Insulin secretion and Ca2+ dynamics in β-cells are regulated by PERK (EIF2AK3) in concert with calcineurin

J Biol Chem. 2013 Nov 22;288(47):33824-33836. doi: 10.1074/jbc.M113.503664. Epub 2013 Oct 10.

Abstract

Protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) (EIF2AK3) is essential for normal development and function of the insulin-secreting β-cell. Although genetic ablation of PERK in β-cells results in permanent neonatal diabetes in humans and mice, the underlying mechanisms remain unclear. Here, we used a newly developed and highly specific inhibitor of PERK to determine the immediate effects of acute ablation of PERK activity. We found that inhibition of PERK in human and rodent β-cells causes a rapid inhibition of secretagogue-stimulated subcellular Ca(2+) signaling and insulin secretion. These dysfunctions stem from alterations in store-operated Ca(2+) entry and sarcoplasmic endoplasmic reticulum Ca(2+)-ATPase activity. We also found that PERK regulates calcineurin, and pharmacological inhibition of calcineurin results in similar defects on stimulus-secretion coupling. Our findings suggest that interplay between calcineurin and PERK regulates β-cell Ca(2+) signaling and insulin secretion, and that loss of this interaction may have profound implications in insulin secretion defects associated with diabetes.

Keywords: Calcineurin; Calcium Dynamics; Calcium Imaging; Diabetes; ER Stress; Insulin Secretion; PERK; Sarcoplasmic Endoplasmic Reticulum-Calcium ATPase; Store-operated Calcium Entry.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Calcineurin / genetics
  • Calcineurin / metabolism*
  • Calcium / metabolism*
  • Calcium Signaling / physiology*
  • Cell Line
  • Humans
  • Insulin / genetics
  • Insulin / metabolism*
  • Insulin Secretion
  • Insulin-Secreting Cells / cytology
  • Insulin-Secreting Cells / metabolism*
  • Mice
  • Mice, Mutant Strains
  • Rats
  • Rats, Sprague-Dawley
  • Sarcoplasmic Reticulum / genetics
  • Sarcoplasmic Reticulum / metabolism
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases / genetics
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases / metabolism
  • eIF-2 Kinase / genetics
  • eIF-2 Kinase / metabolism*

Substances

  • Insulin
  • EIF2AK3 protein, human
  • PERK kinase
  • eIF-2 Kinase
  • Calcineurin
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Calcium