Ueno S, Bambauer H J, Umar H, Ueck M, Ogawa K
Cell Tissue Res. 1984;237(3):479-89. doi: 10.1007/BF00228432.
Ca++-ATPase activity was demonstrated histochemically at light- and electron-microscopic levels in inner and outer segments of retinal photoreceptor cells of the guinea pig with the use of a newly developed one-step lead-citrate method (Ando et al. 1981). The localization of ouabain-sensitive, K+-dependent p-nitrophenylphosphatase (K+-NPPase) activity, which represents the second dephosphorylative step of the Na+-K+-ATPase system, was studied by use of the one-step method newly adapted for ultracytochemistry (Mayahara et al. 1980). In retinal photoreceptor cells fixed for 15 min in 2% paraformaldehyde the electron-dense Ca++-ATPase reaction product accumulated significantly on the inner membranes of the mitochondria but not on the plasmalemma or other cytoplasmic elements of the inner segments. The membranes of the outer segments remained unstained except the membrane arrays in close apposition to the retinal pigment epithelium. The cytochemical reaction was Ca++- and substrate-dependent and showed sensitivity to oligomycin. When Mg++-ions were used instead of Ca++-ions, a distinct reaction was also found on mitochondrial inner membranes. In contrast to the localization of the Ca++-ATPase activity, the K+-NPPase activity was demonstrated only on the plasmalemma of the inner segments, but not on the mitochondria, other cytoplasmic elements or the outer segment membranes. This reaction was almost completely abolished by ouabain or by elimination of K+ from the incubation medium.
利用新开发的一步柠檬酸铅法（安藤等人，1981年），在光镜和电镜水平上对豚鼠视网膜光感受器细胞内段和外段的Ca++-ATP酶活性进行了组织化学证明。采用新适用于超微细胞化学的一步法（马亚哈拉等人，1980年）研究了哇巴因敏感的、依赖钾的对硝基苯磷酸酶（K+-NPP酶）活性的定位，该活性代表钠钾ATP酶系统的第二步去磷酸化步骤。在2%多聚甲醛中固定15分钟的视网膜光感受器细胞中，电子致密的Ca++-ATP酶反应产物显著积聚在线粒体内膜上，而不是内段的质膜或其他细胞质成分上。外段的膜除了与视网膜色素上皮紧密相邻的膜排列外均未染色。细胞化学反应依赖于Ca++和底物，并对寡霉素敏感。当用Mg++离子代替Ca++离子时，在线粒体内膜上也发现了明显的反应。与Ca++-ATP酶活性的定位相反，K+-NPP酶活性仅在内段的质膜上得到证明，而在线粒体、其他细胞质成分或外段膜上未得到证明。哇巴因或从孵育培养基中去除K+几乎完全消除了这种反应。
