate formation and neural crest formation, dnFGFR4a blocks neural patterning without blocking neural crest formation. These results suggest that different signal transduction mechanisms may be used in neural crest induction, and anteroposterior neural patterning. PMID:12783784

  14. Endogenous electric fields in embryos during development, regeneration and wound healing

    Nuccitelli, R


    All embryos that have been investigated drive ionic currents through themselves and these currents will generate internal electric fields. Here, those examples in which such fields have been measured directly are discussed. The first such measurements were made in chick embryos and about 20 mV mm-1 was measured near the posterier intestinal portal in 2-4-day-old embryos. This electric field is important for the development of tail structures because reducing its magnitude results in abnormal tail development. The second embryonic electric field measured directly was in the axolotl, where a rostral-caudal field of about the same magnitude was detected. Modification of this field during neurulation but not gastrulation caused developmental abnormalities. Most recently, the development of left-right asymmetry in frog and chick embryos was found to require a voltage difference between blastomeres at a very early developmental stage. This field was measured in the chick embryo to be 10-20 mV mm-1 across the primitive streak. Mammalian skin wounds generate 150 mV mm-1 fields lateral to the wound and corneal epidermal wounds exhibit lateral fields of 40 mV mm-1. The presence of these endogenous fields would suggest that exposures to external electric fields should be limited to magnitudes of less than 0.1 V m-1. (author)

  15. Immunologic effects of low levels of ochratoxin A in ovo: utilization of a chicken embryo model.

    Harvey, R B; Kubena, L F; Naqi, S A; Gyimah, J E; Corrier, D E; Panigrahy, B; Phillips, T D


    Ochratoxin A (OA) was administered to 13-day-old chicken embryos via the chorioallantoic membrane. The 7-day LD50 value (day 20 incubation) of OA was calculated at 7.9 micrograms of OA. Ochratoxin-treated embryos (2.5 micrograms) had slight but significant changes in numbers of immunoglobulin-bearing cells in the bursa but not in the spleen. Chicks hatched from in ovo-treated eggs were challenged with 9 X 10(4) colony-forming units (CFU) of beta-hemolytic Escherichia coli (O1:K1) at 7 days of age via the thoracic air sac. Lesion scores of OA-treated chicks were equal to or less severe than those of controls. Hatchmates of the above chicks were vaccinated with a homologous killed E. coli bacterin (O1:K1) at both 2 and 4 weeks of age and challenged with 10(4) CFU of E. coli at 7 weeks. Post-challenge lesions were present in three vaccinated untreated controls and no OA-treated chicks. We conclude that although in ovo exposure to OA may marginally suppress immunoglobulin-bearing cells of bursa, chicks hatched from OA-treated eggs respond as well as controls to an antigen and resist infection by a virulent organism. PMID:3327493

  16. Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System

    SUGIMURA, Satoshi; Akai, Tomonori; HASHIYADA, Yutaka; Aikawa, Yoshio; OHTAKE, Masaki; Matsuda, Hideo; KOBAYASHI, Shuji; Kobayashi, Eiji; Konishi, Kazuyuki; Imai, Kei


    Abstract To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 µl medium for 168 h. The blasto...

  17. Hippocampus and medial striatum dissociation during goal navigation by geometry or features in the domestic chick: An immediate early gene study.

    Mayer, Uwe; Pecchia, Tommaso; Bingman, Verner Peter; Flore, Michele; Vallortigara, Giorgio


    We employed a standard reference memory task to study the involvement of the hippocampal formation (HF) of domestic chicks that used the boundary geometry of a test environment to orient to and locate a reward. Using the immediate early gene product c-Fos as a neuronal activity marker, we found enhanced HF activation in chicks that learned to locate rewarded corners using the shape of a rectangular arena compared to chicks trained to solve the task by discriminating local features in a square-shaped arena. We also analyzed neuronal activity in the medial part of the medial striatum (mMSt). Surprisingly, in mMSt we observed a reverse pattern, with higher activity in the chicks that were trained to locate the goal by local features. Our results identify two seemingly parallel, memory systems in chicks, with HF central to the processing of spatial-geometrical information and mMSt important in supporting local feature discrimination. PMID:26135386

  18. Embryos, microscopes, and society.

    Maienschein, Jane


    Embryos have different meanings for different people and in different contexts. Seen under the microscope, the biological embryo starts out as one cell and then becomes a bunch of cells. Gradually these divide and differentiate to make up the embryo, which in humans becomes a fetus at eight weeks, and then eventually a baby. At least, that happens in those cases that carry through normally and successfully. Yet a popular public perception imagines the embryo as already a little person in the very earliest stages of development, as if it were predictably to become an adult. In actuality, cells can combine, pull apart, and recombine in a variety of ways and still produce embryos, whereas most embryos never develop into adults at all. Biological embryos and popular imaginations of embryos diverge. This paper looks at some of the historical reasons for and social implications of that divergence. PMID:26996410

  19. Effects of LED lighting during incubation on layer and broiler hatchability, chick quality, stress susceptibility and post-hatch growth.

    Huth, Jesse C; Archer, Gregory S


    Providing light during incubation has been shown to affect hatchability, but the use of LED lights has not been evaluated. This experiment evaluated the effects of LED lighting during embryogenesis on White Leghorn and commercial broiler eggs. To determine this, two experiments were conducted, the first using White Leghorn eggs (N=3456) and the second using commercial broiler eggs (N=3456) where eggs were incubated 12 h of light and 12 h of darkness (LED) or complete darkness (DARK); the light level was 250 lux. Hatchability, embryo mortality, and chick quality were measured in both studies, and a subset of one of the broiler egg trials were grown out to investigate fear and stress parameters. There was no effect (P>0.05) on hatchability of layer eggs; however, there was a difference (P=0.02) observed in chick quality, with the LED group having more chicks (75.34%) with no defects than the DARK group (56.53%). Broiler eggs exposed to LED light showed an increase in hatchability (90.12%, P=0.03) and an increase in no-defect chick percentage (86.12%, P=0.04) at hatch compared to the DARK chicks (85.76% and 69.43%, respectively). Differences were observed between treatments during the 14 d grow-out. The LED birds had lower (P0.05) observed between treatments in growth, FCR, or fear measures at 14 d. These results indicate that providing LED light during incubation can improve chick quality in both white layer and broiler eggs; however, it only appears to improve hatchability in broilers, which could be related to shell pigmentation. It was also demonstrated that providing LED light during incubation can reduce the stress susceptibility of broilers post-hatch. Utilizing light during incubation may be useful tool for the poultry industry. PMID:26475072

  20. Hydrogen peroxide potentiates organophosphate toxicosis in chicks

    Banan K. Al-Baggou


    Full Text Available Objective: The purpose of the present study was to examine the effect of hydrogen peroxide(H2O2 on the acute toxicity of organophosphate insecticides dichlorvos and diazinon and their inhibitoryactions on plasma, brain and liver cholinesterase activities. Material and Methods: H2O2 was given indrinking water (0.5% v/v for 2 weeks in unsexed day old chicks, a regimen known to induce oxidativestress in this species. A control group received drinking tap water. All experiments were conducted onthe chicks at the age of 15 days after exposure to H2O2. The acute (24 h oral LD50 values of dichlorvosand diazinon in the insecticidal preparations as determined by the up-and-down method in the controlchicks were 9.4 and 15.6 mg/kg, respectively. Results: The poisoned chicks manifested signs ofcholinergic toxicosis within one hour after the dosing including salivation, lacrimation, gasping, frequentdefecation, drooping of wings, tremors, convulsions and recumbency. The acute (24 h oral LD50 valuesof dichlorvos and diazinon in chicks provided with H2O2 were reduced to 3.5 and 6.5 mg/kg, by 63 and58%, respectively when compared to respective control LD50 values. The intoxicated chicks also showedcholinergic signs of toxicosis as described above. Plasma, brain and liver cholinesterase activities of thechicks exposed to H2O2 were significantly lower than their respective control (H2O values by 25, 28 and27%, respectively. Oral dosing of chicks with dichlorvos at 3 mg/kg significantly inhibited cholinesteraseactivities in the plasma, brain and liver of both control (42-67% and H2O2-treated (15-59% chicks.Diazinon at 5 mg/kg, orally also inhibited cholinesterase activities in the plasma, brain and liver of bothcontrol (36-66% and H2O2-treated (15-30% chicks. In the H2O2 groups, dichlorvos inhibition of livercholinesterase activity and diazinon inhibition of liver and brain cholinesterase activities weresignificantly lesser than those of the respective values of

  1. Blastocyst development potential of D3 low quality embryos

    Zhen Jing-ran; Sun Zhen-yi; Yu Qi; Deng Cheng-yan; Zhou Yuan-zheng; He Fang-fang


    Objective:To investigate the potential of blastocyst development of D3 low quality embryos for blastocyst culturing and freezing.Methods:Two thousand one hundred and eighty embryos of 398 IVF/ICSI patients' were observed after D3 embryo transfer.The low-quality embryos were cultured by sequential micro-drops and continuously observed blastocyst formation from D3 to D6.Results:(1) A total of 1,546 low-quality embryos were collected,and 426 blastocysts formed in DS-D6.Total blastocyst formation rate (BFR) was 27.56%,of which embryos with grade 6 Ⅲ/Ⅳ were 40.87 % (318/778),5 Ⅰ /Ⅱ 28.85% (30/104),4Ⅰ/Ⅱ 8.88% (16/180),4Ⅲ/IV 19.72% (56/284),2-3 Ⅰ/Ⅱ 3% (6/200).(2) Embryos with more blastomeres had a higher blastocyst formation rate.The lower qualities of embryos resulted in lower blastocyst formation rate.Conclusions:Low-quality embryos still have the potential to develop into blastocysts.Therefore,they should not be discarded on D3.They should be continually cultured to D6 in order to reduce embryo wastage and get good clinical results.

  2. Increased blastocyst formation of cloned porcine embryos produced with donor cells pre-treated with digitonin and Xenopus egg extract

    Liu, Ying; Østrup, Olga; Li, Juan;


    Pre-treating donor cells before somatic cell nuclear transfer (SCNT, ‘cloning’) may improve the efficiency of the technology. The aim of this study was to evaluate the early development of cloned embryos produced with porcine fibroblasts pre-treated with a permeabilizing agent and extract from...... Xenopus laevis eggs. In Experiment 1, fetal fibroblasts were permeabilized by digitonin, incubated in egg extract and, after re-sealing of cell membranes, cultured for 3 or 5 days before use as donor cells in handmade cloning (HMC). Controls were produced by HMC with non-treated donor cells. The...... blastocyst rate for reconstructed embryos increased significantly when digitonin-permeabilized, extract-treated cells were used after 5 days of culture after re-sealing. In Experiment 2, fetal and adult fibroblasts were treated with digitonin alone before re-sealing the cell membranes, then cultured for 3 or...

  3. Direct localised measurement of electrical resistivity profile in rat and embryonic chick retinas using a microprobe

    Harald van Lintel


    Full Text Available We report an alternative technique to perform a direct and local measurement of electrical resistivities in a layered retinal tissue. Information on resistivity changes along the depth in a retina is important for modelling retinal stimulation by retinal prostheses. Existing techniques for resistivity-depth profiling have the drawbacks of a complicated experimental setup, a less localised resistivity probing and/or lower stability for measurements. We employed a flexible microprobe to measure local resistivity with bipolar impedance spectroscopy at various depths in isolated rat and chick embryo retinas for the first time. Small electrode spacing permitted high resolution measurements and the probe flexibility contributed to stable resistivity profiling. The resistivity was directly calculated based on the resistive part of the impedance measured with the Peak Resistance Frequency (PRF methodology. The resistivity-depth profiles for both rat and chick embryo models are in accordance with previous mammalian and avian studies in literature. We demonstrate that the measured resistivity at each depth has its own PRF signature. Resistivity profiles obtained with our setup provide the basis for the construction of an electric model of the retina. This model can be used to predict variations in parameters related to retinal stimulation and especially in the design and optimisation of efficient retinal implants.

  4. Thermal manipulations of turkey embryos: The effect on thermoregulation and development during embryogenesis.

    Piestun, Y; Zimmerman, I; Yahav, S


    Previous studies conducted on meat-type chickens in our laboratory showed that thermal manipulations (TMS:) of the embryo during the time window of the hypothalamus-hypophysis-thyroid axis development and maturation significantly reduced the metabolic rates of the embryo and the chicken, improving the posthatch feed conversion rate (FCR:). The aim of the present study was to investigate the effect of intermittent TMs during turkey embryogenesis on embryo development. Fertile turkey eggs were divided into three treatments: control; 6H--with TM by elevation of temperature and RH by 1.7°C and 9%, respectively, above the control conditions for 6 h/d, from E10 through E22, i.e., 240 through 552 h of incubation; and 12H--with TM as above, for 12 h/d, during the same time period. From E0 through E10 and from E23 onward all eggs were incubated under control conditions. The embryo growth rate was not negatively affected by TM. During TM eggshell temperature, the embryonic heart rate and oxygen consumption were elevated by the manipulation while the embryos were in their ectothermic phase. However, by the end of the TM period and until hatch (the endothermic phase) these parameters were significantly lower in both TM treatments than in the control, indicating a lower metabolic rate and heat production. The TM embryos hatched approximately 10 h earlier than the controls, without any negative effects on chick body weight or hatchability. Nevertheless, TM treatments resulted in a higher proportion of chicks with unhealed navels. Body temperature at hatch was significantly lower in the TM chicks than in the controls, suggesting lower heat production and metabolic rate, which might affect the energy requirements for posthatch maintenance. It was concluded that TM during turkey embryogenesis might have altered the thermoregulatory set point, and thus lowered the embryo metabolic rate, which might have a long-lasting posthatch effect. PMID:25630674

  5. Amelioration of scopolamine-induced amnesia by phosphatidylserine and curcumin in the day-old chick.

    Barber, Teresa A; Edris, Edward M; Levinsky, Paul J; Williams, Justin M; Brouwer, Ari R; Gessay, Shawn A


    In the one-trial taste-avoidance task in day-old chicks, acetylcholine receptor activation has been shown to be important for memory formation. Injection of scopolamine produces amnesia, which appears to be very similar in type to that of Alzheimer's disease, which is correlated with low levels of acetylcholine in the brain. Traditional pharmacological treatments of Alzheimer's disease, such as cholinesterase inhibitors and glutamate receptor blockers, improve memory and delay the onset of impairments in memory compared with placebo controls. These agents also ameliorate scopolamine-induced amnesia in the day-old chick trained on the one-trial taste-avoidance task. The present experiments examined the ability of two less traditional treatments for Alzheimer's disease, phosphatidylserine and curcumin, to ameliorate scopolamine-induced amnesia in day-old chicks. The results showed that 37.9 mmol/l phosphatidylserine and 2.7 mmol/l curcumin significantly improved retention in chicks administered scopolamine, whereas lower doses were not effective. Scopolamine did not produce state-dependent learning, indicating that this paradigm in day-old chicks might be a useful one to study the effects of possible Alzheimer's treatments. In addition, chicks administered curcumin or phosphatidylserine showed little avoidance of a bead associated with water reward, indicating that these drugs did not produce response inhibition. The current results extend the findings that some nontraditional memory enhancers can ameliorate memory impairment and support the hypothesis that these treatments might be of benefit in the treatment of Alzheimer's disease. PMID:27388114

  6. The adrenocortical response of tufted puffin chicks to nutritional deficits.

    Kitaysky, Alexander S; Romano, Marc D; Piatt, John F; Wingfield, John C; Kikuchi, Motoshi


    In several seabirds, nutritional state of a nest-bound chick is negatively correlated with the activity of its hypothalamus-pituitary-adrenal (HPA) axis. Increased corticosterone (cort) secretion has been shown to facilitate changes in behavior that allow hungry chicks to obtain more food from parents. However, if parents are not willing/able to buffer their young from temporary food shortages, increased cort secretion could be detrimental to undernourished chicks. In a system where parents are insensitive to chick demands, low benefits and high costs of activation of the HPA-axis in hungry chicks should lead to a disassociation of the nutritional state of the young and the activity of its HPA-axis. We tested this novel hypothesis for the tufted puffin (Fratercula cirrhata), a seabird with intermittent provisioning of a nest-bound semi-precocial chick. We examined the HPA-axis activity of captive chicks exposed to the following: (1) a short-term (24 h) food deprivation; and (2) an array of prolonged (3 weeks) restrictions in feeding regimens. We found that in response to a short-term food deprivation chicks decreased baseline levels of cort and thyroid hormones. In response to prolonged restrictions, food-limited chicks exhibited signs of nutritional deficit: they had lower body mass, endogenous lipid reserves, and thyroid hormone titers compared to chicks fed ad libitum. However, baseline and maximum acute stress-induced levels of cort were also lower in food-restricted chicks compared to those of chicks fed ad libitum. These results support a major prediction of the study hypothesis that puffin chicks suppress HPA-axis activity in response to short- and long-term nutritional deficits. This physiological adaptation may allow a chick to extend its development in the nest, while eluding detrimental effects of chronic cort elevation. PMID:15811363

  7. Influence of polychlorinated biphenyls on the growth of chicken embryos

    Hatano, Yasuhiko; Hatano, Akira [Chub Women`s College, Gifu-ken (Japan)


    Incubation of chicken embryos with either 0.01 or 0.03 ppm polychlorinated biphenyls (PCBs) for 12, 15, or 18 d resulted in a significant decrease in liver and body weight associated with enhanced mortality. Teratological examination revealed an increased frequency of malformations including hydrops, eventration, wry neck, and brevicollis. PCB exposure was also found to produce histologic damage to liver and cutaneous tissue. Our data demonstrate that exposure of chicks to PCBs during development results in a retardation of growth, an increased incidence of malformations, and histopathologic damage. 9 refs., 2 figs., 4 tabs.

  8. The influence of low-level radiation and gangliosides on adenylate cyclase activity in thymus and thyroid glands of chicks in ontogenesis

    Adenylate cyclase (AC) activity was studied in thymus and thyroid gland of intact chick embryos and those irradiated with a dose of 0.029 Gy prior to incubation, and newly hatched chocks in the presence of total ganglioside fractions extracted from the same organs. Gangliosides were shown to increase the enzyme activity of thymocytes and thyreocytes during the postnatal development. It is suggested that small radiation doses potentiate the stimulatory effect of ganglioside fractions on AC

  9. The ultrastructure of the mature embryo sac in the natural tetraploid of red clover (Trifolium pratense L.: that has a very low rate of seed formation

    Gönül Algan


    Full Text Available In this study, ultrastructural organization of cells in the mature embryo sac of natural tetraploid Trifolium pratense L. was investigated. The mature embryo sac of this plant contains an egg cell with two synergids at the micropylar end, and a central cell with two polar nuclei. The ultrastructure of these cells agrees with what is known for most angiosperms studied with the electron microscope. The egg cell is a large and highly vacuolate cell, partially surrounded by a wall. Much of the cytoplasm is located around the nucleus at the chalazal end and there are few numbers of channel-shaped endoplasmic reticulum, mitochondria, plastids and numerous ribosomes distributed throughout the cytoplasm. Unlike the egg cell, much of the cytoplasm in synergid cells is located at micropylar part of the cell and the synergid cytoplasm contains especially, large numbers of rough endoplasmic reticulum, free ribosomes, mitochondria and plastids. The central cell of T. pratense L. contains two large polar nuclei which lie close to the egg apparatus. Each polar nucleus has a single, large, dense nucleolus that contains several nucleolar vacuoles. Much of the central cell cytoplasm consisting of granular and agranular endoplasmic reticulum, mitochondria, plastids, ribosomes, dictyosomes and lipid bodies are placed around polar nuclei.

  10. Higher levels of CO2 during late incubation alter the hatch time of chicken embryos.

    Tong, Q; McGonnell, I M; Roulston, N; Bergoug, H; Romanini, C E B; Garain, P; Eterradossi, N; Exadaktylos, V; Bahr, C; Berckmans, D; Demmers, T G M


    1. It has been reported that the increasing CO2 tension triggers the embryo to pip the air cell and emerge from the egg. However, the mechanism by which higher CO2 concentrations during the last few days of incubation affect chick physiology and the hatching process is unclear. This study investigated the effect of CO2 concentrations up to 1% during pipping, on the onset and length of the hatch window (HW) and chick quality. 2. Four batches of Ross 308 broiler eggs (600 eggs per batch) were incubated in two small-scale custom-built incubators (Petersime NV). During the final 3 d of incubation, control eggs were exposed to a lower CO2 concentration (0.3%), while the test eggs experienced a higher CO2 concentration programme (peak of 1%). 3. There were no significant differences in blood values, organ weight and body weight. There was also no difference in hatchability between control and test groups. However, a small increase in the chick weight and the percentage of first class chicks was found in the test groups. Furthermore, plasma corticosterone profiles during hatching were altered in embryos exposed to higher CO2; however, they dropped to normal levels at d 21 of incubation. Importantly, the hatching process was delayed and synchronised in the test group, resulting in a narrowed HW which was 2.7 h shorter and 5.3 h later than the control group. 4. These results showed that exposing chicks to 1% CO2 concentration during pipping did not have negative impacts on physiological status of newly hatched chicks. In addition, it may have a significant impact on the physiological mechanisms controlling hatching and have benefits for the health and welfare of chickens by reducing the waiting time after hatching. PMID:25900009

  11. Metabolomic Assessment of Embryo Viability

    Uyar, Asli; Seli, Emre


    Preimplantation embryo metabolism demonstrates distinctive characteristics associated with the developmental potential of embryos. On this basis, metabolite content of culture media was hypothesized to reflect the implantation potential of individual embryos. This hypothesis was tested in consecutive studies reporting a significant association between culture media metabolites and embryo development or clinical pregnancy. The need for a noninvasive, reliable, and rapid embryo assessment strat...

  12. 人类辅助生殖技术中DAY3胚胎质量与囊胚形成相关性分析%Correlation analysis of the quality of the 3th embryo with the blastocyst formation during human ART

    胡煜; 刘吉; 李宝山; 于洪君; 宫立国


    Objective: To analyze the correlation of the quality of the 3th embryo with the blastocyst formation during human ART. Methods: In this study, All 130 patients accorded with our study standard. The 3th surplus embryos after transferred were cultured to the 6th days for the blastocyst formation. We tried to find the relationship among the patient's age, the 3th day embryo quality score and cell number with blastocyst formation. Result; The patient's age was not related with blastocyst formation. The remarkable result was the positive correlation beween blastulation rate with the day 3 embryo cell number or with the day3 embryo quality score. The blastocyst formation rate of the good quality embryo ( I - II ) was significantly higher than that of the poor quality embryo ( ID -IV ). The blastocyst formation rate of the ≥6 cells embryos in the 3th days was higher than that of the ≤5 cells embryos. By comprehensive analysis about the correlation of the blastocyst formation with day3 embryo cell numbe, the day3 embryo quality score and culture time, the result showed that the ≥6 cells I - IIdegree embryos had the highest blastocyst formation rate, the blastocyst formation rate of the≥6 cells III - IV degree embryos was higher. The blastocyst formation rate of the ≤5 cells embryos was lower than the ≥6 cells embryos. In all four groups embryos, the blastocyst formation rates in the 6th day were higher than in the 5th day. Conclusion; The day3 embryo cell number, quality score and culture time were important factors for the blastocyst formation, in which the 3th day embryo cell number was the most important. The culture to the 6th day brought some chance to blastocyst for surplus embryos which quality score was lower or developed slower. This culture progress effectually selected and saved the embryos which had potentiality to form the bastula.%目的 探讨人类辅助生殖技术中,day3胚胎质量与囊胚形成的相关性.方法


    Nikolaev S. I.; Karapetyan A. K.; Kornilova E. V.; Struk M. V.


    This article presents the results of the chick peas use instead of sunflower cake, in feeding young and adult livestock hens-layers of the cross "Hajseks brown". The researches were carried out in the JSC "Agrofirm Vostok" of the Nikolayevskiy district in the Volgograd region. The sunflower cake replacement with legumes - chickpeas as the part of the experimental animal fodder for young and adult livestock hens-layers had a positive influence on productivity, physiological state of the birds,...

  14. The developmental stage of chicken embryos modulates the impact of in ovo olfactory stimulation on food preferences.

    Bertin, Aline; Calandreau, Ludovic; Arnould, Cécile; Lévy, Frédéric


    Like mammals, bird embryos are capable of chemosensory learning, but the ontogeny of their feeding preferences has not been examined. We tested if the timing of stimulation in chicken embryos modulates the impact of in ovo olfactory stimulation on later food preferences. We exposed chicken embryos to an olfactory stimulus for a 4-day period in the middle or toward the end of the incubation period. The chicks were tested for their preference between foods with and without the olfactory stimulus in 3-min choice tests and on a 24-h time scale. Regardless of the type of food (familiar or novel) or the duration of the test, the control chicks not exposed to the olfactory stimulus consistently showed significant preferences for non-odorized foods. Chicks that were exposed in ovo to the olfactory stimulus did not show a preference for odorized or non-odorized foods. Only those chicks that were exposed to the olfactory stimulus toward the end of the incubation period differed from the controls and incorporated a higher proportion of odorized food into their diets on a 24-h time scale. This result indicates that olfactory stimulation at the end of embryonic development has a stronger impact on later feeding preferences. Our findings contribute to the growing pool of recent data appreciating the impact of olfactory signals on behavior regulation in avian species. PMID:22080043

  15. Comparative analysis of 3D expression patterns of transcription factor genes and digit fate maps in the developing chick wing.

    Malcolm Fisher

    Full Text Available Hoxd13, Tbx2, Tbx3, Sall1 and Sall3 genes are candidates for encoding antero-posterior positional values in the developing chick wing and specifying digit identity. In order to build up a detailed profile of gene expression patterns in cell lineages that give rise to each of the digits over time, we compared 3 dimensional (3D expression patterns of these genes during wing development and related them to digit fate maps. 3D gene expression data at stages 21, 24 and 27 spanning early bud to digital plate formation, captured from in situ hybridisation whole mounts using Optical Projection Tomography (OPT were mapped to reference wing bud models. Grafts of wing bud tissue from GFP chicken embryos were used to fate map regions of the wing bud giving rise to each digit; 3D images of the grafts were captured using OPT and mapped on to the same models. Computational analysis of the combined computerised data revealed that Tbx2 and Tbx3 are expressed in digit 3 and 4 progenitors at all stages, consistent with encoding stable antero-posterior positional values established in the early bud; Hoxd13 and Sall1 expression is more dynamic, being associated with posterior digit 3 and 4 progenitors in the early bud but later becoming associated with anterior digit 2 progenitors in the digital plate. Sox9 expression in digit condensations lies within domains of digit progenitors defined by fate mapping; digit 3 condensations express Hoxd13 and Sall1, digit 4 condensations Hoxd13, Tbx3 and to a lesser extent Tbx2. Sall3 is only transiently expressed in digit 3 progenitors at stage 24 together with Sall1 and Hoxd13; then becomes excluded from the digital plate. These dynamic patterns of expression suggest that these genes may play different roles in digit identity either together or in combination at different stages including the digit condensation stage.

  16. Klhl31 attenuates β-catenin dependent Wnt signaling and regulates embryo myogenesis.

    Abou-Elhamd, Alaa; Alrefaei, Abdulmajeed Fahad; Mok, Gi Fay; Garcia-Morales, Carla; Abu-Elmagd, Muhammad; Wheeler, Grant N; Münsterberg, Andrea E


    Klhl31 is a member of the Kelch-like family in vertebrates, which are characterized by an amino-terminal broad complex tram-track, bric-a-brac/poxvirus and zinc finger (BTB/POZ) domain, carboxy-terminal Kelch repeats and a central linker region (Back domain). In developing somites Klhl31 is highly expressed in the myotome downstream of myogenic regulators (MRF), and it remains expressed in differentiated skeletal muscle. In vivo gain- and loss-of-function approaches in chick embryos reveal a role of Klhl31 in skeletal myogenesis. Targeted mis-expression of Klhl31 led to a reduced size of dermomyotome and myotome as indicated by detection of relevant myogenic markers, Pax3, Myf5, myogenin and myosin heavy chain (MF20). The knock-down of Klhl31 in developing somites, using antisense morpholinos (MO), led to an expansion of Pax3, Myf5, MyoD and myogenin expression domains and an increase in the number of mitotic cells in the dermomyotome and myotome. The mechanism underlying this phenotype was examined using complementary approaches, which show that Klhl31 interferes with β-catenin dependent Wnt signaling. Klhl31 reduced the Wnt-mediated activation of a luciferase reporter in cultured cells. Furthermore, Klhl31 attenuated secondary axis formation in Xenopus embryos in response to Wnt1 or β-catenin. Klhl31 mis-expression in the developing neural tube affected its dorso-ventral patterning and led to reduced dermomyotome and myotome size. Co-transfection of a Wnt3a expression vector with Klhl31 in somites or in the neural tube rescued the phenotype and restored the size of dermomyotome and myotome. Thus, Klhl31 is a novel modulator of canonical Wnt signaling, important for vertebrate myogenesis. We propose that Klhl31 acts in the myotome to support cell cycle withdrawal and differentiation. PMID:25796573

  17. Effect of zona pellucida on porcine parthenogenetically activated embryos

    Li, Rong; Liu, Ying; Li, Juan;


    The need for zona pellucida (ZP) during pre-implantation embryo development is still debated. In porcine parthenogenetically activated (PA) embryos, we have previously shown a different distribution in cell numbers on Day 6 blastocysts cultured with or without ZP (Li et al. 2010 Reprod. Fertil. Dev...... porcine PA embryos, especially at the timing of embryonic genome activation (5-cell stage). Furthermore, the zona pellucida can benefit the blastocyst formation and cryo-tolerance for PA embryos, perhaps by creating a more stable microenvironement....

  18. Obtaining barley haploid embryos and seedlings using anther culture technique

    The effect of three barley genotypes (Igri, Arabi abiad, and Taqa 76), three irradiation doses (0, 5, and 10 Gy), and two media (FW, modified FW), on the number of formed embryos, and the ratio between regenerated embryos to green seedlings and albinos, were studied using anther culture. Also the study involved the compatibility between seedling morphology and chromosome number. results indicated significant differences among the genotypes, and media in callus and embryos formation and also in the ratio and albino seedlings. However, the effect of gamma rays dose was significant only on embryos regeneration. A high percentage of compatibility (90%) was obtained between the seedling morphology and chromosome number. (author)

  19. Bulk elastic properties of chicken embryos during somitogenesis

    Glazier James A


    Full Text Available Abstract We present measurements of the bulk Young's moduli of early chick embryos at Hamburger-Hamilton stage 10. Using a micropipette probe with a force constant k ~0.025 N/m, we applied a known force in the plane of the embryo in the anterior-posterior direction and imaged the resulting tissue displacements. We used a two-dimensional finite-element simulation method to model the embryo as four concentric elliptical elastic regions with dimensions matching the embryo's morphology. By correlating the measured tissue displacements to the displacements calculated from the in-plane force and the model, we obtained the approximate short time linear-elastic Young's moduli: 2.4 ± 0.1 kPa for the midline structures (notocord, neural tube, and somites, 1.3 ± 0.1 kPa for the intermediate nearly acellular region between the somites and area pellucida, 2.1 ± 0.1 kPa for the area pellucida, and 11.9 ± 0.8 kPa for the area opaca.

  20. Novel Two-Step Hierarchical Screening of Mutant Pools Reveals Mutants under Selection in Chicks.

    Yang, Hee-Jeong; Bogomolnaya, Lydia M; Elfenbein, Johanna R; Endicott-Yazdani, Tiana; Reynolds, M Megan; Porwollik, Steffen; Cheng, Pui; Xia, Xiao-Qin; McClelland, Michael; Andrews-Polymenis, Helene


    Contaminated chicken/egg products are major sources of human salmonellosis, yet the strategies used bySalmonellato colonize chickens are poorly understood. We applied a novel two-step hierarchical procedure to identify new genes important for colonization and persistence ofSalmonella entericaserotype Typhimurium in chickens. A library of 182S.Typhimurium mutants each containing a targeted deletion of a group of contiguous genes (for a total of 2,069 genes deleted) was used to identify regions under selection at 1, 3, and 9 days postinfection in chicks. Mutants in 11 regions were under selection at all assayed times (colonization mutants), and mutants in 15 regions were under selection only at day 9 (persistence mutants). We assembled a pool of 92 mutants, each deleted for a single gene, representing nearly all genes in nine regions under selection. Twelve single gene deletion mutants were under selection in this assay, and we confirmed 6 of 9 of these candidate mutants via competitive infections and complementation analysis in chicks.STM0580,STM1295,STM1297,STM3612,STM3615, andSTM3734are needed forSalmonellato colonize and persist in chicks and were not previously associated with this ability. One of these key genes,STM1297(selD), is required for anaerobic growth and supports the ability to utilize formate under these conditions, suggesting that metabolism of formate is important during infection. We report a hierarchical screening strategy to interrogate large portions of the genome during infection of animals using pools of mutants of low complexity. Using this strategy, we identified six genes not previously known to be needed during infection in chicks, and one of these (STM1297) suggests an important role for formate metabolism during infection. PMID:26857572

  1. Migration and Growth of Protoplanetary Embryos. I. Convergence of Embryos in Protoplanetary Disks

    Zhang, Xiaojia; Liu, Beibei; Lin, Douglas N. C.; Li, Hui


    According to the core accretion scenario, planets form in protostellar disks through the condensation of dust, coagulation of planetesimals, and emergence of protoplanetary embryos. At a few AU in a minimum mass nebula, embryos' growth is quenched by dynamical isolation due to the depletion of planetesimals in their feeding zone. However, embryos with masses (Mp ) in the range of a few Earth masses (M ⊕) migrate toward a transition radius between the inner viscously heated and outer irradiated regions of their natal disk. Their limiting isolation mass increases with the planetesimals surface density. When Mp > 10 M ⊕, embryos efficiently accrete gas and evolve into cores of gas giants. We use a numerical simulation to show that despite stream line interference, convergent embryos essentially retain the strength of non-interacting embryos' Lindblad and corotation torques by their natal disks. In disks with modest surface density (or equivalently accretion rates), embryos capture each other in their mutual mean motion resonances and form a convoy of super-Earths. In more massive disks, they could overcome these resonant barriers to undergo repeated close encounters, including cohesive collisions that enable the formation of massive cores.

  2. Migration and growth of protoplanetary embryos. I. Convergence of embryos in protoplanetary disks

    According to the core accretion scenario, planets form in protostellar disks through the condensation of dust, coagulation of planetesimals, and emergence of protoplanetary embryos. At a few AU in a minimum mass nebula, embryos' growth is quenched by dynamical isolation due to the depletion of planetesimals in their feeding zone. However, embryos with masses (Mp ) in the range of a few Earth masses (M ⊕) migrate toward a transition radius between the inner viscously heated and outer irradiated regions of their natal disk. Their limiting isolation mass increases with the planetesimals surface density. When Mp > 10 M ⊕, embryos efficiently accrete gas and evolve into cores of gas giants. We use a numerical simulation to show that despite stream line interference, convergent embryos essentially retain the strength of non-interacting embryos' Lindblad and corotation torques by their natal disks. In disks with modest surface density (or equivalently accretion rates), embryos capture each other in their mutual mean motion resonances and form a convoy of super-Earths. In more massive disks, they could overcome these resonant barriers to undergo repeated close encounters, including cohesive collisions that enable the formation of massive cores.

  3. Migration and growth of protoplanetary embryos. I. Convergence of embryos in protoplanetary disks

    Zhang, Xiaojia; Lin, Douglas N. C. [Department of Astronomy and Astrophysics, University of California, Santa Cruz, CA 95064 (United States); Liu, Beibei [Kavli Institute for Astronomy and Astrophysics and Department of Astronomy, School of Physics, Peking University, Beijing 100871 (China); Li, Hui, E-mail: [Los Alamos National Laboratory, Los Alamos, NM 87545 (United States)


    According to the core accretion scenario, planets form in protostellar disks through the condensation of dust, coagulation of planetesimals, and emergence of protoplanetary embryos. At a few AU in a minimum mass nebula, embryos' growth is quenched by dynamical isolation due to the depletion of planetesimals in their feeding zone. However, embryos with masses (M{sub p} ) in the range of a few Earth masses (M {sub ⊕}) migrate toward a transition radius between the inner viscously heated and outer irradiated regions of their natal disk. Their limiting isolation mass increases with the planetesimals surface density. When M{sub p} > 10 M {sub ⊕}, embryos efficiently accrete gas and evolve into cores of gas giants. We use a numerical simulation to show that despite stream line interference, convergent embryos essentially retain the strength of non-interacting embryos' Lindblad and corotation torques by their natal disks. In disks with modest surface density (or equivalently accretion rates), embryos capture each other in their mutual mean motion resonances and form a convoy of super-Earths. In more massive disks, they could overcome these resonant barriers to undergo repeated close encounters, including cohesive collisions that enable the formation of massive cores.

  4. Effects of 17beta-estradiol on distribution of primordial germ cell migration in male chicks

    Xiu-Mei Jin; Yi-Xiang Zhang; Zan-Dong Li


    Aim: To assess whether exogenous estradiol has any effect on migration of primordial germ cells (PGCs) in the chick. Methods: Fertilized eggs were treated with 17beta-estradiol (Ez) (80 lag/egg) at stage X (day 0 of incubation),stages 8-10 (incubation 30 h) and 13-15 (incubation 55 h). Controls received vehicle (emulsion) only. Changes in PGC number were measured on different days according to developmental stages. Results: In male right gonads,but not in female left gonads, at stages 28-30 (incubation 132 h) significant decreases in the mean number of PGCs aggregating were observed compared with the controls (P 0.05). Conclusion: The present study provides evidence that E2 has significant effects on the localization of PGCs in male fight, but not female left, gonads of chicken embryos at stages28-30, compared with controls.

  5. Adenylyl Cyclase Signaling in the Developing Chick Heart: The Deranging Effect of Antiarrhythmic Drugs

    Lucie Hejnova


    Full Text Available The adenylyl cyclase (AC signaling system plays a crucial role in the regulation of cardiac contractility. Here we analyzed the key components of myocardial AC signaling in the developing chick embryo and assessed the impact of selected β-blocking agents on this system. Application of metoprolol and carvedilol, two commonly used β-blockers, at embryonic day (ED 8 significantly downregulated (by about 40% expression levels of AC5, the dominant cardiac AC isoform, and the amount of Gsα protein at ED9. Activity of AC stimulated by forskolin was also significantly reduced under these conditions. Interestingly, when administered at ED4, these drugs did not produce such profound changes in the myocardial AC signaling system, except for markedly increased expression of Giα protein. These data indicate that β-blocking agents can strongly derange AC signaling during the first half of embryonic heart development.

  6. Heart Myofibrillogenesis Occurs in Isolated Chick Posterior Blastoderm: A Culture Model

    Early cardiogenesis including myofibrillogenesis is a critical event during development. Recently we showed that prospective cardiomyocytes reside in the posterior lateral blastoderm in the chick embryo. Here we cultured the posterior region of the chick blastoderm in serum-free medium and observed the process of myofibrillogenesis by immunohistochemistry. After 48 hours, explants expressed sarcomeric proteins (sarcomeric α-actinin, 61%; smooth muscle α-actin, 95%; Z-line titin, 56%; sarcomeric myosin, 48%); however, they did not yet show a mature striation. After 72 hours, more than 92% of explants expressed I-Z-I proteins, which were incorporated into the striation in 75% of explants or more (sarcomeric α-actinin, 75%; smooth muscle α-actin, 81%; Z-line titin, 83%). Sarcomeric myosin was expressed in 63% of explants and incorporated into A-bands in 37%. The percentage incidence of expression or striation of I-Z-I proteins was significantly higher than that of sarcomeric myosin. Results suggested that the nascent I-Z-I components appeared to be generated independently of A-bands in the cultured posterior blastoderm, and that the process of myofibrillogenesis observed in our culture model faithfully reflected that in vivo. Our blastoderm culture model appeared to be useful to investigate the mechanisms regulating the early cardiogenesis

  7. Measuring the style of chick lit and literature

    Jautze, K.J.


    This paper examines to what extent the distribution of the hundred most frequent function words of two novelistic genres (chick lit and literature) gives insight into the genre styles. The results shows that the literary style is more descriptive and informational, whereas the style of the chick-lit

  8. Gamma irradiation treatment of cereal grains for chick diets

    Wheat (W), triticale (T), hulled barley (HB), hull-less barley (HLB), hulled oats (HO), and hull-less oats (HLO) were gamma irradiated (60Co) at 0, 3, 6 and 9 Mrad to study the effect of irradiation on the nutritional value of cereal grains for chicks. A significant curvilinear relationship between radiation dose and 3-wk body weight of chicks fed irradiated cereals was noted for T, HB, HLB, HO and HLO. Chicks fed W or T showed no effect or lower body weight, respectively, while body weights of chicks fed barley or oat samples were higher with irradiation. The improvement tended to be maximal at the 6 Mrad level. Irradiation significantly improved the gain-to-feed ratio for chicks fed either HO or HLO. Apparent fat retention and tibia ash were higher in chicks fed irradiated HLO than in those fed untreated HLO. In a second experiment chick body weight, apparent amino acid and fat retention, tibia ash, and gain-to-feed ratios were lower in chicks fed autoclaved (121 degrees C for 20 min) barley than in those fed untreated barley. Irradiation (6 Mrad) subsequent to autoclaving barley samples eliminated these effects. Irradiation appears to benefit cereals containing soluble or mucilagenous fiber types as typified by beta-glucan of barley and oats. These fibers appear prone to irradiation-induced depolymerization, as suggested by increased beta-glucan solubility and reduced extract viscosity for irradiated barley and oat samples

  9. Feasibility Study of Ex Ovo Chick Chorioallantoic Artery Model for Investigating Pulsatile Variation of Arterial Geometry.

    Kweon-Ho Nam

    Full Text Available Despite considerable research efforts on the relationship between arterial geometry and cardiovascular pathology, information is lacking on the pulsatile geometrical variation caused by arterial distensibility and cardiomotility because of the lack of suitable in vivo experimental models and the methodological difficulties in examining the arterial dynamics. We aimed to investigate the feasibility of using a chick embryo system as an experimental model for basic research on the pulsatile variation of arterial geometry. Optical microscope video images of various arterial shapes in chick chorioallantoic circulation were recorded from different locations and different embryo samples. The high optical transparency of the chorioallantoic membrane (CAM allowed clear observation of tiny vessels and their movements. Systolic and diastolic changes in arterial geometry were visualized by detecting the wall boundaries from binary images. Several to hundreds of microns of wall displacement variations were recognized during a pulsatile cycle. The spatial maps of the wall motion harmonics and magnitude ratio of harmonic components were obtained by analyzing the temporal brightness variation at each pixel in sequential grayscale images using spectral analysis techniques. The local variations in the spectral characteristics of the arterial wall motion were reflected well in the analysis results. In addition, mapping the phase angle of the fundamental frequency identified the regional variations in the wall motion directivity and phase shift. Regional variations in wall motion phase angle and fundamental-to-second harmonic ratio were remarkable near the bifurcation area. In summary, wall motion in various arterial geometry including straight, curved and bifurcated shapes was well observed in the CAM artery model, and their local and cyclic variations could be characterized by Fourier and wavelet transforms of the acquired video images. The CAM artery model with

  10. Embryo-maternal communication

    Østrup, Esben; Hyttel, Poul; Østrup, Olga


    Communication during early pregnancy is essential for successful reproduction. In this review we address the beginning of the communication between mother and developing embryo; including morphological and transcriptional changes in the endometrium as well as epigenetic regulation mechanisms...... directing the placentation. An increasing knowledge of the embryo-maternal communication might not only help to improve the fertility of our farm animals but also our understanding of human health and reproduction....