REGULATION OF GAMETE USE AND NEURAL PATHWAYS IN REPRODUCTION

项目摘要 有性生殖需要精确地协调雄性和雌性的无数基因的表达， 女性这些基因在分子、细胞、神经和整体上介导两性之间的相互作用。 生物体水平，包括精子的差异生育成功从两个竞争的男性，交配到一个 单身女性在当前的资助期内，我们使用了易于处理的果蝇模型系统， 章鱼胺能神经元中表达的基因在多配偶女性的精子使用差异。在这 更新的应用，我们提出了一套实验，确定基因和量化他们的目标1 对交配栓排出的影响，一种在精子竞争结果中起关键作用的近端表型。 交配栓弹射时间允许女性对未来后代的父权施加控制。早期射血 交配栓的存在会减少雄性可以使用的精子数量，从而使雄性处于不利地位;相反， 插塞的较长保持时间给予男性父权优势。男性和女性都有助于 配合插头及其弹出。完成GWAS后，女性和男性的交配插头决定因素 喷射时间和验证产生的基因，我们将执行网格交叉，以确定是否变化， 交配插塞时间主要由雄性或雌性的遗传变异或相互作用决定 两人之间这些基因的后续扰动将开始解开连接男性和女性的遗传途径。 质量或其感知与配套插头弹出。在目标2中，我们将探讨我们最近的 发现交配诱导女性中大量基因使用差异外显子或启动子 个脑袋有趣的是，其中两个基因，desat 1和觅食，在交配歧视或其他生殖- 相关行为。每一个都强烈激活四种替代启动子中的一种以响应交配。我们将 调查转录组以量化雄性因素的变化如何驱动交配后差异转录物的使用， 后者如何在女性基因型中变化，以及这种调节如何发生。在目标3中，我们扩展和 推广旨在量化和预测成对相互作用结果的模型。经典 Bradley-Terry模型，广泛应用于预测运动比赛的结果， 但竞争，有一个直接的比喻，在精子竞争的“比赛”。评估是否适合此类模型将 测试男性的健康状况是否可以按等级排序，以及这些等级排序是否会产生准确的结果 预测。我们将把这些模型扩展到目标1和目标2的实验结果， 多年来在这个项目中进行的，使用结果来评估男性x女性的整体重要性 每种情况下的互动。我们将研究的许多过程都显示出高水平的进化 这意味着我们的研究结果将扩大我们对男性x女性相互作用的理解， 生殖可能与可能涉及遗传的特发性人类不育症病例有关 合作伙伴之间的不协调。

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.