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Observation of Parkin-Mediated Mitophagy in Pancreatic β-Cells

Protocol
Part of the Methods in Molecular Biology book series

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 

Notes

Acknowledgements

This study was supported in part by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan (MEXT) (KAKENHI 15K09144 to SM).

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Cardiovascular Medicine, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan

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