Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, 333, Taiwan.
Development. 2014 Feb;141(3):563-73. doi: 10.1242/dev.097022. Epub 2014 Jan 8.
The nutritional environment is crucial for Drosophila oogenesis in terms of controlling hormonal conditions that regulate yolk production and the progress of vitellogenesis. Here, we discovered that Drosophila Endophilin B (D-EndoB), a member of the endophilin family, is required for yolk endocytosis as it regulates membrane dynamics in developing egg chambers. Loss of D-EndoB leads to yolk content reduction, similar to that seen in yolkless mutants, and also causes poor fecundity. In addition, mutant egg chambers exhibit an arrest at the previtellogenic stage. D-EndoB displayed a crescent localization at the oocyte posterior pole in an Oskar-dependent manner; however, it did not contribute to pole plasm assembly. D-EndoB was found to partially colocalize with Long Oskar and Yolkless at the endocytic membranes in ultrastructure analysis. Using an FM4-64 dye incorporation assay, D-EndoB was also found to promote endocytosis in the oocyte. When expressing the full-length D-endoB(FL) or D-endoB(ΔSH3) mutant transgenes in oocytes, the blockage of vitellogenesis and the defect in fecundity in D-endoB mutants was restored. By contrast, a truncated N-BAR domain of the D-EndoB only partially rescued these defects. Taken together, these results allow us to conclude that D-EndoB contributes to the endocytic activity downstream of Oskar by facilitating membrane dynamics through its N-BAR domain in the yolk uptake process, thereby leading to normal progression of vitellogenesis.
营养环境对果蝇卵子发生至关重要,它可以控制激素条件,从而调节卵黄的产生和卵黄发生的进程。在这里,我们发现果蝇内收蛋白 B(D-EndoB)作为内收蛋白家族的一员,是卵黄内吞作用所必需的,因为它调节了正在发育的卵囊中膜的动力学。D-EndoB 的缺失会导致卵黄含量减少,这与卵黄缺失突变体相似,也会导致繁殖力下降。此外,突变的卵囊中室在卵黄前发生阶段停滞。D-EndoB 以 Oskar 依赖的方式在卵母细胞后极呈现新月形定位;然而,它不参与极质体的组装。在超微结构分析中,D-EndoB 被发现部分与 Long Oskar 和 Yolkless 共定位于内吞膜上。使用 FM4-64 染料掺入测定法,还发现 D-EndoB 促进了卵母细胞的内吞作用。当在卵母细胞中表达全长 D-endoB(FL)或 D-endoB(ΔSH3)突变基因时,D-endoB 突变体中卵黄发生的阻断和繁殖力的缺陷得到了恢复。相比之下,D-EndoB 的截断 N-BAR 结构域只能部分挽救这些缺陷。总之,这些结果表明,D-EndoB 通过其 N-BAR 结构域促进膜动力学,从而有助于 Oskar 下游的内吞作用,在卵黄摄取过程中导致正常的卵黄发生进程。