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Non-centrosomal MTs play a crucial role in organization of MT array in interphase fibroblasts

机译：非中心体MT在间期成纤维细胞中MT阵列的组织中起关键作用

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Microtubules in interphase fibroblast-like cells are thought to be organized in a radial array growing from a centrosome-based microtubule-organizing center (MTOC) to the cell edges. However, many morphogenetic processes require the asymmetry of the microtubules (MT) array. One of the possible mechanisms of this asymmetry could be the presence of non-centrosomal microtubules in different intracellular areas. To evaluate the role of centrosome-born and non-centrosomal microtubules in the organization of microtubule array in motile 3T3 fibroblasts, we have performed the high-throughput analysis of microtubule growth in different functional zones of the cell and distinguished three subpopulations of growing microtubules (centrosome-born, marginal and inner cytoplasmic). Centrosome as an active microtubule-organizing center was absent in half of the cell population. However, these cells do not show any difference in microtubule growth pattern. In cells with active centrosome, it was constantly forming short (ephemeral) MTs, and ~15–20 MT per minute grow outwards for a distance >1 μm. Almost no persistent growth of microtubules was observed in these cells with the average growth length of 5–6 μm and duration of growth periods within 30 s. However, the number of growing ends increased towards cell margin, especially towards the active edges. We found the peripheral cytoplasmic foci of microtubule growth there. During recovery from nocodazole treatment microtubules started to grow around the centrosome in a normal way and independently in all the cell areas. Within 5 minutes microtubules continued to grow mainly near the cell edge. Thus, our data confirm the negligible role of centrosome as MTOC in 3T3 fibroblasts and propose a model of non-centrosomal microtubules as major players that create the cell asymmetry in the cells with a mesenchymal type of motility. We suggest that increased density of dynamic microtubules near the active lamellum could be supported by microtubule-based microtubule nucleation.
机译：相间成纤维细胞样细胞中的微管被认为以放射状排列，从基于中心体的微管组织中心（MTOC）到细胞边缘。但是，许多形态发生过程需要微管（MT）阵列的不对称性。这种不对称的可能机制之一可能是在不同的细胞内区域中存在非中心体微管。为了评估中心体出生的和非中心体的微管在运动性3T3成纤维细胞中微管阵列的组织中的作用，我们对细胞不同功能区域中的微管生长进行了高通量分析，并区分了生长中的微管的三个亚群（出生于中心体，边缘和内部细胞质）。在一半的细胞中不存在作为活性微管组织中心的中心体。但是，这些细胞在微管生长模式上没有显示任何差异。在具有活性中心体的细胞中，它不断形成短的（短暂的）MT，每分钟约15-20 MT向外生长，距离> 1μm。在这些细胞中几乎没有观察到微管的持续生长，其平均生长长度为5–6μm，并且生长时间在30 s内。但是，生长末端的数量朝向细胞边缘增加，特别是向活性边缘增加。我们发现那里的微管生长的外围细胞质灶。从诺考达唑治疗中恢复期间，微管开始以正常方式在中心体周围生长，并独立于所有细胞区域生长。在5分钟内，微管主要在细胞边缘附近继续生长。因此，我们的数据证实了中心体在3T3成纤维细胞中作为MTOC的作用微不足道，并提出了一种非中心体微管模型，作为在具有间充质运动类型的细胞中产生细胞不对称性的主要参与者。我们建议通过基于微管的微管成核可以支持在活动板层附近增加动态微管的密度。

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