Simon B J, Hill D A
Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston 77550.
Biophys J. 1992 May;61(5):1109-16. doi: 10.1016/S0006-3495(92)81920-3.
Charge movement currents (IQ) and calcium transients (delta[Ca2+]) were measured simultaneously in frog skeletal muscle fibers, voltage clamped in a double vaseline gap chamber, using Antipyrylazo III as the calcium indicator. The rate of release of calcium from the SR (Rrel) was calculated from the calcium transients using the removal model of Melzer, W., E. Rios, and M. F. Schneider (1987. Biophys. J. 51:849-863.). IQ and delta [Ca2+] were calculated for 100 ms depolarizing test pulses to membrane potentials from -30 to +20 mV. To eliminate an inactivating component of Rrel, each test pulse was preceded by a large, fixed prepulse to +20 mV. The resulting Rrel records, which represent the noninactivating component of Rrel, were compared with integral of IQdt.(Q), the total charge that moves. The voltage dependence of the steady state Rrel was steeper then that of Q and shifted to the right. During depolarization, the Rrel waveform was similar to that of Q but was delayed by several ms, while, during repolarization, Rrel preceded Q. All of these results could be explained with a Hodgkin-Huxley type model for E-C coupling in which four voltage sensors in the t-tubule membrane which give rise to IQ must all be in their activating positions for the calcium release channel in the SR membrane to open. his model is consistent with the structural architecture of the triadic junction in which four dihydropyridine receptors (the voltage sensors for E-C coupling) in the t-tubule membrane are closely associated with each ryanodine receptor(the calcium release channel) in the SR membrane [Block, B. A., T. Imagawa, K. P. Campbell, and C. Franzini-Armstrong. 1988. J.Cell. Biol. 107:2587-2600.]). Some aspects of this work have appeared in abstract form (Simon, B. J., and D. Hill. 1991. Biophys. J.59:64a. ([Abstr.]).
在双凡士林间隙室中电压钳制的青蛙骨骼肌纤维中,使用安替比拉宗III作为钙指示剂,同时测量电荷移动电流(IQ)和钙瞬变(δ[Ca2+])。根据钙瞬变,使用梅尔泽、E. 里奥斯和M. F. 施奈德(1987年,《生物物理学杂志》51:849 - 863)的去除模型计算从肌浆网(SR)释放钙的速率(Rrel)。对于从 - 30到 + 20 mV的膜电位的100 ms去极化测试脉冲,计算IQ和δ[Ca2+]。为了消除Rrel的失活成分,每个测试脉冲之前都有一个到 + 20 mV的大的固定预脉冲。将得到的代表Rrel非失活成分的Rrel记录与移动的总电荷IQdt的积分(Q)进行比较。稳态Rrel的电压依赖性比Q更陡峭且向右移动。在去极化期间,Rrel波形与Q相似,但延迟了几毫秒,而在复极化期间,Rrel先于Q。所有这些结果都可以用霍奇金 - 赫胥黎型的兴奋 - 收缩偶联模型来解释,其中横管膜中的四个电压传感器产生IQ,它们都必须处于激活位置,SR膜中的钙释放通道才能打开。该模型与三联体连接的结构架构一致,其中横管膜中的四个二氢吡啶受体(兴奋 - 收缩偶联的电压传感器)与SR膜中的每个兰尼碱受体(钙释放通道)紧密相关[布洛克、B. A.、T. 今川、K. P. 坎贝尔和C. 弗兰齐尼 - 阿姆斯特朗。1988年,《细胞生物学杂志》107:2587 - 2600]。这项工作的一些方面已以摘要形式发表(西蒙、B. J. 和D. 希尔。1991年,《生物物理学杂志》59:64a。[摘要])。