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PubMed1982

Host antigens on avian oncoviruses: presence on viruses produced by quail, duck and rat cells of antigens related to membrane antigens of chick embryo fibroblasts and chicken erythrocytes.

Avian sarcoma viruses (ASV) produced by Japanese quail embryo fibroblasts (QEF) were inactivated to the same degree as ASV produced by chick embryo fibroblasts (CEF), with or without complement, by rabbit antisera to CEF and to membrane antigens of chick embryo and adult chicken erythrocytes, in particular their unique age-specific antigens. ASV produced by duck embryo fibroblasts (DEF) were only inactivated by the antisera to the chicken erythrocyte antigens. Hence, viruses produced by QEF and DEF bear on their envelope antigens related to the chicken antigens picked up by ASV in CEF. These antigens are presumably coded by the quail and duck cells, since antigens related to the chicken antigens were found on QEF and on quail and duck erythrocytes. Antigens related to the chicken antigens have also been found on ASV shed in low amounts by semi-permissive rat sarcoma cells (line 17RBI77) and on the surface of these cells, as well as on that of another semi-permissive cell line (RBH) originating from a hamster sarcoma produced by inoculation of the rat cells. The origin of these antigens, which may play a role in semi-permissiveness, remains to be explained since they do not appear to be normally expressed by rat or hamster cells. It was also found that, contrary to an earlier conclusion and in agreement with a recent report, uninfected CEF bear on their membrane an antigen that is related or identical to the specific antigen of chick embryo erythrocytes. Therefore, only the antigen related to the adult-specific chicken erythrocyte antigen does not pre-exist on CEF.

Animals↗

PubMed1996

Grafts of the third branchial arch in chick embryos.

The parathyroid glands have been classically considered derivatives of the third and fourth pharyngeal pouches in most species, including humans. The presence of neural crest-derived cells in parathyroid glands connective tissue has apparently been established. However, our previous studies have provided a new hypothesis on the origin of these glands in human and chick embryos. To determine the true origin of the third parathyroid (parathyroid III) gland in the chick embryo, pieces of the third branchial arch from donor chick embryos at Hamburger and Hamilton's stage 19 (embryonic day 3) were grafted to host chick embryos at the same stage of development. Starting from Hamburger and Hamilton's stage 27 (embryonic day 5), a structure identified as the parathyroid III appeared in the ectodermal (epipharyngeal) placode of the third branchial arch graft, from which it subsequently became separated at Hamburger and Hamilton's stage 28 (embryonic day 5.5) and continued to develop and mature. Our findings suggest the conclusion that the parathyroid III gland begins to develop from the epipharyngeal placode, so that this gland, from our point of view, could be considered ectodermal in nature.

Animals↗

PubMed1987

Expression of an endogenous galactose-binding lectin in the early chick embryo.

The gastrulating chick blastoderm contains lectin activity specific for beta-D-galactoside groups. The galactose-binding lectin isolated by affinity chromatography on rho-aminophenyl-beta-D-lactoside separates into two bands when studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. One of these LII has a relative molecular mass of 70 (+/- 2) X 10(3) while the other LI is a polypeptide that migrates with the dye front in 10% gels. We have prepared an antiserum against this lectin preparation and have affinity-purified antibodies against LI. When embryos at stages 3-7 were examined by immunofluorescence using the affinity-purified antibodies, lectin was expressed in cells at the lowest portions of the primitive streak as well as in cells migrating laterally from this region to form the endoderm. Lectin was also expressed by the cells of the extra-embryonic endoderm and the primordial germ cells of the proximal area opaca. In transfers of gradient gels stained with affinity-purified antibodies against LI, this lectin had an approximate molecular weight of 6.5 X 10(3). Our results indicate that this lectin is expressed in areas that are undergoing cell spreading.

Animals↗

PubMed1991

Secretion of chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase from cultured chick embryo chondrocytes.

We found that chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase were released into the culture medium from the cultured chick embryo chondrocytes. Since the release of the sulfotransferases was observed not only in serum-supplemented medium but also in serum-free medium, the released sulfotransferases were unlikely to be derived from serum. Addition of ascorbate to the serum-free medium supported the continuous release of the sulfotransferases. Monensin, which is known to cause dilatation of the Golgi apparatus and to inhibit sulfation of proteoglycan, was found to affect the release of the sulfotransferases. In the presence of 10(-6) M monensin, chondroitin 6-sulfotransferase activity in the cell layer was decreased to less than one tenth of the control, and the rate of the release of the activity became much smaller than the control after the initial rapid release. The activity of chondroitin 4-sulfotransferase was also affected by monensin, but the reduction of the chondroitin 4-sulfotransferase activity in the cell layer was not so great as the reduction of chondroitin 6-sulfotransferase activity. Unlike to the microsomal sulfotransferases, both chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase released into the culture medium were retained in the soluble fraction after centrifugation at 100,000 x g for 60 min, and were not activated by detergent. pH optimum and requirements for sulfhydryl compounds of the released sulfotransferases were similar to those observed previously in the chondroitin sulfotransferases from chick embryo cartilage and from cultured chick embryo chondrocytes. These results suggest that chondroitin sulfotransferases, which are localized in the Golgi apparatus, may be secreted to the extracellular space in a soluble form under the culture conditions.

Animals↗

PubMed1956

Latent viral infection of cells in tissue culture. I. Studies on latent infection of chick embryo tissues with psittacosis virus.

Chick embryo tissues maintained for from 11 to 28 days in Hank's balanced salt solution lost their capacity to support the multiplication of psittacosis virus. The virus however infected such cells, as active multiplication of the virus occurred on the addition of beef embryo extract to this nutritionally poor medium at any period up to 28 days of cultivation in balanced salt solution. The virus remained in a state of latency for as long as 15 days in these starved cells in a non-infectious phase. These results obtained in this in vitro model system for the investigation of latent infections of cells with psittacosis virus suggest that cell nutrition as well as an alteration in the immunological defenses of the host may prove to be an important factor in the activation of latent viral infections.

Chlamydophila psittaci↗

PubMed1998

Cytokinetic pattern in the thoracic spinal cord of chick embryos (incubation day 5-13) using PCNA staining and TUNEL method.

For the cytokinetic studies using spinal cords of chick embryos, chronological patterns of cell proliferation and programmed cell death (apoptosis) should be known. Information in the early stages of chick embryos is available while data on later stages are seldom available. To investigate the chronological patterns of cell proliferation and apoptosis in the thoracic spinal cord of normal chick embryos on incubation day 5, 6, 8, 10 and 13 (Hamburger and Hamilton stage 26-40), proliferating cell nuclear antigen (PCNA) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method were used. Cell proliferation was active at the germinal layer on days 5 and 6. It markedly declined on day 8 and became negligible on day 13. TUNEL-positive cells were mainly found in the germinal layer, the ventrolateral part of the mantle layer and the dorsal root ganglion. Compared to PCNA-positive cells, TUNEL-positive cells were sparse, especially after day 10, when only a few positive cells were scattered. These results will be used as a control data for the studies such as an experimental research for neural tube defects.

Animals↗

PubMed1978

Synergistic effect of tumor virus transformation and tumor promoter treatment on the production of plasminogen activator by chick embryo fibroblasts.

Cultures of Rous sarcoma virus-transformed chick embryo fibroblasts (RSVCEF) produce 50-fold more of the protease plasminogen activator (PA), than do normal chick embryo fibroblasts. Treatment of RSVCEF cultures with the tumor promoter phorbol myristate acetate (PMA) further enhances (8- to 12-fold) the level of PA activity. Increased levels of PA activity in RSVCEF are observed as early as 1 to 2 hr after PMA treatment. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrates that the PA produced by PMA-treated cultures has a molecular weight identical to that of the PA produced by untreated cultures. PA induction by PMA cannot be accomplished in cell-free extracts, but requires protein synthesis in intact cells. Under serum-free conditions, PMA-treated RSVCEF secrete high levels of PA for 4 to 6 days and undergo pronounced morphological alterations. Modified culture conditions and the use of PMA to induce PA has allowed for the accumulation of large amounts of RSVCEF culture fluid and the subsequent purification of the enzyme. The sensitivity of transformed CEF to PMA and the generation of enhanced proteolytic activity in the cellular microenvironment may provide a model system to examine the role of both PA in malignant transformation and PMA in tumor promotion.

Avian Sarcoma Viruses↗

PubMed1977

A continuous line of Rous sarcoma virus-transformed chick embryo cells.

A continuous line of Rous sarcoma virus (RSV)-transformed chick embryo cells was established. The cells, designated RTAZ-1, which constitute the only known line of continuously growing RSV-transformed cells of chick embryo origin, grow rapidly, display uniform morphology, and perpetually release large amounts of RSV (Rous-associated virus, type 1). RTAZ-1 cells display a heteroploid chromosome complement with 92-94 chromosomes characteristic of chicken cells. Molecular hybridization studies demonstrated that DNA from these cells hybridized with normal chick DNA but not with human (KB) cell DNA, thus providing additional evidence of the chicken origin of RTAZ-1.

Animals↗

PubMed1986

Effect of activity on the selective stabilization of the motor innervation of fast muscle posterior latissimus dorsi from chick embryo.

The role of neuromuscular activity in the maturation of the motor innervation was investigated in the fast focally innervated posterior latissimus dorsi (PLD) muscle of the chick embryo. The axonal supply in the PLD motor nerve, and the focal multiple innervation of the endplates were described on days 15 and 16 of embryonic life in normal and experimental embryos. In the first series of experiments, chick embryos were paralyzed by repeated injections between days 4 and 10 in ovo of the curare-like agent, flaxedil. Twice more axons in the PLD motor nerve and about twice more nerve terminal profiles at the endplates in the PLD muscles were found in paralyzed than in control embryos. In a second series of experiments, electrodes were implanted around the spinal cord of 7-day-old embryos and electric pulses delivered at 0.5 Hz frequency from day 10 to days 15-16 of incubation. At day 15.5, no change was observed in the axonal supply in the PLD motor nerve of stimulated embryos, while a two-fold decrease was observed in the number of motor nerve terminal profiles per endplate in the corresponding PLD muscle. The statistical distribution of the number of motor nerve terminal profiles per endplate was described from complete semi-serial sections in the PLD muscle from normal, paralyzed and stimulated chick embryos. In these three cases, the distribution of supernumerary nerve terminal profiles followed a Poisson law after one nerve ending had been subtracted from the number of nerve endings counted per endplate.

Animals↗

PubMed1990

The insulin signal initiating cellular differentiation is preserved by chick embryo myoblasts incubated at 2 degrees C.

Insulin pulse treatment, lasting for 1 to 3 min, stimulates differentiation of chick embryo myoblasts to myotubes and myofibers in a serum-free medium. The use of serum-free medium supplemented with 2,2'-thiodiethanol permits terminal differentiation of chick embryo myoblasts to cross-striated, spontaneously contracting myofibers. The experiments carried out showed that incubation of chick embryo myoblasts after the insulin pulse treatment for 10 days at 2 degrees C does not inhibit the progress of their differentiation. The data demonstrate that differentiation can be interrupted at any time by transferring cells to 2 degrees C and resumed without delay after returning to 37 degrees C.

Animals↗

PubMed1976

Development of 1,25-dihydroxycholecalciferol receptor in the duodenal cytosol of chick embryo.

The development of 1,25-(OH)2D3 receptor in the duodenal cytosol of chick embryo was studied by the sucrose density gradient analysis. The binding profile for 1,25-(OH)2D3 in the cytosol of vitamin D-deficient chick duodenum on the sucrose density gradient revealed 3 binding components, and the sedimentation constant was estimated as 2.5, 3.5 and 5.5S respectively. The 3.5S binding component has high affinity and low capacity for 1,25-(OH)2D3 and is thought to be 1,25-(OH)2D3 receptor. During the development of chick embryo, the 3.5S binding component was not detected in 13-day embryonic duodenum, it appeared on 15th day of incubation and then gradually increased to the level of vitamin D-deficient chick on 19th day of incubation. The 5.5S binding component was specific for 25-OH-D3 and it was found even in 13-day embryo, but it did not show any significant change during development. On the other hand, the 2.5S component was not specific for either 1,25-(OH)2D3 or 25-OH-D3. However, it was main binding component in early stages of development and decreased during development. From these results, it is suggested that the receptor for 1,25-(OH)2D3 is available a few days before hatching and the inability to produce CaBP in the duodenum of chick embryo could not be ascribed to the absence of the receptor.

Animals↗

PubMedDate not supplied

[Effect of heterogeneous inductors on the ectoderm of the early gastrula in Rana temporaria in vitro. 5. Biochemical analysis of induction-active extracts from chick embryo retina and brain].

The water extracts from the retina and brain of 7-8-day old chick embryos were centrifuged at 20,000 g; sediments were discarded and supernatants were additionally centrifuged at 110,000 g. The inductive activity of supernatants (20,000 and 110,000 g) and sediments (110,000 g) was estimated in vitro on the Rana temporaria early gastrula ectoderm. The neutralizing activity was related exclusively to the soluble fractions of the extracts from the chick embryo retina and brain. The lens-inducing activity appeared to be characteristic of both the supernatants and the microsome fractions of these extracts. A comparative biochemical analysis of the extracts (isoelectrofocusing, electrophoresis in the presence of sodium dodecylsulfate, electroblotting) has shown that the chick embryo retina and brain are similar by the spectrum and properties of peptides. It is suggested that the similarity of the extracts inducing effect on the early gastrula ectoderm is due to the presence of the same proteins (peptides) in the retina and brain. Peptides with a positive immunohistochemical reaction to vimentin and peptides of neurofilaments were found in trace quantities in the retina and brain extracts by means of immunoelectroblotting.

Animals↗

PubMed1983

Inhibition of uroporphyrinogen decarboxylase by 3,3',4,4'-tetrachlorobiphenyl in chick embryo liver cell culture.

An assay was developed to measure uroporphyrinogen decarboxylase activity in 17-day-old chick embryo liver cells in culture. Decarboxylation products arising from the action of uroporphyrinogen decarboxylase upon uroporphyrinogen III were quantitated using high-pressure liquid chromatography. 3,3',4,4'-Tetrachlorobiphenyl (TCBP) was added to chick embryo liver cells and 24 h later the uroporphyrinogen decarboxylase activity in the cells was measured; a significant inhibition was found. On the other hand, when TCBP was added directly to the crude uroporphyrinogen decarboxylase enzyme no inhibition was noted. The inhibition of uroporphyrinogen decarboxylase observed in this study explains the accumulation of uroporphyrin and heptacarboxylic porphyrin in chick embryo liver cell cultures treated with TCBP.

Animals↗

PubMed1996

[Effect of ethanol on expression of nitric oxide synthases in the cerebral culture cells from chick embryo].

The effect of ethanol on nitric oxide synthase (NOS) activity was examined histochemically and biochemically in cultured cerebral cells of chick embryos. The cells were isolated from 13- to 14-day-old chick embryos to which 10% ethanol (ethanol group) or saline (control) had been injected on the 3rd day of embryogenesis. Expression of NADPH diaphorase in cultured cells was stained using nitro blue tetrazolium (NBT). The activity of NOS was observed by the following 2 assays: NADPH diaphorase activity was determined using the substrate NBT with the cofactor NADPH, and NOS activity was determined by measurement as radiochemical activity of the conversion of [3H]L-arginine to [3H]L-citrulline. The number of isolated cells and viability from one cerebrum of chick embryo were 3 approximately 4 x 10(6) and more than 97%, respectively. In the neuronal cells, moderately positive expression of NADPH diaphorase was first detected on about the 3rd day of culture in both the control and ethanol group. The NADPH diaphorase and NOS activities in the isolated cells were higher in the ethanol group than in the control group. The NADPH diaphorase and NOS activities were significantly higher in the ethanol group than in the control group on the 4th and 2nd day of culture, respectively. These findings suggest that NO-released by elevated NADPH diaphorase activity and NOS activity is responsible for the induction of neuronal cell disorders attributed to chronic ethanol damage.

Animals↗

PubMed1989

Expression of cellular retinoic acid binding protein II (chick-CRABP II) in the chick embryo.

We previously demonstrated the presence of cellular retinoic acid binding protein II, chick-CRABP II, in chick embryos. In the present study, we investigated the distribution of chick-CRABP II in 14-day chick embryos by means of immunoblot analysis. Chick-CRABP II was expressed in skin, muscle, bone with tendon of the embryos, but not expressed in the nervous system. In adult chick tissues, chick-CRABP II was not detected on immunoblotting; Chick-CRABP II in adults amounts to less than 10 ng/mg soluble protein. These observations suggest that chick- CRABP II is an embryonic protein involved in the development of specific tissues, such as bone, muscle and skin.

Animals↗

PubMed1985

The acute and the chronic effects of imipramine on the spontaneous motility in chick embryos.

The effect of imipramine on the spontaneous motility and development of chick embryos was studied from the 4th to the 19th day of incubation. On acute administration (a single dose of 12.5 of 25 mg/kg egg weight), imipramine already induced significant depression of spontaneous motility in 11-day embryos--an effect which increased significantly after the 15th day of incubation. The similar effect of imipramine in spinal embryos testifies to its direct action on the spinal cord and draws attention to certain details of the role of supraspinal structures of the CNS in the acute effect of imipramine. The chronic administration of imipramine showed that it had an almost 100% lethal effect from 4th to the 7th day of incubation. Between the 8th and the 10th day it caused longlasting depression of spontaneous motility. When it was administered between the 11th and 16th day of incubation, no significant effect on the development of spontaneous motor activity was found in chick embryos.

Animals↗

PubMed1997

Proliferative effect in vitro of follicle-stimulating hormone on the left ovary of the chick embryo.

The purpose of the present study was to evaluate the effect of human follicle-stimulating hormone (hFSH) on cellular proliferation in the chick embryo ovary. Left ovaries from 18-day-old chick embryos (Babcock B300) were dissociated by trypsin (0.25%) treatment. In some experiments, dissociated ovarian cells were further separated by a continuous metrizamide gradient (0-20%). Four cellular subpopulations were recovered from the density gradient: (a) typical steroidogenic cells (F1, density 1.026 g/ml), (b) primary oocytes (F2, 1.048 g/ml), (c) pregranulosa cells, and (d) poorly differentiated epithelial cells (both cellular types were found in F3, 1.059 g/ml, and F4, 1.071 g/ml). Samples (5 x 10(5) cells) of dissociated cells of the whole ovary and of the four cellular subpopulations obtained from the density gradient were cultured on polycarbonate membranes in a defined medium with 0.1 microCi of [3H]thymidine for 60 hr. When necessary, 17beta-estradiol, hFSH, recombinant human FSH, and hCG were added to the medium at the begining of the culture. The total number of cells and the incorporation of [3H]thymidine to the ovarian cell aggregate at the end of the culture increased when hFSH and recombinant hFSH (0.5 IU/ml) were added. No changes were produced with hCG (2.0 IU/ml) and 17beta-estradiol (200 ng/ml) treatments. The secretion of 17beta-estradiol by cultured ovarian cells was stimulated in the hFSH-, recombinant hFSH-, and hCG-treated groups. The dose-response curve to hFSH resulted in an ED50 of 0.03 IU/ml. In the temporal curve, the stimulatory effect of hFSH on the total number of cells and the [3H]thymidine incorporation were observed at 36 hr of culture and maintained up to 60 hr. The proliferative effect of hFSH measured as [3H]thymidine incorporation was observed only in the F4 fraction of the density gradient, which includes poorly differentiated epithelial cells and pregranulosa cells from the ovarian medulla and cortex, respectively. Present results demonstrate a dose-dependent stimulation of DNA synthesis by hFSH in the inmature ovary of the 18-day-old chick embryo. Either pregranulosa cells or poorly differentiated epithelial cells or both subpopulations would be the target cells for the proliferative effect of the gonadotropin.

Animals↗

PubMed1992

Effect of lead on lipid peroxidation of the hepatic subcellular organelles of developing chick embryos.

The effect of lead on hepatic mitochondria and microsomes of chick embryo was studied with special attention to the role of lipid peroxidation in the manifestation of lead toxicity. Mitochondrial enzymes such as succinate dehydrogenase, cytochrome C oxidase and ATPase were found to decrease in a dose dependent manner upon lead administration. Further, mitochondrial cytochromes, microsomal cytochrome P-450 and heme levels were reduced considerably with concomitant increase of mitochondrial and microsomal lipid peroxides. In the present investigation an attempt has been made to correlate the lead induced lipid peroxidation, the loss of mitochondrial and microsomal hemoproteins and the inhibition of mitochondrial enzymes in chick embryo.

Animals↗