Actions of Seminal Proteins in Mated Drosophila Females

精液蛋白在交配果蝇雌性中的作用

基本信息

批准号：
10266142
负责人：
Mariana Federica Wolfner
金额：
$ 37.43万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10266142
关键词：
Affect Animals Area Assisted Reproductive Technology Behavior Binding Binding Proteins Biochemical Biochemistry Biological Assay Biological Models Biology CRISPR/Cas technology Cell Culture Techniques Conflict (Psychology)Cysteine Data Dengue Diagnosis Directed Molecular Evolution Disease Drosophila genus Drosophila inturned protein Evolution Female Fertility Funding Future Gene Expression Genetic Genetic Models Genetic Screening Genome Gland Grant Health Human In Vitro Individual Infertility Insecta Invertebrates Investigation Lectin Ligands Location Malaria Male Infertility Mammalian Oviducts Mammals Mass Spectrum Analysis Measures Mediating Methods Modeling Molecular Molecular Conformation Molecular Genetics Mosquito Control Mus Muscle Contraction Mutation Nature Neurons Organism Orthologous Gene Ovulation Partner in relationship Pathway interactions Peptide Hydrolases Peptides Phenotype Physiological Physiology Play Prostate Proteins Proteome Proteomics Reproduction Reproductive Physiology Resources Role Seminal Seminal Plasma Seminal Vesicles Seminal fluid Signal Transduction Site Sterility Structure System Testing Variant Work X-Ray Computed Tomography ZIKA base design experimental study genetic signature human disease improved insect disease vector interest male male fertility microCT neuromuscular offspring protein function receptor reproductive reproductive success reproductive tract response secretory protein sex sperm cell sperm function sperm protein tool

项目摘要

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.

概括从昆虫到哺乳动物，半流体蛋白（SFP）显着影响交配的女性，精子在女性内部的储存和释放，（在小鼠中）甚至表型后代。特定SFP的雄性异常是无菌或子系数，包括人类。一些SFP紧密结合精子；其他人则是半多花体的免费。目的地它们在繁殖中的重要性，对 SFP如何影响女性或精子在女性中的行为。重要的是，许多SFP 迅速发展，与分子/进化性冲突中的作用一致。了解功能对SFPS的演变和与女性相互作用的蛋白质的限制将指导未来调查人类生育中的SFP行动。我们将结合分子遗传和功能方法来研究：（1）SFP如何与女性分子引起生殖反应，（2）SFP与精子如何介导其介导效果以及两种功能的发展方式。我们将使用果蝇是一种用于剖析SFP功能的首要遗传模型系统，并拥有广泛的资源进化比较。重要的是，果蝇SFP具有许多分子和现象学的相似之处给哺乳动物的人。 AIM 1专注于卵蛋白，该卵蛋白通过诱导的神经元章鱼能信号传导刺激排卵。这种信号传导调节女性生殖道的肌肉收缩，放松输卵管和排卵率提高。使用遗传筛选和蛋白质 - 蛋白质共同进化的特征，我们有确定了女性卵蛋白受体（OVR）的强大候选者。我们将测试这些卵蛋白结合和然后确定OVR定位，以查明卵蛋白作用的位点。然后，我们将研究不同的效果物种的卵蛋白介导卵蛋白的作用和OVR结合，以阐明其功能的演变。在AIM 2中，我们将重点关注与精子结合的第二种蛋白或通过蛋白质组学方法。我们最近的数据表明，SFPS Prime精子用于与关键的SFP结合称为性别肽。我们将询问哪种半蛋白在此启动途径中起作用，哪些半蛋白独立行动那条路。我们还将调查女性分泌物是否也参与启动。最后，我们会的确定在相关的相关的SFP子集的子集的功能范围的程度果蝇物种。阐明SFP与分子水平上的女性相互作用并影响女性的方式，以及这些相互作用的演变，对于理解和诊断基于SFP的地下实用性很重要，在考虑辅助生殖技术的策略，这些技术将受益于关键SFP，以及开发新的方法来控制双翅目损伤，从而传播登革热，寨卡病毒和疟疾等严重疾病。