Objective: To develop a Parkinson’s disease (PD) model based on aging, vesicular monoamine transporter 2 (VMAT2) gene susceptibility and chronic rotenone exposure in SK-N-SH cell, in order to better mimic the etiology and pathogenesis of PD.

Background: PD is the second prevailing neurodegenerative disease while the etiology still remains unclear. Though many models have been built to study the pathogenesis of PD, none of them recapitulates the aging-gene-environment interaction pattern, which becomes a conspicuous issue that halts the progression of PD research. Model that could better mimic the etiology and pathogenesis of PD is needed urgently to overcome the bottleneck of PD research. It was indicated that transgenic mice expressing 5% normal VMAT2 levels showed the motor, non-motor symptoms and pathology of α-synuclein accumulation, but only at 28 months old or elder. In this study, we propose to develop a cell model of PD based on ageing, VMAT2 gene susceptibility and chronic rotenone exposure in SK-N-SH cell.

Methods: QRT-PCR was used to evaluate the transfection efficiency. Oxidative stress levels, mitochondrial membrane potentials, aggregations of α-synuclein were compared between different groups with knockdown or overexpression of VMAT2 after rotenone exposure. Results of the qRT-PCR, flow cytometry, and western were analyzed based on SPSS16.0, FlowJo, and Image J respectively.

Results: QRT-PCR and Western blot assays indicated that the levels of VMAT2 mRNA/protein in cultured SK-N-SH cells were significantly reduced to 37.9±17.5% (p<0.01) or raised to 14.83±2.21 (p<0.01) times of control values after VMAT2 shRNA expression plasmids transfection. VMAT2 knockdown enhanced the vulnerability of SK-N-SH cells to the environmental toxin rotenone: obvious cell shrinkage, more accumulation of reactive oxygen species and α-synuclein protein were discovered. Furthermore, though without significant statistical difference, the overexpression group showed a lower oxidative stress level and higher mitochondrial membrane potential than the negative control group, suggesting the protective effect of overexpressing VMAT2 protein in SK-N-SH cells.

Conclusions: In summary, we developed a cell model of PD based on ageing, VMAT2 gene susceptibility and chronic rotenone exposure in SK-N-SH cells, which may better mimic the aging-gene-environment interaction pattern of PD pathogenesis.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/exploring-aging-vmat2-gene-rotenone-interactions-in-a-cell-model-for-parkinsons-disease/