Alpha4 contributes to the dysfunction of the pancreatic beta cell under metabolic stress. (1st November 2022)

• Record Type: Journal Article
• Authors: Hali, Mirabela; Wadzinski, Brian E.; Kowluru, Anjaneyulu
• Is Part Of: Molecular and cellular endocrinology. Volume 557(2022)
• Publication Date: 2022-11-01
• Subjects: Pancreatic islet -- Alpha4 -- PP2A or Protein-serine/threonine phosphatase -- Connexin36 -- Metabolic stress -- Diabetes; Endocrinology -- Periodicals; Molecular biology -- Periodicals; Cytology -- Periodicals; Endocrinology -- Periodicals; Hormones -- Periodicals; Endocrinologie -- Périodiques; Cytology; Endocrinology; Molecular biology; Periodicals; 573.4
• DOI: 10.1016/j.mce.2022.111754
• Languages: English
• ISSNs: 0303-7207

Abstract

Abstract: The current study examined the roles of Alpha4, a non-canonical subunit of protein phosphatase 2A, in the regulation of acute (insulin secretion) and chronic (cell dysfunction) effects of glucose in pancreatic beta cells. Alpha4 is expressed in human islets, rat islets and INS-1832/13 cells. Incubation of INS-1832/13 cells and rat islets with high glucose (HG) significantly increased the expression of Alpha4. C2-Ceramide, a biologically active sphingolipid, also increased the expression of Alpha4 in INS-1832/13 cells and rat islets. Subcellular distribution studies of Alpha4 in low glucose (LG) and HG exposed INS-1832/13 cells revealed that it is predominantly cytosolic, and its expression is significantly increased in the non-nuclear/cytosolic fractions in cells exposed to HG. siRNA-mediated knockdown of Alpha4 exerted minimal effects on glucose- or KCl-induced insulin secretion. siRNA-mediated deletion of Alpha4 significantly increased p38MAPK and JNK1/2 phosphorylation under LG conditions, comparable to the degree seen under HG conditions. Paradoxically, a significant potentiation of HG-induced p38MAPK and JNK2 phosphorylation was noted following Alpha4 deletion. HG-induced CHOP expression (ER stress marker) and caspase-3 activation were markedly attenuated in cells following Alpha4 knockdown. Deletion of Alpha4 in INS-1832/13 cells prevented HG-induced loss in the expression of Connexin36, a gap junction channel protein, which has been implicated in normal beta cell function. Lastly, depletion of endogenous Alpha4 significantly reduced HG-induced cell death in INS-1832/13 cells. Based on these findings we conclude that Alpha4 contributes to HG-induced metabolic dysfunction of the islet beta cell. Graphical abstract: Image 1 Highlights: Alpha4 is expressed in human and rodent islets and clonal beta (INS-1832/13) cells. Alpha4 does not contribute to physiological insulin secretion. Metabolic stress induces expression of Alpha4 in beta cells. Alpha4-KD prevents metabolic stress-induced ER stress, caspase 3 activation, loss in Connexin36 expression and cell death.