Kimura I, Kimura M, Kimura M
Br J Pharmacol. 1985 Oct;86(2):319-26. doi: 10.1111/j.1476-5381.1985.tb08899.x.
A convenient method is described for measuring simultaneously Ca2+-related aequorin luminescence and twitch tension in the isolated diaphragm muscle of the mouse. Forty to fifty fibres were injected intracellularly with aequorin solution and the mechanical and luminescence responses to direct stimulation were recorded. The replacement of Na+ by K+ (to obtain 59 or 143.4 mM K+) in the nutrient solution decreased both aequorin luminescence and twitch tensions, but after a time lag, it produced a contracture. Caffeine (5 or 10 mM) increased both aequorin luminescence and twitch tensions, and after a time lag, it also produced a contracture. Dantrolene (1 and 30 microM) and procaine (10 microM, 300 microM and 1 mM) decreased aequorin luminescence transients and twitch tension. In addition procaine inhibited the caffeine-induced increase of aequorin luminescence, but dantrolene did not have this effect. At concentrations causing neuromuscular block, suxamethonium (130 microM) decreased aequorin luminescence transients and twitch tension. By contrast, (+)-tubocurarine (6.5 microM) did not affect the aequorin luminescence in directly stimulated muscles. These results suggest that Ca+-related aequorin luminescence transients accompanied by twitch tensions reflect the intracellular fast mobilization of compartmentalized Ca2+ from plasma membrane or sarcoplasmic reticulum, and that the increase in resting luminescence caused by a K+- or caffeine-induced contracture may be produced by the slow mobilization of Ca2+ from sarcoplasmic reticulum.
本文描述了一种便捷的方法,用于同时测量小鼠离体膈肌中与Ca2+相关的水母发光蛋白发光和抽搐张力。将40至50根肌纤维细胞内注射水母发光蛋白溶液,并记录对直接刺激的机械和发光反应。在营养液中用K+替代Na+(使K+浓度达到59或143.4 mM)会降低水母发光蛋白发光和抽搐张力,但经过一段时间延迟后,会产生挛缩。咖啡因(5或10 mM)会增加水母发光蛋白发光和抽搐张力,经过一段时间延迟后,也会产生挛缩。丹曲林(1和30 microM)和普鲁卡因(10 microM、300 microM和1 mM)会降低水母发光蛋白发光瞬变和抽搐张力。此外,普鲁卡因会抑制咖啡因诱导的水母发光蛋白发光增加,但丹曲林没有这种作用。在引起神经肌肉阻滞的浓度下,琥珀酰胆碱(130 microM)会降低水母发光蛋白发光瞬变和抽搐张力。相比之下,(+)-筒箭毒碱(6.5 microM)对直接刺激的肌肉中的水母发光蛋白发光没有影响。这些结果表明,伴有抽搐张力的与Ca+相关的水母发光蛋白发光瞬变反映了细胞膜或肌浆网中分隔的Ca2+的细胞内快速动员,而K+或咖啡因诱导的挛缩引起 的静息发光增加可能是由肌浆网中Ca2+的缓慢动员产生的。