Actions of Seminal Proteins in Mated Drosophila Females

总结 从昆虫到哺乳动物，精液蛋白（Sfps）显著影响生殖生理， 交配的雌性，精子在雌性体内的储存和释放，甚至（在小鼠中） 后代特定Sfps异常的男性不育或生育力低下，包括人类。一些Sfps紧密结合 精子;其他人是免费的精浆。然而，尽管它们在繁殖中的重要性， SFPs是如何影响雌性或雌性精子的行为的。重要的是，许多SFPs 进化迅速，与分子/进化性冲突中的角色一致。理解功能 对Sfps进化的限制以及它们在女性中相互作用的蛋白质将指导未来的发展。 调查粮食计划署在人类生育方面的行动。 我们将联合收割机结合分子遗传学和功能学的方法来研究：（1）Sfps如何与 雌性分子引起生殖反应和（2）Sfps如何与精子相关联，以介导它们的生殖反应。 影响，以及这两种类型的功能是如何演变的。我们将调查这些问题使用 果蝇是研究Sfp功能的首要遗传模型系统， 进化比较重要的是，果蝇Sfps有许多分子和现象学上的相似之处， 与哺乳动物的相似 目的1主要关注通过诱导神经元章鱼胺能信号刺激排卵的ovulin。 这种信号调节女性生殖道的肌肉收缩，放松输卵管， 提高排卵率。利用遗传筛选和蛋白质-蛋白质协同进化的特征，我们有 确定了女性排卵素受体（OvR）的强有力候选者。我们将测试这些卵蛋白结合， 然后确定OvR定位，以确定Ovulin作用的位点。然后我们将研究 物种的ovulins介导ovulin作用和OvR结合，以阐明其功能的进化。 在目标2中，我们将关注与精子结合的精液蛋白，我们已经通过它们的共同进化确定了它们 或通过蛋白质组学方法。我们最近的数据表明，Sfp为精子与关键的Sfp（性）结合做好了准备 肽我们将探讨哪些精液蛋白在这条启动通路中起作用，哪些独立起作用 这条路。我们还将调查女性分泌物是否也参与启动。最后我们将 确定精子结合Sfp的子集的功能在相关的基因组中保守的程度。 果蝇属。 阐明Sfps如何在分子水平上与女性相互作用并影响女性，以及这些 相互作用的演变，是重要的理解和诊断Sfp为基础的不孕症，在考虑 将从纳入关键的SFPs中受益的辅助生殖技术战略， 开发新的方法来控制传播登革热、寨卡和疟疾等严重疾病的双翅目昆虫。

SUMMARY From insects to mammals, seminal fluid proteins (Sfps) significantly affect the reproductive physiology of mated females, the storage and release of sperm inside females, and (in mice) even the phenotype of progeny. Males abnormal for specific Sfps are sterile or subfertile, including in humans. Some Sfps bind tightly to sperm; others are free in seminal plasma. Yet despite their importance in reproduction, little is known about exactly how Sfps act to influence the female or the behavior of sperm in females. Importantly, many Sfps evolve rapidly, consistent with roles in molecular/evolutionary sexual conflicts. Understanding functional constraints on the evolution of Sfps and the proteins with which they interact in females will guide future investigations into Sfp actions in human fertility. We will combine molecular genetic and functional approaches to investigate: (1) how Sfps interact with female molecules to elicit reproductive responses and (2) how Sfps associate with sperm to mediate their effects, as well as how both types of function have evolved. We will investigate these questions using Drosophila, a premier genetic model system for dissecting Sfp function, with extensive resources for evolutionary comparisons. Importantly, Drosophila Sfps have many molecular and phenomenological parallels to those of mammals. Aim 1 focuses on ovulin, which stimulates ovulation by inducing neuronal octopaminergic signaling. This signaling regulates muscle contraction in the female reproductive tract, relaxing the oviducts and increasing ovulation rate. Using genetic screens and signatures of protein-protein coevolution, we have identified strong candidates for the female’s receptor for ovulin (OvR). We will test these for ovulin binding and then determine OvR localization, to pinpoint the site of ovulin action. We will then examine how well different species’ ovulins mediate ovulin action and OvR binding, to elucidate the evolution of their function. In Aim 2 we will focus on seminal proteins that bind to sperm, which we have identified by their coevolution or by proteomic methods. Our recent data show that Sfps prime sperm for binding to the critical Sfp called Sex Peptide. We will ask which seminal proteins function within this priming pathway and which act independently of that pathway. We will also investigate whether female secretions are also involved in priming. Finally, we will determine the extent to which the functions of a subset of sperm-bound Sfps are conserved across related Drosophila species. Elucidating how Sfps interact with and affect the female at the molecular level, as well as how these interactions evolve, is important for understanding and diagnosing Sfp-based infertilities, in considering strategies for assisted reproductive technologies that would benefit from inclusion of critical Sfps, and for developing new ways to control dipteran insects that transmit serious diseases like dengue, Zika, and malaria.