Protocol

Part of the series Methods in Molecular Biology pp 1-6

Date:

Observation of Parkin-Mediated Mitophagy in Pancreatic β-Cells

  • Atsushi HoshinoAffiliated withDepartment of Cardiovascular Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine
  • , Satoaki MatobaAffiliated withDepartment of Cardiovascular Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine Email author 

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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 mKeima Pancreatic MIN6 β-cells Parkin Retrovirus