Transduction of yeast NDI1 gene reduces the damages demonstrated by a rotenone-induced differentiated Parkinson's disease cell model

Abstract

Objective To provide a basis for gene therapy of sporadic Parkinson's disease (PD) caused by mitochondrial complex Ⅰ dysfunction, yeast complex Ⅰ (expressed by internal NADH dehydrogenase,NDI1) was tested to replace human complex Ⅰ with functional defects. Methods The recombinant adeno-associated virus (rAAV-NDI1) infected the rotenone-induced differentiated cell model of PD. Three groups (DMSO+vector, rotenone+vector, rotenone+NDI1) were designed. The cytopathology and mitochondrial functions were examined by the methods of Western blot, immunofluorescence staining, measurement of oxygen consumption, ATP content and ROS, etc. Results Compared with rotenone+vector group, the rotenone+NDI1 group showed significantly improved cell morphology, increase in cell survival (P＜0.05), and decrease in level of pS129 α-synuclein and of whole-cell autophagy (P＜0.05, P＜0.001). Compared with rotenone+vector group, the rotenone+NDI1 group also displayed significant increase of complex Ⅰ-dependent oxygen consumption (P＜0.01), significant increase in total cellular ATP synthesis and mitochondrial oxidative phosphorylation-coupled ATP synthesis (P＜0.01),significant decrease in the level of mitochondrial ROS and mitochondrial mitophagy(P＜0.01, P＜0.001). Conclusions Yeast NDI1 can replace and compensate complex Ⅰ-related functional defects in rotenone-induced differentiated PD cell model, and can alleviate the damage of cytopathology and mitochondrial functions.

Keywords

• yeast ndi1|recombinant adeno-associated virus (raav)|rotenone|all-trans retinoic acid|parkinson's disease