Jha Anupma, Traub Linton M
Department of Cell Biology, University of Pittsburgh School of Medicine, 3500 Terrace Street, 15261, Pittsburgh, PA, USA.
Methods Mol Biol. 2014;1174:349-60. doi: 10.1007/978-1-4939-0944-5_24.
In oviparous animals, clathrin-dependent endocytosis is often critical to stockpile a necessary supply of yolk within the maturing oocyte, which enables subsequent embryonic development. In the physically linked chains of maturing egg chambers within the Drosophila melanogaster ovary, a distinct, morphologically discernable subset undergoes a massive burst clathrin-mediated endocytosis to accumulate yolk in a process termed vitellogenesis. Here, we describe how to prepare isolated ovaries to follow endocytosis, and detail approaches to follow live uptake of soluble reporters into vitellogenic Drosophila egg chambers.
在卵生动物中,网格蛋白依赖的内吞作用对于在成熟卵母细胞内储存必要的卵黄供应通常至关重要,这能使后续的胚胎发育得以进行。在果蝇卵巢中成熟卵室的物理连接链中,一个独特的、形态上可辨别的亚群会经历大量网格蛋白介导的内吞作用,在一个称为卵黄生成的过程中积累卵黄。在这里,我们描述了如何制备分离的卵巢以追踪内吞作用,并详细介绍了追踪可溶性报告分子在果蝇卵黄生成卵室中的实时摄取方法。