Gamete Interactions in Caenorhabditis elegans

说明(申请人提供)：受精是一种生物过程，具有重要的医学、社会和经济意义。通过广泛的研究，人们对受精事件有了更详细的了解。然而，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于生化和免疫学方法。我们的工作在将经典基因分析应用于这一重要研究领域方面具有开创性。原理：许多为线虫开发的遗传和分子工具在传统上用于研究受精的其他生物中是不可用的或非常困难的。线虫最显著的优势之一是能够分离和保持影响精子或卵子的突变株，而不影响其他细胞。以前，通过对不育突变体的研究，我们已经确定了线虫中产生配子相互作用所需的一些第一批分子。我们的目标是更好地了解精子分子(SPE-9、SPE-38、SPE-41/Trp-3和SPE-45/Izumo)和卵母细胞分子(卵子-1/2)如何发挥作用以确保成功受精。SPE-45/Izumo基因代表了从蠕虫到人类的保守的精子功能。此外，如果没有更完整的分子组成清单，就不可能全面了解受精的分子机制。因此，我们的目标是通过遗传和分子分析来鉴定新的受精分子。将特别强调识别卵母细胞所需的基因。目的：本研究的目的是通过以下实验目的来加深我们对线虫受精的理解：1)确定受精缺陷突变表型所定义的新配子功能基因的分子性质。2)研究已发现的配子功能分子的机制连接、功能和受精途径。3)利用丰富配子功能突变体的创新遗传筛选策略，对受精突变体进行饱和诱变。这项工作将补充其他生物的生育研究，并为细胞-细胞相互作用的机制和生殖策略的多样性提供见解。

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. RATIONALE: 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 fo studying fertilization. One of the most significant advantages of C. elegans is the 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 molecules required for productive gamete interactions in C. elegans. We aim to get a better understanding how sperm molecules (SPE-9, SPE-38, SPE-41/TRP-3 and SPE- 45/Izumo) and oocyte molecules (EGG-1/2) functions to insure successful fertilization. The SPE-45/Izumo gene represents a conserved sperm function from worms to humans. Further, a full understanding of the molecular mechanisms of fertilization is impossible without a more complete inventory of molecular components. Therefore we aim to identify new fertilization molecules through genetic and molecular analysis. Particular emphasis will be placed on the identification of genes required in oocytes. OBJECTIVES: The goal of this proposal is to further our understanding of fertilization in C. elegans by conducting the following experimental aims: 1) Determine the molecular nature of new gamete function genes defined by a fertilization defective mutant phenotype. 2) Investigate mechanistic connections, functions and fertilization pathways of previously identified gamete function molecules. 3) Conduct a saturation mutagenesis for fertilization mutants taking advantage of an innovative genetic screening strategy that enriches for gamete function 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