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雏鸡离体前庭毛细胞中的机械电转导电流

Mechano-electrical transduction currents in isolated vestibular hair cells of the chick.

作者信息

Ohmori H

出版信息

J Physiol. 1985 Feb;359:189-217. doi: 10.1113/jphysiol.1985.sp015581.

Abstract

Properties of a mechano-electrical transduction channel were studied in enzymatically dissociated chick vestibular hair cells by using a whole-cell recording variation of the patch voltage-clamp technique. The apical hair bundle was stimulated by a glass rod which moved along a one-dimensional axis when stimulated by either a triangular or a trapezoidal command voltage. The motion of the glass rod was monitored optically using a photodiode. In response to triangular stimuli, the hair cell generated a current of triangular wave form with occasional step-like spiky or zigzag-appearing events. Control experiments confirmed that the current was generated only when the hair bundle was displaced towards the tallest stereocilium. The mechano-sensitive current was blocked by streptomycin and by neomycin. The blockage by streptomycin was clearly voltage dependent: the reduction of the current became larger with hyperpolarization of the membrane. This suggests that the positively charged antibiotic molecules plug the mechanically gated channels. From the evidence presented in 3 and 4 above, the mechano-sensitive current recorded here was identified as the mechano-electrical transduction (m-e.t.) current. The permeability of the m-e.t. channel to various monovalent cations was determined from reversal potential measurements. Since a CsCl-EGTA intracellular medium was used, all the permeabilities were calculated relative to PCs. The sequence of permeabilities was Li greater than Na greater than or equal to K greater than or equal to Rb greater than Cs greater than choline greater than TMA greater than TEA. External Ca ions were indispensable for the recording of transduction current and Sr ions could replace Ca ions without loss of the transduction activity. The minimum [Ca]o for stable generation of the m-e.t. current was 20 microM in Cs saline. The addition of 50-200 microM-Ca to the isotonic Ba saline could maintain the m-e.t. current. The m-e.t. current was observed in isotonic Ca and in Sr salines. Isotonic Ba, Mg and Mn salines were enriched with 1-2 mM-Ca in order to generate the m-e.t. current. The permeabilities of the divalent cations relative to Cs were calculated from the reversal potentials, and the sequence of permeabilities among divalent cations was Ca greater than Sr greater than Ba greater than Mn greater than Mg. Step-like m-e.t. currents were observed in Cs saline. The smallest step amplitude with clear resolution had a conductance of 49.7 +/- 4.5 pS (mean +/- S.D., n = 7 cells). This is likely to be an elementary m-e.t. channel conductance.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

采用膜片电压钳技术的全细胞记录变体,在酶解的鸡前庭毛细胞中研究了机械电转导通道的特性。用一根玻璃棒刺激顶端毛束,当受到三角波或梯形指令电压刺激时,玻璃棒会沿一维轴移动。使用光电二极管对玻璃棒的运动进行光学监测。响应三角波刺激时,毛细胞产生三角波形电流,偶尔会出现阶梯状尖峰或锯齿状事件。对照实验证实,只有当毛束朝着最高的静纤毛位移时才会产生电流。机械敏感电流被链霉素和新霉素阻断。链霉素的阻断明显依赖电压:随着膜的超极化,电流的减小变得更大。这表明带正电荷的抗生素分子堵塞了机械门控通道。根据上述3和4中的证据,此处记录的机械敏感电流被确定为机械电转导(m-e.t.)电流。通过反转电位测量确定了m-e.t.通道对各种单价阳离子的通透性。由于使用了CsCl-EGTA细胞内介质,所有通透性都是相对于Cs计算的。通透性顺序为Li>Na≥K≥Rb>Cs>胆碱>TMA>TEA。外部钙离子对于转导电流的记录是不可或缺的,锶离子可以替代钙离子而不丧失转导活性。在Cs盐溶液中,稳定产生m-e.t.电流的最小[Ca]o为20μM。向等渗Ba盐溶液中添加50 - 200μM - Ca可以维持m-e.t.电流。在等渗Ca盐溶液和Sr盐溶液中观察到了m-e.t.电流。等渗Ba、Mg和Mn盐溶液中添加了1 - 2 mM - Ca以产生m-e.t.电流。根据反转电位计算了二价阳离子相对于Cs的通透性,二价阳离子之间的通透性顺序为Ca>Sr>Ba>Mn>Mg。在Cs盐溶液中观察到了阶梯状m-e.t.电流。具有清晰分辨率的最小阶梯幅度的电导为49.7±4.5 pS(平均值±标准差,n = 7个细胞)。这可能是一个基本的m-e.t.通道电导。(摘要截取自400字)

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cea/1193371/fcf0018e29bf/jphysiol00580-0224-a.jpg

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