Electrical responses in hair cells located in the peripheral regions and in the central region of the frog crista ampullaris were investigated in thin slice preparations by using the whole-cell configuration of the patch-clamp technique. 2. Hair cells from the peripheral regions exhibited mostly a club-like shape and had an average resting potential of -46 mV, whereas cells from the central region had mostly a cylindrical shape and a more negative resting potential (-57 mV). 3. Voltage-clamp recordings revealed that ionic conductances differed in the two epithelial regions. Cells from the peripheral regions exhibited a transient K+ current of A-type (IA) in conjunction with a slow rectifier outward K+ current (IK). Cells from the central region showed little or no IA and generated an IK together with an inward rectifier K+ current (IIR). In both regions, hair cells showed a rapidly activating Ca(2+)-dependent outward K+ current (IK(Ca)) that rapidly inactivated to reach a steady-state level during 150-ms test pulses. 4. IA activated close to -60 mV and was inhibited by 12 mM 4-aminopyridine (4-AP). The time course of this current showed time to peak values of 3-4 ms at 0 mV. Inactivation was fast and almost voltage-independent. The decay time constant was approximately 35 ms at 0 mV. 5. IK was recruited close to -60 mV and activated slowly, reaching peak values in approximately 100 ms at 0 mV. It showed no evidence of inactivation during 150-ms test pulses and it was insensitive to 4-AP. 6. IIR activated at membrane potentials more negative than -90 mV and was blocked by exposure to 6 mM Cs+ or to a K(+)-free medium. This current showed an outward relaxation at potentials more negative than -140 mV, an effect that disappeared after exposure to a Na(+)-free medium. 7. IK(Ca) was recruited close to -40 mV and was inhibited by exposure to a Ca(2+)-free external medium or to 0.5 mM Cd2+. The time to peak of this current was approximately 3 ms at 0 mV and inactivation was very fast and almost independent from the membrane potential. The decay time constant was approximately 4 ms at 0 mV. 8. IK and IA were prominent in hair cells from the peripheral regions, whereas IK accounted for most of the membrane conductance in cells from the central region. The contribution of IK(Ca) was comparable in cells from both epithelial regions.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
采用膜片钳技术的全细胞模式,在薄片标本中研究了位于青蛙壶腹嵴外周区域和中央区域的毛细胞的电反应。2. 外周区域的毛细胞大多呈棒状,平均静息电位为-46 mV,而中央区域的细胞大多呈柱状,静息电位更负(-57 mV)。3. 电压钳记录显示,两个上皮区域的离子电导不同。外周区域的细胞表现出A型瞬时钾电流(IA)以及缓慢整流外向钾电流(IK)。中央区域的细胞几乎没有或没有IA,产生IK以及内向整流钾电流(IIR)。在两个区域,毛细胞都表现出快速激活的钙依赖性外向钾电流(IK(Ca)),在150毫秒的测试脉冲期间迅速失活至稳态水平。4. IA在接近-60 mV时激活,被12 mM 4-氨基吡啶(4-AP)抑制。该电流的时间进程显示在0 mV时达到峰值的时间为3-4毫秒。失活很快且几乎与电压无关。在0 mV时衰减时间常数约为35毫秒。5. IK在接近-60 mV时被募集并缓慢激活,在0 mV时约100毫秒达到峰值。在150毫秒的测试脉冲期间没有失活的迹象,并且对4-AP不敏感。6. IIR在膜电位比-90 mV更负时激活,暴露于6 mM Cs+或无钾培养基中会被阻断。该电流在电位比-140 mV更负时表现出外向松弛,这种效应在暴露于无钠培养基后消失。7. IK(Ca)在接近-40 mV时被募集,暴露于无钙外部培养基或0.5 mM Cd2+中会被抑制。该电流在0 mV时达到峰值的时间约为3毫秒,失活非常快且几乎与膜电位无关。在0 mV时衰减时间常数约为4毫秒。8. IK和IA在外周区域的毛细胞中占主导,而IK在中央区域的细胞中占膜电导的大部分。IK(Ca)在两个上皮区域的细胞中的贡献相当。(摘要截断于400字)