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Biomedical subjects

D F Newgreen

Publications and source records attributed to D F Newgreen.

41 records · Page 3Linked to original sources

Morphology and behaviour of neural crest cells of chick embryo in vitro.

Neural primordia of chick embryos were cultured for three days and the behaviour of migrating neural crest cells studied. Somite cells were used as a comparison. Crest cells were actively multipolar with narrow projections which extended and retracted rapidly, contrasting to the gradual extension of somite-cell lamelleae. On losing cell contact, somite cells were also more directionally persistent. The rate of displacement of isolated crest cells was particularly low when calculated over a long time base. Both crest and somite cells were monolayered; contact paralysis occurred in somite cell collisions but was not ascertained for crest cells. However, crest cells in a population were far more directionally persistent than isolated cells. Contact duration between crest cells increased with time and they formed an open network. Eventually, retraction clumping occurred, initially and chiefly at the periphery of the crest outgrowth. Crest cells did not invade cultured embryonic mesenchymal or epithelial populations but endoderm underlapped them. No effects were observed on crest cells prior to direct contact. Substrate previously occupied by endoderm of ectoderm caused crest cells to flatten while substrate previously occupied by the neural tube caused them to round up and clump prematurely.

Animals↗

A procedure for the rapid freezing of whole embryos.

A procedure for the rapid freezing of whole chick embryos for histochemical treatment is described. The problems of deformation during preparation for quenching and orientation for sectioning have been largely overcome by placing embryos inside lengths of chicken trachea. The subsequent disorientation of tissues that follows cracking and shattering due to the rapid freezing of whole embryos is avoided. The method permitted a more precise identification of the position and time of appearance of formaldehyde-induced fluorescence and myosin antibody immunofluorescence in serially sectioned embryos.

Animals↗

Catecholamine accumulation in neural crest cells and the primary sympathetic chain.

Catecholamine accumulation in chick embryos of stages 16 to 24 was investigated using formaldehyde-induced fluorescence. Fluorescence first appeared at stage 21 in the anterior sympathetic chain. After L-DOPA treatment, this fluorescence appeared at stage 18. Noradrenaline could not advance the onset of fluorescence or reconstitute fluorescence after its depletion by reserpine at stages 22 to 24. Under no conditions could fluorescence be identified in neural crest cells prior to their aggregation to form the primary sympathetic chain. Noradrenaline induced fluorescence in the neural tube, notochord, myotome, sclerotome, gut mesenchyme and suprarenal cortical cells. In addition to these structures, the dorsal pancreas and some blood cells were fluorescent after l-DOPA treatment. The implication of the results for the neural crest origin of APUD (Amine Precursor Uptake Decaboxylase) cells is considered.

Animals↗

Extraneuronal effects of 6-hydroxydopamine and extraneuronal uptake of noradrenaline. In-vivo and in-vitro studies on adrenocortical cells of lizards and rats.

6-hydroxydopamine (6-OHDA) was shown to cause ultrastructural changes in adrenocortical cells of lizards and rats. These changes comprised the formation of dense bodies with lamellar and crystalloid patterns, a decrease in the number of mitochondria and structural alterations of mitochondria. Alterations in adrenomedullary cells were not affected as a rule. Only in young animals did 6-OHDA cause deposits of an electron-dense material in medullary cells. An attempt was made to obtain information on amine uptake into cortical cells using the Falck-Hillarp technique to analyse the in-vivo and in-vitro uptake of noradrenaline (NA) into the adrenal cortex in adult rats. Extraneuronal uptake into heart and spleen was studied as well. Our results suggest that NA is taken up into cortical cells, particularly into nuclei, after exposure to 10(-4) gm/ml in-vitro indicating that uptake of 6-OHDA is also likely. Investigations using labelled 6-OHDA is also likely. Investigations using labelled 6-OHDA are required for further elucidating its extraneuronal uptake.

Adrenal Cortex↗

Differentiation in vitro of sympathetic cells from chick embryo sensory ganglia.

This study was carried out in order to determine what factors control the differentiation of certain neural crest cells in the chick embryo. Emphasis was placed on the morphologically and biochemically divergent sensory and sympathetic pathways of differentiation. Embryos were precisely stage according to Hamburger & Hamilton (1951) and it was observed that sensory ganglia with somites, explanted at stages 21-24, gave rise to cells showing formaldehyde-induced fluorescence in more than 25% of explants. These cells were identical in properties to the fluorescent cells of the sympathetic system of embryos of similar age, and appeared by 12 days in vitro. These fluorescent cells did not appear when somites and sensory ganglia explants were maintained separately. The incidence of fluorescent cells in combined explants was considerably reduced or absent when cultures were maintained for 7 days or less, or when the explants were obtained from stage 25-26 embryos. Furthermore, when neural tube was also included in the cultures, the appearance of fluorescent cells was markedly inhibited. The requirement for somitic tissue to induce fluorescent cells in combined explants can be repalced by forelimb-bud tissue. The origin of these cells and the factors that control their differentiation in vitro are discussed with reference to the neural crest origin of the sensory ganglion, and the possible conditions pertaining in vivo in this region.

Animals↗