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A Method for Mouse Pancreatic Islet Isolation and Intracellular cAMP Determination
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Restoring connexin-36 function in diabetogenic environments precludes mouse and human islet dysfunction.

Joshua R St Clair1, Matthew J Westacott1, Jose Miranda1

  • 1Department of Bioengineering, University of Colorado Denver, Anschutz Medical Campus, Aurora, Colorado, USA.

The Journal of Physiology
|August 14, 2023
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Summary

Impaired connexin-36 (Cx36) gap junction coupling disrupts islet function in type 2 diabetes (T2D). Enhancing Cx36 coupling recovers calcium dynamics and insulin secretion, offering a potential therapeutic strategy for T2D.

Keywords:
Ca2+diabetesgap junctionsislet of Langerhanspeptide mimetic

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Area of Science:

  • Endocrinology
  • Cell Biology
  • Diabetes Research

Background:

  • Insulin secretion from pancreatic β-cells is crucial for glucose homeostasis.
  • Type 2 diabetes (T2D) is characterized by impaired β-cell function and insulin secretion.
  • β-cell electrical coupling via connexin-36 (Cx36) gap junctions coordinates islet activity.

Purpose of the Study:

  • To investigate the role of Cx36 gap junction coupling in T2D.
  • To determine if disrupted coupling can be restored to improve islet function.
  • To evaluate novel therapeutic strategies targeting Cx36 coupling.

Main Methods:

  • Examined islets from human T2D donors and db/db mice.
  • Treated islets with pro-inflammatory cytokines and free fatty acids.
  • Modulated Cx36 coupling via overexpression, modafinil, and a novel peptide mimetic (S293).
  • Assessed gap junction permeability, calcium dynamics, and apoptosis.

Main Results:

  • Cx36 coupling and coordinated calcium dynamics were reduced in T2D islets and in islets exposed to inflammatory stimuli or palmitate.
  • Cx36 overexpression, modafinil, and S293 peptide enhanced coupling and protected calcium dynamics.
  • S293 peptide and Cx36 overexpression reduced cytokine-induced apoptosis.
  • S293 peptide restored coupling and calcium dynamics in db/db mouse islets and some T2D human islets.

Conclusions:

  • Cx36 gap junction coupling is impaired in T2D, contributing to β-cell dysfunction.
  • Restoring Cx36 coupling improves islet electrical coordination and function.
  • The Cx36 peptide mimetic S293 shows therapeutic potential for T2D by enhancing gap junction function.