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PubMed1978

Cleavage in the chick embryo.

Chick embryos ranging from the stage of first cleavage to that of about 700 cells were removed from the oviduct and examined by transmission electron microscopy. Beneath the cell membrane is yolk-free cortical region containing microfilaments. Beneath this lies cytoplasm which contains yolk spheres which are graded in size, the dorsal ones being smaller than the ventral ones. The subgerminal periblast possesses a greater proportion of yolk to cytoplasm than do the cells proper, but it merges with the cytoplasm at the incomplete borders of the 'open' cells. Specialized accumulations of membranes lie in the marginal periblast, and it is suggested that they play a role in cell membrane formation.

Animals↗
PubMed1988

Extragonadal distribution of primordial germ cells in the early chick embryo.

Chick primordial germ cells (PGCs), after separation from the endoderm in early embryonic development, temporarily circulate via the blood vascular system and finally migrate into the gonadal anlagen. It has been noted by some authors that some PGCs are present in extragonadal sites in some vertebrates. In the present study, we examined the distribution and localization of PGCs in extragonadal sites in the chick embryo. PGCs were identified by periodic acid-Schiff staining with light microscopy. In embryos at stages 20-24 (PGCs are in the settlement stage in the gonadal primordium), approximately 20% of the total number of PGCs were observed in extragonadal regions. Approximately 90% of these ectopic PGCs were found in the head, mainly in the mesenchyme surrounding the neural tube. Even at stage 14 when PGCs were usually circulating in the blood vessels, some of the PGCs had emerged from the blood vessels and were detected in the extragonadal site. This pattern of distribution of ectopic PGCs in the head area is probably attributed to the earlier, dominant development of the capillary network, and to the sluggish capillary blood flow in that region, which allows intravascular PGCs to escape into the tissue.

Animals↗
PubMed1979

Development of the basal lamina and extracellular materials in the early chick embryo.

Chick embryos at developmental stages up to primitive streak formation were fixed in a mixture of tannic acid and glutaraldehyde. A basal lamina was present in the unincubated embryo and consisted of a lucent lamina interna and a lamina densa. At the primitive streak stage the lamina densa showed a periodicity of stained elements. Densely stained materials were present on the cell surfaces lining the cavity between the epiblast and endoblast, and on the mesoderm cells within this cavity. Considerable amounts of extracellular material were observed in the cavity. Hyaluronidase treatment removed the cell surface and extracellular material, indicating that hyaluronic acid is a major component. This enzyme disrupted the basal lamina, leaving a fibrillar remnant with no periodic structure. It is therefore suggested that the dense periodicities consist of glycosaminoglycan built on an enzyme-resistant framework which is probably collagen. Enzyme-resistant fibrils, presumably collagen precursors, are present elsewhere within the tissue spaces.

Animals↗
PubMed1975

The retrograde transport of horseradish peroxidase from the developing limb of the chick embryo.

Chick embryos ranging in age from 4.0 to 18 days of incubation and 1-2-day-old hatchlings received injections of HRP solutions directly into the leg musculature. After survival periods of from 5 to 25 h motoneurons and cells in the spinal sensory ganglia were found to be stained with the HRP reaction product. It was found that the first appearance of a positive HRP reaction coincided with the time when nerve processes are first detected in the limb-bud by silver techniques (i.e. at 4.5 days of incubation). Only neurons with processes in the region of injection showed a positive reaction. The demonstration of a retrograde transport mechanism in neurons and axons which are still undergoing growth and differentiation provides a possible mechanism for the peripheral regulation of certain features of CNS neurogenesis. The application of this technique during development may also allow one to map neuroanatomical pathways during their formation.

Animals↗
PubMed1986

Scanning electron microscopical observations on the differentiating mesonephros of the chick embryo.

Chick embryos were staged according to the method of Hamburger and Hamilton [1951] and fixed. Cross sections through the cephalic fourth of the mesonephric ridges were examined by scanning electron microscopy. The steps in glomerular differentiation could be observed with ease. The first foot processes to appear in podocytes arose directly from the basal surface of the cell body. In a second step, lateral branches appeared and gave off secondary or even tertiary branches that interdigitated with those from neighbouring podocytes, following a pattern that was very similar to the one previously described by other authors in metanephric nephrons. Endothelial pores appeared in the glomerular capillaries at very early stages of the glomerular differentiation. The differentiation of the epithelium of proximal tubules was characterized by the growth of apical microvilli and of finger-like evaginations from the lateral membranes. At stages 20 and 21, the most differentiated glomeruli had only basal foot processes; only after stage 25 did the first generation nephrons reach full maturity. Because during this period the mesonephros is known to produce urine, our results indicate that nephrons start to function before they have completed their differentiation.

Animals↗
PubMed1994

Agonist administration in ovo down-regulates cerebellar GABAA receptors in the chick embryo.

Chick embryos with an undeveloped blood-brain barrier were used to examine the down-regulation of GABAA receptors in vivo. The GABAA receptor agonist isoguvacine (5 mumol) was applied to the vascularized chorioallantoic membrane of 8 day embryos. This treatment was repeated on embryonic days 11, 14, and 17, and the embryos were sacrificed on day 18 (stage 42). Isoguvacine administration reduced the clonazepam-displaceable binding of [3H]flunitrazepam to washed cerebellar membranes by 34.0 +/- 3.0% compared to vehicle-treated controls. Binding reductions of lower magnitude were found in membranes from the cerebrum and optic lobes. Administration of isoguvacine had no significant effect on the wet weights of whole embryos or cerebella, the yield of cerebellar membranes, or the binding of [3H]N-methylscopolamine. The reduction of [3H]flunitrazepam binding to cerebellar membranes was dose-dependent, allowing a half saturation value of 8 microM isoguvacine to be estimated. Scatchard analysis showed that the Bmax for [3H]flunitrazepam binding was reduced by 28.3 +/- 6.7% compared to controls, without a change in the Kd. Embryonic exposure to isoguvacine also caused a reduction of 43.6 +/- 6.0% in the binding of the GABAA receptor channel ligand [35S]t-butylbicyclophosphorothionate to washed cerebellar membranes. Taken together, these results indicate that isoguvacine induces a down-regulation of the receptor subunits in vivo. However, measurements of cerebellar GABAA receptor mRNAs for the alpha 1, beta 2L, beta 2S, beta 4, gamma 1, gamma 2L, and gamma 2S subunits by reverse transcriptase-polymerase chain reaction (RT-PCR) revealed no significant alterations by isoguvacine administration. The data suggest that translational or post-translational mechanisms, rather than those modulating the synthesis or stability of subunit mRNAs, take precedence in establishing GABAA receptor down-regulation.

Allantois↗
PubMed1988

Relative growth rates of maxillary mesenchyme in the chick embryo.

Chick embryos were injected with [3H]-thymidine at days 3-7 of incubation and were fixed and embedded in plastic. The embryos were divided into three stage groupings of development [Hamburger and Hamilton: J Morphol 88:49-92, 1951], and labeling indices were determined for each of the following delineated regions within the maxillary process at each stage: region 1, subepithelial mesenchyme located at the medial side of the maxillary process adjacent to the roof of the stomodeum; region 2, subepithelial mesenchyme at the ventral tip of the maxillary process (as seen on cross section); region 3, subepithelial mesenchyme at the lateral portion of the maxillary process below the eye; and region 4, interior mesenchyme defined as the central portion of the maxillary process and separated from the epithelium by the three other regions. Results indicated that differences exist among the regions examined and that these differences were stage specific. At stages 19-21 and stages 24-25 1/2, growth rates were higher in subepithelial regions than interiorly. At stages 28-29, however, a statistically significant difference among the regions was not found. These results suggested that there is an association between growth rates in the maxillary process mesenchyme and its proximity to the overlying epithelium and that these effects are related to the stage of development.

Animals↗
PubMed1987

Relationship between genital ridge formation and settlement site of primordial germ cells in chick embryos.

Chick embryos from stage 15 to stage 18, which is the most frequent extravasation period, were investigated by means of serial sections and light microscopy in order to learn the detailed relationship between the settlement sites of the primordial germ cells (PGCs) and the forming genital ridge. PGCs circulating in the vascular system came out of the small vessels in the splanchnopleure posterior to the vitelline artery. This PGC extravasation was limited to an area about 1.2 mm caudal to the vitelline artery. After the extravasation, the PGCs entered the neighboring thickened coelomic epithelium of the splanchnopleure. This thickened epithelium, which had incorporated the PGCs, changed the location toward the future gonadal site with the advance of development. At stage 16, the thickened portion of the epithelium was located in the splanchnopleure; then it moved toward the somatopleure via the coelomic angle; finally, at stage 18, this epithelium occupied the region between coelomic angle and the mesonephros which corresponded to the future genital ridge. This means that the thickened epithelium of the splanchnopleure which initially incorporated the PGCs becomes the superficial epithelium of the genital ridge in more advanced stages. The thickened portion of the epithelium of the splanchnopleure at stage 16 is thought to be the definitive gonadal anlage.

Animals↗
PubMed1981

The effects of insulin-glucose on microappendages of the plasmalemma in the early chick embryo.

Chick embryos incubated for 18-24 hours (stages 4-6) were immersed for 3-5 minutes in Hanks' Balanced Salt Solution containing either insulin (10(-9) M) or D-glucose (10(-1) M) or a mixture of both. The ventral and dorsal surfaces of the entoderm and the ventral aspect of the mesoblast were examined in a scanning electron microscope. D-glucose alone or insulin alone produced negligible change. When combined these ingredients generated highly pleomorphic microappendages of the plasmalemma. The ventral surface of the entoderm was characterized by a large number of microvilli, both long and short. Concentrations of microvilli and blebs appeared in recesses at cell margins. Some of these represented mesoblastic protrusions through intercellular apertures in the entoderm. The dorsal entodermal surface developed stellate clusters of microvilli and blebs. The ventral aspect of the mesoblast became the most pleomorphic area of all. It possessed a variety of microappendages including large populations of microvilli and blebs that resembled the protrusions observed from the subblastodermic cavity. There were also stellate clusters similar to those on the dorsal entodermal surface. The same mesoblastic area in controls was virtually free of microappendages. The mesoblastic protrusions, occurring as they do prior to an established circulation, are interpreted as a "feeding" phenomenon.

Animals↗
PubMed1990

Cytoskeletal organization and synthesis in substrate-independent and -dependent myogenesis in chick embryos.

Chick embryo myoblasts were fused in suspension culture to form myoballs by modification of previous procedures that excluded the use of divalent ion chelators and antimitotic drugs and included the continuous presence of serum in order to analyze the organized appearance and synthesis of major cytoskeletal proteins during cell attachment and spreading. The organization of the major cytoskeletal proteins actin, tubulin, and vimentin was assessed by fluorescence microscopy under these conditions as well as under conditions in which the myoballs were allowed to attach and spread on a collagen-coated substrate. Actin, detected by fluorescence microscopy, stained myoballs diffusely and was reorganized to form stress fibers in the attached and spreading myoball. Nuclei were segregated to a centrally located lattice of microtubules. The microtubule-specific drugs nocodazole and taxol prevented myoball spreading and the establishment of myotube polarity, respectively. Vimentin appeared as wavy ribbons in a perinuclear position around attached and spreading myoballs. In parallel studies, the synthesis of these cytoskeletal proteins was analyzed by radioisotopic labeling and polyacrylamide gel electrophoresis. These studies showed that myoballs possess altered ratios of actin and tubulin isoforms and of phosphorylated and nonphosphorylated vimentin compared to myotubes. These ratios rapidly change to the myotube pattern when myoballs are allowed to attach to solid substrata. Thus, although both myoballs and myotubes undergo muscle-specific differentiation, their cytoskeletal proteins are morphologically and biochemically distinct.

Actin Cytoskeleton↗
PubMed1976

Collagen glucosyltransferase. Partial purification and characterization of the enzyme from whole chick embryos and chick-embryo cartilage.

A purification of over 2000-fold is reported for collagen glucosyltransferase from Triton X-100 extract of whole chick embryos and one of about 160-fold from similar extract of chick embryo cartilage. The addition of the detergent more than doubled the enzyme activity in the homogenates. The purified enzyme preparations from whole chick embryos showed one major band and two or three minor bands in polyacrylamide gel electrophoresis and were entirely free of collagen galactosyltransferase activity. The molecular weight of collagen glucosyltransferase from both sources was about 52000 -- 54000, as determined by gel filtration. In some enzyme preparations an additional form was observed, with an elution position corresponding to a molecular weight of about 130000. Manganese was the most effective metal co-factor for the purified enzyme, but partial replacement could be obtained with Co2+, Mg2+ and Ca2+, whereas no replacement was found with other metals. The activity of the purified enzyme was stimulated by the addition of dithiothreitol to the incubation system and inhibited by preincubation with p-mercuribenzoate. UDP-glucose or the collagen substrate partially protected the enzyme against p-mercuribenzoate inactivation in the presence of Mn2+ but not in its absence. Some protection was also noted with Mn2+ alone.

Animals↗
PubMed1982

Development of the articular cavity in paralyzed chick embryos and in chick embryo limb buds cultured on chorioallantoic membranes.

The development of digital joints of chick embryo paralyzed with dacamethonium bromide and of joints in chick embryo limb buds cultured on chorioallantoic membranes was studied. The digital joints of 45 paralyzed and 40 control embryos and the joints of 37 grafts and 30 control legs were examined histologically. Six preselected grafts were studied by electron microscope. Our results demonstrate that paralysis does not completely inhibit the early stages of joint clefting. In many paralysis does not completely inhibit the early stages of joint clefting. In many paralyzed embryos, small clefts appeared in the periphery and occasionally in the central part of the interzone. Unlike in normal embryos, they rapidly disappeared, inducing fusions across the joints. Early, 2-day paralysis during a period of normal clefting inhibited irreversibly the development of the articular cavity that failed to appear after a cessation of the drug. Paralysis initiated in older embryos caused rapid regression and partial fibrous ankylosis of previously fully differentiated joints. Grafts from 4- to 6-day-old embryos usually failed to develop a full range of skeletal segments. Normal development of skeletal segments occurred in grafts from older donors. The articular cavity failed to develop in almost all grafts. Early cavitation accompanied by characteristic cell changes was found in a few joints. These cellular changes are suggestive of an intrinsic mechanism of early cavitation, but joint motion is required for the full differentiation and maintenance of the joint cavity.

Allantois↗
PubMed1987

Histochemical studies of the differentiation of microglial cells in the cerebral hemispheres of chick embryos and chicks.

Using histochemical procedures to reveal the presence of nucleoside diphosphatase (NDPase), thiamine pyrophosphatase (TPPase) and acid phosphatase (AcPase), we investigated the appearance, distribution and ultrastructure of amoeboid and microglial cells in the cerebral hemispheres of chick embryos and young chicks, in order to elucidate the relationship between these two cell populations. On day 6 of incubation, a few round cells exhibiting NDPase, TPPase and AcPase activity were first detected in the thin mantle layer of the cerebral hemisphere. In the corpus striatum, these round cells increased rapidly in abundance until day 13 of incubation, after which their numbers gradually decreased, so that, on day 19 of incubation, they had entirely disappeared. Between day 10 and day 17 or 18 of incubation, round cells were located mainly in the zone of the mantle layer closest to the lumen. On day 10 of incubation, NDPase-, TPPase- and AcPase-positive cells that had a few short cytoplasmic processes (poorly ramified cells) were detected in the intermediate and basal zones of mantle layer. They increased in abundance until day 17 or 18 of incubation and thereafter rapidly decreased in number. Round and poorly ramified cells exhibited NDPase activity on their plasma membranes and in their cytoplasmic vacuoles, with TPPase and AcPase activity being localized within their vacuoles. On day 19 of incubation, NDPase- and TPPase-positive cells with long, well-ramified cytoplasmic processes (well-ramified cells) were observed in the corpus striatum, these being mainly localized in the basal zone. After hatching, these cells increased rapidly in abundance and were distributed throughout the corpus striatum.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Anhydride Hydrolases↗
PubMed1979

[Serotoninergic innervation of the paraventricular organ in chick embryo and chick; histofluorescence and autoradiography study optic and electron microscopy].

Fluorescence microscopy (Falck and Hillarp technique) of 11th day old chick embryo shows aminergic neurons and processes in the paraventricular organ area. Administration of tritiated amines permits us to observe uptake and storage of serotonin only as early as the 10th day; this suggests a very early hypothalamic serotoninergic nervous system differentiation.

Animals↗
PubMed1976

[Effect of stimulating doses of ionizing radiation on the stability of the liver mitochondria of chick embryos and chicks].

Treatments at the rate of 3 rad of chick embryos and young birds have led to the stabilizing of the mitochondrial structures and to lowering the access of enzymes to their substrates. The free activity of malate dehydrogenase drops. Electron microscopic studies reveal that the number of mitochondria rises following irradiation with stimulating doses (3 rad). Mitochondrial cristae remain intact and the matrix presents higher electronic density.

Age Factors↗
PubMedDate not supplied

[Organization of impulse flows and selective neuronal death in the hyperstriatum of chick embryos and chicks].

Studies have been made on changes in the number of neurones per volume unit of the tissue of hyperstriatum tissue in chick embryos from the 18th day of incubation to the 3rd day after hatching. Significant interhemispheric asymmetry in migration rate and cell death was revealed in the dorsal hyperstriatum from the 19th day of embryogenesis, which may be due to asymmetry in the intensity of visual afferentation which depends on the stationary position of the embryo in the egg. In the activity of neuronal populations within the dorsal hyperstriatum at a stage which immediately precedes mass death of cells (up to 40%), a structure of impulse volley was found which is characterised by the onset of impulse discharges following each other in a form of short series. Sometimes it is possible to reveal correlation between the duration of dominating interdischarge intervals and the extremes in the recovery of excitability of neuronal pools, which indirectly indicates putative reverberative origin of impulse cyclic phenomenon.

Animals↗
PubMed2006

The effect of beta-bungarotoxin, or geniculate ganglion lesion on taste bud development in the chick embryo.

Chick taste bud (gemmal) primordia normally appear on embryonic day (E) 16 and incipient immature, spherical-shaped buds at E17. In ovo injection of beta-bungarotoxin at E12 resulted in a complete absence of taste buds in lower beak and palatal epithelium at developmental ages E17 and E21. However, putative gemmal primordia (solitary clear cells; small, cell groupings) remained, lying adjacent to salivary gland duct openings as seen in normal chick gemmal development. Oral epithelium was immunonegative to neural cell adhesion molecule (NCAM) suggesting gemmal primordia are nerve-independent. Some NCAM immunoreactivity was evident in autonomic ganglion-like cells and nerve fibers in connective tissue. After unilateral geniculate ganglion/otocyst excision on E2.5, at developmental ages E18 and posthatching day 1, approximately 12% of surviving ipsilateral geniculate ganglion cells sustained approximately 54% of the unoperated gemmal counts. After E18, proportional stages of differentiation in surviving developing buds probably reflect their degree of innervation, as well as rate of differentiation. Irrespective of the degree of geniculate ganglion damage, the proportion of surviving buds can be sustained at the same differentiated bud stage as on the unoperated side, or may differentiate to a later bud stage, consistent with the thesis that bud maturation, maintenance, and survival are nerve-dependent.

Animals↗