Glucose-regulated glucagon secretion requires insulin receptor expression in pancreatic α-cells

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  • The insulin receptor (IR) and its signaling appear to be essential for insulin secretion from pancreatic β-cells. However, much less is known about the role of the IR in α-cells. To assess the role of the IR in glucagon and insulin secretion, we engineered adeno-viruses for high efficiency small interference RNA (siRNA)-IR expression in isolated mouse pancreatic islets and lentiviruses for siRNA-IR expression in pancreatic α- and β-cell lines (α-TC6 and MIN6) with specific, long term stable IR knockdown. Western blot analysis showed that these strategies resulted in 60-80% reduction of IR protein in islets and α- and β-cell lines. Cell growth was reduced by 35-50% in α-TC and MIN6 cells stably expressing siRNA-IR, respectively. Importantly, glucagon secretion, in response to glucose (25 to 2.8 mM), was completely abolished in islets expressing siRNA-IR, whereas secretion increased 1.7-fold in islets expressing control siRNA. In contrast, there was no difference in glucose-stimulated insulin secretion when comparing siRNA-IR and siRNA control, with both groups showing a 1.7-fold increase. Islet glucagon and insulin content were also unaffected by IR knockdown. To further explore the role of the IR, siRNA-IR was stably expressed in pancreatic cell lines, which dramatically suppressed glucose-regulated glucagon secretion in α-TC6 cells (3.4-fold) but did not affect GSIS in MIN6 cells. Defects in siRNA-IR-expressing α-cells were associated with an alteration in the activity of Akt and p70S6K where insulin-induced phosphorylation of protein kinase B/AKt was greatly reduced while p70S6K activation was enhanced, suggesting that the related pathways play important roles in α cell function. This study provides direct evidence that appropriate expression of the IR in α-cells is required for glucose-dependent glucagon secretion.

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    © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.
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    • Diao, J., Asghar, Z., Chan, C. B., & Wheeler, M. B. (2005). Glucose-regulated glucagon secretion requires insulin receptor expression in pancreatic α-cells. Journal of Biological Chemistry, 280(39), 33487-33496.
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