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自动化多维图像分析揭示 Abl 在胚胎伤口修复中的作用。

Automated multidimensional image analysis reveals a role for Abl in embryonic wound repair.

机构信息

Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada M5S 3G9.

Developmental Biology Program, Sloan-Kettering Institute, New York, NY 10065, USA.

出版信息

Development. 2014 Jul;141(14):2901-11. doi: 10.1242/dev.106898. Epub 2014 Jun 19.

Abstract

The embryonic epidermis displays a remarkable ability to repair wounds rapidly. Embryonic wound repair is driven by the evolutionary conserved redistribution of cytoskeletal and junctional proteins around the wound. Drosophila has emerged as a model to screen for factors implicated in wound closure. However, genetic screens have been limited by the use of manual analysis methods. We introduce MEDUSA, a novel image-analysis tool for the automated quantification of multicellular and molecular dynamics from time-lapse confocal microscopy data. We validate MEDUSA by quantifying wound closure in Drosophila embryos, and we show that the results of our automated analysis are comparable to analysis by manual delineation and tracking of the wounds, while significantly reducing the processing time. We demonstrate that MEDUSA can also be applied to the investigation of cellular behaviors in three and four dimensions. Using MEDUSA, we find that the conserved nonreceptor tyrosine kinase Abelson (Abl) contributes to rapid embryonic wound closure. We demonstrate that Abl plays a role in the organization of filamentous actin and the redistribution of the junctional protein β-catenin at the wound margin during embryonic wound repair. Finally, we discuss different models for the role of Abl in the regulation of actin architecture and adhesion dynamics at the wound margin.

摘要

胚胎表皮具有快速修复伤口的惊人能力。胚胎伤口修复是由进化保守的细胞骨架和连接蛋白在伤口周围的重新分布驱动的。果蝇已成为筛选与伤口闭合相关因子的模型。然而,遗传筛选受到手动分析方法的限制。我们引入了 MEDUSA,这是一种新颖的图像分析工具,可从延时共聚焦显微镜数据中自动量化多细胞和分子动力学。我们通过量化果蝇胚胎的伤口闭合来验证 MEDUSA,结果表明我们的自动分析结果与手动描绘和跟踪伤口的结果相当,同时显著减少了处理时间。我们还证明 MEDUSA 也可用于研究三维和四维中的细胞行为。使用 MEDUSA,我们发现保守的非受体酪氨酸激酶 Abelson (Abl) 有助于快速的胚胎伤口闭合。我们证明 Abl 在胚胎伤口修复过程中在丝状肌动蛋白的组织和连接蛋白β-catenin 在伤口边缘的重新分布中发挥作用。最后,我们讨论了 Abl 在调节伤口边缘处肌动蛋白结构和黏附动力学中的不同作用模型。

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