Transient increases in cytosolic free calcium appear to be required for the migration of adherent human neutrophils.
Transient increases in cytosolic free calcium appear to be required for the migration of adherent human neutrophils.
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DOI：
10.1083/jcb.110.1.43
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发表时间：
1990-01
期刊：
影响因子：
--
通讯作者：
Maxfield FR
中科院分区：
文献类型：
--
作者：
Marks PW;Maxfield FR
人中性粒细胞自发和对趋化剂N-甲酰基-L-甲硫基-L-亮氨基-L-苯丙氨酸(Jaconi，M.E.，R.W.Rivest，W.Schlegel，C.B.Wollheim，D.Pittet和P.D.Lew)的响应，表现出胞浆游离钙浓度的多次增加。1988年。J.Biol.化学。263：10557-10560)。这些重复性的[Ca~(2+)]i升高的作用，以及Ca~(2+)在人中性粒细胞迁移中的作用，仍未得到解决。我们使用微量荧光分光光度法测量了单个Fura-2负载的人中性粒细胞在有血清存在的情况下在多聚D-赖氨酸涂层玻璃上移动时的[Ca~(2+)]i。为了研究钙离子在人中性粒细胞迁移中的作用，我们研究了细胞外钙离子存在和不存在的细胞，以及细胞内钙(+)缓冲和钙(+)耗尽的细胞。在细胞外Ca~(2+)存在的情况下，细胞内[Ca~(2+)]i经常出现多次升高和降低，在细胞的趋化、趋化和吞噬过程中，每个运动细胞都至少发生一次这种短暂的[Ca~(2+)]_i升高。此外，即使在没有化学诱导剂的情况下，中性粒细胞在电镀到表面后延伸伪足并呈现极化形态的中性粒细胞总是表现出[Ca+]i瞬变。相反，只有一个未极化的静止细胞(n=15)观察到[Ca~(2+)]i瞬变。虽然一些细胞表现出相对周期性的[Ca~(2+)]i的上升和下降，类似于在某些细胞类型中观察到的有规律的振荡，但许多其他细胞的[Ca~(2+)]i的上升和下降在时间、幅度和持续时间上有所不同。缓冲[Ca~(2+)]i或清除细胞外Ca~(2+)可抑制或阻断[Ca~(2+)]i的一过性升高，减少或抑制中性粒细胞的迁移。在这种情况下，极化的细胞经常被观察到反复尝试迁移，但它们仍然固定在它们的后部。这些数据表明，在血清存在的情况下，人中性粒细胞在多聚D-赖氨酸涂层玻璃上的迁移可能需要短暂的[Ca~(2+)]i增加，因为它允许中性粒细胞从先前的附着位置释放出来。
Human neutrophils exhibit multiple increases in cytosolic free calcium concentration [( Ca2+]i) spontaneously and in response to the chemoattractant N-formyl-L-methionyl-L-leucyl-L-phenylalanine (Jaconi, M. E. E., R. W. Rivest, W. Schlegel, C. B. Wollheim, D. Pittet, and P. D. Lew. 1988. J. Biol. Chem. 263:10557-10560). The function of these repetitive increases in [Ca2+]i, as well as the role of Ca2+ in human neutrophil migration, remain unresolved. We have used microspectrofluorometry to measure [Ca2+]i in single fura-2-loaded human neutrophils as they moved on poly-D-lysine-coated glass in the presence of serum. To investigate the role of Ca2+ in human neutrophil migration, we examined cells in the presence and absence of extracellular Ca2+, as well as intracellular Ca2(+)-buffered and Ca2(+)- depleted cells. In the presence of extracellular Ca2+, multiple increases and decreases in [Ca2+]i were frequently observed, and at least one such transient increase in [Ca2+]i occurred in every moving cell during chemokinesis, chemotaxis, and phagocytosis. In addition, neutrophils that extended pseudopodia and assumed a polarized morphology after plating onto a surface were always observed to exhibit [Ca2+]i transients even in the absence of chemoattractant. In contrast, a [Ca2+]i transient was observed in only one of the nonpolarized stationary cells that were examined (n = 15). Although some cells exhibited relatively periodic increases and decreases in [Ca2+]i, resembling the regular oscillations that have been observed in some cell types, many others exhibited increases and decreases in [Ca2+]i that varied in their timing, magnitude, and duration. Buffering of [Ca2+]i or removal of extracellular Ca2+ damped out or blocked transient increases in [Ca2+]i and reduced or inhibited the migration of neutrophils. Under these conditions, polarized cells were often observed to make repeated attempts at migration, but they remained anchored at their rear. These data suggest that transient increases in [Ca2+]i may be required for the migration of human neutrophils on poly- D-lysine-coated glass in the presence of serum by allowing them to release from previous sites of attachment.