The Zinc Transporter Slc30a8/ZnT8 Is Required in a Subpopulation of Pancreatic α-Cells for Hypoglycemia-induced Glucagon Secretion

J Biol Chem. 2015 Aug 28;290(35):21432-42. doi: 10.1074/jbc.M115.645291. Epub 2015 Jul 15.

Abstract

SLC30A8 encodes a zinc transporter ZnT8 largely restricted to pancreatic islet β- and α-cells, and responsible for zinc accumulation into secretory granules. Although common SLC30A8 variants, believed to reduce ZnT8 activity, increase type 2 diabetes risk in humans, rare inactivating mutations are protective. To investigate the role of Slc30a8 in the control of glucagon secretion, Slc30a8 was inactivated selectively in α-cells by crossing mice with alleles floxed at exon 1 to animals expressing Cre recombinase under the pre-proglucagon promoter. Further crossing to Rosa26:tdRFP mice, and sorting of RFP(+): glucagon(+) cells from KO mice, revealed recombination in ∼ 30% of α-cells, of which ∼ 50% were ZnT8-negative (14 ± 1.8% of all α-cells). Although glucose and insulin tolerance were normal, female αZnT8KO mice required lower glucose infusion rates during hypoglycemic clamps and displayed enhanced glucagon release (p < 0.001) versus WT mice. Correspondingly, islets isolated from αZnT8KO mice secreted more glucagon at 1 mm glucose, but not 17 mm glucose, than WT controls (n = 5; p = 0.008). Although the expression of other ZnT family members was unchanged, cytoplasmic (n = 4 mice per genotype; p < 0.0001) and granular (n = 3, p < 0.01) free Zn(2+) levels were significantly lower in KO α-cells versus control cells. In response to low glucose, the amplitude and frequency of intracellular Ca(2+) increases were unchanged in α-cells of αZnT8KO KO mice. ZnT8 is thus important in a subset of α-cells for normal responses to hypoglycemia and acts via Ca(2+)-independent mechanisms.

Keywords: calcium; diabetes; glucagon; secretion; zinc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cation Transport Proteins / analysis
  • Cation Transport Proteins / genetics
  • Cation Transport Proteins / metabolism*
  • Cells, Cultured
  • Female
  • Gene Deletion
  • Glucagon / metabolism*
  • Glucagon-Secreting Cells / cytology
  • Glucagon-Secreting Cells / metabolism*
  • Glucose / metabolism
  • Hypoglycemia / genetics
  • Hypoglycemia / metabolism*
  • Insulin Resistance
  • Mice, Inbred C57BL
  • Zinc / metabolism
  • Zinc Transporter 8

Substances

  • Cation Transport Proteins
  • Slc30a8 protein, mouse
  • Zinc Transporter 8
  • Glucagon
  • Glucose
  • Zinc