Observation of Parkin-Mediated Mitophagy in Pancreatic β-Cells

Methods Mol Biol. 2018:1759:41-46. doi: 10.1007/7651_2017_39.

Abstract

Mitophagy is a cellular process of autophagy-based mitochondrial degradation that eliminates dysfunctional mitochondria and ensures cellular homeostasis. In pancreatic islet β-cells, mitochondria play a pivotal role in glucose-stimulated insulin secretion through ATP production from glucose oxidation. Recent studies have shown that impaired mitophagy and the subsequent mitochondrial compromise contribute to β-cell dysfunction and glucose intolerance. In this chapter, we describe a protocol to monitor Parkin-mediated mitophagy in pancreatic MIN6 β-cells using flow cytometry and a pH-sensitive fluorophore, mKeima.

Keywords: Flow cytometry; Mitophagy; Pancreatic MIN6 β-cells; Parkin; Retrovirus; mKeima.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Flow Cytometry
  • Genetic Vectors / genetics
  • Humans
  • Insulin-Secreting Cells / metabolism*
  • Mice
  • Mitochondria / genetics*
  • Mitochondria / metabolism*
  • Mitophagy*
  • Retroviridae / genetics
  • Transduction, Genetic
  • Ubiquitin-Protein Ligases / genetics*
  • Ubiquitin-Protein Ligases / metabolism*

Substances

  • Ubiquitin-Protein Ligases
  • parkin protein