Department of Biochemistry, Brandeis University, Waltham, MA 02453, USA.
J Gen Physiol. 2013 Mar;141(3):309-21. doi: 10.1085/jgp.201210909. Epub 2013 Feb 11.
The number of voltage-gated sodium (Na(V)) channels available to generate action potentials in muscles and nerves is adjusted over seconds to minutes by prior electrical activity, a process called slow inactivation (SI). The basis for SI is uncertain. Na(V) channels have four domains (DI-DIV), each with a voltage sensor that moves in response to depolarizing stimulation over milliseconds to activate the channels. Here, SI of the skeletal muscle channel Na(V)1.4 is induced by repetitive stimulation and is studied by recording of sodium currents, gating currents, and changes in the fluorescence of probes on each voltage sensor to assess their movements. The magnitude, voltage dependence, and time course of the onset and recovery of SI are observed to correlate with voltage-sensor movements 10,000-fold slower than those associated with activation. The behavior of each voltage sensor is unique. Development of SI over 1-160 s correlates best with slow immobilization of the sensors in DI and DII; DIII tracks the onset of SI with less fidelity. Showing linkage to the sodium conduction pathway, pore block by tetrodotoxin affects both SI and immobilization of all the sensors, with DI and DII significantly suppressed. Recovery from SI correlates best with slow restoration of mobility of the sensor in DIII. The findings suggest that voltage-sensor movements determine SI and thereby mediate Na(V) channel availability.
电压门控钠离子 (Na(V)) 通道的数量可在数秒至数分钟内通过先前的电活动进行调节,这个过程称为缓慢失活 (SI)。SI 的基础尚不清楚。Na(V) 通道有四个结构域(DI-DIV),每个结构域都有一个电压传感器,该传感器会响应毫秒级的去极化刺激而移动,从而激活通道。这里,通过重复刺激诱导骨骼肌通道 Na(V)1.4 的 SI,并通过记录钠离子电流、门控电流以及每个电压传感器上探针的荧光变化来研究其运动,以评估它们的运动情况。观察到 SI 的幅度、电压依赖性以及起始和恢复的时程与电压传感器的运动相关,其速度比与激活相关的运动慢 10,000 倍。每个电压传感器的行为都是独特的。在 1-160 秒内观察到 SI 的发展与 DI 和 DII 中传感器的缓慢固定相关性最佳;DIII 以较低的保真度跟踪 SI 的起始。与钠传导途径的连接表明,河豚毒素对所有传感器的 SI 和固定均有影响,其中 DI 和 DII 受到显著抑制。从 SI 中恢复与 DIII 中传感器缓慢恢复运动相关性最佳。研究结果表明,电压传感器的运动决定了 SI,从而介导 Na(V) 通道的可用性。