Shrub (CG8055) encodes the vps32/snf7 protein, a filament‐forming subunit of the ESCRT (endosomal sorting complexes required for transport)‐III complex involved in inward membrane budding. It was reported that shrub was required for abscission in female germline stem cells. In this study, we showed that the expression level of shrub in the testis was significantly higher than that in the ovary of 1‐day‐old Drosophila melanogaster, suggesting a role in male reproduction. Then we used nosGal4 driver to knockdown shrub specifically in the fly testis and found that this resulted in a significantly lower paternal effect egg hatch rate relative to the control group. Immunofluorescence staining showed that shrub knockdown in fly testes caused an accumulation of early‐stage germ cells and lack of spectrin caps. In the late stages (spermiogenesis), the control testis contained multiple compacted spermatid bundles and individualization complexes (ICs) consisting of actin cones, whereas there were scattered spermatid nuclei and only a few ICs with disorganized actin cones in the shrub knockdown testis. Finally, the control seminal vesicle was full of mature sperms with needle‐like heads, but in shrub knockdown testis 75% of seminal vesicles had no mature sperms. We also found that knockdown of shrub in fly testes led to upregulated expression of several cytoskeleton‐associated genes, and an accumulation of ubiquitylated proteins. These results suggest that knockdown of shrub in fly testes might damage spermatogenesis by affecting transportability.

中文翻译：

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果蝇的生精需要灌木

灌木（CG8055）编码vps32 / snf7蛋白，该蛋白是ESCRT（运输所需的内体分拣复合物）-III内膜发芽的细丝形成亚基。据报道，雌性种系干细胞的脱落需要灌木。在这项研究中，我们表明灌木在睾丸中的表达水平显着高于1日龄果蝇的卵巢中的表达水平，表明其在雄性生殖中的作用。然后我们使用nosGal4驱动程序关闭灌木特别是在苍蝇的睾丸中，发现与对照组相比，这导致了父本卵孵化率明显降低。免疫荧光染色显示，果蝇睾丸中的灌木敲除导致早期生殖细胞的积累和缺乏血影蛋白的帽。在后期（生精），对照睾丸包含多个紧密的精子束和由肌动蛋白视锥组成的个体化复合物（IC），而在灌木击倒的睾丸中，散布的精子细胞核和仅有少数具有杂乱的肌动蛋白视锥的IC 。最后，对照精囊中充满了成熟的精子，具针状头，但处于灌木丛中击倒睾丸的精囊中有75％没有成熟的精子。我们还发现，在果蝇睾丸中敲除灌木会导致一些细胞骨架相关基因的表达上调，以及泛素化蛋白的积累。这些结果表明，果蝇睾丸中的灌木敲除可能通过影响转运性而损害精子发生。