Laser ablation and fluid flows reveal the mechanism behind spindle and centrosome positioning
Laser ablation and fluid flows reveal the mechanism behind spindle and centrosome positioning
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DOI：
10.1038/s41567-023-02223-z
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发表时间：
2023-11-02
期刊：
影响因子：
19.6
通讯作者：
Needleman，Daniel J.
中科院分区：
文献类型：
--
作者：
Wu，Hai-Yin;Kabacaoglu，Gokberk;Needleman，Daniel J.
很少有技术可以用来研究驱动亚细胞动力学的力的性质。在这里，我们开发了两个互补的。第一种是飞秒立体定向激光消融术，它可以迅速对亚细胞结构进行复杂的切割，并能够精确地剖析产生作用力的时间、地点和方向。第二个是通过比较使用微注入荧光纳米钻石的直接流动测量与不同力传递模型的大规模流体结构模拟来评估亚细胞流体流动。我们应用这些技术研究了秀丽线虫早期胚胎中纺锤体和中心体的定位，并探讨了微管推动、细胞质拉动和皮质拉动对中心体微管的贡献。基于我们的结果，我们构建了一个生物物理模型来解释中心体的动力学。我们证明，皮质牵引力为中心体介导的许多行为提供了一般解释，包括原核迁移和定心、旋转、中期纺锤体定位、不对称纺锤体伸长和纺锤体振荡。这项工作建立了解开导致细胞生物学现象的力量的方法论。
Few techniques are available for studying the nature of forces that drive subcellular dynamics. Here we develop two complementary ones. The first is femtosecond stereotactic laser ablation, which rapidly creates complex cuts of subcellular structures and enables precise dissection of when, where and in what direction forces are generated. The second is an assessment of subcellular fluid flows by comparison of direct flow measurements using microinjected fluorescent nanodiamonds with large-scale fluid-structure simulations of different force transduction models. We apply these techniques to study spindle and centrosome positioning in earlyCaenorhabditis elegansembryos and to probe the contributions of microtubule pushing, cytoplasmic pulling and cortical pulling upon centrosomal microtubules. Based on our results, we construct a biophysical model to explain the dynamics of centrosomes. We demonstrate that cortical pulling forces provide a general explanation for many behaviours mediated by centrosomes, including pronuclear migration and centration, rotation, metaphase spindle positioning, asymmetric spindle elongation and spindle oscillations. This work establishes methodologies for disentangling the forces responsible for cell biological phenomena.