半滑舌鳎脑组织中神经型一氧化氮合成酶免疫组织化学定位研究

Immunocytochemical Localization of Neuronal Nitric Oxide Synthase in Brain Tissue of Semi-smooth Sole Cynoglossus semilaevis Günther

为探明神经型一氧化氮合成酶在半滑舌鳎脑组织中的分布情况,利用免疫组织化学SP法以及NADPH-d组织化学染色法对脑组织中神经型一氧化氮合成酶进行定位研究。免疫组织化学试验结果表明,大脑中的神经型一氧化氮合成酶位于神经元胞浆内,阳性神经元多呈梭形、三角形、圆形等多种形状。反应呈阳性的神经纤维,相互交织呈网状,分布在大脑皮质浅层。小脑中反应呈阳性的神经元分布在皮质的颗粒层中,分子层内有许多呈阳性的神经纤维和浦肯野细胞。NADPH-d组织化学染色试验结果显示,大脑中有许多蓝色的神经元,小脑中神经元、神经纤维、浦肯野细胞呈蓝色。利用蛋白免疫印迹技术对脑组织中神经型一氧化氮合成酶进行分析,结果显示分子量分别为118ku和80ku的两条蛋白条带。研究表明,在半滑舌鳎脑组织中存在大量的神经型一氧化氮合成酶,由此推测,神经型一氧化氮合成酶在鱼类神经系统的信息传导和生理活动中起着重要调节作用。

The localization of neuronal nitric oxide synthase （nNOS） in the brain tissue of semi-smooth sole Cynoglossus semilaevis Günther was studied using immunohistochemical SP and NADPH-diaphorase staining method .The results showed that the nNOS was found in the cytoplasm of neurons in the cerebra , mostly positive neurons being fusiform , triangle , or round in shape . T he positive nerve fibers in the superficial layer of cerebral cortex were interwoven into a reticulation ,and the positive neurons of cerebel were distributed in the particle layer of cortex ,with many positive nerve fibers and Purkinje cells in the molecular layer .NADPH-d staining showed that there were numerous blue neurons in the cerebra ,and blue neurons ,nerve fibers and Purkinje cells in the cerebel .Western-Blot revealed that there were two protein bands with molecular weight of 118 ku and 80 ku in nNOS in the brain tissue .The findings showed that there was plenty of neuronal nitric oxide synthase in the brain tissue of the sole ,indicating that nNOS can play an important modulatory role in the signal transduction and physiological activity of the fish nervous system .