The effects of minoxidil on the development of the chick embryo

The effects of minoxidil were studied on chick embryos of 24 and 48 hours of incubation. Minoxidil (3%) was injected into the air sacs of the eggs at doses of 20, 30, 40, and 50 microliters per egg. The controls received 100 microliters of physiological saline. All the embryos, including controls, were examined at day 13. The total number of eggs used in this study was 300. At 24 hours incubation, the percentage of survival ranged from 87 to 21 as the dosages of minoxidil were increased from 20 microliters to 50 microliters per egg (controls = 87%). The survival of the embryos ranged from 79% to 9% after the 48-hour treatment with the similar dosages of minoxidil utilized for the 24-hour group (controls = 83%). A low incidence of gross malformations such as twisted limbs, abnormal beak, short neck and everted viscera were observed; however, the increased incidence was not statistically significant when compared to controls. Body hemorrhage and edema were of high occurrence among the treated embryos. These effects are probably secondary to the known pharmacological effects of minoxidil. The frequency and types of gross malformations did not vary much in the 24 or 48-hour treated groups.     

The effects of L- and D-carnitine administration on cardiovascular development of the chick embryo

A single 1.0-ml volume of L- or D-carnitine solution, at several selected mmole concentrations, was applied to the extraembryonic membranes of 3- and 4-day chick embryos in ovo. Hamburger-Hamilton stages of chick development ranged from 17 to 23. During the 17-18th days of incubation, embryos were dissected, and both survival and intracardiac anomaly rates were determined. Only at extremely high doses, both stereoisomers of carnitine exhibited a statistically significant toxigenic effect (p less than 0.001) as measured by a sharp decrease in survival rate when compared to chick Ringer's saline controls. Furthermore, since the anomaly rates became significant only near the LD50's, this indicated that intracardiac anomalies were induced only at toxic doses. Therefore, it is suggested that cardiovascular teratogenicity may be the result of toxicity. Below the LD50, anomaly rates were not significantly different from those of control embryos. In comparison, L- and D-carnitine were significantly different from one another (p less than 0.001) both in survival rate and percent affected embryos at a dose of 0.5 mmole. In summary, exogenous carnitine administration to the chick embryo does not appear to be deleterious to the developing cardiovascular system.     

Molecular characterization of the rostral-most somites in early somitic stages of the chick embryo

Segmentation consists on the progressive formation of repetitive embryonic structures, named somites, which are formed from the most rostral part of the presomitic mesoderm. Somites are subdivided into anterior and posterior compartments and several genes are differentially expressed in either compartment. This has provided evidence for the importance of establishing the anterior-posterior polarity within each somite, which is critical for the correct segmented pattern of the adult vertebrate body. Although all somites appear morphologically similar, fate map studies have shown that the first 4 somites do not give rise to segmented structures, in contrast to more posterior ones. Moreover, in several somitogenesis-related mutants the anterior somites are not affected while posterior somites present clear defects or do not form at all. Altogether these data suggest relevant differences between rostral and caudal somites. In order to check for molecular differences between anterior and posterior somites, we have performed a detailed expression pattern analysis of several Notch signalling related genes. For the first time, we show that the somitic expression pattern profile is not the same along the anterior-posterior axis and that the differences are not observed always at the same somite level.     

Segmental lineage restrictions in the chick embryo spinal cord depend on the adjacent somites.

We have investigated whether the developing spinal cord is intrinsically segmented in its rostrocaudal (anteroposterior) axis by mapping the spread of clones derived from single labelled cells within the neural tube of the chick embryo. A single cell in the ventrolateral neural tube of the trunk was marked in situ with the fluorescent tracer lysinated rhodamine dextran (LRD) and its descendants located after two days of further incubation. We find that clones derived from cells labelled before overt segmentation of the adjacent mesoderm do not respect any boundaries within the neural tube. Those derived from cells marked after mesodermal segmentation, however, never cross an invisible boundary aligned with the middle of each somite, and tend to be elongated along the mediolateral axis of the neural tube. When the somite pattern is surgically disturbed, neighbouring clones derived from neuroectodermal cells labelled after somite formation behave like clones derived from younger cells: they no longer respect any boundaries, and are not elongated mediolaterally. These results indicate that periodic lineage restrictions do exist in the developing spinal cord of the chick embryo, but their maintenance requires the presence of the adjacent somite mesoderm.     

Early development of the chick embryo

The ultrastructure of the early chick embryo was investigated, using scanning (SEM) and transmission electron microscopy (TEM). Eggs were obtained from the shell gland by injecting hens intravenously with a synthetic prostaglandin or arginine vasopressin. Embryos were examined during late cleavage (stages IV–VI, Eyal-Giladi and Kochav, '76), formation of the area pellucida (stages VII–XI), and formation of the hypoblast (stages X–XIV). SEM highlighted the reduction in cell number at the underside of the embryo during formation of the area pellucida although it became apparent that the thickness of the embryo is not reduced to a single layer of cells at stage X. In addition, blastomeres at the perimeter of embryos (stages V–VI) project filopodial extensions onto a smooth membrane that separates the sub-embryonic cavity from the yolk. During hypoblast formation, epiblast cells generate stellate projections at their basal aspect, thus providing a meshwork for the advancing secondary hypoblast cells. By stage XII the epiblast was one cell thick and reminiscent of a columnar epithelium when viewed transversely. Cells of the deep portion of the posterior marginal zone were distinguished morphologically in the stage XII embryo by their many cell surface projections and ruffled appearance. Blastomeres at the perimeter of stage V–VI embryos projected filopodial extensions onto a smooth membrane which separates the sub-embryonic cavity from the yolk. This membrane is presumed to be confluent with the cytolemma. Evidence is presented demonstrating the presence of intracellular membrane-bound droplets which are hypothesised to contain sub-embryonic fluid. © 1993 Wiley-Liss, Inc.     

Consequences of neural tube and notochord excision on the development of the peripheral nervous system in the chick embryo

Notochordectomy and neuralectomy were carried out either in one- or in two-step experiments on the chick embryo. The aim of this operation was to study the influence of the axial organs (notochord and neural tube) on the development of the ganglia of the peripheral nervous system. The neural crest cells from which most peripheral ganglion cells arise were labeled through the quail-chick marker system and their fate was followed under various experimental conditions. It appeared that the development of the dorsal root and sympathetic ganglia depends on survival and differentiation of somite-derived structures. In the absence of neural tube and notochord, somitic cells die rapidly, and so do the neural crest cells that are present in the somitic mesenchyme at that time. In contrast, those crest cells which can reach the mesenchymal wall of the aorta, the suprarenal glands, or the gut survive and develop normally into nerve and paraganglion cells. Differentiation of the neural crest- and placode-derived sensory ganglia of the head which develop in the cephalic mesenchyme is not affected by removal of notochord and encephalic vesicles. These results show that the peripheral ganglia are differentially sensitive to the presence of the neural tube and the notochord. Among the various ganglia of the peripheral nervous system, spinal and sympathetic ganglia are the only ones which require the presence of these axial structures. The neural tube allows both the spinal and the sympathetic ganglia to develop in the absence of the notochord. In contrast, if the notochord is left in situ and the neural tube removed, the spinal ganglia fail to differentiate and only sympathetic ganglia can develop.     

Kynurenine metabolism in the development chick embryo

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1. In the embryonic chick liver during the midperiod of development (8–12 days), kynureninase is the primary enzyme involved in the catabolism of kynurenine. Kynurenine aminotransferase was not found.     

Electron microscope study on the development of ciliary components of the neural epithelium of the chick embryo

Summary Ciliary development was studied in the cells of the neural canal of chick embryos incubated from 60 hours to 7 days.It was found that centrioles move after the last mitosis to the cell periphery where one of them enters into contact (terminal contact) with the cell membrane; the other centriole remains close by, its axis aligned along the axis of the former.The cell membrane was seen afterwards bulging at the contact point, and the content of the ciliary bud thus formed is only constituted at the beginning of a varied number of vesicles of about 140 Å diameter.The ciliary bud becomes elongated shortly after filaments start becoming organized in the bud matrix.Roughly coinciding with the initiation of filament organization the centrioles move inward and the cilium becomes deeply invaginated in the cell. At the end of ciliary growth the centriole moves again toward the surface and the cilium emerges in the neural canal lumen.     

Methylcarbamate effects on Meckel's cartilages in the chick embryo

On the basis of previous observations on the teratogenic effects of a variety of organophosphorus and methylcarbamate compounds on the avian skeletal apparatus, the Meckel's cartilage shape and structure were analyzed in carbaryl (1-naphthyl N-methylcarbamate) treated and control chick embryos of 9, 10, 12 days of incubation. The results indicate that both during normal development and under experimental conditions these cartilages undergo similar deformities, apparently subsequent to chondroblast death and regressive processes in the extracellular matrix. Since the macro- and microscopical cartilage alterations are significantly more frequent in the treated embryos than in the controls, a hypothesis is advanced that the methylcarbamate may increase the spontaneous tendency of the above mentioned cartilaginous anlagen to be affected by degenerative processes during embryogenesis.