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The Female Post-Mating Response Requires Genes Expressed in the Secondary Cells of the Male Accessory Gland in Drosophila melanogaster

机译:女性交配后反应需要在果蝇的男性附件腺的次级细胞中表达的基因。

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摘要

Seminal proteins from the Drosophila male accessory gland induce post-mating responses (PMR) in females. The PMR comprise behavioral and physiological changes that include increased egg laying, decreased receptivity to courting males, and changes in the storage and use of sperm. Many of these changes are induced by a “sex peptide” (SP) and are maintained by SP’s binding to, and slow release from, sperm. The accessory gland contains two secretory cell types with distinct morphological and developmental characteristics. Products of these “main” and “secondary” cells work interdependently to induce and maintain the PMR. To identify individual genes needed for the morphology and function of secondary cells, we studied iab-6cocu males, whose secondary cells have abnormal morphology and fail to provide products to maintain the PMR. By RNA-seq, we identified 77 genes that are downregulated by a factor of >5× in iab-6cocu males. By functional assays and microscopy, we tested 20 candidate genes and found that at least 9 are required for normal storage and release of SP in mated females. Knockdown of each of these 9 genes consequently leads to a reduction in egg laying and an increase in receptivity over time, confirming a role for the secondary cells in maintaining the long-term PMR. Interestingly, only 1 of the 9 genes, CG3349, encodes a previously reported seminal fluid protein (Sfp), suggesting that secondary cells may perform essential functions beyond the production and modification of known Sfps. At least 3 of the 9 genes also regulate the size and/or abundance of secondary cell vacuoles, suggesting that the vacuoles’ contents may be important for the machinery used to maintain the PMR.
机译:果蝇雄性副腺的精蛋白诱导雌性交配后反应(PMR)。 PMR包括行为和生理变化,包括产卵增加,男性对求爱的接受力下降以及精子的存储和使用发生变化。其中许多变化是由“性肽”(SP)引起的,并通过SP与精子的结合和从精子中的释放而得以维持。副腺含有两种具有不同形态和发育特征的分泌细胞。这些“主要”和“次要”细胞的产物相互依存,以诱导和维持PMR。为了鉴定次级细胞的形态和功能所需的单个基因,我们研究了iab-6 cocu 雄性,其次级细胞形态异常且无法提供维持PMR的产物。通过RNA-seq,我们鉴定了77个基因,这些基因在iab-6 cocu 雄性中被下调了> 5倍。通过功能测定和显微镜检查,我们测试了20个候选基因,发现在交配雌性中正常存储和释放SP至少需要9个。因此,这9个基因中的每一个基因的敲除都可导致卵生减少和随时间推移的接受性增加,从而确认了次级细胞在维持长期PMR中的作用。有趣的是,这9个基因中只有1个基因CG3349编码先前报道的精液蛋白(Sfp),这表明,除了已知Sfps的产生和修饰之外,次生细胞可能还具有基本功能。 9个基因中的至少3个也调节次级细胞液泡的大小和/或丰度,这表明液泡的含量对于维持PMR的机制可能很重要。

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