Masuzawa T, Ohta T, Kawamura M, Nakahara N, Sato F
Brain Res. 1984 Jun 8;302(2):357-62. doi: 10.1016/0006-8993(84)90250-6.
To determine if canine and rat choroid plexus Na+,K+-ATPase can be localized by immunoperoxidase staining after fixation and embedding, we prepared rabbit antiserum to purified canine kidney medulla Na+,K+-ATPase. When sodium dodecylsulfate polyacrylamide electrophoretic gels of purified canine kidney Na+,K+-ATPase and canine kidney microsomes were treated with antiserum followed by [125I]protein A and autoradiography, the canine microsomes and purified Na+,K+-ATPase showed a prominent radioactive band coincident with the alpha-, beta- and gamma-subunits of the purified canine kidney enzyme. When the rabbit immunoglobulin that was purified from the Na+,K+-ATPase antiserum through DEAE-cellulose ion exchange chromatography was used for immunoperoxidase staining of the choroid plexus fixed with Bouin's fixative, intense immunoreactive staining was present on the epithelial cells of both choroid plexuses but was not found in the tissue around the vessel. The staining was especially confined to apical surfaces of the epithelial cells. The same procedure was performed in the canine kidney, and immunostaining was obtained in the tubules where Baskin and Stahl described the enzyme localization. No staining was seen with pre-immune serum of the normal rabbit. We concluded that both the canine and rat choroid plexus are rich in Na+,K+-ATPase, which plays an important role in cerebrospinal fluid (CSF) secretion.
为了确定犬和大鼠脉络丛的Na⁺,K⁺-ATP酶在固定和包埋后能否通过免疫过氧化物酶染色进行定位,我们制备了针对纯化的犬肾髓质Na⁺,K⁺-ATP酶的兔抗血清。当用抗血清处理纯化的犬肾Na⁺,K⁺-ATP酶和犬肾微粒体的十二烷基硫酸钠聚丙烯酰胺电泳凝胶,然后用[¹²⁵I]蛋白A和放射自显影处理时,犬微粒体和纯化的Na⁺,K⁺-ATP酶显示出一条与纯化的犬肾酶的α、β和γ亚基一致的明显放射性带。当通过DEAE-纤维素离子交换色谱从Na⁺,K⁺-ATP酶抗血清中纯化的兔免疫球蛋白用于对经Bouin固定液固定的脉络丛进行免疫过氧化物酶染色时,两个脉络丛的上皮细胞均出现强烈的免疫反应性染色,但在血管周围组织中未发现。染色特别局限于上皮细胞的顶端表面。在犬肾中进行了相同的操作,并在Baskin和Stahl描述该酶定位的肾小管中获得了免疫染色。正常兔的免疫前血清未见染色。我们得出结论,犬和大鼠脉络丛均富含Na⁺,K⁺-ATP酶,其在脑脊液(CSF)分泌中起重要作用。