Retention of Ejaculate by Drosophila melanogaster Females Requires the Male-Derived Mating Plug Protein PEBme
Retention of Ejaculate by Drosophila melanogaster Females Requires the Male-Derived Mating Plug Protein PEBme
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DOI：
10.1534/genetics.115.176669
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发表时间：
2015-08-01
期刊：
影响因子：
3.3
通讯作者：
Wolfner, Mariana F.
中科院分区：
文献类型：
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作者：
Avila, Frank W.;Cohen, Allie B.;Wolfner, Mariana F.
在许多分类群的雌性交配生殖道内，精液蛋白（SFPs）凝结成一种称为交配塞（MP）的结构。MPs有多种作用，包括阻止雌性交配，改变雌性交配后的接受能力，以及成功储存精子的必要条件。黑腹果蝇（Drosophila melanogaster）的mps在交配后在雌性体内维持数小时，由交配开始后迅速形成的后mps （PMP）和随后形成的前mps （AMP）组成。PMP是由来自男性生殖道射精球（EB）的精液蛋白组成的。为了研究PMP蛋白PEBme在黑腹龙繁殖中的作用，我们鉴定了EB GAL4驱动因子，并利用它靶向PEBme进行RNA干扰（RNAi）敲除。雄性PEBme敲低会损害其配偶的PMP凝血，导致雌性生育能力显著降低，对交配后子宫形态、精子储存、交配难耐性、产卵和后代产生不利影响。这些缺陷是由于雌性在交配后无法将射精保留在生殖道中。未凝固的MP被击倒的雄虫破坏了解偶联，当雄虫最终解偶联时，射精通常会从雌虫体内抽出。因此，PEBme和MP凝固是黑胃d.m anogaster最佳生育能力所必需的。鉴于PMP对生育的重要性，我们通过质谱法鉴定了额外的MP蛋白，并发现了其中两种蛋白的生育功能。我们的研究结果强调了PMP和组成它的蛋白质在生殖中的重要性，并表明在果蝇中，PMP是将射精保留在雌性生殖道内，确保交配雌性储存精子所必需的。
Within the mated reproductive tracts of females of many taxa, seminal fluid proteins (SFPs) coagulate into a structure known as the mating plug (MP). MPs have diverse roles, including preventing female remating, altering female receptivity postmating, and being necessary for mated females to successfully store sperm. The Drosophila melanogaster MP, which is maintained in the mated female for several hours postmating, is comprised of a posterior MP (PMP) that forms quickly after mating begins and an anterior MP (AMP) that forms later. The PMP is composed of seminal proteins from the ejaculatory bulb (EB) of the male reproductive tract. To examine the role of the PMP protein PEBme in D. melanogaster reproduction, we identified an EB GAL4 driver and used it to target PEBme for RNA interference (RNAi) knockdown. PEBme knockdown in males compromised PMP coagulation in their mates and resulted in a significant reduction in female fertility, adversely affecting postmating uterine conformation, sperm storage, mating refractoriness, egg laying, and progeny generation. These defects resulted from the inability of females to retain the ejaculate in their reproductive tracts after mating. The uncoagulated MP impaired uncoupling by the knockdown male, and when he ultimately uncoupled, the ejaculate was often pulled out of the female. Thus, PEBme and MP coagulation are required for optimal fertility in D. melanogaster. Given the importance of the PMP for fertility, we identified additional MP proteins by mass spectrometry and found fertility functions for two of them. Our results highlight the importance of the MP and the proteins that comprise it in reproduction and suggest that in Drosophila the PMP is required to retain the ejaculate within the female reproductive tract, ensuring the storage of sperm by mated females.