title: Phe-met-arg-phe (FMRF)-amide is a substrate source of NO synthase in the gastropod nervous system.
abstract: The possible involvement of the L-arginine-containing Phe-met-arg-phe (FMRF)-amide (FMRFa) in neuronal nitric oxide (NO) biosynthesis was studied in a gastropod species. We found NADPH-diaphorase-positive neurons and FMRFa-containing fibers in close proximity in the enteric nervous system. Administration of L-arginine and FMRFa induced quantitatively similar nitrite production in both intact intestinal tissues and tissue homogenates. These changes could be prevented by the presence of NOARG (an NO synthase inhibitor). Neither chemically modified FMRFa (D-arginine instead of L-arginine) nor amino acid constituents of FMRFa (methionine, phenylalanine) affected basal nitrite production. FMRFa-induced alterations were reduced in the presence of Na+ channel blockers (tetrodotoxin, amiloride, lidocaine), the Na+/K+ATPase inhibitor ouabain, or protease inhibitors (leupeptine, pepstatine-a). FMRFa and its amino acid constituents were analyzed by paper chromatography. When FMRFa was added to tissue homogenates, the peptide was eliminated within 1-2 min, whereas methionine, phenylalanine, arginine, and citrulline levels were elevated simultaneously. We tested the effects of FMRFa, L-arginine, and NOARG on intestinal contractile activity. FMRFa relaxed the intestine for 1-2 min and then induced contractions for 20-40 min. In the presence of NOARG, no relaxant effect of FMRFa was recorded. As administration of L-arginine strongly inhibits the mechanical activity of the intestinal muscle, NO production presumably plays a substantial role in the action of FMRFa, at least in the initial phase. Our biochemical data indicate a direct involvement of FMRFa in NO biosynthesis. FMRFa might be hydrolyzed by extracellular peptidases and then the locally released arginine might be transported into the cells and broken-down to produce NO. Depolarization-induced NO production attributable to the activation of amiloride-sensitive Na+ channels might also be involved.
authors.affiliation: Department of Animal Anatomy & Physiology, Faculty of Science, Debrecen University, P.O. Box 15, H-4010 Debrecen, Hungary.
authors.forename: Tamás
authors.initials: T
authors.lastname: Roszer
authors.forename: Eva
authors.initials: E
authors.lastname: Kiss-Tóth
authors.forename: Mihály
authors.initials: M
authors.lastname: Petkó
authors.forename: A József
authors.initials: AJ
authors.lastname: Szentmiklósi
authors.forename: Gáspár
authors.initials: G
authors.lastname: Bánfalvi
mesh_terms: D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D019835:FMRFamide / Q000032:analysis / Q000378:metabolism*; D049851:Gastropoda / Q000378:metabolism*; D007150:Immunohistochemistry; D008012:Lidocaine / Q000494:pharmacology; D009119:Muscle Contraction / Q000187:drug effects; D009252:NADPH Dehydrogenase / Q000378:metabolism; D009420:Nervous System / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism*; D009479:Neuropeptides / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D026941:Sodium Channel Blockers / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology
chemical_list: D009479:Neuropeptides; D026941:Sodium Channel Blockers; D013779:Tetrodotoxin; D019835:FMRFamide; D000584:Amiloride; D008012:Lidocaine; D019001:Nitric Oxide Synthase; D009252:NADPH Dehydrogenase