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: 10736814
• 负责人: DAVID Joel MILLER
• 金额: $ 34.1万
• 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 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736814
• 关键词: Abnormal placentation; Adhesions; Affect; Affinity; Affinity Chromatography; Assisted Reproductive Technology; Binding; CRISPR/Cas technology; CatSper; Cells; Characteristics; Cholesterol; Citric Acid Cycle; Coenzyme A; Complex; CpG Islands; Deposition; Development; Electron Transport; Embryo; Embryonic Development; Epithelial Cells; Family suidae; Fertility; Fertilization; Fertilization in Vitro; Frequencies; Fumarates; Funding; Genes; Genetic; Germ Cells; Glycocalyx; Goals; Health; Human; Immobilization; In Vitro; Infertility; Insemination; Intracytoplasmic Sperm Injections; Knowledge; Lead; Longevity; Low Birth Weight Infant; Mammalian Oviducts; Membrane; Metabolic; Metabolism; Methods; Microscope; Modeling; Molecular; Mus; Mutate; Mutation; Oocytes; Ovary; Oxidative Phosphorylation; Phosphotransferases; Polysaccharides; Premature Birth; Process; Production; Progesterone; Proteins; Reactive Oxygen Species; Research; Risk; Seminal fluid; Signal Transduction; Site; Slide; Sperm Count Procedure; Sperm-Ovum Interactions; Sulfate; System; Techniques; Tissues; Translating; Trisaccharides; Ubiquinone; Ubiquitin; Uterus; cell motility; cost; cytotoxic; differential expression; diminished oxidative phosphorylation; fertility improvement; fundamental research; improved; in vivo; insight; methylome; methylomics; offspring; promoter; receptor; response; sialylation; sperm cell; sperm viability; stillbirth; success; tool; transcriptome; transcriptomics

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 the 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 or lead to the development of improved therapies (i.e. improvements in IVF). 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 but our studies have begun to fill some of these gaps. We have used a glycomic approach to screen hundreds of glycans and found that glycans with affinity for porcine sperm have either of two motifs, sulfated Lewis X trisaccharide or branched 6- sialylated complex glycans. We also identified two candidate receptors for both glycans on the sperm membrane, PKDREJ and ADAM5, that were not known to bind glycans. 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 or microscope slides, they can extend sperm lifespan, much like binding to oviduct cells prolongs the lifespan of sperm. Finally, we found that COCs secrete progesterone that signals sperm release from the lower oviduct by inducing hyperactivation so sperm can move toward the site of fertilization. The Specific Aims of this renewal will provide a mechanistic understanding of how sperm bind the oviduct, how binding prolongs sperm lifespan, and how these results may be translated to improve IVF. 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 have mutations in these genes have been produced. Sperm that are deficient in each of these proteins will be examined to determine if their ability to bind oviduct cells and their fertility are affected. Aim 2. To determine if sperm binding to glycans diminishes oxidative phosphorylation and the citric acid cycle to lengthen sperm lifespan. Sperm bound to immobilized glycans will be examined to ascertain the specific metabolic changes that are induced and the intracellular signaling that leads to these changes. Aim 3. To determine if oviduct glycans select superior sperm for storage and in vitro fertilization. We will examine whether sperm selected by glycan adhesion have improved characteristics themselves and also produce embryos that more closely resemble in vivo-produced embryos by comprehensive analysis of embryo transcriptomes and methylomes. The completion of these Specific Aims will provide important keys to resolving how sperm bind to the oviduct, are stored in the oviduct, and are released in response to the COC. This fundamental information could be used to develop simpler, safer, and less costly assisted reproductive technologies.

项目摘要 生育能力取决于成功的受精和早期发育，这是输卵管中发生的过程。常见疗法 对于人类不育症，例如体外受精和胞质内的精子注射，很昂贵，并且增加了 各种问题的风险。更多了解输卵管如何与精子相互作用，积云 - 元素复合物 （COC），发展中的胚胎可能会提高生育能力，并减少对疗法的需求或导致发展 改进的疗法（即IVF的改进）。椭圆形充当精子的储层，精液沉积和 受精之前。与输卵管的结合保持精子生存能力并抑制运动性。精子被释放以移动 到上卵形（ampulla）以施肥卵母细胞。这种精子 - 植物相互作用的模型中有很多差距，但是我们 研究已经开始填补其中一些空白。我们已经使用糖胶方法来筛选数百种聚糖并发现 对猪精子亲和力的聚糖具有两个基序中的含有硫化的刘易斯X trisaccharide或分支6-- 尖晶的复合物聚糖。我们还确定了两个聚糖在精子膜上的两个候选受体， PKDREJ和ADAM5，尚不知道会结合聚糖。值得注意的是，鼠标精子缺乏PKDREJ和其他 亚当斯不会累积在子宫大管交界处，但尚不清楚这是否是由于问题 输卵管中的结合和保留。值得注意的是，如果将这些聚糖固定在珠子或显微镜载玻片上，则 可以延长精子寿命，就像结合与卵形细胞的结合可以延长精子的寿命。最后，我们发现COC 通过诱导过度激活来分泌孕激素，该孕酮信号从下输卵管释放出来，以便精子可以移动 走向受精部位。这种续签的具体目的将提供对精子的机械理解 结合输卵管，结合如何延长精子寿命，以及如何翻译这些结果以改善IVF。目标1： 通过阻断每种蛋白质并突变每个基因，确定PKDREJ和ADAM5在精子中的功能 猪。已经产生了这些基因突变的猪的精子。这些精子不足 将检查蛋白质，以确定其结合输卵管细胞的能力及其生育能力。目标2 确定精子与甘氨酸的结合是否会减少氧化磷酸化和柠檬酸循环以延长 精子寿命。将检查与固定聚糖的精子，以确定特定的代谢变化 被诱导并导致这些变化的细胞内信号传导。目的3。确定卵形甘油是否选择 用于储存和体外受精的优质精子。我们将检查通过聚糖粘附选择的精子是否具有 改善特征本身，还产生胚胎，与体内生产的胚胎更相似 胚胎转录组和甲基组的综合分析。这些特定目标的完成将提供 解决精子如何与输卵管结合的重要关键，存储在输卵管中，并释放以响应于 COC。这些基本信息可用于开发更简单，更安全且成本较低的辅助生殖 技术。