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Biomedical subjects

S Komazaki

Publications and source records attributed to S Komazaki.

44 records · Page 3Linked to original sources

Epibolic extension of the presumptive ectodermal layer of embryos of the newt Cynops pyrrhogaster before and during gastrulation.

Epibolic extension of the presumptive ectodermal layer (PEL) was investigated in embryos of the newt Cynops pyrrhogaster before and during gastrulation. The PEL was composed of only one layer of columnar cells at all stages examined. The cells of the PEL became elongated from the blastula to the early gastrula stage. They were most elongated at the early gastrula stage and then shortened during gastrulation. Present observations suggest that changes in cell shape of the PEL play an important role in the control of the epibolic extension of the newt embryos. The morphology and movement of the isolated cells from the PEL were examined in an attempt to elucidate the role of cell movement in epibolic extension of the PEL. Blebbing and vermiform cells which showed active cell movement appeared at the early blastula stage. The blebbing cells, which formed large hyaline blebs that moved around the circumference of each cell, appeared in large numbers at the early blastula stage. The frequency of the blebbing cells decreased from the early blastula to the early gastrula stage and increased again during gastrulation. The vermiform cells, which had an elongated cell body and moved in a worm-like manner, increased in frequency from the early blastula to the early gastrula stage. The relative number of such vermiform cells was maximal at the early gastrula stage and decreased abruptly during gastrulation. These results suggest that the elongation of the cells of the PEL is controlled by the active cell movement which resembles that of a worm.

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Ultrastructural localization of calcium in the chick chorioallantoic membrane as revealed by cytochemistry and X-ray microanalysis.

The chorioallantoic membrane (CAM) of the chick embryo actively transports calcium from the egg shell into the embryonic circulation. To investigate the intracellular pathway of calcium transport across the CAM, ultrastructural localization of intracellular calcium in cells of the chorionic ectoderm (CE) was determined using cytochemical methods and X-ray microanalysis. Treatment of the CE with potassium oxalate, potassium ferricyanide or potassium pyroantimonate revealed large numbers of electron-dense granules (EDGs) in the ectodermal cells. These measure 30-40 nm in diameter, and are not membrane-bound. These granules were seen in all three cell types of the CE. The presence of calcium in the EDG was directly confirmed by X-ray microanalysis. When strontium or barium ions were applied to the shell membrane side of the CAM, the cells of the CE incorporated these divalent cations and sequestered them in granules (25-40 nm in diameter) in cytoplasm and mitochondria. This study indicates that calcium enters the CE cells by means other than endocytosis, as the EDGs are not membrane-bound, that all three types of the CE cells appear to function in transport of calcium from shell to embryo during embryogenesis, and that the EDG plays important roles in intracellular accumulation of calcium during the process of calcium transport across the chorioallantoic membrane.

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Development of cell surface activity and cell surface adhesiveness in early embryos of the newt, Cynops pyrrhogaster.

The development of cell surface activity and adhesiveness was examined in relation to cleavage number in early embryos of the newt, Cynops pyrrhogaster. Both large hyaline bleb formation and surface adhesiveness to substratum were manifested in presumptive ectodermal cells isolated from embryos after the eleventh cleavage (mid-blastula stage). Scanning electron microscopy of the inner surface of the blastocoelic wall (presumptive ectodermal cell layer) revealed the formation of large blebs after the eleventh cleavage. Treatment with alcian blue and lanthanum nitrate demonstrated the accumulation of an extracellular matrix (ECM) on the surface of large blebs.

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Effects of salts in promoting the adhesion of amphibian gastrula cells.

Presumptive ectodermal and endodermal cells were isolated from the gastrula of the newt Cynops pyrrhogaster, and the effects of salts on embryonic cell adhesion were examined. When the concentrations of NaCl, KCl, MgSO4 and Ca (NO3)2 in the culture medium were increased individually, they all effectively promoted ectodermal cell adhesion. MgSO4 and Ca(NO3)2 promoted endodermal cell adhesion, but NaCl and KCl did not. The culture medium containing increased NaCl (90 mM) nonspecifically promoted ectodermal cell adhesion to the four substrata studied--polystyrene dish, glass, collagen, and fibronectin. The effects of drugs that influence the membrane permeability of Na+ and Ca2+ and the replacement of Na+, Cl-, and Ca2+ with other ions were studied. The results show that the flow of Na+ and Ca2+ into the cytoplasm promotes ectodermal cell adhesion but exerts no influence on endodermal cell adhesion.

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Effect of prolactin on transcutaneous Na transport in the Japanese newt, Cynops pyrrhogaster.

The effect of ovine prolactin (PRL) on the electric parameters related to active Na transport across the skin was investigated in the Japanese newt, Cynops pyrrhogaster. Spring newts living in a pond or stream for breeding and autumn newts migrating from water to land for hibernation were collected and used for the experiments. Autumn newt skin was found to have a higher transepithelial potential difference (TEP) and higher resistances to the active Na current (RNa) and to the shunt pathway (RS) than spring newt skin. Injection of PRL, 20 micrograms/g body wt, into autumn newts every other day for 2 weeks induced an increase in RNa, resulting in decreases in TEP and the short-circuit current (SCC), but barely changed the skin resistance (RM), the electromotive force of the active Na current (ENa), or RS. On the other hand, such injections of PRL into spring newts induced decreases in TEP and SCC, but had no effect on RM, ENa, RNa, or RS. Autumn newts appear to be more sensitive to PRL than spring newts. Insofar as autumn newts are concerned, it is concluded that the effect of PRL is probably due to blockade of the Na channel rather than inhibition of Na pump activity.

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Factors related to the initiation of cell migration along the inner surface of the blastocoelic wall during amphibian gastrulation.

Changes in the quantity of extracellular matrix on the inner surface of the blastocoelic wall (BW) and the adhesiveness of cells to fibronectin were examined in relation to the timing of initiation of cell migration along the BW during gastrulation of the newt, Cynops pyrrhogaster. Extracellular matrix fibrils were already abundantly present all over the inner surface of the BW prior to the beginning of cell migration. On the other hand, the adhesiveness of cells to fibronectin rapidly increased in the cells of the dorsal region at the same time as the cell migration initiated along the BW.

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Early cardiogenesis in the newt embryo.

The migration of cardiogenic cells and the formation of a tubular heart in newt embryos were examined mainly by scanning electron microscopy (SEM). Cardiogenic cells are known to localize at the border region of lateral mesoderm migrating in the space between the ectoderm and the endoderm. They initially (before stage 20 or mid-neurula) appeared to attach to the basal surface of the ectoderm, whereas later (after stage 22 or late neurula) they changed their scaffold to the endoderm. On the scaffold cell surface, very fine fibrils of extracellular matrix (ECM) were found. These fibrils were proved to be composed partly of fibronectin by the immunofluorescence method as well as by immunoSEM using latex bead-labeled antibody, suggesting their seemingly important role in migration of cardiogenic cells. At stage 26 or the early tail bud stage, when the tips of bilateral cardiogenic areas begin to fuse under the foregut, several free vasoformative cells are seen there and the mesodermal sheet itself splits into two layers to produce a coelomic cavity. The splanchnic wall of the coelomic or pericardial cavity was recognized to form a though consisting of cobblestone-like myocardial cells not yet covered with the epicardium.

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Seasonal and geographical changes of spontaneous skin papillomas in the Japanese newt Cynops pyrrhogaster.

The occurrence of spontaneous skin papillomas in Japanese newts (Cynops pyrrhogaster) from Niigata prefecture in Northern Japan was monitored over four seasons covering a period of 2 years. Of 13,613 newts, 249 were found to possess papillomas. The percentage of newts with papillomas was highest in autumn, ranging from 1.93 to 5.45% of the total average. These values were more than four times as high as values obtained in the other three seasons (0.16 to 0.50%). A total of 12,167 newts were collected from their natural environment in 10 prefectures of Japan in the autumn of 1980 and 1981. Newts collected from the northern, seaside prefectures had higher papilloma rates (1.00 to 5.45%) than did newts from the Southern, Pacific Ocean prefectures of Japan (0 to 0.27%). Male and female newts were affected equally often by the epitheliomas. Virus-like bodies, resembling herpes-type virus, were found in the cytoplasm of the epitheliomas. These virus-like bodies were not seen in the control epithelium of newts with normal skin and without papilloma or in nonaffected regions of newts with papilloma. It is suggested that tumor and virus are causally related.

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