Carney-Anderson L, Donaldson S K
Department of Physiology, School of Medicine, University of Minnesota, Minneapolis 55455.
Am J Physiol. 1994 Oct;267(4 Pt 1):C1087-94. doi: 10.1152/ajpcell.1994.267.4.C1087.
Skeletal muscle excitation-contraction (EC) coupling may involve secondary mechanisms, such as those involving G proteins. The aim of this study was to identify possible G protein effects on sarcoplasmic reticulum (SR) Ca2+ release, in general, and on voltage-dependent EC coupling, in particular. Effects of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) were studied using a single peeled rabbit skeletal muscle fiber preparation that is capable of releasing SR Ca2+ in response to transverse tubule (TT) depolarization. Because of possible nonspecific and residual effects of 200 microM GTP gamma S, a lower concentration of 50 microM GTP gamma S was used to stimulate G proteins in the peeled fiber system. Under conditions for steady-state (resting) polarization of TT, GTP gamma S rarely elicited SR Ca2+ release. When the TTs are in steady-state (resting) depolarization, 50 microM GTP gamma S or GTP elicited SR Ca2+ release and associated tension transients in only 69% of fibers tested. In contrast GTP gamma S always augmented Ca2+ release during TT depolarization-induced EC coupling. These results indicate the presence of at least two excitatory G proteins for SR Ca2+ release, only one of which is a modulator, albeit nonessential, of peeled fiber EC coupling.
骨骼肌兴奋-收缩(EC)偶联可能涉及二级机制,比如那些涉及G蛋白的机制。本研究的目的是确定G蛋白对肌浆网(SR)Ca2+释放的可能影响,总体而言,特别是对电压依赖性EC偶联的影响。使用单个去膜家兔骨骼肌纤维标本研究了5'-O-(3-硫代三磷酸)鸟苷(GTPγS)的作用,该标本能够响应横管(TT)去极化释放SR Ca2+。由于200μM GTPγS可能存在非特异性和残留效应,因此在去膜纤维系统中使用较低浓度的50μM GTPγS来刺激G蛋白。在TT处于稳态(静息)极化的条件下,GTPγS很少引发SR Ca2+释放。当TT处于稳态(静息)去极化时,50μM GTPγS或GTP仅在69%的测试纤维中引发SR Ca2+释放和相关的张力瞬变。相比之下,在TT去极化诱导的EC偶联过程中,GTPγS总是增强Ca2+释放。这些结果表明存在至少两种促进SR Ca2+释放的兴奋性G蛋白,其中只有一种是去膜纤维EC偶联的调节剂,尽管并非必需。