Gancz Dana, Gilboa Lilach
Department of Biological Regulation, Weizmann Institute of Science, Rehovot, 76100, Israel.
Methods Mol Biol. 2017;1463:75-83. doi: 10.1007/978-1-4939-4017-2_5.
In the fruit fly Drosophila melanogaster, ovarian germline stem cells (GSCs) and their niches form during larval development. This process is poorly studied partly due to technical difficulties in isolating early larval ovaries. In addition, purifying RNA from larval ovaries proves to be more challenging than purifying it from other organs. Here we describe a technique for dissecting ovaries from early larvae and advise on how to extract RNA with maximum yield and purity. RNA isolation allows assaying gene expression in a direct and quantitative manner, which is invaluable for understanding molecular events underlying ovarian niche formation and GSC establishment.
在果蝇黑腹果蝇中,卵巢生殖系干细胞(GSCs)及其微环境在幼虫发育过程中形成。由于分离早期幼虫卵巢存在技术困难,这一过程的研究较少。此外,从幼虫卵巢中纯化RNA比从其他器官中纯化RNA更具挑战性。在这里,我们描述了一种从早期幼虫中解剖卵巢的技术,并就如何以最大产量和纯度提取RNA提供建议。RNA分离允许以直接和定量的方式测定基因表达,这对于理解卵巢微环境形成和GSC建立背后的分子事件非常宝贵。