Dascal N, Chilcott G, Lester H A
Department of Physiology and Pharmacology, Sackler School of Medicine, Tel Aviv University, Israel.
J Neurosci Methods. 1991 Aug;39(1):29-38. doi: 10.1016/0165-0270(91)90090-m.
We describe a method for internal perfusion of Xenopus laevis oocytes that allows control of the composition of intracellular and extracellular solutions, including the possibility of sequential introduction of different substances inside and outside the cell. Using this method, it was possible to record Ca2+ dependent Cl- current and to inhibit it by intracellular perfusion of EGTA-containing solution. With a high BA2+ solution at the external surface of the perfused oocyte, Ba2+ currents through voltage-dependent Ca2+ channels were observed in native and in cardiac RNA-injected oocytes. Finally, a delayed rectifier K+ current was recorded and blocked by internally perfused Cs+ in oocytes injected with mRNA of a cloned (MBK1) K+ channel. The method is expected to be useful for the study of function and modulation of ion channels and transporters in the oocyte, which is an important and widely used model system.
我们描述了一种非洲爪蟾卵母细胞内部灌注的方法，该方法可控制细胞内和细胞外溶液的成分，包括在细胞内外顺序引入不同物质的可能性。使用这种方法，能够记录Ca2+依赖性Cl-电流，并通过灌注含EGTA的溶液在细胞内抑制该电流。在灌注卵母细胞的外表面使用高Ba2+溶液时，在天然卵母细胞和注射了心脏RNA的卵母细胞中观察到Ba2+通过电压依赖性Ca2+通道的电流。最后，在注射了克隆的（MBK1）K+通道mRNA的卵母细胞中记录到延迟整流K+电流，并被内部灌注的Cs+阻断。该方法有望用于研究卵母细胞中离子通道和转运体的功能及调节，卵母细胞是一个重要且广泛使用的模型系统。
