Schulze E, Kirschner M
J Cell Biol. 1986 Mar;102(3):1020-31. doi: 10.1083/jcb.102.3.1020.
The sites of microtubule growth and the kinetics of elongation have been studied in vivo by microinjection of biotin-labeled tubulin and subsequent visualization with immunocytochemical probes. Immunofluorescence and immunoelectron microscopy demonstrate that injected biotin-labeled subunits are incorporated into new segments of growth which are contiguous with unlabeled microtubules. Rapid incorporation occurs by elongation of existing microtubules and new nucleation off the centrosome. The growth rate is 3.6 micron/min and is independent of the concentration of injected labeled tubulin. This rate of incorporation together with turnover of existing microtubules leads to approximately 80% exchange in 15 min. The observed kinetics and pattern of microtubule turnover allow for an evaluation of the relevance of several in vitro models for steady-state dynamics to the in vivo situation. We have also observed a substantial population of quasi-stable microtubules that does not exchange subunits as rapidly as the majority of microtubules and may have specialized functions in the cell.
通过向体内显微注射生物素标记的微管蛋白并随后用免疫细胞化学探针进行可视化,研究了微管生长的位点和伸长动力学。免疫荧光和免疫电子显微镜显示,注射的生物素标记的亚基被整合到与未标记微管相邻的新生长段中。通过现有微管的伸长和从中心体的新成核作用快速整合。生长速率为3.6微米/分钟,且与注射的标记微管蛋白的浓度无关。这种整合速率连同现有微管的周转导致在15分钟内约80%的交换。观察到的微管周转动力学和模式有助于评估几种体外稳态动力学模型与体内情况的相关性。我们还观察到大量准稳定微管,其亚基交换速度不如大多数微管快,可能在细胞中具有特殊功能。