Ariano M A, Lewicki J A, Brandwein H J, Murad F
Proc Natl Acad Sci U S A. 1982 Feb;79(4):1316-20. doi: 10.1073/pnas.79.4.1316.
The immunohistochemical localization of guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] has been examined in rat neocortex, caudate-putamen, and cerebellum by using specific monoclonal antibodies. Immunofluorescence could be seen within somata and proximal dendrites of neurons in the these regions. A nuclear immunofluorescence reaction to guanylate cyclase was characteristically absent. The staining pattern for guanylate cyclase was coincident with previously described localizations of cyclic GMP immunofluorescence within medium spiny neurons of the caudate-putamen and pyramidal cells of the neocortex. Cerebellar guanylate cyclase immunoreactivity was primarily confined to Purkinje cells and their primary dendrites, similar to the pattern reported for cyclic GMP-dependent protein kinase localization. Guanylate cyclase immunofluorescence was abolished when the monoclonal antibodies were exposed to purified enzyme prior to incubation of the tissue slices or when control antibody was substituted for the primary antibody. Immunohistochemical localization of cyclic AMP in these same tissues was readily distinguished from that of guanylate cyclase or cyclic GMP, showing uniform fluorescence throughout the cell bodies of neurons and glial elements.
利用特异性单克隆抗体,已在大鼠新皮层、尾状核 - 壳核和小脑中检测了鸟苷酸环化酶[GTP焦磷酸裂解酶(环化),EC 4.6.1.2]的免疫组织化学定位。在这些区域的神经元胞体和近端树突内可见免疫荧光。对鸟苷酸环化酶的核免疫荧光反应典型地不存在。鸟苷酸环化酶的染色模式与先前描述的尾状核 - 壳核的中等棘状神经元和新皮层的锥体细胞内的环鸟苷酸免疫荧光定位一致。小脑鸟苷酸环化酶免疫反应性主要局限于浦肯野细胞及其初级树突,类似于报道的环鸟苷酸依赖性蛋白激酶定位模式。当在组织切片孵育前将单克隆抗体与纯化酶接触,或用对照抗体替代一抗时,鸟苷酸环化酶免疫荧光消失。这些相同组织中环磷酸腺苷的免疫组织化学定位很容易与鸟苷酸环化酶或环鸟苷酸的定位区分开来,在神经元和神经胶质细胞的整个细胞体中显示出均匀的荧光。