Saeki Y, Kurihara S, Hongo K, Tanaka E
Department of Physiology, Tsurumi University School of Dental Medicine, Yokohama, Japan.
Adv Exp Med Biol. 1993;332:639-47; discussion 647-8. doi: 10.1007/978-1-4615-2872-2_57.
To elucidate the effects of mechanical constraints on the (Ca2+) affinity of cardiac troponin C, we studied the relationships among the myoplasmic Ca2+ concentration ([Ca2+]i), tension and length in steadily activated intact cardiac muscle. The Ca2+ sensitive photoprotein, aequorin, was micro-injected into cells of ferret right ventricular papillary muscles to monitor the [Ca2+]i. The muscle was then steadily activated with ouabain (10(-4) M)(ouabain contracture) or high frequency stimuli in the presence of ryanodine (5 microM)(tetanic contraction); the tension and aequorin light (AL) transients in response to a step length change were then analyzed. The tension transient response to either the stretch or release in length was oscillatory: tension decreased rapidly during the release and then increased, after which it lapsed into a new steady level in a series of damped oscillations. The opposite was true for the stretch. The oscillatory responses were conspicuous and less damped in the ouabain contracture. The transient AL response was also oscillatory, the time course of which corresponded exactly to that of the tension transient response, though no detectable changes in AL were observed at the initial phase of the stretch response. The increase in AL corresponded exactly to the decrease in tension, likewise the decrease in AL to the increase in tension. The steady level of AL after release was decreased in ouabain contracture, but was increased in tetanic contraction. These results suggest that the Ca2+ affinity of cardiac troponin C is increased with an increase in tension (i.e., the cross-bridge attachment) and decreased with a decrease in tension (i.e., the cross-bridge detachment), and that the myoplasmic calcium concentration is lowered by release, at least in a Ca(2+)-overloaded condition, mainly through the sarcoplasmic reticulum.
为阐明机械性限制对心肌肌钙蛋白C(Ca2+)亲和力的影响,我们研究了稳定激活的完整心肌中肌浆Ca2+浓度([Ca2+]i)、张力和长度之间的关系。将Ca2+敏感光蛋白水母发光蛋白微量注射到雪貂右心室乳头肌细胞中,以监测[Ca2+]i。然后用哇巴因(10(-4)M)(哇巴因挛缩)或在存在雷诺丁(5 microM)的情况下用高频刺激(强直收缩)稳定激活肌肉;随后分析对步长变化的张力和水母发光蛋白光(AL)瞬变。长度拉伸或释放时的张力瞬变响应是振荡性的:释放过程中张力迅速下降,然后上升,之后在一系列衰减振荡中进入新的稳定水平。拉伸时情况相反。在哇巴因挛缩中,振荡响应明显且衰减较小。瞬态AL响应也是振荡性的,其时间进程与张力瞬变响应完全对应,尽管在拉伸响应的初始阶段未观察到AL有可检测到的变化。AL的增加与张力的降低完全对应,同样,AL的降低与张力的增加对应。释放后AL的稳定水平在哇巴因挛缩中降低,但在强直收缩中升高。这些结果表明,心肌肌钙蛋白C的Ca2+亲和力随张力增加(即横桥附着)而增加,随张力降低(即横桥脱离)而降低,并且至少在Ca(2+)过载状态下,肌浆钙浓度通过释放而降低,主要通过肌浆网。