Overexpression of Gjb4 impairs cell proliferation and insulin secretion in primary islet cells

Mol Metab. 2020 Nov:41:101042. doi: 10.1016/j.molmet.2020.101042. Epub 2020 Jun 18.

Abstract

Objective: Altered gene expression contributes to the development of type 2 diabetes (T2D); thus, the analysis of differentially expressed genes between diabetes-susceptible and diabetes-resistant mouse models is an important tool for the determination of candidate genes that participate in the pathology. Based on RNA-seq and array data comparing pancreatic gene expression of diabetes-prone New Zealand Obese (NZO) mice and diabetes-resistant B6.V-ob/ob (B6-ob/ob) mice, the gap junction protein beta 4 (Gjb4) was identified as a putative novel T2D candidate gene.

Methods: Gjb4 was overexpressed in primary islet cells derived from C57BL/6 (B6) mice and INS-1 cells via adenoviral-mediated infection. The proliferation rate of cells was assessed by BrdU incorporation, and insulin secretion was measured under low (2.8 mM) and high (20 mM) glucose concentration. INS-1 cell apoptosis rate was determined by Western blotting assessing cleaved caspase 3 levels.

Results: Overexpression of Gjb4 in primary islet cells significantly inhibited the proliferation by 47%, reduced insulin secretion of primary islets (46%) and INS-1 cells (51%), and enhanced the rate of apoptosis by 63% in INS-1 cells. Moreover, an altered expression of the miR-341-3p contributes to the Gjb4 expression difference between diabetes-prone and diabetes-resistant mice.

Conclusions: The gap junction protein Gjb4 is highly expressed in islets of diabetes-prone NZO mice and may play a role in the development of T2D by altering islet cell function, inducing apoptosis and inhibiting proliferation.

Keywords: Beta cell; Connexins; Gap junction; Insulin secretion; Proliferation; Type 2 diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Proliferation / physiology
  • Connexins / genetics
  • Connexins / metabolism*
  • Diabetes Mellitus, Type 2 / metabolism
  • Diabetes Mellitus, Type 2 / physiopathology
  • Gene Expression
  • Glucose / metabolism
  • Glucose Tolerance Test
  • Insulin / metabolism
  • Insulin Secretion / genetics*
  • Insulin Secretion / physiology
  • Insulin-Secreting Cells / physiology
  • Islets of Langerhans / metabolism*
  • Islets of Langerhans / physiology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Obese
  • Obesity / metabolism
  • Pancreas / metabolism

Substances

  • Connexins
  • Insulin
  • connexin 30.3
  • Glucose