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牛离体气管平滑肌细胞中电压门控性钙通道的特性

The properties of voltage-operated Ca(2+)-channels in bovine isolated trachealis cells.

作者信息

Green K A, Small R C, Foster R W

机构信息

Department of Physiological Sciences, University of Manchester, UK.

出版信息

Pulm Pharmacol. 1993 Mar;6(1):49-62. doi: 10.1006/pulp.1993.1008.

Abstract

Freshly-dispersed bovine trachealis cells were used for recording by the patch clamp technique of whole-cell and unitary currents through Ca(2+)-channels. Whole-cell Ca(2+)-current (ICa) activated at -40 mV and appeared to be carried by a single type of Ca(2+)-channel. Inactivation of ICa was increased by increasing the concentration of free Ca2+ within the recording pipette but reduced by using Ba2- as the charge carrier. Steady-state inactivation studies showed that the Ca(2+)-channels were half-maximally available following a conditioning depolarization to -35 mV. A two-pulse protocol showed that ICa induced by the step to a test potential was inversely related to ICa induced by the step to the conditioning potential. Unitary Ba(2+)-currents were activated at a threshold of -30 mV and had a reversal potential of +41 mV. The channel carrying the unitary Ba(2+)-currents had a slope conductance of 23 pS. Steady-state inactivation studies showed that the unitary Ba(2+)-currents were half-maximally available at a holding potential of -28 mV. ICa and unitary Ba(2+)-currents were inhibited by nifedipine (10 nM-1 microM) but augmented by Bay K 8644 (10 microM). It is concluded that the plasmalemma of bovine trachealis muscle contains a single population of voltage-dependent Ca(2+)-channels of the L-type. These channels may be subject to inactivation primarily by an increase in the concentration of free Ca2+ on the cytosolic side of the plasmalemma and secondarily by a voltage-dependent mechanism. Overlap of the inactivation and activation curves of ICa may allow the passage of 'window current' through the Ca(2+)-channels during sustained depolarization.

摘要

采用膜片钳技术,利用新鲜分离的牛气管平滑肌细胞记录通过钙通道的全细胞电流和单通道电流。全细胞钙电流(ICa)在-40 mV时被激活,似乎由单一类型的钙通道携带。通过增加记录电极内游离Ca2+的浓度可增强ICa的失活,但使用Ba2+作为载流子则可降低失活。稳态失活研究表明,钙通道在经-35 mV的预处理去极化后,其可用性达到一半最大值。双脉冲方案显示,由阶跃至测试电位诱发的ICa与由阶跃至预处理电位诱发的ICa呈负相关。单通道Ba2+电流在-30 mV的阈值时被激活,反转电位为+41 mV。携带单通道Ba2+电流的通道斜率电导为23 pS。稳态失活研究表明,单通道Ba2+电流在-28 mV的钳制电位下可用性达到一半最大值。ICa和单通道Ba2+电流被硝苯地平(10 nM - 1 μM)抑制,但被Bay K 8644(10 μM)增强。结论是,牛气管平滑肌的质膜含有单一群体的L型电压依赖性钙通道。这些通道可能主要通过质膜胞质侧游离Ca2+浓度的增加而失活,其次通过电压依赖性机制失活。ICa失活和激活曲线的重叠可能允许在持续去极化期间“窗电流”通过钙通道。

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