Proinsulin mRNA and peptide are present in beta-cells of diabetic BB rats.

Abstract

Previous studies have demonstrated that islets isolated from newly diabetic BB rat pancreata retain the ability to release insulin in culture, although in vivo the insulin response to stimulation is absent. The purpose of this study was to determine whether the beta-cells in these newly diabetic animals were releasing stored insulin or whether they were still capable of insulin biosynthesis, since secretory defects may reflect abnormalities in insulin synthetic capacity. Insulin gene transcription was examined using in situ hybridization to detect preproinsulin mRNA (ppImRNA) at the level of the single cell since this technique provides a valid semiquantitative index of insulin biosynthesis. In situ hybridization with digoxigenin-labeled rat insulin probes resulted in strong labeling of beta-cells in normal Wistar rat pancreata; other islet and acinar cells were negative. Double labeling of sections with an antibody to insulin confirmed that the labeled cells were beta-cells only. The intensity of the staining was variable between different islets within the same section, and sometimes within an islet. Nondiabetic and diabetic BB islets were also positive for ppImRNA not only in normal islets but also in islets affected by insulitis. Islets that contained very few beta-cells also contained ppImRNA. A consistent finding was that the intensity of the hybridization signal in many islets from the diabetic BB rats was stronger than in controls, suggesting that there is more ppImRNA in these islets. beta-Cells that were positive for ppImRNA but negative for insulin peptide were also observed; these were in islets that were affected by insulitis.(ABSTRACT TRUNCATED AT 250 WORDS)