Berlin J R
Bockus Research Institute, Graduate Hospital, Philadelphia, Pennsylvania 19146, USA.
Am J Physiol. 1995 Sep;269(3 Pt 2):H1165-70. doi: 10.1152/ajpheart.1995.269.3.H1165.
Spatial and temporal changes of intracellular calcium ion concentration ([Ca2+]i) during stimulated contractions were observed by confocal microscopy in rat ventricular and guinea pig atrial myocytes. Fluorescence intensity profiles in fluo 3-acetoxymethyl ester (fluo 3-AM)-loaded cells were collected from the entire cell, selected regions of the cell, or along a single scanned line across the cell. In rat ventricular myocytes, the increase of [Ca2+]i after a single stimulus from field electrodes occurred synchronously across the cell whether fluo 3 fluorescence was monitored in a narrow region aligned with the long axis of the cell or in line-scan images of a single z-line across the cell. However, during the onset of Ca2+ channel blockade by nifedipine (5 microM), electrical stimulation produced spatially nonuniform, focal increases of [Ca2+]i. In guinea pig atrial myocytes, stimulated increases of [Ca2+]i first appeared in focal regions at the cell periphery before spreading to the cell interior. Line-scan images showed the peripheral rise of [Ca2+]i led that at the center of the cell by 34 +/- 4 ms (mean +/- SE, n = 3). These data demonstrate that the t-tubular network ensures synchronous increases of [Ca2+]i throughout the cell during an action potential. In the absence of t tubules or when the number of sarcolemmal Ca2+ channels opened by membrane depolarization is greatly reduced, stimulated increases of [Ca2+]i can be observed to arise in focal regions of the cell.
通过共聚焦显微镜观察大鼠心室肌细胞和豚鼠心房肌细胞在刺激收缩过程中细胞内钙离子浓度([Ca2+]i)的时空变化。从加载了fluo 3-乙酰甲酯(fluo 3-AM)的细胞的整个细胞、选定的细胞区域或沿着细胞的单条扫描线收集荧光强度分布图。在大鼠心室肌细胞中,无论在与细胞长轴对齐的狭窄区域监测fluo 3荧光还是在穿过细胞的单条z线的线扫描图像中监测,来自场电极的单次刺激后[Ca2+]i的增加在整个细胞中同步发生。然而,在硝苯地平(5 microM)阻断Ca2+通道开始时,电刺激产生了[Ca2+]i的空间不均匀、局灶性增加。在豚鼠心房肌细胞中,刺激引起的[Ca2+]i增加首先出现在细胞周边的局灶区域,然后扩散到细胞内部。线扫描图像显示细胞周边[Ca2+]i的升高比细胞中心早34±4毫秒(平均值±标准误,n = 3)。这些数据表明,横管网络可确保在动作电位期间整个细胞内[Ca2+]i同步增加。在没有横管或当膜去极化打开的肌膜Ca2+通道数量大幅减少时,可观察到刺激引起的[Ca2+]i增加出现在细胞的局灶区域。