边缘细胞迁移：集体细胞运动实时成像与遗传分析的模型系统。
Border Cell Migration: A Model System for Live Imaging and Genetic Analysis of Collective Cell Movement.
作者信息
Prasad Mohit, Wang Xiaobo, He Li, Cai Danfeng, Montell Denise J
机构信息
Department of Biological Chemistry, Johns Hopkins School of Medicine, Baltimore, MD, 21205, USA.
出版信息
Methods Mol Biol. 2015;1328:89-97. doi: 10.1007/978-1-4939-2851-4_6.
Border cell migration in the Drosophila ovary has emerged as a genetically tractable model for studying collective cell movement. Over many years border cell migration was exclusively studied in fixed samples due to the inability to culture stage 9 egg chambers in vitro. Although culturing late-stage egg chambers was long feasible, stage 9 egg chambers survived only briefly outside the female body. We identified culture conditions that support stage 9 egg chamber development and sustain complete migration of border cells ex vivo. This protocol enables one to compare the dynamics of egg chamber development in wild-type and mutant egg chambers using time-lapse microscopy and taking advantage of a multiposition microscope with a motorized imaging stage. In addition, this protocol has been successfully used in combination with fluorescence resonance energy transfer biosensors, photo-activatable proteins, and pharmacological agents and can be used with wide-field or confocal microscopes in either an upright or an inverted configuration.
果蝇卵巢中的边界细胞迁移已成为研究集体细胞运动的一个易于进行遗传操作的模型。多年来，由于无法在体外培养9期卵室，边界细胞迁移仅在固定样本中进行研究。尽管长期以来培养晚期卵室是可行的，但9期卵室在雌性体外只能短暂存活。我们确定了支持9期卵室发育并维持边界细胞在体外完全迁移的培养条件。该方案使人们能够使用延时显微镜，并利用带有电动成像载物台的多位置显微镜，比较野生型和突变型卵室中卵室发育的动态过程。此外，该方案已成功与荧光共振能量转移生物传感器、光激活蛋白和药剂结合使用，并且可用于正置或倒置配置的宽视野或共聚焦显微镜。
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