培养细胞中PINK1/Parkin介导的线粒体自噬的免疫细胞化学监测
Immunocytochemical Monitoring of PINK1/Parkin-Mediated Mitophagy in Cultured Cells.
作者信息
Fujimaki Motoki, Saiki Shinji, Sasazawa Yukiko, Ishikawa Kei-Ichi, Imamichi Yoko, Sumiyoshi Katsuhiko, Hattori Nobutaka
机构信息
Department of Neurology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, 113-8421, Japan.
Department of Nutrition, Tokiwa University, Mito, Ibaraki, Japan.
出版信息
Methods Mol Biol. 2018;1759:19-27. doi: 10.1007/7651_2017_20.
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.
PINK1和帕金蛋白分别是家族性早发性帕金森病(PD)的致病基因(分别为PARK6和PARK2)。多项证据表明,线粒体功能障碍与PD的发病机制有关。路易小体是PD的病理标志之一,含有α-突触核蛋白,这是一种家族性PD(PARK1/4)基因产物,可通过巨自噬清除,而PINK1和帕金蛋白协同介导线粒体自噬(以下称为PINK1/帕金蛋白介导的线粒体自噬),这一发现最早由尤尔团队报道。线粒体质量控制系统专门用于清除受损线粒体,尤其是在用线粒体解偶联剂(如CCCP或FCCP)处理诱导线粒体膜电位丧失的情况下。在本章中,我们总结了使用培养细胞监测PINK1/帕金蛋白介导的线粒体自噬的免疫细胞化学方法。