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以高时间分辨率测量青蛙骨骼肌纤维中的细胞内钙离子瞬变和张力。

The intracellular Ca2+ transient and tension in frog skeletal muscle fibres measured with high temporal resolution.

作者信息

Claflin D R, Morgan D L, Stephenson D G, Julian F J

机构信息

Department of Anesthesia Research Laboratories, Harvard Medical School, Brigham and Women's Hospital, Boston, MA 02115-6195.

出版信息

J Physiol. 1994 Mar 1;475(2):319-25. doi: 10.1113/jphysiol.1994.sp020072.

Abstract
  1. The purpose of this study was to determine, with high temporal resolution, the relationship between the intracellular Ca2+ transient (ICT) and the mechanical responses of intact, single skeletal muscle fibres of frogs following stimulation by a single, brief depolarization. 2. The time course of the ICT was monitored using the Ca(2+)-sensitive fluorescent dyes mag-fura-2 (furaptra) and mag-fura-5. The mag-fura dyes have a low affinity for Ca2+ and have been shown to track the ICT with no appreciable kinetic delay. Continuous records of mag-fura fluorescence, tension and stiffness responses were obtained simultaneously at high time resolution at a sarcomere length of 2.9 microns. Experimental temperature was 3 degrees C. 3. When a delay of 0.4 ms due to the low-pass filter associated with the photodetector was included, the onset of the fluorescence response preceded the onset of latency relaxation (the small fall in tension that precedes positive tension generation) by 3.1 +/- 0.2 ms (mean +/- S.E.M., n = 8). After its onset, the mag-fura fluorescence signal continued to change rapidly (indicating increasing intracellular [Ca2+]) to an extreme level that occurred 1.5 +/- 0.5 ms (mean +/- S.E.M., n = 7) before tension had recovered to its resting level following latency relaxation. The time delay from the extreme of the fluorescence signal to the peak of the tension signal was 239 +/- 27 ms (mean +/- S.E.M., n = 6).(ABSTRACT TRUNCATED AT 250 WORDS)
摘要
  1. 本研究的目的是在高时间分辨率下,确定蛙完整单根骨骼肌纤维在单次短暂去极化刺激后,细胞内钙离子瞬变(ICT)与机械反应之间的关系。2. 使用钙离子敏感荧光染料mag-fura-2(furaptra)和mag-fura-5监测ICT的时间进程。mag-fura染料对钙离子亲和力低,已证明可追踪ICT且无明显动力学延迟。在2.9微米的肌节长度下,以高时间分辨率同时获得mag-fura荧光、张力和刚度反应的连续记录。实验温度为3摄氏度。3. 当计入与光电探测器相关的低通滤波器导致的0.4毫秒延迟时,荧光反应的起始比潜伏性松弛(正张力产生之前的小张力下降)的起始提前3.1±0.2毫秒(平均值±标准误,n = 8)。起始后,mag-fura荧光信号继续快速变化(表明细胞内[Ca2+]增加),直至达到一个极值水平,该极值水平在潜伏性松弛后张力恢复到静息水平之前1.5±0.5毫秒(平均值±标准误,n = 7)出现。从荧光信号极值到张力信号峰值的时间延迟为239±27毫秒(平均值±标准误,n = 6)。(摘要截于250字)

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Fura-2 calcium transients in frog skeletal muscle fibres.蛙骨骼肌纤维中的fura-2钙瞬变
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Fura-2 calcium transients in frog skeletal muscle fibres.蛙骨骼肌纤维中的fura-2钙瞬变
J Physiol. 1988 Sep;403:151-92. doi: 10.1113/jphysiol.1988.sp017244.
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A fluorescent indicator for measuring cytosolic free magnesium.一种用于测量胞质游离镁的荧光指示剂。
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