Persistently enhanced sensitivity of pancreatic islets from ob/ob mice to PKC-stimulated insulin secretion.

Islets from 2-wk-old ob/ob and lean littermate mice were cultured for 4-12 days and then perifused or statically incubated to identify early-onset differences in their regulation of insulin secretion. Islets from these young ob/ob and lean mice increased insulin secretion similarly in response to glucose (10 or 20 mM), whereas responsiveness to glucose plus acetylcholine (10 microM) was greater in islets from ob/ob mice than lean mice. This phenotype-specific effect of acetylcholine was mimicked by phorbol 12-myristate 13-acetate (PMA, 100 nM), a protein kinase C (PKC) agonist, whereas prior downregulation of PKC abolished this phenotype-specific effect of acetylcholine. A high concentration of PMA (1 microM) equally and substantially increased insulin secretion from islets of ob/ob and lean mice, suggesting an enhanced regulatory sensitivity rather than altered responsiveness of the PKC system in islets of ob/ob mice. Addition of BAY K 8644, a Ca2+ channel agonist, to the perifusate enhanced acetylcholine-induced insulin secretion from islets of lean mice to attain the high rates observed in islets from ob/ob mice exposed to acetylcholine alone. We propose that acetylcholine-induced PKC regulation of insulin secretion is altered in islets from ob/ob mice, that this alteration may directly or indirectly involve Ca2+ channels, and that it persists even when islets are cultured for up to 12 days.