Krueger J W, Forletti D, Wittenberg B A
J Gen Physiol. 1980 Nov;76(5):587-607. doi: 10.1085/jgp.76.5.587.
We have observed the dynamics of sarcomere shortening and the diffracting action of single, functionally intact, unattached cardiac muscle cells enzymatically isolated from the ventricular tissue of adult rats. Sarcomere length was measured either (a) continuously by a light diffraction method or (b) by direct inspection of the cell's striated image as recorded on videotape or by cinemicroscopy (120--400 frames/s). At physiological levels of added CaCl2 (0.5--2.0 mM), many cells were quiescent (i.e., they did not beat spontaneously) and contracted in response to electrical stimulation (less than or equal to 1.0-ms pulse width). Sarcomere length in the quiescent, unstimulated cells (1.93 +/- 0.10 [SD] micrometers), at peak shortening (1.57 +/- 0.13 micrometers, n = 49), and the maximum velocity of sarcomere shortening and relengthening were comparable to previous observations in intact heart muscle preparations. The dispersion of light diffracted by the cell remained narrow, and individual striations remained distinct and laterally well registered throughout the shortening-relengthening cycle. In contrast, appreciable nonuniformity and internal buckling were seen at sarcomere lengths < 1.8 micrometers when the resting cell, embedded in gelatin, was longitudinally compressed These results indicate (a) that shortening and relengthening is characterized by uniform activation between myofibrils within the cardiac cell and (b) that physiologically significant relengthening forces in living heart muscle originate at the level of the cell rather than in extracellular connections. First-order diffracted light intensity, extremely variable during sarcomere shortening, was always greatest during midrelaxation preceding the onset of a very slow and uniform phase of sarcomere relengthening.
我们观察了从成年大鼠心室组织中酶解分离出的单个、功能完整、未附着的心肌细胞的肌节缩短动力学和衍射作用。肌节长度通过以下两种方法测量:(a) 采用光衍射法连续测量;(b) 直接观察录像带或电影显微镜(120 - 400帧/秒)记录的细胞横纹图像。在添加生理水平的氯化钙(0.5 - 2.0 mM)时,许多细胞处于静止状态(即它们不自发搏动),并在电刺激(脉冲宽度小于或等于1.0毫秒)下收缩。静止、未受刺激的细胞中的肌节长度(1.93±0.10[标准差]微米)、缩短峰值时的肌节长度(1.57±0.13微米,n = 49)以及肌节缩短和再延长的最大速度与之前在完整心肌制剂中的观察结果相当。细胞衍射光的色散保持狭窄,并且在整个缩短 - 再延长周期中,单个横纹保持清晰且横向对齐良好。相比之下,当嵌入明胶中的静止细胞受到纵向压缩时,在肌节长度<1.8微米时会出现明显的不均匀性和内部弯曲。这些结果表明:(a) 缩短和再延长的特征是心肌细胞内肌原纤维之间的均匀激活;(b) 活心肌中具有生理意义的再延长力起源于细胞水平而非细胞外连接。在肌节缩短过程中变化极大的一级衍射光强度,在肌节非常缓慢且均匀的再延长阶段开始前的中期松弛期间总是最大。