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PubMed1982

Separation of the mRNAs coding for different forms of chick embryo gizzard tropomyosin.

The fractionation of gizzard tropomyosin from 17-day-old chick embryos by electrophoresis in polyacrylamide gels indicates that this protein is composed of three different polypeptides that were designated as alpha, beta, and gamma. Each one of these polypeptides is resolved into two components with different isoelectric points when tropomyosin is fractionated by two-dimensional electrophoresis; the major components were designated as tropomyosins alpha 1, beta 1, and gamma 1 and the minor ones as tropomyosins alpha 2, beta 2, and gamma 2. The results of in vitro translation of gizzard RNA indicate that only tropomyosins alpha 1, beta 1, and gamma 1 are synthesized in the cell-free system. A partial separation of the mRNAs coding for tropomyosins alpha 1 and gamma 1 was obtained by fractionation of gizzard RNA by zonal centrifugation in sucrose gradients. Altogether these results indicate that in gizzard tissue from 17-day-old chick embryos three genes coding for different forms of tropomyosin are expressed.

Animals↗

PubMed1981

Regulation of collagen post-translational modification in transformed human and chick-embryo cells.

Changes in the regulation of collagen post-translational modification in transformed cells were studied in three established human sarcoma cell lines and in chick-embryo fibroblasts freshly transformed by Rous sarcoma virus. The collagens synthesized by all but one of these and by all the control human and chick-embryo cell lines were almost exclusively of types I and/or III. The relative rate of collagen synthesis and the amounts of prolyl hydroxylase activity and immunoreactive protein were markedly low in all the transformed human cell lines. The other enzymes studied, lysyl hydroxylase, hydroxylysyl galactosyltransferase and galactosylhydroxylysyl glucosyltransferase, never showed as large a decrease in activity as did prolyl hydroxylase, suggesting a more efficient regulation of the last enzyme than of the three others. The chick-embryo fibroblasts freshly transformed by Rous sarcoma virus differed from the human sarcoma cells in that prolyl hydroxylase activity was distinctly increased, whereas the decreases in immunoreactive prolyl hydroxylase protein and the three other enzyme activities were very similar to those in the simian-virus-40-transformed human fibroblasts. It seems possible that this increased prolyl hydroxylase activity is only a temporary phenomenon occurring shortly after the transformation, and may be followed by a decrease in activity later. The newly synthesized collagens of all the transformed cells that produced almost exclusively collagen types I and/or III had high extents of lysyl hydroxylation, and there was also an increase in the ratio of glycosylated to non-glycosylated hydroxylysine. The data suggest that one critical factor affecting modification is the rate of collagen synthesis, which affects the ratio of enzyme to substrate in the cell.

Animals↗

PubMedDate not supplied

[Positive inotropic action of acetylcholine on the heart ventricles of the chick embryo].

Acetylcholine increased twitch tension in the whole ventricle or in ventricular strips from 2-10-day chick embryos. The effect of acetylcholine was mediated by muscarinic receptors, since it was prevented by atropine, but not by tubocurarine or propranolol. Prostigmine significantly increased the sensitivity to acetylcholine in the strips from 7-day embryos, being almost ineffective in the strips from 3-day embryos. The decrease in acetylcholine sensitivity in the developing chick embryo is presumably associated with the increase in cholinesterase activity of the myocardial tissue.

Acetylcholine↗

PubMed1979

Prebursal stem cells in the intraembryonic mesenchyme of the chick embryo at 7 days of incubation.

Cyclophosphamide-treated 18-day-old chick embryos were transplanted with cells from 7-day intraembryonic mesenchyme; the recipients and donors were identical at the major histocompatibility locus. At the age of 35 days, the cell recipients were studied to determine the reconstitution capacity of the transplanted cells. The transplantation resulted in a complete restoration of IgM and IgG class antibody production against human gammaglobulin and Brucella abortus, and of microscopic morphology of the bursa of Fabricius and of the germinal center formation in the spleen. These findings demonstrate that 7-day intraembryonic mesenchyme of the chick embryo harbor prebursal stem cells. These findings confirm our previous observations in the yolk sac-embryo chimeras indicating that lymphoid stem cells originate in the intraembryonic hematopoietic sites.

Aging↗

PubMed2006

Continuous delivery of a monoclonal antibody against Reissner's fiber into CSF reveals CSF-soluble material immunorelated to the subcommissural organ in early chick embryos.

The subcommissural organ (SCO) is an ependymal differentiation located in the dorsal midline of the caudal diencephalon under the posterior commissure. SCO cells synthesize and release glycoproteins into the cerebrospinal fluid (CSF) forming a threadlike structure known as Reissner's fiber (RF), which runs caudally along the ventricular cavities and the central canal of the spinal cord. Numerous monoclonal antibodies have been raised against bovine RF and the secretory material of the SCO. For this study, we selected the 4F7 monoclonal antibody based on its cross-reactivity with chick embryo SCO glycoproteins in vivo. E4 chick embryos were injected with 4F7 hybridoma cells or with the purified monoclonal antibody into the ventricular cavity of the optic tectum. The hybridoma cells survived, synthesized and released antibody into the CSF for at least 13 days after the injection. E5 embryos injected with 4F7 antibody displayed precipitates in the CSF comprising both the monoclonal antibody and anti-RF-positive material. Such aggregates were never observed in control embryos injected with other monoclonal antibodies used as controls. Western blot analysis of CSF from E4-E6 embryos revealed several immunoreactive bands to anti-RF (AFRU) antibody. We also found AFRU-positive material bound to the apical surface of the choroid plexus primordia in E5 embryos. These and other ultrastructural evidence suggest the existence of soluble SCO-related molecules in the CSF of early chick embryos.

Animals↗

PubMed1987

Protective role of calcium in lead toxicity during development of chick embryo.

In the present study the interaction of lead (Pb) and calcium (Ca) during embryogenesis was investigated. Lead (50 micrograms) alone and in combination with Ca (100 micrograms, 500 micrograms and 1000 micrograms) was injected on seventh day of incubation into the yolk sac of developing chick embryos. Treatment with lead alone caused stunted growth in chick embryos with several deformities e.g. defective beak and legs, hydrocephalus, microphthalmia and anophthalmia. The combined treatment of Pb with Ca showed a remarkable protective effect against lead induced deformities characterized by development of chick embryos with a low incidence of deformities.

Abnormalities, Drug-Induced↗

PubMed1995

3,4-Diaminopyridine induced hydrolysis of phosphoinositide in cultured neurons from embryo chick forebrain.

The effect of 3,4-diaminopyridine (DAP) on phosphoinositide hydrolysis in cultured neurons from embryo chick forebrain has been studied. DAP produced a dose- and time-dependent accumulation of inositol phosphates. At 1 mM DAP a maximal effect was obtained. In Ca2+ free medium, DAP-activated turnover of phosphoinositide was reduced, but was still significant. Blocking Ca2+ entry with 200 microM Cd2+ also did not abolish the DAP-induced accumulation of inositol phosphates. As a comparison the effect of high K+ exposure was investigated. High K+ enhanced phosphoinositide hydrolysis, and this effect was also reduced by excluding Ca2+ influx. Moreover, DAP had no additional effect on the high K(+)-induced hydrolysis of phosphoinositide. Using oxonol-V, a depolarization of the membrane potential was seen in the neurons bathed in DAP containing medium. It is suggested that the depolarization may play a role in DAP-activated phosphoinositide turnover in cultured neurons of the embryo chick forebrain, but that Ca2+ entry is not necessary for this effect.

4-Aminopyridine↗

PubMed1981

On the mechanism by which insulin stimulates protein synthesis in chick embryo fibroblasts.

It has been known that insulin raises the rate of incorporation of [3H]leucine into the total protein of hormone-deficient chick embryo fibroblasts by approximately 1.5-fold. The elevation is not dependent upon the production of new messenger ribonucleic acids (mRNAs). Evidence is now presented in support of the following points: the greater labelling is due to more rapid polypeptide synthesis, not to an increase in the specific activity of leucyl-tRNA; the enhanced synthesis derives largely or entirely from a speeding up of the process of initiation, rather than that of elongation or termination; and the 1.5-fold stimulation is due to the elevated rates of formation of at least many of the fibroblast proteins. The hormone was shown before to stimulate posttranscriptionally and highly preferentially for formation of ribosomal proteins in the resting chick embryo cells. The question has been asked here whether insulin increases the production of total cell ribosomal protein by chemically altering preformed mRNAs. Results obtained by translating messages from deprived and hormone-treated cells in wheat germ and reticulocyte preparations do not support a mechanism involving covalent modification of preformed mRNAs. The observations, coupled with those previously made with inhibitors of translation, lead us to suggest that insulin stimulates protein synthesis in the resting chick embryo cells by activating limiting components of the initiation system. The effects of the hormone are greatest with messages, such as those for the ribosomal proteins, that have low affinities for the limiting initiation components.

Animals↗

PubMed1996

Induction effects of polychlorinated biphenyls, polycyclic aromatic hydrocarbons and other widespread aromatic environmental pollutants on microsomal monooxygenase activities in chick embryo liver.

Cytochrome P450-dependent 7-ethoxyresorufin O-deethylase (EROD), 7-pentoxyresorufin O-dealkylase (PROD) and 7-ethoxycoumarin O-deethylase (ECOD) activities in 14-day-old chick embryo livers were determined 24 h after pretreatment with selected widespread aromatic environmental contaminants, including polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), hexachlorobenzene, and dialkylesters of phthalic acid, and compared with the inducing potencies of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the coplanar and mono-o-chlorinated PCBs. The effects of other model inducers, i.e. phenobarbital and pyrazole, were also examined. Specificity of EROD induction was estimated with regard to contaminants frequently present in environmental samples and dose-response curves for EROD induction were determined. A strong induction (comparable with that by mono-o-chlorinated biphenyl treatment) by dibenzo[a,h]anthracene, benzo[k]fluoranthene or benzo[b]fluoranthene was found, but the maximal level of EROD activity inducible by TCDD was not achieved, partly due to the high toxicity of the tested PAHs. 3-Methylcholanthrene showed moderate inducing potencies; benz[a]anthracene, benzo[a]pyrene, chrysene and 2,2',3,4,4',5'-hexachlorobiphenyl appeared to be weak inducers. Other PAHs and PCBs tested, as well as hexachlorobenzene, dialkyl phthalates, phenobarbital and pyrazole had no marked effects on the EROD level. ECOD activities were increased non-specifically by TCDD, 3-methylcholanthrene, hexachlorobenzene and phenobarbital. A significant enhancement of PROD activity by TCDD and related inducers was observed, while phenobarbital induced the PROD activity only weakly; SDS-PAGE analysis showed that the chicken phenobarbital-inducible cytochromes P4502H with apparent molecular weights 50 kDa were not markedly induced by the TCDD- or 3-methylcholanthrene treatments. Inhibition of EROD and PROD by 9-hydroxyellipticine, a specific inhibitor of rat hepatic cytochrome P4501A1, revealed that PROD induction by TCDD and other P4501A-inducers was probably a result of a broader substrate specificity of chick embryo P4501A. Measurement of EROD activities in chick embryo liver is highly sensitive, specific and suitable for the determination of TCDD-type toxicity of new drugs, agrochemicals, and industrial pollutants.

7-Alkoxycoumarin O-Dealkylase↗

PubMed1975

Incidence and origin of heteroploidy, especially haploidy, in chick embryos from intraline and interline matings.

Preparations for chromosomal analysis were made from 2107 chick embryos at 16 hours of incubation. The embryos resulted from intraline and reciprocal interline matings of two genetically different stocks (AG and D6). The two stocks had been previously characterized as producers of high (AG) and low (D6) frequencies of chromosomally aberrant embryos. The overall frequency of aberrant embryos was 4.0 plus or minus 0.42%. The types and frequencies of abnormalities were: haploidy and haploid-euploid mosaics, 57%; polyploidy and polyploid-diploid mosaics, 19%; aneuploidy, 17%; aneuploid-diploid mosaics, 5%; and structural aberrations, 2%. Although there were no significant differences among the four types of matings in the overall frequency of heteroploid embryos (P greater than 0.1), a significant difference (P smaller than 0.01) in the frequency of haploid and haploid-euploid mosaic embryos was found. The difference was entirely attributable to the line of dam; D6 dams had 1.2% and AG dams had 2.7% haploid and haploid-euploid mosaic embryos. The difference between lines of sires was not significant. In addition, there was evidence of nonrandom distribution, among dams of both lines, of haploid and haploid-euploid mosaic embryos. It was concluded that the genotype of dam is an important influencing factor in the production of haploid cell lines in embryos. The superfluous genome in triploid embryos is usually maternal in origin, resulting from retention of the second polar body in the ovum. Sex-chromosome aneuploidy in chick embryos apparently derives from nondisjunction at meiosis I of oogenesis. The sex proportion of 2023 chromosomally sexed, diploid embryos was 50.2 plus or minus 1.1% male. No significant heterogeneity was observed among the types of matings. Sex proportion was not significantly influenced by any of a number of nongenetic variables.

Aneuploidy↗

PubMed2003

Retinal growth hormone in the chick embryo.

The presence of GH and GH mRNA in the eyes of embryonic chicks is controversial and has, therefore, been further examined. In this study, GH cDNAs identical in size and sequence to the full-length pituitary transcript were generated by RT-PCR from mRNA extracted from the neural retinas of embryonic day (ED) 7 chick embryo eyes. GH immunoreactivity in the neural retina of embryos was primarily associated with proteins of 15 and 16 kDa, whereas only trace amounts of monomer (22-25 kDa) GH, the most abundant form in the pituitary gland, were present. GH immunoreactivity was also present in the vitreous humor, although this was associated only with the 15-kDa protein. After hatch, retinal proteins with GH immunoreactivity of 15 and 16 kDa were present neonatally but not after 42 d of age. The GH immunoreactivity in the neural retina of ED8 embryos was widespread, although GH staining was particularly abundant in retinal ganglion cells (RGCs). Full-length GH mRNA was similarly located, by in situ hybridization, throughout the neural retina and concentrated in cells in the RGC layer. The neural retina is also a site of GH action because 10-6 m chicken GH greatly increased (4- to 5-fold) the content of IGF-1 mRNA in 48-h cultured ED8 neural retinas. These results demonstrate the presence of full-length GH mRNA in the neural retina of chick embryos, in which GH immunoreactivity is primarily associated with RGCs and submonomer GH proteins of 15-16 kDa. These results also demonstrate GH action in the neural retina of embryos and suggest hitherto unsuspected roles for GH in retinal development and/or ocular function.

Amino Acid Sequence↗

PubMed1980

Alterations of chondroitin sulfate synthesized by chick embryo cartilage cultured in the presence of 6-aminonicotinamide.

Treatment of day-4 chick embryos with 6-aminonicotinamide (6-AN) impairs limbs chondrogenesis and produces micromelia. Interference with limb cartilage development may be related to decreased NAD-dependent synthesis of ATP due to the fact that chondrogenesis is dependent upon anaerobic metabolism. To better understand the effect of 6-AN on chondrogenesis, isolated cartilage epiphyses from day-11 chick embryos were treated in vitro. Sulfate incorporation into total glycosaminoglycans of treated epiphyses was 30% of control. Incorporation of [3H]glucosamine was normal. Fractionation by gel chromatography showed that 40% of the glycosaminoglycans synthesized by treated cells had a molecular weight of less than 15000 compared with 5% of that of the control. A decrease in amount of chondroitin 6-sulfate, an increase of chondroitin 4-sulfate and no change in amount of unsulfated polysaccharide were observed. These results suggest that, upon exposure to 6-AN, chondrocytes produce shorter than normal chondroitin sulfate chains that are preferentially sulfated in the 4 position. Since endochondral bone formation plays an integral role in growth and development of the limb, a defect in production of chondroitin sulfate, a major constituent of cartilage matrix, appears to be involved in 6-AN-induced micromelia.

6-Aminonicotinamide↗

PubMed1987

T3 stimulation of 2-deoxy-D-glucose uptake in cultured chick embryo carcass derived cells, requires neosynthesis of proteins.

T3 stimulated, in a dose dependent manner, the uptake of 2-deoxy-D-glucose (2-DOG) into cultured chick embryo carcass derived cells. A significant increase in sugar uptake was seen already at a T3 concentration of 1 pmol/l. The stimulation of 2-DOG uptake increased with time during the 6 h of exposure to T3. The hormone also stimulated, within 45 min, the incorporation of [3H]leucine and [3H]uridine into the trichloroacetic acid precipitable material of these cells. Actinomycin-D (100 micrograms/l) and cycloheximide (1 mg/l) each were capable of blocking the stimulatory effect of 10(-8) mol/l T3 on sugar uptake and on uridine and leucine incorporation. Thus, T3 in these cultured chick embryo cells stimulated sugar transport through processes dependent on neosynthesis of proteins. In this respect the effect of T3 is different from that seen in cultured chick embryo cardiomyocytes.

Animals↗

PubMed1993

Cloning and sequencing of a chick embryo cDNA encoding the 20-kDa hypusine-containing protein, eIF-5A.

Chick embryo contains 18- and 20-kDa isoforms of eukaryotic translation initiation factor 5A (eIF-5A). cDNA clones corresponding to the 20-kDa eIF-5A were isolated and sequenced. A full-length cDNA clone encodes a 153-amino-acid (aa) protein. The deduced aa sequence exactly matches with the partial aa sequence determined for this protein and shows high identity to that of human or rabbit eIF-5A. The results of Southern and Northern hybridization provide evidence for multiple transcripts for chick embryo eIF-5A or an eIF-5A-like protein that presumably derive from more than one gene.

Amino Acid Sequence↗

PubMed1996

Developmental changes in the levels of molecular species of triacylglycerol that contain docosahexaenoic acid in adipose tissue of the chick embryo.

Triacylglycerols from the initial yolk and from the adipose tissue of the chick embryo were subjected high-performance liquid chromatography in the silver ion mode to resolve molecular species of triacylglycerol on the basis of the degree of unsaturation. A total of 12 such species was resolved from the yolk samples, with only one of these species containing docosahexaenoic acid. In contrast, 17 species were resolved from adipose tissue at day 12 of embryo development, of which 8 contained docosahexaenoic acid. The major species containing this fatty acid consisted of docosahexaenoic acid in combination with palmitic and oleic acids. The amounts of the species consisting of C16 and C18 fatty acids in the adipose tissue increased continuously from day 12 until just before hatching at day 19 and then decreased slightly over the hatching period. In contrast, the amounts within the adipose depot of the species containing docosahexaenoic acid reached a maximal level by day 16 and then decreased dramatically to almost undetectable levels during the hatching period. The possibility that adipose tissue triacylglycerol in the chick embryo may function as a temporary store of docosahexaenoic acid for subsequent delivery to the developing neural tissues is suggested.

Adipose Tissue↗

PubMed1983

Survival of HRP-labeled spinal motoneurons of chick embryo in tissue and cell cultures.

Motoneurons in the lumbar spinal cord of chick embryos 6 days old, the age when their cell death starts, were labeled in vivo by retrograde axonal transport of horseradish peroxidase (HRP) and then cultured as cord fragments or isolated cells. In Eagle's minimum essential medium with no additives, spinal cord cells survived well, but among them, the survival of HRP-labeled motoneurons was poor. Muscle-conditioned medium and a mixture of horse serum (HS) and whole chick embryo extract (CEE) promoted motoneuron survival in both suspension culture of spinal cord fragments and monolayer culture of spinal cord cells. It appeared that multiple factors may be involved in the increased extent of motoneuron survival, because HS or CEE alone showed only weak activity in contrast with the combination of HS and CEE.

Animals↗

PubMed1992

N-glycosylated proteins interfere with the first cellular migrations in early chick embryo.

Cell adhesion and migration properties which are known to play a crucial role in developmental events seem to be modulated by variations in glycosylation of glycoproteins. In the chick embryo, the extracellular matrix (ECM) appears as a loose meshwork of fibrillar material in the space between the epiblast and the hypoblast shortly before the first major cell migrations start. Chick embryos treated with tunicamycin (TN), a specific inhibitor of N-linked glycosylation of proteins, show little or no ECM, diminished cell adhesion and a dramatic alteration in the architecture of the epiblast and of the hypoblast. The first major cell migrations which signal the onset of PS and gastrula formation are inhibited irreversibly in these embryos. Tunicamycin induces a substantial change in the labeling pattern with change in mobility of some polypeptides and with the induction or marked accentuation of multiple charged species (isoforms) of polypeptides different from these already present in the control blastoderm. The N-linked glycosylation of protein(s) that are synthesized during the interaction of the epiblast and of the hypoblast seem to play a critical role in cell adhesion and in the morphogenetic movements of gastrulation in the early chick embryo.

Animals↗

PubMed1977

The effect of propranolol upon chick embryo cardiogenesis.

The teratogenic effects of propranolol HCl on cardiac development were studied in chick embryos of days 3 and 4 of incubation. Propranolol was injected into the yolk sac at doses ranging from 0.05 to 0.6 mg per egg. All the treated and control embryos were examined on day 7. The LD50 for the embryos treated on the 3rd and 4th day was 0.15 and 0.35 mg per embryo, respectively. Cardiac anomalies such as aortic stenosis ventricular septal defects and common truncus arteriosus were observed. Other malformations included atrial septal defects, thin atrial wall and defects of the pulmonic, aortic and atrioventricular valves. The incidence of cardiac anomalies in the controls was very low. Propranolol was observed to slow the heart rate in the experimental embryos. It is suggested that slowing of heart rate at the early stages of heart development caused aberrant bloodstream flow patterns which probably resulted in the genesis of cardiac anomalies. The results of this study indicate that propranolol has teratogenic effects on chick embryo cardiogenesis.

Abnormalities, Drug-Induced↗