The neurons of the retina have electrical properties that are different from those of most of the other neurons of the central nervous system. To identify the voltage-gated ion channels found in the retina, we screened mouse retinal cDNA libraries with oligonucleotide probes homologous to the mammalian K+ channel MBK1 (Kv1.1) and ligated two partial clones to produce a full-length clone with no significant differences from MBK1. 2. Expression of MBK1 mRNA was determined by RNAse protection. MBK1 mRNA was detected in retinal RNA and was also detected in brain, liver, and heart RNAs. 3. We transcribed the full-length clone, injected it into oocytes of Xenopus laevis, and measured the membrane currents 2 to 6 days later. Depolarization from a holding voltage of -90mV induced a slowly activated outward current with a peak value as large as 20 microA. The current inactivated very slowly with a single exponential time course [mean time constant, 6.5 +/- 0.4 sec (SEM) for activation voltage of -10mV]. 4. The outward current was reduced to half-maximal by 0.42 mM tetraethylammonium, 1.1 mM 4-aminopyridine, and 3.2 mM Ba2+ but was not significantly attenuated by Co2+ (1 mM). 5. The reversal potential (measured with tail currents) changed by 53mV per decade change of [K+] from 1 to 77 mM. 6. The voltage for half-maximal activation of the conductance was -26.6mV (+/- 1.7mV), and the voltage required for an e-fold increase in conductance was 6.9mV (+/- 0.5mV). 7. Thus, the mRNA for MBK1 found in the mouse retina causes the expression of a voltage-dependent K+ current which has properties suitable for may retinal neurons.
摘要
视网膜神经元具有与中枢神经系统大多数其他神经元不同的电特性。为了鉴定视网膜中发现的电压门控离子通道,我们用与哺乳动物K+通道MBK1(Kv1.1)同源的寡核苷酸探针筛选小鼠视网膜cDNA文库,并连接两个部分克隆以产生一个与MBK1无显著差异的全长克隆。2. 通过RNA酶保护测定MBK1 mRNA的表达。在视网膜RNA中检测到MBK1 mRNA,在脑、肝和心脏RNA中也检测到。3. 我们转录全长克隆,将其注射到非洲爪蟾的卵母细胞中,并在2至6天后测量膜电流。从-90mV的保持电压去极化诱导出一个缓慢激活的外向电流,峰值高达20微安。电流以单指数时间进程非常缓慢地失活[-10mV激活电压下的平均时间常数为6.5±0.4秒(标准误)]。4. 外向电流在0.42 mM四乙铵、1.1 mM 4-氨基吡啶和3.2 mM Ba2+作用下降低至最大值的一半,但在1 mM Co2+作用下没有显著减弱。5. 反转电位(用尾电流测量)随[K+]从1 mM到77 mM每十倍变化改变53mV。6. 电导半最大激活电压为-26.6mV(±1.7mV),电导增加e倍所需电压为6.9mV(±0.5mV)。7. 因此,在小鼠视网膜中发现的MBK1 mRNA导致表达一种电压依赖性K+电流,其特性适合许多视网膜神经元。