Dougherty T J, Driska S P
Physiology Department, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
Am J Physiol. 1991 Mar;260(3 Pt 1):C449-56. doi: 10.1152/ajpcell.1991.260.3.C449.
The relationship between unloaded shortening velocity and Ca2+ concentration was determined for hog carotid arterial smooth muscle cells, freshly isolated by digestion with papain. Cells were exposed to various [Ca2+] for 60 s at 37 degrees C and then stimulated with 10 microM histamine. Cell length was measured by a video analysis system. Shortening velocity was expressed as an exponential rate constant by fitting the cell lengths to the following equation: length = Lmin + (Lmax-Lmin)exp[-v (time-latency)], where Lmax is length before contraction, Lmin is shortest length reached, time is time elapsed after addition of agonist, latency is time from addition of agonist until contraction starts, and v is the exponential rate constant (s-1). Cells shortened substantially, usually reaching one-fourth to one-third of their initial length within 1 min. Shortening velocities of the cells were much faster than published values of maximum shortening velocity in muscle strips from this same tissue. At 1.6 mM Ca2+, v was 0.173 +/- 0.015 s-1. When Ca2+ was increased to 5 or 10 mM, v was not significantly different. However, when Ca2+ was decreased to 0.5 and 0.16 mM, v increased to 0.288 and 0.258 s-1, respectively. The difference between 0.5 and 1.6 mM was significant. The unexpected increase in shortening velocity at low Ca2+ was also seen when 10 mM caffeine was used as a stimulus: v at 1.6 mM Ca2+ was 0.156 s-1, whereas v at 0.16 mM Ca2+ was 0.272 s-1. The high shortening velocities we measured suggest that measurements made on multicellular tissues seriously underestimate the potential shortening velocity of isolated individual cells.
用木瓜蛋白酶消化新鲜分离的猪颈动脉平滑肌细胞,测定了无负荷缩短速度与Ca2+浓度之间的关系。将细胞在37℃下暴露于不同的[Ca2+]浓度60秒,然后用10微摩尔组胺刺激。通过视频分析系统测量细胞长度。缩短速度通过将细胞长度拟合到以下方程表示为指数速率常数:长度 = Lmin + (Lmax - Lmin)exp[-v(时间 - 延迟)],其中Lmax是收缩前的长度,Lmin是达到的最短长度,时间是添加激动剂后经过的时间,延迟是从添加激动剂到收缩开始的时间,v是指数速率常数(秒-1)。细胞大幅缩短,通常在1分钟内达到其初始长度的四分之一到三分之一。这些细胞的缩短速度比来自同一组织的肌肉条带中公布的最大缩短速度值快得多。在1.6毫摩尔Ca2+时,v为0.173±0.015秒-1。当Ca2+增加到5或10毫摩尔时,v没有显著差异。然而,当Ca2+降低到0.5和0.16毫摩尔时,v分别增加到0.288和0.258秒-1。0.5和1.6毫摩尔之间的差异是显著的。当使用10毫摩尔咖啡因作为刺激时,在低Ca2+时也观察到缩短速度意外增加:在1.6毫摩尔Ca2+时v为0.156秒-1,而在0.16毫摩尔Ca2+时v为0.272秒-1。我们测量的高缩短速度表明,在多细胞组织上进行的测量严重低估了分离的单个细胞的潜在缩短速度。