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首页> 美国卫生研究院文献>Blood >Hemostasis Thrombosis and Vascular Biology: Visualization of microtubule growth in living platelets reveals a dynamic marginal band with multiple microtubules
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Hemostasis Thrombosis and Vascular Biology: Visualization of microtubule growth in living platelets reveals a dynamic marginal band with multiple microtubules

机译：止血血栓形成和血管生物学：活血小板中微管生长的可视化揭示了具有多个微管的动态边缘带

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The marginal band of microtubules maintains the discoid shape of resting blood platelets. Although studies of platelet microtubule coil structure conclude that it is composed of a single microtubule, no investigations of its dynamics exist. In contrast to previous studies, permeabilized platelets incubated with GTP-rhodamine-tubulin revealed tubulin incorporation at 7.9 (± 1.9) points throughout the coil, and anti-EB1 antibodies stained 8.7 (± 2.0) sites, indicative of multiple free microtubules. To pursue this result, we expressed the microtubule plus-end marker EB3-GFP in megakaryocytes and examined its behavior in living platelets released from these cells. Time-lapse microscopy of EB3-GFP in resting platelets revealed multiple assembly sites within the coil and a bidirectional pattern of assembly. Consistent with these findings, tyrosinated tubulin, a marker of newly assembled microtubules, localized to resting platelet microtubule coils. These results suggest that the resting platelet marginal band contains multiple highly dynamic microtubules of mixed polarity. Analysis of microtubule coil diameters in newly formed resting platelets indicates that microtubule coil shrinkage occurs with aging. In addition, activated EB3-GFP–expressing platelets exhibited a dramatic increase in polymerizing microtubules, which travel outward and into filopodia. Thus, the dynamic microtubules associated with the marginal band likely function during both resting and activated platelet states.
机译：微管的边缘带保持静止血小板的盘状形状。尽管对血小板微管线圈结构的研究得出的结论是它由单个微管组成，但尚无关于其动力学的研究。与以前的研究相比，与GTP-若丹明-微管蛋白一起孵育的透化血小板显示，在整个线圈中微管蛋白的掺入量为7.9（±1.9）点，抗EB1抗体染色的位置为8.7（±2.0），表明存在多个游离微管。为了追求这一结果，我们在巨核细胞中表达了微管末端标记EB3-GFP，并检查了它们在从这些细胞释放的活血小板中的行为。静息血小板中的EB3-GFP延时显微镜显示了线圈内的多个组装位点和双向组装模式。与这些发现一致的是，酪氨酸微管蛋白（一种新组装的微管的标志物）位于静止的血小板微管线圈上。这些结果表明静止的血小板边缘带包含多个极性混合的高动态微管。对新形成的静息血小板中微管线圈直径的分析表明，微管线圈的收缩随着老化而发生。此外，活化的表达EB3-GFP的血小板在聚合微管中显着增加，这些微管向外传播并进入丝状伪足。因此，与边缘带相关的动态微管可能在静止状态和激活状态下均起作用。

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