Role of intercellular and intracellular communication by nitric oxide in coupling of muscarinic receptors to activation of guanylate cyclase in neuronal cells.
Muscarinic receptor-mediated cyclic GMP formation and release of nitric oxide (NO) (or a precursor thereof) were compared in mouse neuroblastoma N1E-115 cells. [3H]Cyclic GMP was assayed in cells prelabeled with [3H]guanine. Release of NO upon the addition of muscarinic agonists to unlabeled neuroblastoma cells (NO donor cells) was quantitated indirectly by its ability to increase the [3H]cyclic GMP level in labeled cells whose muscarinic receptors were inactivated by irreversible alkylation (NO detector cells). Carbachol increased NO release in a concentration-dependent manner, with half-maximal stimulation at 173 microM (compared to 96 microM for direct activation of cyclic GMP formation). The maximal effect of carbachol in stimulating release of NO when measured indirectly was lower than that in elevating [3H]cyclic GMP directly in donor cells. Hemoglobin was more effective in blocking the actions of released NO than in attenuating direct stimulation of [3H]cyclic GMP synthesis. There was a good correlation between the ability of a series of muscarinic agonists to release NO or to activate [3H]cyclic GMP formation directly, and the potency of pirenzepine in inhibiting the two responses. Furthermore, there was a similar magnitude of desensitization of both responses by prolonged receptor activation or stimulation of protein kinase C. NO release was also regulated in relation to the cellular growth phase. A model is proposed in which a fraction of NO generated upon receptor activation does not diffuse extracellularly and stimulates cyclic GMP synthesis within the same cell where it is formed (locally acting NO). The remainder of NO that is extruded extracellularly might travel to neighboring cells (neurotransmitter NO) or might be taken back into the cells of origin (homing NO).
Hu
J
J
Division of Neuroscience Research in Psychiatry, University of Minnesota Medical School, Minneapolis 55455.
el-Fakahany
E E
EE
D000478:Alkylation; D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002450:Cell Communication; D006152:Cyclic GMP / Q000096:biosynthesis; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000378:metabolism*; D006454:Hemoglobins / Q000494:pharmacology; D051379:Mice; D009447:Neuroblastoma; D009474:Neurons / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism*; D010890:Pirenzepine / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000502:physiology*; D013482:Superoxide Dismutase / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D006454:Hemoglobins; D011976:Receptors, Muscarinic; D009569:Nitric Oxide; D010890:Pirenzepine; D002217:Carbachol; D013482:Superoxide Dismutase; D011493:Protein Kinase C; D006162:Guanylate Cyclase; D006152:Cyclic GMP