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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Redundant mechanisms for anaphase chromosome movements: crane-fly spermatocyte spindles normally use actin filaments but also can function without them
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Redundant mechanisms for anaphase chromosome movements: crane-fly spermatocyte spindles normally use actin filaments but also can function without them

机译：后期染色体移动的冗余机制：鹤蝇精子纺锤体通常使用肌动蛋白丝，但如果没有它们，也可以发挥作用

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Actin inhibitors block or slow anaphase chromosome movements in crane-fly spermatocytes, but stopping of movement is only temporary; we assumed that cells adapt to loss of actin by switching to mechanism(s) involving only microtubules. To test this, we produced actin-filament-free spindles: we added latrunculin B during prometaphase, 9-80 min before anaphase, after which chromosomes generally moved normally during anaphase. We confirmed the absence of actin filaments by staining with fluorescent phalloidin and by showing that cytochalasin D had no effect on chromosome movement. Thus, actin filaments are involved in normal anaphase movements, but in vivo, spindles nonetheless can function normally without them. We tested whether chromosome movements in actin-filament-free spindles arise via microtubules by challenging such spindles with anti-myosin drugs. Y-27632 and BDM (2,3-butanedione monoxime), inhibitors that affect myosin at different regulatory levels, blocked chromosome movement in normal spindles and in actin-filament-free spindles. We tested whether BDM has side effects on microtubule motors. BDM had no effect on ciliary and sperm motility or on ATPase activity of isolated ciliary axonemes, and thus it does not directly block dynein. Nor does it block kinesin, assayed by a microtubule sliding assay. BDM could conceivably indirectly affect these microtubule motors, though it is unlikely that it would have the same side effect on the motors as Y-27632. Since BDM and Y-27632 both affect chromosome movement in the same way, it would seem that both affect spindle myosin; this suggests that spindle myosin interacts with kinetochore microtubules, either directly or via an intermediate component.
机译：肌动蛋白抑制剂阻止或减慢鹤蝇精子细胞的后期染色体运动，但停止运动只是暂时的。我们假设细胞通过切换到仅涉及微管的机制来适应肌动蛋白的损失。为了测试这一点，我们生产了无肌动蛋白丝的纺锤体：我们在中期前（后期前9-80分钟）添加了latrunculin B，此后染色体通常在后期后期正常移动。我们通过用荧光鬼笔环肽染色并显示细胞松弛素D对染色体运动没有影响来证实肌动蛋白丝不存在。因此，肌动蛋白丝参与正常的后期运动，但是在体内，纺锤体仍然可以在没有它们的情况下正常运行。我们通过用抗肌球蛋白药物挑战此类纺锤体，测试了无肌动蛋白丝纺锤体中的染色体运动是否通过微管发生。 Y-27632和BDM（2,3-丁二酮单肟）是在不同调节水平上影响肌球蛋白的抑制剂，可阻止正常纺锤体和无肌动蛋白丝纺锤体中的染色体运动。我们测试了BDM是否对微管马达有副作用。 BDM对睫状和精子运动没有影响，也对孤立的睫状轴蛋白的ATPase活性没有影响，因此它不直接阻断动力蛋白。通过微管滑动测定法测定，它也不会阻断驱动蛋白。可以想象，BDM可能间接影响这些微管电机，尽管它对电机的副作用不太可能与Y-27632相同。由于BDM和Y-27632都以相同的方式影响染色体运动，因此似乎都影响纺锤体肌球蛋白。这表明纺锤体肌球蛋白直接或通过中间成分与动粒微管相互作用。

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