Pinto L H, Kaneko A
Department of Neurobiology and Physiology, Northwestern University, Evanston, IL 60208.
Vis Neurosci. 1991 Jan;6(1):69-74. doi: 10.1017/s0952523800000912.
Total RNA was purified from freshly isolated retinas of adult carp and injected into oocytes of Xenopus laevis (stages 5-6). Two to six days after injection, depolarizing voltage-clamp steps evoked a slowly activated outward current as large as 3 microA. This current inactivated slowly with a single time constant (tau = 3.1 +/- 0.24 S.E.M., for Vm = +30 mV). The current was inhibited by tetraethylammonium (3.8 mM for half-maximal inhibition). In the presence of Co2+ (1 mM) or barium methanesulfonate (40 mM), the current-voltage relationship shifted to slightly more depolarized values (5-10 mV); the maximal value of the current that was sensitive to Co2+ or Ba2+ treatments was only a small fraction (about 10%) of the TEA-sensitive current, and its current-voltage relationship was similar to that for uninjected oocytes. The reversal potential of the membrane current was studied with [K+]0 of 1-77 mM. For [K+]0 greater than 20 mM, the reversal potential changed with a slope of 63 mV (+/- 2 mV S.E.M.) per 10-fold change in [K+]0. The conductance was induced half-maximally at 17 mV (+/- 0.9 mV S.E.M.). The depolarization required for an e-fold increase in conductance was 13 mV (+/- 0.6 mV S.E.M.). From these results, we conclude that the injection of total RNA from carp retinas induces the formation of a membrane K+ channel in Xenopus oocytes. The channel formed has many of the properties reported for the maintained outward current of goldfish horizontal and bipolar cells.
从成年鲤鱼新鲜分离的视网膜中纯化总RNA,并将其注射到非洲爪蟾(5-6期)的卵母细胞中。注射后2至6天,去极化电压钳制步骤诱发了高达3微安的缓慢激活外向电流。该电流以单一时间常数缓慢失活(对于Vm = +30 mV,τ = 3.1 +/- 0.24 S.E.M.)。该电流被四乙铵抑制(半最大抑制浓度为3.8 mM)。在存在Co2+(1 mM)或甲磺酸钡(40 mM)的情况下,电流-电压关系向稍去极化的值(5-10 mV)移动;对Co2+或Ba2+处理敏感的电流的最大值仅为TEA敏感电流的一小部分(约10%),并且其电流-电压关系与未注射卵母细胞的相似。在[K+]0为1-77 mM的条件下研究了膜电流的反转电位。对于[K+]0大于20 mM,反转电位随[K+]0每10倍变化以63 mV(+/- 2 mV S.E.M.)的斜率变化。电导在17 mV(+/- 0.9 mV S.E.M.)时被诱导至半最大。电导增加e倍所需的去极化是13 mV(+/- 0.6 mV S.E.M.)。从这些结果,我们得出结论,注射鲤鱼视网膜的总RNA可诱导非洲爪蟾卵母细胞中形成膜钾通道。所形成的通道具有许多报道的金鱼水平细胞和双极细胞持续外向电流的特性。