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The anatomical and functional relationship between neurons expressing nitric oxide (NO) synthase and molluscan cardioexcitatory (FMRFamide)-like neuropeptides was studied in the central ganglia of Helix lucorum (Pulmonata, Gastropoda), applying NADPHdiaphorase (NADPHd) histochemistry to visualize NO synthase and immunocytochemistry to demonstrate FMRFamide (FMRFa) at the light microscopic level. The NO production of the ganglia was detected by the colorimetric Griess determination of nitrite, a breakdown product of NO. Effects of the NO synthase substrate amino acid L-arginine, the NO synthase inhibitor Nomega-nitro-L-arginine (NOARG), synthetic FMRFa and the FMRFa sensitive ion channel blocker amiloride hydrochloride on nitrite production were also tested. NADPHd reaction labeled nerve cells and fibers in the procerebra, mesocerebra and metacerebra within the cerebral ganglia, and cell clusters in the postcerebral ganglia. FMRFa immunolabeling could be observed within subpopulations of NADPHd positive cells and in pericellular varicose fibers surrounding NADPHd stained neurons. Nitrite production of the ganglia was stimulated by L-arginine (10- 20 mM) but was decreased by NOARG (1-2 mM). Synthetic FMRFa (0.830-3.340 mM) increased the nitrite production in a dose dependent manner, but was ineffective in the presence of NOARG. Amiloride hydrochloride (7.890 mM) reduced the FMRFa evoked nitrite production in all ganglia. This is the first description of an anatomical relationship between putative NO producing and FMRFa containing cells, suggesting a possible regulatory role of FMRFa in the NO mediated signaling in an invertebrate nervous system., (Copyright 2004 S. Karger AG, Basel)