SPREADING OF HUMAN-NEUTROPHILS IS IMMEDIATELY PRECEDED BY A LARGE INCREASE IN CYTOPLASMIC FREE CALCIUM
SPREADING OF HUMAN-NEUTROPHILS IS IMMEDIATELY PRECEDED BY A LARGE INCREASE IN CYTOPLASMIC FREE CALCIUM
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DOI：
10.1073/pnas.83.9.2919
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发表时间：
1986-05-01
影响因子：
11.1
通讯作者：
MAXFIELD, FR
中科院分区：
文献类型：
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作者：
KRUSKAL, BA;SHAK, S;MAXFIELD, FR
当人类多形核白细胞（PMN）被放置在不同的表面时，它们附着并迅速扩散，其直径增加数倍。扩散与细胞骨架的广泛改变有关。由于许多细胞骨架事件是由Ca2+调节的，我们测量了个体PMN扩散时的胞质游离钙浓度（[Ca2+]i）。[Ca2+]i是用荧光Ca2+敏感染料fura-2在单细胞中测定的。在扩散之前，PMN的[Ca2+]i从69 .+-迅速增加。51 nM到547 +-。190nm(平均+-。SD, n = 12)。在接下来的一分钟内，随着细胞的扩散，[Ca2+]i恢复到接近静止水平。细胞外钙的清除、维拉帕米、钙调素拮抗剂或线粒体或微管毒物都不能阻断扩散和[Ca2+]i尖峰。扩散，但不是[Ca2+]i增加，被微丝抑制剂细胞松弛素b阻止。扩散和[Ca2+]i尖峰都被ATP耗尽阻止，并通过将细胞置于含有高渗蔗糖或氯化钠的培养基中可逆地阻止。这些数据强烈表明，来自非线粒体胞内池的[Ca2+]i的增加在微丝介导的PMN扩散过程中起重要作用。
When human polymorphonuclear leukocytes (PMN) are placed on various surfaces, they attach and spread rapidly, increasing their diameter severalfold. The spreading is associated with extensive changes in the cytoskeleton. Since many cytoskeletals events are regulated by Ca2+, we measured the cytosolic free calcium concentration ([Ca2+]i) in individual human PMN as they spread. [Ca2+]i was measured in single cells by microspectrofluorometry using the fluorescent Ca2+-sensitive dye fura-2. Immediately before spreading, PMN exhibit a rapid increase in [Ca2+]i from 69 .+-. 51 nM to 547 .+-. 190 nM (Mean .+-. SD, n = 12). [Ca2+]i returns to near resting levels during the next minute, as the cells spread. Neither the spreading nor the [Ca2+]i spike is blocked by removal of extracellular calcium, by verapamil, by calmodulin antagonists, or by mitochondrial or microtubule poisons. Spreading, but not the [Ca2+]i increase, is blocked by the microfilament inhibitor cytochalasin B. Both spreading and the [Ca2+]i spike are blocked by ATP depletion and reversibly blocked by placing the cells in medium containing hypertonic sucrose or sodium chloride. These data strongly suggest that an increase in [Ca2+]i derived from nonmitchondrial intracellular pools, plays an important role in the microfilament-mediated process of PMN spreading.