cytochrome-c-t and 2-2--(hydroxynitrosohydrazono)bis-ethanamine

Nerve growth factor (NGF) can induce apoptosis by signaling through the p75 neurotrophin receptor (p75(NTR)) in several nerve cell populations. Cultured embryonic motor neurons expressing p75(NTR) are not vulnerable to NGF unless they are exposed to an exogenous flux of nitric oxide (*NO). In the present study, we show that p75(NTR)-mediated apoptosis in motor neurons involved neutral sphingomyelinase activation, increased mitochondrial superoxide production, and cytochrome c release to the cytosol. The mitochondria-targeted antioxidants mitoQ and mitoCP prevented neuronal loss, further evidencing the role of mitochondria in NGF-induced apoptosis. In motor neurons overexpressing the amyotrophic lateral sclerosis (ALS)-linked superoxide dismutase 1(G93A) (SOD1(G93A)) mutation, NGF induced apoptosis even in the absence of an external source of *NO. The increased susceptibility of SOD1(G93A) motor neurons to NGF was associated to decreased nuclear factor erythroid 2-related factor 2 (Nrf2) expression and downregulation of the enzymes involved in glutathione biosynthesis. In agreement, depletion of glutathione in nontransgenic motor neurons reproduced the effect of SOD1(G93A) expression, increasing their sensitivity to NGF. In contrast, rising antioxidant defenses by Nrf2 activation prevented NGF-induced apoptosis. Together, our data indicate that p75(NTR)-mediated motor neuron apoptosis involves ceramide-dependent increased mitochondrial superoxide production. This apoptotic pathway is facilitated by the expression of ALS-linked SOD1 mutations and critically modulated by Nrf2 activity.

Topics: Animals; Animals, Genetically Modified; Apoptosis; Cells, Cultured; Cytochromes c; Embryo, Mammalian; Enzyme Activation; Enzyme Inhibitors; Gene Expression Regulation; Mitochondria; Motor Neurons; Nerve Growth Factor; NF-E2-Related Factor 2; Nitric Oxide Donors; Nitroso Compounds; Oligodeoxyribonucleotides, Antisense; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Receptor, Nerve Growth Factor; Sphingomyelin Phosphodiesterase; Spinal Cord; Superoxide Dismutase