Larson Matthew E, Bement William M
Medical Scientist Training Program, University of Wisconsin-Madison, Madison, WI 53706
Cellular and Molecular Biology Graduate Program, University of Wisconsin-Madison, Madison, WI 53706.
Mol Biol Cell. 2017 Mar 15;28(6):746-759. doi: 10.1091/mbc.E16-06-0355. Epub 2017 Jan 18.
Proper spindle positioning at anaphase onset is essential for normal tissue organization and function. Here we develop automated spindle-tracking software and apply it to characterize mitotic spindle dynamics in the embryonic epithelium. We find that metaphase spindles first undergo a sustained rotation that brings them on-axis with their final orientation. This sustained rotation is followed by a set of striking stereotyped rotational oscillations that bring the spindle into near contact with the cortex and then move it rapidly away from the cortex. These oscillations begin to subside soon before anaphase onset. Metrics extracted from the automatically tracked spindles indicate that final spindle position is determined largely by cell morphology and that spindles consistently center themselves in the -plane before anaphase onset. Finally, analysis of the relationship between spindle oscillations and spindle position relative to the cortex reveals an association between cortical contact and anaphase onset. We conclude that metaphase spindles in epithelia engage in a stereotyped "dance," that this dance culminates in proper spindle positioning and orientation, and that completion of the dance is linked to anaphase onset.
在后期开始时纺锤体的正确定位对于正常组织的结构和功能至关重要。在此,我们开发了自动纺锤体追踪软件,并将其应用于表征胚胎上皮细胞有丝分裂纺锤体的动力学。我们发现中期纺锤体首先经历持续旋转,使其与最终方向共轴。这种持续旋转之后是一系列引人注目的定型旋转振荡,使纺锤体与皮质几乎接触,然后迅速将其从皮质移开。这些振荡在后期开始前不久开始减弱。从自动追踪的纺锤体中提取的指标表明,最终纺锤体位置在很大程度上由细胞形态决定,并且纺锤体在后期开始前始终在平面内居中。最后,对纺锤体振荡与纺锤体相对于皮质的位置之间关系的分析揭示了皮质接触与后期开始之间的关联。我们得出结论,上皮细胞中的中期纺锤体进行一种定型的“舞蹈”,这种舞蹈最终导致纺锤体的正确定位和定向,并且舞蹈的完成与后期开始相关联。