Rubin Thomas, Huynh Jean-René
Department of Genetics and Developmental Biology, Institut Curie, 26 rue d'Ulm, 75248, Paris, France.
Methods Mol Biol. 2015;1328:29-55. doi: 10.1007/978-1-4939-2851-4_3.
Drosophila melanogaster oogenesis is a versatile model system used to address many important questions of cell and developmental biology such as stem cell regulation, cell determination, cell polarization, cell-cell signaling, cell-cell adhesion, and cell-cycle regulation. The ovary is composed of germline and somatic cells of different origins and functions. Mosaic analysis using the powerful genetic tools available in Drosophila melanogaster allows deciphering the contribution of each cell type in the different processes leading to the formation of a mature egg. Germ cells and follicle cells are produced by actively dividing stem cells, which permit the use of recombinases, such as FLP, to generate genetic mosaics using mitotic recombination. This chapter summarizes the different methods used to create genetic mosaics in the germline and in somatic cells of adult ovaries. We briefly introduce the morphology and development of the adult female ovary. We then describe in practical terms how to generate mosaics with examples of cross schemes and recombining strains. We also explain how to identify the appropriate progeny and how to prepare clonal tissues for phenotypic analysis.
黑腹果蝇卵子发生是一个通用的模型系统，用于解决细胞和发育生物学中的许多重要问题，如干细胞调控、细胞决定、细胞极化、细胞间信号传导、细胞间黏附以及细胞周期调控。卵巢由不同起源和功能的生殖细胞和体细胞组成。利用黑腹果蝇中可用的强大遗传工具进行嵌合体分析，可以解读每种细胞类型在导致成熟卵子形成的不同过程中的作用。生殖细胞和卵泡细胞由活跃分裂的干细胞产生，这使得可以使用重组酶，如FLP，通过有丝分裂重组来产生遗传嵌合体。本章总结了在成年卵巢的生殖细胞和体细胞中创建遗传嵌合体的不同方法。我们简要介绍成年雌性卵巢的形态和发育。然后我们通过杂交方案和重组菌株的例子实际描述如何生成嵌合体。我们还解释了如何鉴定合适的后代以及如何制备用于表型分析的克隆组织。
