Insulin release from isolated islets of Langerhans of the rat in organ culture.
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Biomedical subjects
Publications and source records attributed to B Ziegler.
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We investigated in vitro the pancreatic B-cell response of hyperinsulinemic animals compared to that of controls with the aim of clarifying if the hyperinsulinemia is related to an alteration of the islet insulin secretion. Hyperinsulinemia was induced by pregnancy (observation at 19.5 days) and electrolytic lesions of the hypothalamic nucleus ventromedialis (2 days after lesion). The kinetics of the glucose dose/response curves (measured during a 60 min incubation period) differed markedly in the two hyperinsulinemic conditions. Whereas the islets from pregnant rats were characterized by a lowered glucose threshold, an enhanced half-maximal and maximal secretion rate, those of lesioned rats showed only an increased half-maximal and maximal secretion response in spite of a comparable islet insulin content. When culturing islets of hyperinsulinemic animals up to 6 days, the insulin secretion of those prepared from VMH-lesioned rats behaved identically to controls, whereas the islets of pregnant rats maintained their enhanced sensitivity at a glucose concentration of 5 mmol/l. These results confirm that the pancreatic B-cell response contributed significantly to the hyperinsulinemia, but different mechanisms are likely to be responsible for the altered B-cell secretion.
The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) promoted the formation of monolayers in cultured pancreatic islets isolated from neonatal rats. Immunofluorescence with specific antisera to insulin and glucagon revealed B-cells and A-cells in these monolayers. Glucose-mediated insulin release was increased by raising the glucose concentration from 5 to 10 mmoles/l. Addition of IBMX (0.1 mmoles/l) to medium containing 10 moles/l glucose produced a further increase in insulin release. Recovery of total insulin, i.e. intracellular insulin plus insulin secreted, was also increased by approximately 50% after 8 days of culture. The B-cells showed a marked biosynthetic response to an acute glucose challenge after prior culture with 10 mmoles/l glucose. Although both unstimulated (1.5 mmoles/l glucose) and stimulated rates (1.5 mmoles/l glucose) of [3H]leucine incorporation into (pro)insulin were significantly higher following culture in 10 mmoles/l glucose plus IBMX (0.1 mmoles/l) than after prior culture with 10 mmoles/l glucose alone, the percentage of (pro)insulin synthesized in relation to total protein synthesis was only increased at the low concentration of glucose. These studies demonstrate that monolayer cultures of neonatal B-cells can be readily produced by IBMX and maintained in a functional state, as defined by their secretory and biosynthetic response. It is suggested that the phosphodiesterase inhibitor exerts a sensitizing effect on the responsiveness of the B-cell to glucose. Moreover, the culture system employed in the present study may prove to be useful for further studies of various agents affecting the B-cell function.
An analysis by scanning electron microscopy was undertaken to investigate the effects of bleaching on enamel-cementum surface morphology. Teeth were investigated in five groups: control; vital bleached for 5, 15, and 30 days; and 37 percent phosphoric acid gel etched for 20 seconds. Only the gel-etched group exhibited etching patterns. Shade change in vital bleached teeth was noted in the group vital bleached for 30 days.
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