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Both insulin and glucagon stimulate transcription of the proglucagon gene, resulting in increased synthesis of proglucagon mRNA, proglucagon immunoreactive (PROG-IR) peptide and pro-insulin. The actions of insulin and glucagon on the transcription and translation of the proglucagon gene were examined in islets and intestinal L cells. Islets were dispersed from the pancreata of neonatal rat or adult rat by collagenase digestion, separated into perifused fractions containing L-cells only, L-cells only, or islets, and cultured for 6 h in the presence or absence of glucose. Glucagon increased levels of proglucagon mRNA, PROG-IR peptide and pro-insulin in islets. This was inhibited by wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3-K), and by small interfering RNA (siRNA) targeted against the p85 regulatory subunit of PI3-K. The increase in proglucagon mRNA, PROG-IR peptide and pro-insulin were maintained in islets cultured in the absence of glucose or at 0.5 and 1.0 mM glucose. Glucagon also increased proglucagon mRNA, PROG-IR peptide and pro-insulin in intestinal L cells in islet-depleted culture. Wortmannin inhibited the increase in proglucagon mRNA, PROG-IR peptide and pro-insulin induced by glucagon in intestinal L cells. siRNA targeted against the p85 regulatory subunit of PI3-K inhibited the increase in proglucagon mRNA, PROG-IR peptide and pro-ins 0b46394aab
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