Gene expression and functional evolution in the Drosophila female reproductive tract

项目总结 果蝇雌性生殖道在形态和功能上都很复杂，作为 多个男性射精和女性产品之间的高风险互动上演，这也是可能的主题 强大且相互冲突的选择压力。此外，生殖道是级联反应的起始点。 雌性生物学中的交配后反应，从隔离精子和产卵到神经变化 基因表达和行为。这些过程中的许多过程可能会经历性对抗选择，因为 结果会影响到两性不同的生育兴趣。尽管如此，研究投入较低 女性生殖道，包括精子储存器官和女性附腺，严重滞后 对男性生殖组织的投资。例如，精囊、子宫和雌性附件 腺体已被排除在所有主要社区的转录组分析之外；资源，如Gal-4驱动程序 对于这些组织，通常不存在标记基因标记；以及几个器官的功能和进化 仍然没有得到充分的描述。这些遗漏限制了对一系列重要问题的洞察，包括 雌性-射精相互作用的机制和进化，生殖组织中快速进化的程度 和它的驱动因素，以及性冲突的结果。然而，这些遗漏也突显了新的 在领先的模式生物中使用成熟的技术进行低障碍、高回报的调查， 这项提议将继续下去。首先，对交配和未交配的雌性从两个不同的自然物种中进行单细胞测序 将首次使用种群来表征女性生殖道中的细胞类型多样性 2)测量5个体细胞雌性生殖组织的基因表达和差异 决议(具体目标1)。这些数据还将检验这样一种假设，即女性体内的基因表达 由于与精液蛋白的共同进化相互作用，生殖组织迅速分化。接下来，基因 作为重要功能进化的候选者将被淘汰并评估其生殖能力 效果。特异性目标2测试7个受体的零等位基因，这些受体在精囊中表达并显示 快速分化的迹象，以检验这些受体介导雌性交配后交配的假设 回应。特定目标3使用一组在精囊中有新表达的从头基因来 探索基因诞生和功能整合的神秘过程。具体地说，从头基因 根据文字记录长度、表达水平 结构复杂性和其他指标将被剔除，并评估对女性生殖的影响 功能。

PROJECT SUMMARY The Drosophila female reproductive tract is morphologically and functionally complex, and as the arena where high-stakes interactions among multiple male ejaculates and female products play out, it is also likely subject to strong and conflicting selection pressures. Moreover, the reproductive tract is the initiation site for a cascade of post-mating responses in female biology, from sequestering sperm and ovipositing to changes in neural gene expression and behavior. Many of these processes may undergo sexually antagonistic selection, as outcomes affect the sexes’ divergent reproductive interests. Nevertheless, research investment in the lower female reproductive tract, including the sperm storage organs and the female accessory glands, severely lags investment in male reproductive tissues. For example, the seminal receptacle, uterus, and female accessory glands have been excluded from every major community transcriptome analysis; resources like Gal-4 drivers or marker gene labels generally do not exist for these tissues; and the function and evolution of several organs remain poorly described. These omissions limit insight into a range of important questions, including the mechanisms and evolution of female-ejaculate interactions, the extent of rapid evolution in reproductive tissues and its drivers, and the outcomes of sexual conflict. Yet these omissions also highlight an opportunity for new low-barrier, high return investigations using well-established technologies in a leading model organism, which this proposal pursues. First, single-cell sequencing of mated and unmated females from two diverged natural populations will be used to 1) characterize cell-type diversity in the female reproductive tract for the first time and 2) measure gene expression and divergence in the 5 somatic female reproductive tissues at cellular resolution (Specific Aim 1). These data will also test the hypothesis that gene expression in female reproductive tissues diverges rapidly owing to coevolving interactions with seminal fluid proteins. Next, genes that are candidates for important functional evolution will be knocked out and evaluated for reproductive effects. Specific Aim 2 tests null alleles of 7 receptors that are expressed in the seminal receptacle and show indications of rapid divergence, to test the hypothesis that these receptors mediate female post-mating responses. Specific Aim 3 uses a set of de novo genes with novel expression in the seminal receptacle to explore the mysterious process of gene birth and functional integration. Specifically, the de novo gene candidates that appear most likely to be functionally integrated based on transcript length, expression level, structural complexity, and other metrics, will be knocked out and evaluated for effects on female reproductive function.