首页> 外文期刊>Cell motility and the cytoskeleton >Detection of the centriole tyr- or acet-tubulin changes in endothelial cells treated with thrombin using microscopic immunocytochemistry.
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Detection of the centriole tyr- or acet-tubulin changes in endothelial cells treated with thrombin using microscopic immunocytochemistry.

机译:使用显微镜免疫细胞化学技术检测凝血酶处理后的内皮细胞中中心酪氨酸或乙酰微管蛋白的变化。

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We used electron microscopic immunocytochemistry to examine the pattern of centriolar staining for tyrosinated or acetylated alpha-tubulin in endothelial cells during short-term incubation with thrombin. Endothelial cells isolated from human aorta (HAEC) and those isolated from umbilical vein (HUVEC) displayed an increase in the intensity of centriolar staining for acet-tubulin within 1 min after thrombin addition. A decrease in the intensity of centriolar staining for tyr-tubulin was detected in HUVEC within 1 min after thrombin addition, while in HAEC centriolar staining for tyr-tubulin became less intense only 5 min later. Mother and daughter centrioles of HUVEC cells displayed different intensity of immunostaining for acet-tubulin and showed no significant variation in the number of subdistal appendages after thrombin addition. Differently, HAEC cells had the same staining pattern of mother and daughter centrioles in both thrombin-untreated and thrombin-treated cultures. A sharp increase in the number of subdistal appendages of mother centriole occurred in HAEC within 5 min of incubation with thrombin. Our findings provided the direct evidence for centrosome involvement in the ligand-mediated signaling events and showed for the first time that ligand-dependent centrosome reorganization includes the centriole per se. Furthermore, based on our observations we would like to propose that MT-nucleating/anchoring properties of the centrosome are subject to rapid regulation by external signals such as thrombin. Cell Motil. Cytoskeleton 62:1-12, 2005. (c) 2005 Wiley-Liss, Inc.
机译:我们使用电子显微镜免疫细胞化学检查了凝血酶短期孵育过程中内皮细胞中酪氨酸或乙酰化的α-微管蛋白的中心粒染色模式。从人主动脉(HAEC)和从脐静脉(HUVEC)分离的内皮细胞在添加凝血酶后1分钟内显示出乙酰微管蛋白的中心粒染色强度增加。在添加凝血酶后1分钟内,HUVEC中检测到的酪氨酸微管的中心丝染色强度降低,而在HAEC中,仅5分钟后,对酪蛋白的中心微丝染色强度降低。 HUVEC细胞的母子中心和子中心显示出不同的乙酰微管蛋白免疫染色强度,并且在添加凝血酶后,近端附肢的数量没有明显变化。不同的是,在未经凝血酶处理和经凝血酶处理的培养物中,HAEC细胞的母子中心和子中心均具有相同的染色模式。在与凝血酶温育的5分钟内,HAEC中母体次近距离附件的数量急剧增加。我们的发现提供了中心体参与配体介导的信号转导事件的直接证据,并首次显示了依赖配体的中心体重组包括中心体本身。此外,根据我们的观察,我们想提出中心体的MT成核/锚定特性会受到诸如凝血酶之类的外部信号的快速调节。细胞动力。 Cytoskeleton 62:1-12,2005.(c)2005 Wiley-Liss,Inc.

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