Manning Lathiena, Starz-Gaiano Michelle
Department of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD, 21250, USA.
Methods Mol Biol. 2015;1328:73-88. doi: 10.1007/978-1-4939-2851-4_5.
Drosophila oogenesis provides many examples of essential processes in development. A myriad of genetic tools combined with recent advances in culturing egg chambers ex vivo has revealed several surprising mechanisms that govern how this tissue develops, and which could not have been determined in fixed tissues. Here we describe a straightforward protocol for dissecting ovaries, culturing egg chambers, and observing egg development in real time by fluorescent microscopy. This technique is suitable for observation of early- or late-stage egg development, and can be adapted to study a variety of cellular, molecular, or developmental processes. Ongoing analysis of oogenesis in living egg chambers has tremendous potential for discovery of new developmental mechanisms.
果蝇卵子发生为发育过程中的许多基本过程提供了诸多实例。大量的遗传工具与近期离体培养卵室的进展相结合，揭示了几种令人惊讶的机制，这些机制决定了该组织的发育方式，而这些机制在固定组织中是无法确定的。在此，我们描述了一种简单的方案，用于解剖卵巢、培养卵室，并通过荧光显微镜实时观察卵子发育。该技术适用于观察早期或晚期卵子发育，并且可以进行调整以研究各种细胞、分子或发育过程。对活卵室中卵子发生的持续分析对于发现新的发育机制具有巨大潜力。
