Röszer Tamás, Jenei Zsolt, Gáll Tamás, Nagy Olivér, Czimmerer Zsolt, Serfözö Zoltán, Elekes Károly, Bánfalvi Gáspár
Department of Animal Anatomy and Physiology, Faculty of Natural Sciences, Debrecen University, Debrecen, Hungary.
Brain Behav Evol. 2004;63(1):23-33. doi: 10.1159/000073757.
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.
运用NADPH黄递酶（NADPHd）组织化学方法在光镜水平观察一氧化氮合酶，并用免疫细胞化学方法检测FMRF酰胺（FMRFa），研究了表达一氧化氮（NO）合酶的神经元与软体动物心脏兴奋（FMRF酰胺样）神经肽之间的解剖学和功能关系。通过比色法格里斯测定法检测神经节中NO的分解产物亚硝酸盐，来检测神经节中NO的产生。还测试了NO合酶底物氨基酸L-精氨酸、NO合酶抑制剂Nω-硝基-L-精氨酸（NOARG）、合成FMRFa以及FMRFa敏感离子通道阻滞剂盐酸阿米洛利对亚硝酸盐产生的影响。NADPHd反应标记了脑神经节中的前脑、中脑和后脑内的神经细胞和纤维，以及后脑神经节中的细胞簇。在NADPHd阳性细胞亚群内以及围绕NADPHd染色神经元的细胞周曲张纤维中可观察到FMRFa免疫标记。神经节中的亚硝酸盐产生受到L-精氨酸（10 - 20 mM）的刺激，但被NOARG（1 - 2 mM）降低。合成FMRFa（0.830 - 3.340 mM）以剂量依赖方式增加亚硝酸盐产生，但在存在NOARG时无效。盐酸阿米洛利（7.890 mM）降低了所有神经节中FMRFa诱发的亚硝酸盐产生。这是对假定产生NO的细胞与含FMRFa的细胞之间解剖学关系的首次描述，表明FMRFa在无脊椎动物神经系统的NO介导信号传导中可能具有调节作用。
