Centrosome Behavior in Motile HGF-Treated PtK2 Cells Expressing GFP-Gamma Tubulin
Centrosome Behavior in Motile HGF-Treated PtK2 Cells Expressing GFP-Gamma Tubulin
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DOI：
10.1002/cm.1041
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发表时间：
2001-10-01
影响因子：
--
通讯作者：
Wadsworth, P
中科院分区：
文献类型：
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作者：
Danowski, BA;Khodjakov, A;Wadsworth, P
为了响应运动线索，许多运动细胞已被证明将其中心体重新定位到细胞核前方的位置，朝向细胞迁移的方向。我们研究了中心体的位置在PtK 2上皮细胞与肝细胞生长因子（HGF）处理。其刺激运动性，但不像趋化剂或单层的创伤，其不提供方向线索。直接观察中心体运动。编码与绿色荧光蛋白融合的人γ微管蛋白的质粒在HGF处理的细胞中表达。在运动不受邻近细胞约束的细胞中，我们发现中心体的位置与细胞运动的方向不相关。此外，在观察期间运动方向改变的细胞中，中心体没有重新定向到新的运动方向。对中心体和细胞核运动的分析表明，中心体的运动往往滞后于细胞核的运动。用γ微管蛋白抗体染色的249个固定细胞的分析证实了我们在活细胞中的观察结果：69%的细胞在细胞核后面有中心体，远离运动方向。其中，41%的中心体位于回缩尾。共聚焦显微镜观察显示HGF处理的PtK 2细胞中的微管阵列主要是非中心体的。由于微管是需要有效的细胞运动，我们建议，非中心体微管稳定的运动方向，而不需要重新定位的中心体。(C)2001 Wiley-Liss，Inc.
In response to locomotory cues, many motile cells have been shown to reposition their centrosome to a location in front of the nucleus, towards the direction of cell migration. We examined centrosome position in PtK2 epithelial cells treated with hepatocyte growth factor (HGF). which stimulates motility but, unlike chemotactic agents or wounding of a monolayer, provides no directional cues. To observe centrosome movement directly. a plasmid encoding human gamma tubulin fused to the green fluorescent protein was expressed in HGF-treated cells. In cells whose movements were unconstrained by neighboring cells, we found that the position of the centrosome was not correlated with the direction of cell locomotion. Further, in cells where the direction of locomotion changed during the observation period, the centrosome did not reorient toward the new direction of locomotion. Analysis of centrosome and nuclear movement showed that motion of the centrosome often lagged behind that of the nucleus. Analysis of 249 fixed cells stained with an antibody to gamma tubulin confirmed our observations in live cells: 69% of the cells had centrosomes behind the nucleus, away from the direction of locomotion. Of these, 41% had their centrosome in the retraction tail. Confocal microscopy showed that the microtubule array in HGF treated PtK2 cells was predominantly non-centrosomal. Because microtubules are required for efficient cellular locomotion, we propose that non-centrosomal microtubules stabilize the direction of locomotion without a requirement for reorientation of the centrosome. (C) 2001 Wiley-Liss, Inc.