REGULATION OF GAMETE USE AND NEURAL PATHWAYS IN REPRODUCTION

项目总结 有性繁殖需要精准地协调雄性和 女性。这些基因在分子、细胞、神经和整体上调节两性之间的相互作用。 生物体水平，包括来自两个相互竞争的雄性的精子与 单身女性。在当前的资助期，我们使用了易驯化的果蝇模型系统来剖析其作用 在雌性多配偶的不同精子使用中，八胺能神经元表达的基因的比例。在这 更新申请，我们建议作为目标1的一组实验，识别基因并量化它们 对交配塞排出的影响，这是一种在精子竞争结果中起关键作用的近端表型。 交配插头排出计时允许雌性对其未来后代的父子关系施加控制。早期射血 交配插头的使用会减少雄性精子的数量，从而使雄性处于不利地位；反之亦然 插头的保留时间更长，使雄性具有亲子优势。男性和女性都对 配合插头及其弹出。在完成配对插头的雌性和雄性决定因素的GWA之后 射出时间和验证产生的基因，我们将进行网格交叉以确定是否存在变异 交配插头的时间主要由雄性、雌性或交互作用的遗传变异决定 在这两者之间。这些基因的后续扰动将开始解开连接男性的遗传途径 质量或其与配合插头顶出的感觉。在目标2中，我们将探讨我们最近的 发现交配导致雌性的一大套基因使用的不同外显子或启动子 大脑。有趣的是，其中的两个基因，desat1和觅食，在交配歧视或其他生殖方面起作用。 相关行为。每一个都强烈激活四个可选启动子中的一个，以响应交配。我们会 研究转录组以量化雄性因素的变异如何驱动交配后差异转录本的使用， 后者如何在女性基因型别中变化，以及这种调节是如何发生的。在目标3中，我们扩展和 推广旨在量化和预测成对互动结果的模型。古典的 布拉德利-特里模型，广泛应用于预测没有预测结果的球队之间的体育比赛 然而竞争，已直接类比于精子竞赛中的“竞赛”。评估是否适合这一类模型将 测试男性的适合度是否可以进行排序，以及这些排序是否能产生准确的结果 预测。我们将把这些模型扩展到目标1和目标2中的实验结果，以及 多年来在这个项目中执行，使用结果来评估男性x女性的总体重要性 每种情况下的相互作用。我们将研究的许多过程都显示出高度的进化 这意味着我们的结果将扩大我们对男性和女性在 生殖可能与人类特发性不孕症有关，可能涉及基因 合作伙伴之间的不相容。

PROJECT SUMMARY Sexual reproduction requires precise orchestration of expression of myriad genes in males and females. These genes mediate interactions between the sexes at molecular, cellular, neural and whole- organism levels, including the differential fertility success of sperm from two competing males that mate to a single female. In the current funding period, we used the tractable Drosophila model system to dissect the role of genes expressed in octopaminergic neurons in differential sperm use by a multiply-mated female. In this renewal application, we propose as Aim 1 a set of experiments that identify genes and quantify their effects on mating plug ejection, a proximal phenotype with a key role in sperm competition outcomes. Mating plug ejection timing allows females to exert control over paternity of their future offspring. Early ejection of a mating plug disadvantages the male by decreasing the number of his sperm that can be used; conversely longer retention of the plug gives the male a paternity advantage. Both males and females contribute to the mating plug and its ejection. After completing a GWAS for female and male determinants of mating plug ejection timing and validating the resulting genes, we will perform a grid cross to determine whether variation in mating plug timing is mostly determined by genetic variation in the male, or the female, or an interaction between the two. Follow-up perturbation of those genes will begin to unravel genetic pathways linking male quality or its perception with mating plug ejection. In Aim 2, we will pursue the implications of our recent discovery that mating induces differential exon or promoter use by a large suite of genes in the female brain. Intriguingly, two of these genes, desat1 and foraging, act in mating discrimination or other reproductive- relevant behaviors. Each strongly activates one of four alternative promoters in response to mating. We will survey the transcriptome to quantify how variation in male factors drive post-mating differential transcript use, how the latter varies across female genotypes, and how this regulation occurs. In Aim 3 we extend and generalize models designed to quantify and predict outcomes of pairwise interactions. The classical Bradley-Terry model, widely applied in predicting outcomes of sporting contests between teams that have not yet competed, has a direct analogy in sperm competition “contests.” Assessing fit to models of this class will test whether the males’ fitness can be rank-ordered, and whether those rank orders produce accurate outcome predictions. We will extend these models to the outcomes of the experiments in Aims 1 and 2, and those performed in this project over the years, using the results to assess the overall importance of male x female interactions in each case. Many of the processes that we will study show high levels of evolutionary conservation, implying that our results will expand our understanding of male x female interactions in reproduction that may have relevance to cases of idiopathic human infertility that may involve genetic incompatibility between the partners.