Wolf G, Würdig S, Henschke G
Institute of Medical Neurobiology, Faculty of Medicine, University of Magdeburg, Federal Republic of Germany.
J Neural Transm Suppl. 1994;43:105-12.
The recent discovery of the identify of nitric oxide synthase with the reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) has powerfully stimulated the anatomical localization of sites of nitric oxide synthesis in the nervous system. In the present study the widely used light microscopical technique for NADPH-d staining was adapted to the electron microscopical level by applying the tetrazolium salt 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl)tetrazolium chloride (BSPT) which produces an electron-dense reaction product, BSPT-formazan. Predominantly membranes of the endoplasmic reticulum were stained. Apart from singular heavily labeled neurons, a majority of nerve cells, light microscopically "unstained", shows sporadically formazan deposits, and, likewise, but regionally different, a few astroglial cells. Lesions induced by the glutamate agonists quinolinic acid and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) display surviving neurons, which are predominantly stained for NADPH-d. Astroglial cells within lesioned areas exhibit increased amounts of reaction product, apparently as a consequence of enzyme induction.
一氧化氮合酶与还原型烟酰胺腺嘌呤二核苷酸磷酸黄递酶（NADPH-d）的同一性最近被发现，这有力地推动了神经系统中一氧化氮合成部位的解剖学定位研究。在本研究中，通过应用能产生电子致密反应产物BSPT-甲臜的四氮唑盐2-（2'-苯并噻唑基）-5-苯乙烯基-3-（4'-邻苯二甲酰肼基）四氮唑氯化物（BSPT），将广泛使用的用于NADPH-d染色的光学显微镜技术应用于电子显微镜水平。主要是内质网的膜被染色。除了个别标记强烈的神经元外，大多数在光学显微镜下“未染色”的神经细胞偶尔会出现甲臜沉积物，同样，一些星形胶质细胞也有，但区域不同。由谷氨酸激动剂喹啉酸和α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）诱导的损伤显示存活的神经元，这些神经元主要被NADPH-d染色。损伤区域内的星形胶质细胞显示出反应产物的量增加，这显然是酶诱导的结果。
