Nitric oxide delays the death of trophic factor-deprived PC12 cells and sympathetic neurons by a cGMP-mediated mechanism.
We have used cultured PC12 cells and rat sympathetic neurons as model systems to examine the regulation of neuronal cell death and survival. Because nitric oxide (NO) may be involved in nerve growth factor (NGF) signaling in PC12 cells, we tested NO-generating compounds for their ability to protect PC12 cells and sympathetic neurons from death after withdrawal of trophic support. Three such agents, S-nitroso-N-acetylpenicillamine (SNAP), diethylenetriamine NO adduct (DETA-NO), and sodium nitroprusside provide (SNP), were found to promote complete short-term survival after removal of serum from naive PC12 cells and of NGF from neuronally differentiated PC12 cells and sympathetic neurons. One major target of NO action is guanylate cyclase, which is activated by nitrosylation of its heme prosthetic group. We observed that inhibition of guanylate cyclase blocks the protective effects of the NO generators on trophic factor-deprived PC12 cells and sympathetic neurons without preventing NGF-induced survival. We also found that permeant cGMP analogs and an inhibitor of cGMP-specific phosphodiesterase enhance cell survival, suggesting that the protective effects of NO are mediated by activation of guanylate cyclase and increased intracellular cGMP. N-Nitro-L-arginine methyl ester, a NO synthase inhibitor, did not block NGF-promoted PC12 cell or sympathetic neuron survival. These findings indicate that like NGF, NO has survival-promoting actions on neurons but that the two agents work by initially independent mechanisms.
Farinelli
S E
SE
Department of Pathology, Columbia University College of Physicians and Surgeons, New York, 10032, USA.
Park
D S
DS
Greene
L A
LA
D000320:Adrenergic Fibers / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D001798:Blood Proteins / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors; D019331:NG-Nitroarginine Methyl Ester; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D016716:PC12 Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D051381:Rats
D001798:Blood Proteins; D004791:Enzyme Inhibitors; D009414:Nerve Growth Factors; D010726:Phosphodiesterase Inhibitors; D009569:Nitric Oxide; D001120:Arginine; D019001:Nitric Oxide Synthase; D006162:Guanylate Cyclase; D006152:Cyclic GMP; D019331:NG-Nitroarginine Methyl Ester