Linke W A, Bartoo M L, Pollack G H
Center for Bioengineering, University of Washington, Seattle 98195.
Circ Res. 1993 Oct;73(4):724-34. doi: 10.1161/01.res.73.4.724.
Spontaneous oscillations observed in various heart muscle preparations are widely thought to be triggered by spontaneous release of Ca2+ from the sarcoplasmic reticulum (SR). Here, we report undamped propagated oscillations that occur in the absence of SR. In single cardiac myofibrils treated with Triton X-100 to remove SR and held isometrically, partial activation initiated periodic fluctuations of sarcomere length persisting up to 1 hour. Oscillation characteristics could be readily quantitated by virtue of the small size of the preparation. In an individual sarcomere, the oscillation cycle generally consisted of a slow shortening phase, followed by a phase of rapid lengthening. Oscillations usually propagated along the myofibril--frequently along the entire specimen--in a wavelike fashion (average velocity, 12.3 microns/s at 10 degrees C; Q10, approximately 1.3). The oscillation period was 2.30 and 1.72 seconds at 10 degrees and 20 degrees C, respectively, and was insensitive to stretch. The average oscillation amplitude, which was temperature independent, decreased with stretch from more than 20% of the mean sarcomere length at lengths below 2 microns to zero beyond a sarcomere length of 3 microns. Stiffening of the Z line by labeling with anti-alpha-actinin resulted in a dose-dependent decrease of oscillation amplitude, while the period was not affected. Tension oscillations could not be detected in single myofibrils but were frequently detectable in myofibril doublets, where the oscillation magnitude (approximately 1 microgram) was above the noise floor. Addition of 10 mumol/L ryanodine to the activating solution did not alter oscillation characteristics, as expected, since the oscillations are unrelated to SR calcium release. On the basis of our results, we consider a mechanism for the oscillations in which a length dependence of myofibrillar Ca2+ sensitivity and a dynamic Z-line structure are essential.
在各种心肌制备物中观察到的自发振荡,普遍认为是由肌浆网(SR)自发释放Ca2+引发的。在此,我们报告了在没有SR的情况下发生的无阻尼传播振荡。在用Triton X-100处理以去除SR并等长固定的单个心肌肌原纤维中,部分激活引发了肌节长度的周期性波动,持续长达1小时。由于制备物尺寸小,振荡特征很容易被量化。在单个肌节中,振荡周期通常包括一个缓慢缩短阶段,随后是一个快速延长阶段。振荡通常以波状方式沿着肌原纤维传播——通常沿着整个标本——(在10℃时平均速度为12.3微米/秒;Q10,约为1.3)。在10℃和20℃时,振荡周期分别为2.30秒和1.72秒,并且对拉伸不敏感。平均振荡幅度与温度无关,在肌节长度低于2微米时,从超过平均肌节长度的20%随着拉伸而减小,在肌节长度超过3微米时降至零。用抗α-辅肌动蛋白标记使Z线变硬,导致振荡幅度呈剂量依赖性降低,而周期不受影响。在单个肌原纤维中未检测到张力振荡,但在双肌原纤维中经常可检测到,其中振荡幅度(约1微克)高于本底噪声。正如预期的那样,向激活溶液中添加10微摩尔/升的兰尼碱不会改变振荡特征,因为这些振荡与SR钙释放无关。基于我们的结果,我们考虑了一种振荡机制,其中肌原纤维Ca2+敏感性的长度依赖性和动态Z线结构是必不可少的。