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The Journal of Neuroscience, May 15, 1998, 18(10):3708-3714
Nitric Oxide-Dependent Production of cGMP Supports the Survival of Rat Embryonic Motor Neurons Cultured with Brain-Derived Neurotrophic Factor
Alvaro G. Estévez1, 6, 8, Nathan Spear1, 6, J. Anthony Thompson2, 4, 6, Trudy L. Cornwell5, Rafael Radi7, Luis Barbeito8, 9, and Joseph S. Beckman1, 2, 3, 6 Departments of 1 Anesthesiology, 2 Biochemistry and Molecular Genetics, 3 Neuroscience, 4 Surgery, and 5 Pathology,
Division of Molecular and Cellular Pathology, and 6 The University of Alabama at Birmingham Center for Free Radical Biology, The University of Alabama at Birmingham, Birmingham, Alabama 35233, and 7 Departamento de Bioquímica, Facultad de Medicina, 8 Sección Neurociencias, Facultad de Ciencias, Universidad de la República 11200 Montevideo, Uruguay, and 9 División Neurobiología Celular y Molecular, Instituto Clemente Estable, 11600 Montevideo, Uruguay Trophic factor deprivation induces neuronal nitric oxide synthase (NOS) and apoptosis of rat embryonic motor neurons in culture. We report here that motor neurons constitutively express endothelial NOS that helps support the survival of motor neurons cultured with brain-derived neurotrophic factor (BDNF) by activating the nitric oxide-dependent soluble guanylate cyclase. Exposure of BDNF-treated motor neurons to nitro-L-arginine methyl ester (L-NAME) decreased cell survival 40-50% 24 hr after plating. Both low steady-state concentrations of exogenous nitric oxide (<0.1 µM) and cGMP analogs protected BDNF-treated motor neurons from death induced by L-NAME. Equivalent concentrations of cAMP analogs did not affect cell survival. Inhibition of nitric oxide-sensitive guanylate cyclase with 2 µM 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) reduced the survival of BDNF-treated motor neurons by 35%. cGMP analogs also protected from ODQ-induced motor neuron death, whereas exogenous nitric oxide did not. In all cases, cell death was prevented with caspase inhibitors. Our results suggest that nitric oxide-stimulated cGMP synthesis helps to prevent apoptosis in BDNF-treated motor neurons.
Key words: motor neurons; BDNF; endothelial nitric oxide synthase; nitric oxide; apoptosis; guanylate cyclase soluble; cGMP
Copyright © 1998 Society for Neuroscience 0270-6474/98/18103708-07$05.00/0
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