Regulation of insulin synthesis in an insulin-producing cell line (RINm5F): Long-term experiments |
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Authors: | Adrian S. Dobs Christiane Broussolle M. Daniel Lane |
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Affiliation: | (1) Department of Medicine, The Johns Hopkins University School of Medicine, Traylor Building, Room 721, 21205 Baltimore, Maryland;(2) Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Traylor Building, Room 721, 21205 Baltimore, Maryland;(3) Present address: Centre Hospitalier, Lyon Sud, 69310, 01133 Pierre Benite, France |
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Abstract: | Summary The insulin-producing cell line RINm5F, has been used in short-term experiments to evaluate insulin secretion. We sought to maintain the responsiveness of these cells to stimuli for up to 2 days. We examined the course of new insulin synthesis over this period by measuring at intervals immunoreactive insulin (IRI) in two parts: IRI in the medium (M) and IRI extracted from the cells (C). Control cells were incubated in RPMI 1640/2.8 mM glucose/10% fetal bovine serum/200 μg/ml bacitracin (to prevent insulin degradation). The addition of dibutyryl cAMP 10 mM to the experimental dishes significantly increased total (M+C) IRI at 48 hr to 37% above the insulin content of the control dishes (p<0.01). Theophylline 10 mM increased total (M+C) IRI by 24% over control (p<0.05) after 24 hrs. Glucose, glyceraldehyde, leucine, arginine, glucagon and tolbutamide, other stimulants of insulin production, had no effect. Under the experimental conditions reported here, including the use of bacitracin, IRI synthesis can be studied for up to 48 hr. Portions of this study have been published in abstract form for the 47th Annual Meeting of the American Diabetes Association, Indianapolis, Indiana, 1987. Supported in part by the American Diabetic Association, Maryland Affiliate. |
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Keywords: | RINm5F cells tissue culture insulin synthesis |
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