Vesicle accumulation and exocytosis at sites of plasma membrane disruption.
Vesicle accumulation and exocytosis at sites of plasma membrane disruption.
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在质膜破坏部位的囊泡积累和胞吐作用。
DOI：
10.1083/jcb.131.6.1737
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发表时间：
1995-12
影响因子：
7.8
通讯作者：
McNeil, PL
中科院分区：
文献类型：
--
作者：
Miyake, K;McNeil, PL
质膜的破坏是通过一种活性的分子机制重新密封的，被认为部分由激动素、CaM激酶、Snap-25和Synaptobrevin组成。我们用HRP标记了机械诱导的质膜破裂的细胞质部位。透射电子显微镜显示，大小不一的小泡在破裂部位周围的细胞质中大量聚集(S)，微绒毛样表面突起覆盖在该区域。扫描电子显微镜证实，损伤细胞上迅速出现一簇簇微绒毛。使用膜特异性染料FM1-43的三种检测方法提供了定量证据，证明干扰诱导钙依赖的胞吐作用涉及一个或多个内体/溶酶体隔室。共聚焦显微镜显示损伤细胞的皮质区域存在明显的FM染料荧光耗竭，这表明局部胞吐是由创伤而不是整体胞吐诱导的。最后，流式细胞术记录到损伤后细胞前向散射增加，表明损伤后细胞大小增加。这些结果首次直接支持了这样的假设，即一个或多个内膜室聚集在破裂部位并在那里与质膜融合，导致膜局部增加到机械损伤细胞的表面。
Plasma membrane disruptions are resealed by an active molecular mechanism thought to be composed, in part, of kinesin, CaM kinase, snap- 25, and synaptobrevin. We have used HRP to mark the cytoplasmic site of a mechanically induced plasma membrane disruption. Transmission electron microscopy revealed that vesicles of a variety of sizes rapidly (s) accumulate in large numbers within the cytoplasm surrounding the disruption site and that microvilli-like surface projections overlie this region. Scanning electron microscopy confirmed that tufts of microvilli rapidly appear on wounded cells. Three assays, employing the membrane specific dye FM1-43, provide quantitative evidence that disruption induces Ca(2+)-dependent exocytosis involving one or more of the endosomal/lysosomal compartments. Confocal microscopy revealed the presence in wounded cells of cortical domains that were strikingly depleted of FM dye fluorescence, suggesting that a local bolus of exocytosis is induced by wounding rather than global exocytosis. Finally, flow cytometry recorded a disruption-induced increase in cell forward scatter, suggesting that cell size increases after injury. These results provide the first direct support for the hypothesis that one or more internal membrane compartments accumulate at the disruption site and fuse there with the plasma membrane, resulting in the local addition of membrane to the surface of the mechanically wounded cell.
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DOI：
10.1016/0005-2736(90)90135-b
发表时间：
1990-04-30
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
FISHMAN, HM;TEWARI, KP;STEIN, PG
通讯作者：
STEIN, PG
影响因子：
7.8
作者：
MILLER, TM;HEUSER, JE
通讯作者：
HEUSER, JE
DOI：
10.1083/jcb.128.4.589
发表时间：
1995-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Muallem S;Kwiatkowska K;Xu X;Yin HL
通讯作者：
Yin HL
影响因子：
7.8
作者：
EDDY, EM;SHAPIRO, BM
通讯作者：
SHAPIRO, BM
影响因子：
2.4
作者：
JOHNSON, RM
通讯作者：
JOHNSON, RM