Ishiwata S, Okamura N
Department of Physics, School of Science and Engineering, Waseda University, Tokyo, Japan.
Biophys J. 1989 Dec;56(6):1113-20. doi: 10.1016/S0006-3495(89)82759-6.
Diffraction rings corresponding to the first, second, and third order were obtained by laser light illumination from a suspension of rabbit glycerinated psoas myofibrils (diameter, 1-2 microns; average length of the straight region, 44 microns; average sarcomere length, 2.2-2.6 microns) of which the optical thickness was appropriately chosen. Dispersed myofibrils were nearly randomly oriented in two dimensions, so that the effects of muscle volume were minimized; these effects usually interfere significantly with a quantitative analysis of laser optical diffraction in the fiber system. The diameters of diffraction rings represented the average sarcomere length. By using this system, we confirmed the ability of the unit cell (sarcomere) structure model to explain the intensity change of diffraction lines accompanying the dissociation from both ends of thick filaments in a high salt solution. The length of an A-band estimated from the relative intensity of diffraction rings and that directly measured on phase-contrast micrographs coincided well with each other. Also, we found that myofibrils with a long sarcomere length shorten to a slack length accompanying the decrease in overlap between thick and thin filaments produced by the dissociation of thick filaments.
通过用激光照射兔甘油化腰大肌肌原纤维(直径1 - 2微米;直区平均长度44微米;平均肌节长度2.2 - 2.6微米)的悬浮液,获得了对应于一阶、二阶和三阶的衍射环,其光学厚度经过适当选择。分散的肌原纤维在二维中几乎随机取向,从而使肌肉体积的影响最小化;这些影响通常会严重干扰纤维系统中激光光学衍射的定量分析。衍射环的直径代表平均肌节长度。通过使用该系统,我们证实了单位细胞(肌节)结构模型能够解释在高盐溶液中伴随着粗肌丝两端解离时衍射线强度的变化。根据衍射环的相对强度估计的A带长度与在相差显微镜照片上直接测量的长度非常吻合。此外,我们发现,随着粗肌丝解离导致粗细肌丝重叠减少,肌节长度较长的肌原纤维会缩短至松弛长度。