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[Effects of taurine on the heart defects in chick embryos which induced by homocysteine].

OBJECTIVE: To explore the effects of taurine on the heart defects in chick embryos induced by homocysteine (HCY). METHODS: Through the teratogenicity test of Chick Embryos, the taurine and HCY were given to yolk sac of 3 day's chick embryos. Histopathological examination after another 2 days culture. RESULTS: The occurrence of heart defects on HCY group were significantly increased. The growth of vitelline vessels was decreased. The prevention of tautine on the heart defects in chick embryos induced by HCY was very significantly effective. CONCLUSION: Taurine can prevent the developmental disturbance of hearts induced by HCY in this experiment.

Animals↗

Evidence that 2-allyl-2-isopropylacetamide, phenobarbital and 3,5-diethoxycarbonyl-1,4-dihydrocollidine induce the same cytochrome P450 mRNA in chick embryo liver.

The induction of cytochrome P450 in chick embryo liver has been studied using three different porphyrinogenic drugs, 2-allyl-2-isopropylacetamide, 3,5-diethoxycarbonyl-1,4-dihydrocollidine and phenobarbital. Pulse-labelling studies have shown that for each drug the cytochrome P450 synthesized either in ovo or in a wheat germ translation system reacted immunologically with antibody raised against the purified 2-allyl-2-isopropylacetamide-induced enzyme (Mr = 50000). To investigate whether this is due to the three drugs inducing the same protein or different proteins with common immunological determinants, nucleic acid hybridization studies have been carried out using a recently characterised 2-allyl-2-isopropylacetamide-induced cytochrome P450 cloned cDNA probe [Brooker, J. D. et al. (1982) Eur. J. Biochem. 129, 325-333]. It has been shown that the mRNA induced by each drug hybridizes with this probe and all are of similar size. The melting profile of the mRNA . cDNA hybrids indicates that the mRNAs induced by the three drugs have at least 98% homology with the cDNA probe. Restriction endonuclease digestions of total chick embryo genomal DNA and a chick cytochrome P450 genomal clone indicates that the cytochrome P450 gene homologous with the cDNA probe is represented in the genome only once. These results strongly suggest that the three drugs cause increased levels of the same cytochrome P450 mRNA, possibly due to enhanced expression of the same gene. Results are also presented which show that other cytochrome-P450-inducing drugs, 3-methylcholanthrene, beta-naphthoflavone or pregnenolone-16 alpha-carbonitrile do not increase the level of the 2-allyl-2-isopropylacetamide-inducible mRNA but rather reduce it to a level which was lower than that of the untreated controls.

Acetamides↗

Effects of mammalian hypophyseal hormones: gonadotropins, thyrotropin and adrenocorticotropin, on the embryonic development of 8-days chick embryo gonads.

When injected into chick eggs on the 8th day of incubation, gonadotropins (FSH + LH) were shown to stimulate the gonadal development on the 20th day of embryogenesis in both sexes. An increase in the testicular weight, in the seminiferous cord diameter as well as in germ and Sertoli cell number was observed in males. Similarly an increase in ovarian weight, in ovarian width, in cortical thickness and in the number of oocytes was reported in females. By contrast, thyrotropin (TSH) and adrenocorticotropin (ACTH) caused a noticeable decrease in gonadal development in both males and females as compared to the controls or to the gonadotropin-treated groups. It is concluded that the gonads of 8 days chick embryo have the capacity to respond to gonadotropic stimulation and not to the other hypophyseal hormones (TSH or ACTH). This suggests the possibility that at this early stage (8 days of incubation) the external specific hormone could induce its future hormone receptors.

Adrenocorticotropic Hormone↗

Side effects of Ricinus lectin (RCA 120) on nucleic acid synthesis in chick embryo fibroblasts.

When fibroblasts from chick embryos were treated with Ricinus lectin, the effects observed depended on the stage of development of the embryo from which the cells were prepared. Thus, in 16-day fibroblasts, which have a weak proliferative capacity, nucleic acid synthesis was less sensitive to the effect of this lectin than that in 8-day fibroblasts, whose proliferative capacity is high.

Animals↗

Germ cell death in the degenerating right ovary of the chick embryo.

The right ovary of chick embryos is known to degenerate during embryogenesis, while the left one advanced its normal growth. The present study demonstrates germ cell death in the degenerating right ovary. In the process of germ cell death, chromatin condensation, reduction of cell size, and formation of apoptotic bodies were observed. The cell organelles were almost normal in the early stages of the process. Fragments of broken germ cells were phagocytosed by neighboring cells or macrophages. From these observations, it may be assumed that germ cell death in the right ovary of chicks is related to apoptosis. In the normal left ovary, similar cell death was only observed in the germ cells in the medulla, but not in the cortex. Therefore, germ cell death seems to be a phenomenon common to the medullae of both the right and left ovaries.

Animals↗

The effect of chick embryo hyperflexion on tracheoesophageal development.

Chick embryos were hyperflexed by placement of a ligature at two critical stages in the development of the foregut, in order to test the theory that esophageal atresia and tracheoesophageal fistula result from embryonic hyperflexion. Only one "H-type" tracheoesophageal fistula was found. There were no significant disturbances of either tracheal or esophageal growth.

Animals↗

Role of the thyroid in the involution of the mesonephric Malpighian corpuscles in the chick embryo.

The mesonephros of the chick embryo normally begins to regress during the second half of embryonic life. Experimental methods, such as adenohypophysis grafting, hypophysectomy or use of antithyroid drugs, which stimulate or depress the thyroid function of the embryo, modified accordingly the regressive processes occurring in the mesonephric Malpighian corpuscles, particularly at the level of the glomerular basement laminae. These results as well as the known sensitivity of the mesonephros to thyroxine and the concordance between the steps of embryonic thyroid development and the mesonephric modifications show that the thyroid normally plays a major determining role in this phenomenon.

Animals↗

Developmental fall in whole body protein turnover of chick embryos during incubation.

Whole body protein turnover was measured in chick embryos during incubation to investigate whether or not there is a fall in fractional rates of protein synthesis and degradation during development. Stable isotopically labelled [15N]phenylalanine was injected intraperitoneally into embryos on days 12 and 19. From 60 to 90 min after injection the isotope enrichment in free and protein-bound phenylalanine was measured with a selected-ion gas-chromatograph mass-spectrometer. The results showed that from days 12 to 19 of incubation, there was a remarkable reduction in fractional rates of protein synthesis and degradation in the whole body of chick embryos. During embryonic growth, protein synthesis per unit of RNA that is, the minimum amino acid translation rate of RNA, did not change significantly, whereas the RNA:protein ratio was reduced to one-third from days 12 to 19 of incubation. It was concluded, therefore, that the dramatic fall in fractional synthesis rate in chick embryos would be entirely attributable to the rapid increase in protein content, thereby changing the RNA:protein ratio in parallel with the fractional synthesis rate.

Animals↗

Extent of intestinal damage in the developing chick embryo after repetitive hypoxia under normoxic or hyperoxic conditions.

BACKGROUND: Episodes of hypoxia and reperfusion play an important role in the development of intestinal damage during perinatal development. The aim of this study was to investigate the histopathology of the intestine in the developing chick embryo after exposure to repetitive hypoxia and recovery under two different conditions: normoxic and hyperoxic (60% O2). METHODS: Chick embryos were exposed to 5 minutes of hypoxia. This was repeated six times with a recovery period of 15 minutes under normoxic conditions (21% O2) for chick embryos in test group 1 (TG1) and under hyperoxic conditions (100% O2) for chick embryos in test group 2 (TG2), from day 11 until day 20. Chick embryos that recovered under hyperoxic conditions (100% O2) were previously incubated under hyperoxic conditions (60% O2 for 24 hours). Histologic evaluation of the ileum was performed at different times after the interventions (2, 4, 8, 16, and 24 hours). RESULTS: In both test groups, only chick embryos older than 19 days showed intestinal damage. Intestinal histology on day 19 showed vasodilation of villus capillaries (10% in TG1 and 15% in TG2), necrosis in the top of the villi (29% in TG1 and 30% in TG2), and necrosis with preservation of base of the crypts (2% in TG1) and transmucosal necrosis (2% in TG2). CONCLUSIONS: Significant histologic changes, compared with the control group, were only found in chick embryos that were studied 2 hours after the interventions. Furthermore, recovery under hyperoxic conditions did not cause more intestinal damage compared with recovery under normoxic conditions.

Age Factors↗

Skull growth after partial prospective lower beak extirpation in chick embryos.

On one side of chick embryos in stage 28, part of the prospective lower beak or the entire prospective lower beak anterior to the buccal corner was removed surgically (CLB-A and CLB-B embryos, respectively). In another group the prospective lower beak was split in the midsagittal plane (CMS embryos). The controls were shame-operated embryos (CS). The skulls of stage 39 CLB-A and CLB-B embryos showed changes in the lower beak as well as in the skull base, the pterygoid, the quadrate, the palatine, and the upper beak. Stage 39 CMS embryos showed shortening of the dentary and Meckel's cartilage. At the tip of the CMS mandible, an excessive amount of osseous material was found. The results support Dullemeijer's (1974) views concerning the issue of dependency of functional components. The value of a combined developmental and functional approach to morphology, i.e., integrated investigation of development of structure and function of the beaks and related jaw musculature, is discussed.

Animals↗

Strong electrical currents leave the primitive streak of chick embryos.

The electrical fields above chick embryos were explored with a vibrating probe. These fields indicate that steady currents with exit densities of the order of 100 microamperes per square centimeter leave the whole streak and return elsewhere through the epiblast. The epicenter of these strong exit currents lies near Hensen's node. They are probably pumped into the intraembryonic space by the epiblast and then leak out of the streak because it is a zone of junctional disruption.

Animals↗

Protection against group B meningococcal disease. I. Comparison of group-specific and type-specific protection in the chick embryo model.

Protection against group B meningococcal infection was examined using the chick embryo. 12-day-old embryos were challenged intravenously with various meningococcal strains. The chick embryo has an active reticuloendothelial system but lacks functional complement. In this model we found that protection against group B infection was primarily group specific. The group B polysaccharide antibody is an effective opsonin, but is a very poor bactericidal antibody. In contrast, the serotype antibody was bactericidal but only slightly protective in the chick embryo where protection is primarily phagocytic in nature. The group-specific and type-specific antibodies are strongly synergistic. Minute amounts of group B polysaccharide antibody caused a very significant increase in the protective effects of the serotype antibody.

Animals↗

NAD(P)H: quinone oxidoreductase (DT-diaphorase) in chick embryo liver. Comparison to activity in rat and guinea pig liver and differences in co-induction with 7-ethoxyresorufin deethylase by 2,3,7,8-tetrachlorodibenzo-p-dioxin.

NAD(P)H:quinone oxidoreductase (EC 1.6.99.2; DT-diaphorase) was present in the liver of 18- and 19-day-old chick embryos as assayed both by reduction of resorufin and by the more traditional assay, reduction of 2,6-dichlorophenolindophenol (DCPIP). Both reductions had the classic characteristics of DT-diaphorase: they were equally supported by NADPH and NADH and almost entirely inhibited by dicumarol. Chick embryo liver DT-diaphorase was entirely cytosolic. It was undetectable in the microsomal and mitochondrial fractions. Chick embryo liver cytosol and mitochondrial fractions contained an enzyme oxidizer of resorufin but not of DCPIP. The Km for NADPH for resorufin reductase was an order of magnitude higher in chick embryo than in rat or guinea pig cytosol (1 mM vs 0.1 mM). Resorufin reductase activity was higher for chick embryo than for rat or guinea pig cytosols: Vmax (nmol resorufin reduced per mg cytosolic protein per min +/- SEM) 355 +/- 28 for chick embryo, 159 +/- 10 for guinea pig and 68 +/- 28 for rat. The Vmax for DCPIP reduction was also twice as high in chick embryo as rat liver cytosol. In the chick embryo, 7 days after treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at 6.4 micrograms/kg egg (1 nmol/egg) mortality was increased 2.4-fold, hepatic DT-diaphorase 1.3-fold, and 7-ethoxyresorufin deethylase (7-EROD) 72-fold over control levels. At 32 micrograms/kg, mortality was increased 4.2-fold, DT-diaphorase 2.3-fold and 7-EROD 100-fold. In the guinea pig, 5 days after treatment with TCDD at 10 micrograms/kg, TCDD toxicity was also evident (loss of body weight and thymus weight); there was no change in DT-diaphorase as measured by resorufin reduction, confirming by a different assay the observation of Beatty and Neal (Biochem Pharmacol 27: 505-510, 1978) that TCDD does not induce DT-diaphorase in guinea pig liver, and 7-EROD was increased 8-fold. In contrast, in the rat, 7 days after exposure to TCDD at 10 micrograms/kg, there was no evidence of toxicity, DT-diaphorase was increased close to 7-fold and 7-EROD, 100-fold. The results demonstrate that avian liver contains DT-diaphorase and show that the extent to which DT-diaphorase is part of the pleiotypic response of the liver to an Ah (aryl hydrocarbon) receptor ligand is species dependent. They also suggest that DT-diaphorase induction and TCDD toxicity may be inversely related. The possibility that DT-diaphorase protects against TCDD toxicity and participates in species differences in sensitivity to TCDD toxicity warrants further investigation.

2,6-Dichloroindophenol↗

Relationship between cardiac cycle length and ventricular relaxation rate in the chick embryo.

We hypothesized that during chick embryo cardiac development, cycle length decrease (heart rate increase) may be associated with developmental changes in the ventricular relaxation rate constant, tau, which can be estimated from ventricular pressure decrease by the function P(t) = P0e-t/tau, where P0 = pressure at the time of minimum time derivative of ventricular pressure and t = time. Natural logarithm conversion results in a linear relation between In P(t) and t with slope of -1/tau. We determined tau in Hamburger-Hamilton stage 17-27 (d 3-5) embryos (n = 35) at intrinsic cycle length and in stage 24 embryos (n = 5) during cardiac cycle length perturbations with the hot-cold probe technique. Regression analysis of In P(t) from the minimum dP/dt until t = 30 ms was used to estimate tau. Intrinsic cycle length decreased during development from stage 17 to 27 and tau decreased linearly as cycle length decreased (r = 0.39, p less than 0.005). Thus, during early development, both tau and cycle length decrease (relaxation rate and heart rate increase). In stage 24 embryos, cycle length perturbation yielded a nonlinear cycle length-dependent relation with tau, with a plateau at cycle lengths less than intrinsic, i.e. tau decreased with cycle length decrease to intrinsic cycle length, then tau plateaued with further cycle length decrease. These findings demonstrate that ventricular relaxation rate is both maturation and cycle length dependent. The plateau effect of the tau-cycle length relationship may be due to developmental limitations of the calcium transport system.

Animals↗

Acetylcholinesterase activity in the myotome of the early chick embryo.

The myotome of early chick embryos was investigated histochemically by means of the acetylcholinesterase (AChE) reaction. Light-microscopically, at the cervical level, the myotome was first recognized and AChE activity demonstrated at stage 13 (2-day-old embryo). Subsequently, the myotome elongated ventro-laterally along the inner surface of the dermomyotome and reached the ventro-lateral end of the dermomyotome at stage 17 to 18 (3 day-old embryo). AChE activity in the myotome showed subsequent increase in intensity during the course of development. The myotome consisted mainly of AChE-positive cells displaying enzymatic activity along the nuclear membrane and within the cytoplasm. In contrast, almost all cells of the dermomyotome and the interstitial cells were AChE-negative. Electron-microscopically, the myotome cells of the 2 day-old embryo and the cells in the dorso-medial portion of the myotome of the 3 day-old embryo were morphologically undifferentiated; AChE activity was detected in the nuclear envelope and in single short profiles of the endoplasmic reticulum (ER). On the other hand, in the 3 day-old embryo the cells in the ventro-lateral portion of the myotome showed AChE activity in the nuclear envelope, numerous profiles of the ER and some Golgi complexes. These AChE-positive cells were regarded as developing myogenic cells based on their morphological characteristics. The present findings indicate (i) that the appearance of AChE activity in the cytoplasm is the first sign of the differentiation of myogenic cells, and (ii) that in these myogenic cells the increase in AChE activity is based on the development of the ER.

Acetylcholinesterase↗

Hydrolysis of adenylyl(2'-5')adenosine in chick embryo sera.

Similar to adult chick sera in chick embryo sera the enzyme activity hydrolyzing the core of the dimer of A-5A - adenylyl(2' -5')adenosine is present. The enzyme activity possesses the same properties in both sera. Heating of the serum to 60 degrees C for 30 minutes does not affect the enzyme activity. A marked decrease in activity occurs after heating of the serum to above 65 degrees C. Hydrolysis of A2' p5' A in the serum is more efficient under alkaline conditions (pH 9-10) than at physiological pH 7.2-7.4.

Adenosine Monophosphate↗

Effects of nerve growth factor on sympathetic neuron development in normal and limbless chick embryos.

Wing bud removal in chick embryos has been shown to affect the generation of sympathetic neurons prior to the normal period of limb innervation [Saltis and Rush (1995) J. auton. nerv. Sys., 51, 117-127.]. Pyknotic activity occurred earlier within the peripherally deprived ganglion, suggesting that a precocious cell death of dividing sympathoblasts led to the reduced neuronal population. We have now sought to test whether the effect of limb bud extirpation can be overcome by the administration of nerve growth factor (NGF). Specifically, the peripherally deprived ganglion has been examined for mitotic activity and total neuronal numbers. In brachial ganglia from the operated side, neuron numbers decreased by 67% by embryonic day (E) 13, but by only 28% when NGF was administered from E9. Ganglia on the unoperated side were unaffected by the NGF treatment. In contrast, in embryos receiving NGF from E5 to E9, neuron numbers in the ganglia increased by more than 100%, on both the intact and operated side. This increase was accompanied by a greater proportion of 3H-thymidine-labelled neurons. We therefore conclude that NGF, in addition to its previously described role of preventing naturally occurring neuron death, can also affect the generation of sympathetic neurons. This ability of NGF to affect gangliogenesis is most likely achieved by increasing the survival of dividing neuroblasts, although a direct effect on mitosis has not been excluded.

Analysis of Variance↗

Scanning electron microscopical and histochemical study of the endoderm in the early chick embryo.

The endoderm of gastrulating chick embryos shows regional variations in cell shape and size. These were studied by scanning electron microscopy, histochemistry and immunofluorescence. Particular attention was given to the distribution of the cytoskeleton. Four zones of differing morphology were observed. The changing size and shape of these zones could be correlated with the entry of the definitive endoblast through the primitive streak, displacing existing primary hypoblast to the edges of the area pellucida. Endodermal cells were shown to have a well organised cytoskeleton. The cytoskeletons of individual cells were linked to give a cytoskeletal network extending across the endoderm as a whole.

Animals↗