Protocol

Part of the series Methods in Molecular Biology pp 1-9

Date:

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

  • Motoki FujimakiAffiliated withDepartment of Neurology, Juntendo University School of Medicine
  • , Shinji SaikiAffiliated withDepartment of Neurology, Juntendo University School of Medicine Email author 
  • , Yukiko SasazawaAffiliated withDepartment of Neurology, Juntendo University School of Medicine
  • , Kei-Ichi IshikawaAffiliated withDepartment of Neurology, Juntendo University School of Medicine
  • , Yoko ImamichiAffiliated withDepartment of Neurology, Juntendo University School of Medicine
  • , Katsuhiko SumiyoshiAffiliated withDepartment of Nutrition, Tokiwa University
  • , Nobutaka HattoriAffiliated withDepartment of Neurology, Juntendo University School of Medicine

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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 PINK1 Parkin Mitophagy Immunocytochemistry Parkinson’s disease Membrane potential Mitochondrial uncoupler