Parkinson disease (PD) is a multifactorial disease resulting in preferential death of the dopaminergic neurons in the substantia nigra. Studies of PD-linked genes and toxin-induced models of PD have implicated mitochondrial dysfunction, oxidative stress, and the misfolding and aggregation of α-synuclein (α-syn) as key factors in disease initiation and progression. Many of these features of PD may be modeled in cells or animal models using the neurotoxin 1-methyl-4-phenylpyridinium (MPP(+)). Reducing oxidative stress and nitric oxide synthase (NOS) activity has been shown to be protective in cell or animal models of MPP(+) toxicity. We have previously demonstrated that siRNA-mediated knockdown of α-syn lowers the activity of both dopamine transporter and NOS activity and protects dopaminergic neuron-like cells from MPP(+) toxicity. Here, we demonstrate that α-syn knockdown and modulators of oxidative stress/NOS activation protect cells from MPP(+)-induced toxicity via postmitochondrial mechanisms rather than by a rescue of the decrease in mitochondrial oxidative phosphorylation caused by MPP(+) exposure. We demonstrate that MPP(+) significantly decreases the synthesis of the antioxidant and obligate cofactor of NOS and TH tetrahydrobiopterin (BH4) through decreased cellular GTP/ATP levels. Furthermore, we demonstrate that RNAi knockdown of α-syn results in a nearly twofold increase in GTP cyclohydrolase I activity and a concomitant increase in basal BH4 levels. Together, these results demonstrate that both mitochondrial activity and α-syn play roles in modulating cellular BH4 levels.

Original publication

DOI

10.1016/j.freeradbiomed.2013.10.008

Type

Journal article

Journal

Free Radic Biol Med

Publication Date

02/2014

Volume

67

Pages

58 - 68

Keywords

Bioenergetics, DAT, Free radicals, GTPCH, MPP(+), Parkinson disease, Tetrahydrobiopterin, α-Synuclein, 1-Methyl-4-phenylpyridinium, Adenosine Triphosphate, Biopterin, Cell Line, Tumor, Dopaminergic Neurons, GTP Cyclohydrolase, Gene Expression Regulation, Guanosine Triphosphate, Humans, Mitochondria, Models, Biological, Oxidative Phosphorylation, Parkinson Disease, RNA, Small Interfering, alpha-Synuclein