Accumulation, Storage, and Release of Sperm in the Oviduct

精子在输卵管中的积累、储存和释放

• 批准号: 10179435
• 负责人: DAVID Joel MILLER
• 金额: $ 26.58万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-04 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10179435
• 关键词: Abnormal placentation; Adhesions; Affect; Affinity; Affinity Chromatography; Animals; Assisted Reproductive Technology; Behavior; Binding; CRISPR/Cas technology; Cell membrane; Cells; Complex; Conditioned Culture Media; Deposition; Development; Embryo; Epithelial Cells; Estrus; Family suidae; Fertility; Fertilization; Fertilization in Vitro; Genes; Genetic; Germ Cells; Glycocalyx; Goals; Human; Immobilization; Infertility; Insemination; Intracytoplasmic Sperm Injections; Knowledge; Lead; Longevity; Low Birth Weight Infant; Mammalian Oviducts; Membrane; Methods; Modeling; Molecular; Mus; Mutate; Oocytes; Polysaccharides; Premature Birth; Process; Production; Progesterone; Prostaglandins; Proteins; Research; Risk; Seminal fluid; Signal Transduction; Signaling Molecule; Site; Sperm Count Procedure; Sperm-Ovum Interactions; Sulfate; System; Techniques; Tissues; Trisaccharides; Tube; Uterus; cell motility; cost; experimental study; fertility improvement; improved; insight; receptor; reproductive; response; sperm cell; sperm viability; stillbirth; success; tool; zygote

Project Summary Fertility depends on successful fertilization and early development, processes that occur in the oviduct. Common therapies for human infertility, such as in vitro fertilization and intracytoplasmic sperm injection, are expensive and increase risks of a variety of problems. More knowledge of how the oviduct interacts with sperm, the cumulus-oocyte complex (COC) and the developing embryo may improve fertility and reduce the need for therapies. The oviduct serves as a reservoir for sperm, after semen deposition and before fertilization. Binding to the oviduct maintains sperm viability and suppresses motility. Sperm are released to move to the upper oviduct (ampulla) to fertilize oocytes. There are many gaps in this model of sperm-oviduct interaction. Our studies have begun to fill some of these gaps. We have used a glycomic approach to screen 377 glycans and found that all glycans with affinity for porcine sperm have either of two motifs, sulfated Lewis X trisaccharide or branched 6-sialylated complex glycans. We have identified two candidate receptors for both glycans on the sperm membrane, PKDREJ and ADAM5. Notably, mouse sperm deficient in PKDREJ and other ADAMs do not accumulate beyond the utero-tubal junction, but it is not known if this is due to a problem in binding and retention in the oviduct. Remarkably, if these glycans are immobilized on beads, they can extend sperm lifespan, much like binding to oviduct cells prolongs the lifespan of sperm. Finally, we found that cumulus-oocyte complexes (COCs) secrete molecules that signal sperm release from the lower oviduct so they can move toward the site of fertilization. The Specific Aims will provide a mechanistic understanding of how sperm bind the oviduct, how binding prolongs sperm lifespan and how sperm are released from the oviduct by the COC. Aim 1: To determine the function of PKDREJ and ADAM5 in sperm by blocking each protein and mutating each gene in swine. Sperm from pigs that are deficient in each of these proteins will be examined to determine if their ability to bind oviduct cells and their fertility is affected. Aim 2. To determine if sperm binding to immobilized oviduct glycans suppresses ROS production, conserves ATP, and stabilizes the plasma membrane, which extends sperm lifespan and fertility. Sperm bound to immobilized glycans will be examined to ascertain what changes adhesion induces. The second group of experiments will determine which changes are sufficient to prolong lifespan by adding agents that scavenge ROS and stabilize the plasma membrane. Aim 3. To determine how the cumulus-oocyte complex signals release of sperm from oviduct cells and glycans. We will examine whether progesterone, prostaglandins, or proteins that may be secreted from COCs might induce porcine sperm release from oviduct cells and immobilized glycans. The completion of these Specific Aims will elucidate how sperm bind to the oviduct, are stored in the oviduct and are released in response to the cumulus-oocyte complex. This fundamental information could be used to develop simpler, safer and less costly assisted reproductive technologies.

项目摘要 生育力取决于成功的受精和早期发育，这些过程发生在输卵管中。共同 用于人类不育症的疗法，例如体外受精和胞浆内精子注射，是昂贵的， 增加了各种问题的风险。更多关于输卵管如何与精子、卵丘-卵母细胞相互作用的知识 复合体（COC）和发育中的胚胎可以提高生育能力并减少对治疗的需求。输卵管作为 在精子沉积后和受精前储存精子。与输卵管结合可维持精子活力， 抑制运动。精子被释放到输卵管上部（壶腹）使卵母细胞受精。存在不少差距 在这个精子输卵管相互作用的模型中。我们的研究已经开始填补其中的一些空白。我们使用了一种糖分析剂 筛选377种聚糖的方法，并发现所有对猪精子具有亲和力的聚糖都具有两种基序中的任一种， 硫酸化刘易斯X三糖或支链6-唾液酸化复合聚糖。我们已经确定了两个候选受体， 精子膜上的两种聚糖，PKDREJ和ADAM 5。值得注意的是，小鼠精子缺乏PKDREJ和其他 亚当斯不会在子宫输卵管交界处以外积聚，但尚不清楚这是否是由于 结合并滞留在输卵管中。值得注意的是，如果这些聚糖固定在珠子上，它们就可以延伸精子 寿命，就像与输卵管细胞结合一样，延长了精子的寿命。最后，我们发现卵丘-卵母细胞 复合体（COCs）分泌分子，信号精子从输卵管下部释放，使它们能够向生殖器部位移动。 受精具体目标将提供精子如何结合输卵管的机械理解，如何结合 精子的寿命以及COC如何从输卵管释放精子。目标1：确定 通过阻断猪精子中的PKDREJ和ADAM 5蛋白质并使每个基因突变。猪的精子 将检查这些蛋白质中的每一种都缺乏的小鼠，以确定它们结合输卵管细胞的能力和它们的生育能力。 受到影响。目标2.为了确定精子与固定的输卵管聚糖结合是否抑制ROS的产生， 保存ATP，稳定质膜，延长精子寿命和生育能力。精子结合到 将检查固定化的聚糖以确定粘附诱导的变化。第二组实验将 通过添加抑制ROS和稳定血浆的试剂，确定哪些变化足以延长寿命 膜的目标3.为了确定卵丘-卵母细胞复合体是如何从输卵管细胞释放精子的信号， 聚糖我们将研究孕酮、肾上腺素或可能从COCs分泌的蛋白质是否可能 诱导猪精子从输卵管细胞和固定聚糖中释放。这些具体目标的实现将 阐明精子如何与输卵管结合，储存在输卵管中，并在卵丘卵母细胞的反应中释放 复杂.这些基本信息可用于开发更简单，更安全，更便宜的辅助生殖技术。 技术.