ER stress increases expression of intracellular calcium channel RyR1 to modify Ca2+ homeostasis in pancreatic beta cells
Irina X. Zhang, Andrea Herrmann, Juan Leon, Sivakumar Jeyarajan, Anoop Arunagiri, Peter Arvan, Patrick Gilon, Leslie S. Satin
Abstract
Pancreatic beta cells maintain glucose homeostasis by secreting pulses of insulin in response to a rise in plasma glucose. Pulsatile insulin secretion occurs as a result of glucose-induced oscillations in beta-cell cytosolic Ca 2+ . The endoplasmic reticulum (ER) helps regulate beta-cell cytosolic Ca 2+ , and ER stress can lead to ER Ca 2+ reduction, beta-cell dysfunction, and an increased risk of type 2 diabetes. However, the mechanistic effects of ER stress on individual calcium channels is not well understood. To determine the effects of tunicamycin-induced ER stress on ER inositol 1,4,5-triphosphate receptors (IP3Rs) and ryanodine receptors (RyRs) and their involvement in subsequent Ca 2+ dysregulation, we treated INS-1 832/13 cells and primary mouse islets with ER stress inducer tunicamycin (TM). We showed TM treatment increased RyR1 mRNA without affecting RyR2 mRNA, and decreased both IP3R1 and IP3R3 mRNA. Furthermore, we found stress reduced ER Ca 2+ levels, triggered oscillations in cytosolic Ca 2+ under subthreshold glucose conditions, and increased apoptosis, and that these changes were prevented by cotreatment with the RyR1 inhibitor dantrolene. In addition, we demonstrate silencing RyR1 suppressed TM-induced subthreshold cytosolic Ca 2+ oscillations, but silencing RyR2 did not affect these oscillations. In contrast, inhibiting IP3Rs with xestospongin-C failed to suppress the TM-induced cytosolic Ca 2+ oscillations and did not protect beta cells from TM-induced apoptosis, although xestospongin-C inclusion did prevent ER Ca 2+ reduction. Taken together, these results show changes in RyR1 play a critical role in ER stress-induced Ca 2+ dysfunction and beta-cell apoptosis.