Krafte D S, Lester H A
Division of Biology, California Institute of Technology, Pasadena 91125.
J Neurosci Methods. 1989 Jan;26(3):211-5. doi: 10.1016/0165-0270(89)90118-0.
We have injected mRNA from rabbit brain into stage II-III Xenopus oocytes to determine whether they will translate exogenous RNA and incorporate functional ion channels into the membrane. Our results show that 48 h after RNA injection, functional voltage-dependent Na channels are present at sufficient densities to allow quantitative electrophysiological recording. The smaller oocytes have at least 2 experimental advantages over the stage V-VI oocytes normally used for electrophysiological experiments: (1) the smaller membrane capacitance (approximately 5-fold) allows a faster settling time following a voltage step and a more detailed kinetic analysis of membrane currents than was previously possible with the 2-microelectrode technique, and (2) roughly 8-fold less RNA is needed for each injection. Thus, the stage II-III Xenopus oocyte is a suitable preparation for the study of ion channels.
我们已将来自兔脑的mRNA注入II - III期非洲爪蟾卵母细胞,以确定它们是否会翻译外源RNA并将功能性离子通道整合到细胞膜中。我们的结果表明,RNA注射后48小时,功能性电压依赖性钠通道以足够的密度存在,从而能够进行定量电生理记录。较小的卵母细胞与通常用于电生理实验的V - VI期卵母细胞相比至少具有两个实验优势:(1)较小的膜电容(约为5倍)使得电压阶跃后的稳定时间更快,并且与之前的双微电极技术相比,能够对膜电流进行更详细的动力学分析;(2)每次注射所需的RNA大约少8倍。因此,II - III期非洲爪蟾卵母细胞是研究离子通道的合适标本。