Gamete Interactions in Caenorhabditis elegans

秀丽隐杆线虫配子相互作用

• 批准号: 8081162
• 负责人: ANDREW W. SINGSON
• 金额: $ 5.81万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-14 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8081162
• 关键词: Address; Affect; Animals; Area; Binding; Biochemical; Biological; Biological Assay; Biological Models; Biological Process; Biology; Caenorhabditis elegans; Calcium Channel; Cell Communication; Cell membrane; Cells; Complement; Contraceptive Agents; Defect; Epidermal Growth Factor; Equipment and supply inventories; Event; Extracellular Domain; Fertility; Fertility Study; Fertilization; Genes; Genetic; Germ Cells; Goals; Immunofluorescence Immunologic; Infertility; Lead; Ligands; Link; Low Density Lipoprotein Receptor; Low-Density Lipoproteins; Medical; Modeling; Molecular; Molecular Genetics; Molecular Models; Movement; Mutagenesis; Nature; Nematoda; Oocytes; Organism; Pathway interactions; Phenotype; Process; Proteins; Reagent; Recombinants; Research; Role; Solid; Sperm-Ovum Interactions; Sterility; Surface; Testing; Vesicle; Work; base; design; economic implication; egg; egg surface sperm receptor; genetic analysis; in vivo; insight; molecular modeling; mutant; novel; public health relevance; receptor; reproductive; reproductive success; social; sperm cell; sperm function; tool

DESCRIPTION (provided by applicant): Fertilization is biological process with important medical, social and economic implications. From extensive study, the events of fertilization are known in some detail. However, the molecular underpinnings of these events generally remain elusive. Most previous work on fertilization has relied on biochemical and immunological approaches. Our work is groundbreaking in the application of classic genetic analysis to this vital area of research. My lab has been helping to pioneer the use of C. elegans for addressing the mechanisms of sperm-egg interactions. Many of the genetic and molecular tools developed for C. elegans are not available or are very difficult to utilize in other organisms traditionally used for studying fertilization. One of the most significant advantages of C. elegans is our ability to isolate and maintain mutants that affect sperm or eggs and no other cells. Previously, through the study of sterile mutants, we have identified some of the first sperm molecules required for productive gamete interactions in C. elegans. These sperm molecules include SPE-9 and SPE-38. SPE-9 is a sperm surface transmembrane molecule with an extracellular domain that contains ten epidermal growth factor (EGF)-like repeats. We hypothesize that SPE-9 functions as a ligand for an egg surface sperm receptor. The spe-38 gene encodes a novel four pass transmembrane molecule. Based on protein localization and genetic interaction studies, we hypothesize that SPE-38 regulates the localization and/or activity of a calcuim channel (SPE-41/TRP-3) and other sperm molecules. We recently identified the first egg molecules required for fertilization in C. elegans. The egg-1 and egg-2 genes encode egg surface Low Density Lipoprotein (LDL) receptor repeat-containing proteins. We hypothesize that EGG-1 and EGG-2 function semi-redundantly as egg surface receptors for sperm during fertilization. The goal of this proposal is to further our understanding of fertilization in C. elegans by conducting the following experimental aims: 1) to further characterize the roles of SPE-9 and EGG-1/EGG-2 in fertilization by developing reagents to test for potential ligand-receptor interactions between them and/or other molecules. 2) to gain a better understanding SPE-38 and SPE-41/TRP-3 function in fertilization by investigating the connection between them through protein interaction studies and mutagenesis. 3) to clone the sperm function gene spe-36 in order to determine its molecular nature and role in fertilization. The spe-36 gene is defined by a sterile mutant phenotype that is identical to spe-9, spe-38 and spe-41/trp-3 mutants. This work will complement fertility studies in other organisms as well as provide insights into the mechanisms of cell-cell interactions and the diversity of reproductive strategies. PUBLIC HEALTH RELEVANCE: Although the events of fertilization are fairly well described for a number of species, the molecular underpinnings of the process are not well understood. We are identifying the molecules and molecular mechanisms of fertilization in the model system Caenorhabditis elegans. This work will complement studies of fertilization in other species and could eventually lead to a better understanding of the causes of infertility, the diversity of reproductive strategies and the design of new contraceptives.

描述（由申请人提供）：受精是具有重要医学，社会和经济影响的生物学过程。从广泛的研究中，详细了解受精的事件。但是，这些事件的分子基础通常仍然难以捉摸。以前关于受精的大多数工作都依赖生化和免疫学方法。我们的工作在将经典遗传分析应用于这一重要的研究领域中是开创性的。我的实验室一直在帮助开拓秀丽隐杆线虫来解决精子互动的机制。为秀丽隐杆线虫开发的许多遗传和分子工具都不可用，或者在传统上用于研究施肥的其他生物中很难使用。秀丽隐杆线虫最重要的优势之一是我们能够分离和维持影响精子或鸡蛋而没有其他细胞的突变体。以前，通过对无菌突变体的研究，我们已经确定了秀丽隐杆线虫中生产性配子相互作用所需的一些第一个精子分子。这些精子分子包括SPE-9和SPE-38。 SPE-9是一种精子表面跨膜分子，其细胞外结构域包含十个表皮生长因子（EGF）类似重复序列。我们假设SPE-9充当卵表面精子受体的配体。 SPE-38基因编码一个新型的四个通过跨膜分子。基于蛋白质的定位和遗传相互作用研究，我们假设SPE-38调节了Calcuim通道（SPE-41/TRP-3）和其他精子分子的定位和/或活性。我们最近确定了秀丽隐杆线虫中受精所需的第一个卵子分子。 Egg-1和Egg-2基因编码鸡蛋表面低密度脂蛋白（LDL）受体重复蛋白。我们假设Egg-1和Egg-2在受精过程中将Egg-1和Egg-2作为精子的卵表面受体发挥作用。该提案的目的是通过进行以下实验目的来进一步了解秀丽隐杆线虫中的施肥：1）通过开发试剂来测试试剂，以进一步表征SPE-9和Egg-1/Egg-2在受精中的作用。它们与/或其他分子之间的潜在配体受体相互作用。 2）通过通过蛋白质相互作用研究和诱变来研究它们之间的联系，以更好地了解SPE-38和SPE-41/TRP-3功能。 3）克隆精子基因SPE-36，以确定其分子性质和在受精中的作用。 SPE-36基因由与SPE-9，SPE-38和SPE-41/TRP-3突变体相同的无菌突变表型定义。这项工作将补充其他生物体的生育研究，并提供对细胞 - 细胞相互作用机制以及生殖策略多样性的见解。公共卫生相关性：尽管对许多物种的施肥事件得到了很好的描述，但该过程的分子基础尚不清楚。我们正在识别模型系统秀丽隐杆线虫中受精的分子和分子机制。这项工作将补充其他物种的施肥研究，并最终可能会更好地理解不育的原因，生殖策略的多样性和新避孕药的设计。