Saeki Y, Kawai M, Zhao Y
Department of Physiology, Tsurumi University School of Dental Medicine, Yokohama, Japan.
Circ Res. 1991 Mar;68(3):772-81. doi: 10.1161/01.res.68.3.772.
This study compares the crossbridge kinetics of intact and skinned preparations from ferret cardiac muscles at 20 degrees C to determine whether skinning causes any alteration in the crossbridge response to an imposed length change. A papillary or trabecular muscle was isolated from the right ventricle, the muscle length adjusted to give the maximum twitch tension (Lmax), and the preparation was subjected to Ba2+ contracture. When steady tension developed, the length of the preparation was perturbed sinusoidally in 19 discrete frequencies, ranging from 0.13 to 135 Hz, and at a small peak-to-peak amplitude (0.25% Lmax). We identified three exponential processes in the sinusodial force-response to the imposed length oscillation, and these were labeled processes B, C, and D in order of increasing speed. A slow process, A, normally present in fast-twitch skeletal muscles, is very small or absent in cardiac muscles. Process B is an exponential delay, and the muscle produces oscillatory work on the forcing apparatus; processes C and D are exponential advances in which the muscle absorbs work. The preparation was chemically skinned and activated in the presence of (mM) CaEGTA 6 (pCa 4.55), MgATP 5, magnesium propionate 1, and phosphate 1, pH 7.0, with ionic strength adjusted to 200 mM with potassium propionate. We found that the crossbridge kinetics were not altered by the skinning procedure. The apparent rate constants extracted from the sinusoidal analysis were nearly identical in Ba2+ contracture (intact preparation) and in Ca2+ activation (skinned preparation), and the Nyquist plots were similar. Because the rate constants changed sensitively with the substrate (MgATP) concentrations, we concluded that the substrate is adequately supplied during Ba2+ contracture in the intact preparation. Our study demonstrates the compatibility of results obtained from an intact and from a skinned preparation.
本研究比较了雪貂心肌完整和去皮标本在20摄氏度时的横桥动力学,以确定去皮是否会使横桥对施加的长度变化的反应发生任何改变。从右心室分离出乳头肌或小梁肌,调整肌肉长度以产生最大抽搐张力(Lmax),然后使标本经历Ba2+挛缩。当达到稳定张力时,以19个离散频率(范围从0.13至135 Hz)对标本长度进行正弦扰动,峰峰值幅度较小(0.25%Lmax)。我们在对施加的长度振荡的正弦力响应中识别出三个指数过程,这些过程按速度增加的顺序标记为过程B、C和D。一个通常存在于快肌骨骼肌中的缓慢过程A,在心肌中非常小或不存在。过程B是指数延迟,肌肉在施力装置上产生振荡功;过程C和D是指数提前,肌肉在其中吸收功。标本经化学去皮,并在含有(mM)CaEGTA 6(pCa 4.55)、MgATP 5、丙酸镁1和磷酸盐1、pH 7.0的条件下激活,用丙酸钾将离子强度调节至200 mM。我们发现去皮过程并未改变横桥动力学。从正弦分析中提取的表观速率常数在Ba2+挛缩(完整标本)和Ca2+激活(去皮标本)中几乎相同,并且奈奎斯特图相似。由于速率常数随底物(MgATP)浓度敏感变化,我们得出结论,在完整标本的Ba2+挛缩过程中底物得到了充分供应。我们的研究证明了从完整标本和去皮标本获得的结果具有兼容性。