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首页> 外文期刊>Molecular biology of the cell >Direct visualization of microtubule flux during metaphase and anaphase in crane-fly spermatocytes
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Direct visualization of microtubule flux during metaphase and anaphase in crane-fly spermatocytes

机译：鹤蝇精细胞中期和后期的微管通量的直接可视化

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摘要

Microtubule flux in spindles of insect spermatocytes, long-used models for studies on chromosome behavior during meiosis, was revealed after iontophoretic microinjection of rhodamine-conjugated (rh)-tubulin and fluorescent speckle microscopy. In time-lapse movies of crane-fly spermtocytes, fluorescent speckles generated when rh-tubulin incorporated at microtubule plus ends moved poleward through each half-spindle and then were lost from microtubule minus ends at the spindle poles. The average poleward velocity of similar to0.7 mum/min for speckles within kinetochore microtubules at metaphase increased during anaphase to similar to0.9 mum/min. Segregating half-bivalents had an average poleward velocity of similar to0.5 mum/min, about half that of speckles within shortening kinetochore fibers. When injected during anaphase, rhtubulin was incorporated at kinetochores, and kinetochore fiber fluorescence spread poleward as anaphase progressed. The results show that tubulin subunits are added to the plus end of kinetochore microtubules and are removed from their minus ends at the poles, all while attached chromosomes move poleward during anaphase A. The results cannot be explained by a Pac-man model, in which 1) kinetochore-based, minus end-directed motors generate poleward forces for anaphase A and 2) kinetochore microtubules shorten at their plus ends. Rather, in these cells, kinetochore fiber shortening during anaphase A occurs exclusively at the minus ends of kinetochore microtubules.
机译：在对若丹明缀合的（rh）-微管蛋白进行离子电渗析显微注射和荧光斑点显微镜后，揭示了昆虫精母细胞纺锤体中的微管通量，这是用于研究减数分裂过程中染色体行为的长期模型。在白鹤飞行的精细胞的延时摄影中，当掺入微管正端的rh-微管蛋白向两极移动时，会产生荧光斑点，然后从纺锤极的负端丢失。在后期，动粒体内微管内的斑点的平均极点速度接近0.7毫米/分钟，在后期增加到接近0.9毫米/分钟。分离的半二价分子的平均极点传播速度接近0.5毫米/分钟，大约是短动发丝纤维内斑点的一半。当在后期进行注射时，rhtubulin掺入了动植物，并且随着后期的进行，线粒体纤维的荧光向极传播。结果表明，微管蛋白亚基被添加到动粒微管的正端，并从极的负端移出，而在后期A期间所有附着的染色体都向极移。结果无法用Pac-man模型解释，其中1）基于动线轴的负向电动机会为后期A产生极向力，并且2）线粒体微管的正端会变短。而是，在这些细胞中，后期A期间的线粒体纤维缩短仅发生在线粒体微管的负端。

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