Immunocytochemical Monitoring of PINK1/Parkin-Mediated Mitophagy in Cultured Cells

Methods Mol Biol. 2018:1759:19-27. doi: 10.1007/7651_2017_20.

Abstract

Both PINK1 and parkin are the responsible genes (PARK6 and PARK2, respectively) for familial early-onset Parkinson's disease (PD). Several lines of evidences have suggested that mitochondrial dysfunction would be associated with PD pathogenesis. Lewy body, one of PD pathological hallmarks, contains alpha-synuclein, a familial PD (PARK1/4)-gene product, which is eliminated by macroautophagy, while PINK1 and parkin coordinately mediate mitophagy (hereafter called as PINK1/parkin-mediated mitophagy) reported firstly by Youle's group. The mitochondrial quality control system is specific for elimination of damaged mitochondria especially in the loss of mitochondrial membrane potential induced by treatment with mitochondrial uncoupler like CCCP or FCCP. In this chapter, we summarized immunocytochemical methods to monitor the PINK1/parkin-mediated mitophagy using cultured cells.

Keywords: Familial Parkinson’s disease; Immunocytochemistry; Membrane potential; Mitochondrial uncoupler; Mitophagy; PINK1; Parkin; Parkinson’s disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy*
  • Cells, Cultured
  • Fluorescent Antibody Technique
  • Gene Expression Regulation / drug effects
  • HeLa Cells
  • Humans
  • Immunohistochemistry*
  • Membrane Potential, Mitochondrial
  • Mitochondria / drug effects
  • Mitochondria / genetics
  • Mitochondria / metabolism*
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*
  • Valinomycin / pharmacology

Substances

  • Valinomycin
  • Ubiquitin-Protein Ligases
  • parkin protein
  • Protein Kinases
  • PTEN-induced putative kinase