A nonenzymatic preparation of epithelial basolateral membrane for patch clamp.
A nonenzymatic preparation of epithelial basolateral membrane for patch clamp.
复制标题
用于膜片钳的上皮基底外侧膜的非酶制剂。
DOI：
10.1152/ajpcell.1990.258.6.c1159
复制
发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Reuss,L
中科院分区：
文献类型：
--
作者：
Wehner,F;Garretson,L;Dawson,K;Segal,Y;Reuss,L
已经开发了一种制剂，其允许以高成功率对Necturus胆囊上皮细胞的基底外侧膜进行膜片钳。将上皮与下层组织机械分离，无需酶处理。将其顶面附着在塑料盖玻片上，并在显微镜观察下用吸移管清洁基底外侧表面（面朝上）。通过这种清洁程序，获得千兆欧姆密封的成功率从不到1%增加到大约10%。细胞似乎保留其结构和功能的完整性，证明了电子显微镜的外观和细胞膜电压的大小。制备的主要优点是基底外侧膜结构域被保留，并且不需要可能改变膜蛋白的酶处理。
A preparation has been developed that permits patch clamping of the basolateral membrane of Necturus gallbladder epithelial cells with a high success rate. The epithelium is separated from the underlying tissues mechanically, without enzymatic treatment. Its apical surface is attached to a plastic cover slip, and the basolateral surface, facing up, is cleaned with a suction pipette under microscopic observation. With this cleaning procedure, the success rate in obtaining gigaohm seals increases from less than 1% to approximately 10% of the attempts. The cells appear to retain their structural and functional integrity, as evidenced by electron-microscopic appearance and magnitude of cell membrane voltages. Major advantages of the preparation are that the basolateral membrane domain is preserved and that enzymatic treatment, which could potentially alter membrane proteins, is not necessary.