Activation of cyclic GMP formation in mouse neuroblastoma cells by a labile nitroxyl radical. An electron paramagnetic resonance/spin trapping study.

Abstract

The receptor-mediated generation of an endothelial-derived relaxing factor (EDRF)-free radical intermediate in a neuronal cell line detected by spin trapping techniques has been reported. Here we report the time course of the appearance of the 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) spin adduct and cyclic GMP formation following addition of carbamylcholine to suspensions of cultured mouse neuroblastoma cells (clone N1E-115). The time course of the appearance of the DBNBS spin adduct shows that spin adduct formation decreases possibly reaching a minimum approximately between 35 and 40 s. This is inversely proportional to cGMP formation which reaches a maximum at approximately 40 s after carbamylcholine activation. In addition, the inhibitory effect of NG-monomethyl-L-arginine (NMMA), potassium ferricyanide, K3Fe(CN)6 and methylene blue in cytosol preparation was investigated. A mechanism is proposed that essentially accounts for the combined results observed by spin trapping/electron paramagnetic resonance (EPR) study providing direct evidence for the muscarinic receptor-mediated formation of a labile, diffusible precursor of nitric oxide (NO.) derived from L-arginine that activates soluble guanylate cyclase.

• Authors: C M Arroyo, C Forray
• Journal: European journal of pharmacology (Eur J Pharmacol) Vol. 208 Issue 2 Pg. 157-61 (Oct 14 1991) ISSN: 0014-2999 [Print] Netherlands
• PMID: 1686865 (Publication Type: Journal Article)

Chemical References

• Benzenesulfonates
• Ferricyanides
• Free Radicals
• Nitrogen Oxides
• Nitroso Compounds
• Receptors, Muscarinic
• omega-N-Methylarginine
• NADP
• 3,5-dibromo-4-nitrosobenzenesulfonate
• Arginine
• Cyclic AMP
• Guanylate Cyclase
• nitroxyl
• Methylene Blue
• potassium ferricyanide

Topics

• Animals
• Arginine (analogs & derivatives, metabolism, pharmacology, physiology)
• Benzenesulfonates (metabolism)
• Cyclic AMP (biosynthesis)
• Electron Spin Resonance Spectroscopy
• Enzyme Activation
• Ferricyanides (pharmacology)
• Free Radicals (pharmacology)
• Guanylate Cyclase (metabolism)
• Methylene Blue (pharmacology)
• Mice
• NADP (physiology)
• Neuroblastoma (metabolism, pathology)
• Nitrogen Oxides (pharmacology)
• Nitroso Compounds (metabolism)
• Receptors, Muscarinic (physiology)
• Tumor Cells, Cultured
• omega-N-Methylarginine