Zhu Xiaodong, Kaverina Irina
Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN, USA.
Methods Mol Biol. 2011;777:235-44. doi: 10.1007/978-1-61779-252-6_17.
Cell polarization is important for multiple physiological processes. In polarized cells, microtubules (MTs) are organized into a spatially polarized array. Generally, in nondifferentiated cells, it is assumed that MTs are symmetrically nucleated exclusively from centrosome [microtubule organizing center (MTOC)] and then reorganized into the asymmetric array. We have recently identified the Golgi complex as an additional MTOC that asymmetrically nucleates MTs toward one side of the cell. Methods used for alternative MTOC identification include microtubule regrowth after complete drug-induced depolymerization and tracking of growing microtubules using fluorescently labeled MT +TIP binding proteins in living cells. These approaches can be used for quantification of MT nucleation sites at diverse subcellular structures.
细胞极化对多种生理过程至关重要。在极化细胞中,微管(MTs)被组织成空间极化阵列。一般来说,在未分化细胞中,假定微管仅从中心体[微管组织中心(MTOC)]对称成核,然后重新组织成不对称阵列。我们最近已将高尔基体复合体鉴定为另一种MTOC,它向细胞的一侧不对称地使微管成核。用于鉴定替代MTOC的方法包括完全药物诱导解聚后的微管再生,以及在活细胞中使用荧光标记的MT +TIP结合蛋白追踪生长中的微管。这些方法可用于定量不同亚细胞结构处的微管成核位点。