Nitric oxide production and regulation of neuronal NOS in tyrosine hydroxylase containing neurons

Experimental Neurology
Qing XuC A Colton

Abstract

CAD cells are a murine CNS catecholaminergic (tyrosine hydroxylase-positive; TH+) neuronal cell line that undergoes morphological differentiation to resemble CNS catecholaminergic neurons upon serum deprivation. We show here that CAD cells also express neuronal nitric oxide synthase (nNOS) mRNA and protein and produce readily measurable levels of NO. Since both NO and catecholamines (L-DOPA; dopamine; norepinephrine) are redox active molecules, their production within the same cell may affect the cell's vulnerability to insult. Thus, we examined the regulation of NO production by CAD cells and the effect of NO on cell survival. NO is generated in a dose-dependent fashion by treatment with agents (ionomycin; A23817; KCl) known to increase calcium entry across the cell membrane. The NO level can be increased further by pretreatment with sepiapterin, a membrane permeable precursor for BH4 synthesis, suggesting that the BH4 levels or access required for nNOS activation is limited in CAD cells. Reducing mitochondrial Ca2+ uptake using ruthenium red (RuR) increased ionomycin-mediated NO production over ionomycin alone and indicates a critical role for mitochondria in nNOS regulation. Cell death was significantly increased by ionomyci...Continue Reading

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Citations

Aug 15, 2014·Experimental Neurology·Lais Takata WalterAlexandre Hiroaki Kihara
Nov 17, 2009·CNS Neuroscience & Therapeutics·George B Stefano, Richard M Kream
Aug 9, 2005·Biochimica Et Biophysica Acta·S Y BaeC A Colton
Dec 7, 2007·Movement Disorders : Official Journal of the Movement Disorder Society·Juliane WinkelmannBertram Müller-Myhsok

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