Grenfell Andrew W, Strzelecka Magdalena, Crowder Marina E, Helmke Kara J, Schlaitz Anne-Lore, Heald Rebecca
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720
J Cell Biol. 2016 Apr 11;213(1):127-36. doi: 10.1083/jcb.201509079. Epub 2016 Apr 4.
Imaging datasets are rich in quantitative information. However, few cell biologists possess the tools necessary to analyze them. Here, we present a large dataset of Xenopusextract spindle images together with an analysis pipeline designed to assess spindle morphology across a range of experimental conditions. Our analysis of different spindle types illustrates how kinetochore microtubules amplify spindle microtubule density. Extract mixing experiments reveal that some spindle features titrate, while others undergo switch-like transitions, and multivariate analysis shows the pleiotropic morphological effects of modulating the levels of TPX2, a key spindle assembly factor. We also apply our pipeline to analyze nuclear morphology in human cell culture, showing the general utility of the segmentation approach. Our analyses provide new insight into the diversity of spindle types and suggest areas for future study. The approaches outlined can be applied by other researchers studying spindle morphology and adapted with minimal modification to other experimental systems.
成像数据集包含丰富的定量信息。然而，很少有细胞生物学家拥有分析这些数据集所需的工具。在此，我们展示了一个非洲爪蟾提取物纺锤体图像的大型数据集，以及一个旨在评估一系列实验条件下纺锤体形态的分析流程。我们对不同纺锤体类型的分析说明了动粒微管如何放大纺锤体微管密度。提取物混合实验表明，一些纺锤体特征会发生滴定，而另一些则经历类似开关的转变，多变量分析显示了调节关键纺锤体组装因子TPX2水平的多效性形态学效应。我们还应用我们的流程来分析人类细胞培养中的核形态，展示了分割方法的普遍实用性。我们的分析为纺锤体类型的多样性提供了新的见解，并提出了未来研究的方向。概述的方法可供其他研究纺锤体形态的研究人员应用，并可在最少修改的情况下适用于其他实验系统。
