Csernoch Laszlo, Pouvreau Sandrine, Ronjat Michel, Jacquemond Vincent
Department of Physiology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary.
J Membr Biol. 2008 Nov-Dec;226(1-3):43-55. doi: 10.1007/s00232-008-9138-0. Epub 2008 Nov 18.
The elementary Ca(2+)-release events underlying voltage-activated myoplasmic Ca(2+) transients in mammalian muscle remain elusive. Here, we looked for such events in confocal line-scan (x,t) images of fluo-3 fluorescence taken from isolated adult mouse skeletal muscle fibers held under voltage-clamp conditions. In response to step depolarizations, spatially segregated fluorescence signals could be detected that were riding on a global increase in fluorescence. These discrete signals were separated using digital filtering in the spatial domain; mean values for their spatial half-width and amplitude were 1.99 +/- 0.09 microm and 0.16 +/- 0.005 DeltaF/F(0) (n = 151), respectively. Under control conditions, the duration of the events was limited by the pulse duration. In contrast, in the presence of maurocalcine, a scorpion toxin suspected to disrupt the process of repolarization-induced ryanodine receptor (RyR) closure, events uninterrupted by the end of the pulse were readily detected. Overall results establish these voltage-activated low-amplitude local Ca(2+) signals as inherent components of the physiological Ca(2+)-release process of mammalian muscle and suggest that they result from the opening of either one RyR or a coherently operating group of RyRs, under the control of the plasma membrane polarization.
哺乳动物肌肉中电压激活的肌质Ca(2+)瞬变所基于的基本Ca(2+)释放事件仍然难以捉摸。在这里,我们在共聚焦线扫描(x,t)图像中寻找此类事件,这些图像是从处于电压钳制条件下的分离成年小鼠骨骼肌纤维中采集的fluo-3荧光图像。响应于阶跃去极化,可以检测到空间上分离的荧光信号,这些信号叠加在荧光的整体增加之上。这些离散信号在空间域中使用数字滤波进行分离;其空间半高宽和幅度的平均值分别为1.99±0.09微米和0.16±0.005ΔF/F(0)(n = 151)。在对照条件下,事件的持续时间受脉冲持续时间限制。相比之下,在存在毛罗钙素的情况下,毛罗钙素是一种疑似破坏复极化诱导的兰尼碱受体(RyR)关闭过程的蝎毒素,很容易检测到在脉冲结束时未中断的事件。总体结果确定这些电压激活的低幅度局部Ca(2+)信号是哺乳动物肌肉生理Ca(2+)释放过程的固有组成部分,并表明它们是由单个RyR或一组协同运作的RyR在质膜极化的控制下开放产生的。