Drosophila Genes Causing Male X Female Interactions in Reproduction

描述（由申请人提供）：精子和卵子的特定组合成功形成受精卵的决定因素涉及无数的分子、细胞、神经和整个生物体行为属性。我们过去的工作主要集中在多交配雌性果蝇精子差异成功的现象学上，并且筛选影响这种精子竞争的基因已经阐明了这个过程的许多方面。我们持续努力的第一个目标是确定女性生殖道中表达的基因在精子成功中的作用。为了剖析这些基因在精子使用中的功能作用，我们将应用高度靶向的RNAi敲除，并评估这些基因在女性生殖道中表达缺失的影响。这很容易用UAS-GAL 4系统完成，使用在雌性生殖道中表达的GAL 4驱动因子。明智的标准将被应用于选择敲除的候选人，我们特别感兴趣的是在女性中表达的一组基因，具有神经功能。将在一组精子竞争试验中针对一组16个近交序列已知的雄性和精液蛋白质空值测试雌性应答基因的敲除，以尝试量化雄性x雌性相互作用以及测试特定相互作用。我们的第二个目标是量化男性之间的遗传差异，在他们的能力，影响交配后转录组反应的女性，和这些表达变化的精子竞争的作用。雌性将与从果蝇遗传参考组（Drosophila Genetic Reference Panel）（其基因组序列和基因表达数据均已可用）中抽取的雄性交配，将解剖生殖道并进行RNA测序。分析将提供一个详细的图片，哪些基因的转录响应交配，哪些基因不同的男性基因型。这些基因将依次在女性中被敲除，以观察它们是否在精子竞争中发挥作用。由于我们研究的基因变异涉及基本的生殖功能，我们的研究结果将扩大理解特发性人类不育症病例的基础，特别是那些无法确定不育症的性别特异性原因的病例，因此可能反映了伴侣之间的遗传不相容性。

DESCRIPTION (provided by applicant): The determinants of which particular combinations of sperm and egg are successful in forming a zygote involve a myriad of molecular, cellular, neurological and whole-organism behavioral attributes. Our past work has focused on the phenomenology of differential success of the sperm in multiply mated female Drosophila, and screens for genes that impact this sperm competition have illuminated many aspects of the process. The first aim of our continuing effort is to determine the role(s) in sperm success of genes expressed by the female in her reproductive tract. To dissect the functional role of these genes in sperm use, we will apply highly targeted RNAi knockdown and assess the impact of loss of expression of these genes in the female reproductive tract. This is readily accomplished with the UAS-GAL4 system, using GAL4 drivers that express in the female reproductive tract. Judicious criteria will be applied to select candidates for knockdown and we are especially interested in the set of genes expressed in females that have neurological function. Knockdowns of female response genes will be tested in a battery of sperm competition tests against a set of 16 inbred sequence-known males and seminal protein nulls to try to quantify male x female interaction as well as testing specific interactions. Our second aim is to quantify genetic differences among males in their ability to influence post-mating transcriptome responses in females, and the roles of those expression changes on sperm competition. Females will be mated with males drawn from the Drosophila Genetic Reference Panel (for which both genome sequence and gene expression data are already available) and reproductive tracts will be dissected and subjected to RNA-sequencing. Analysis will provide a detailed picture of which genes respond transcriptionally to mating, and which genes respond differentially to different male genotypes. These genes will in turn be knocked down in females to see if they play a role in sperm competition. Because the genes whose variation we examine involve basic reproductive functions, our results will expand the foundation for understanding cases of idiopathic human infertility, specifically those in which no sex-specific cause for the infertility can be identified and may thus reflect genetic incompatibility between the partners.