Gamete Interactions in Caenorhabditis elegans

描述：(申请人提供) 受精是一个细胞生物学过程，具有重要的医学、社会和社会意义 对经济的影响。配子的基本细胞生物学功能 相互作用在所有细胞中都是保守的，无论它们是体细胞还是 生殖系起源。尽管进行了密集的研究，精子和卵子的相互作用仍然 在分子水平上知之甚少。大多数以前关于受精的工作 依赖于生化和免疫学方法，而遗传方法 一直以来都很缺乏。 我们率先使用线虫线虫秀丽线虫 阐述精子-卵子相互作用的分子机制。有权势的 为蠕虫开发的经典和分子遗传学工具不是 传统上在其他生物体中可用或很难利用 用于研究受精。线虫的生殖生物学 有助于识别影响精子的突变，而不是其他 细胞。这些突变为确定精子成分提供了一个独特的机会 精子和卵子相互作用所必需的。具有两个相互作用的突变的蠕虫 Spe-9和spe-13基因产生具有野生型形态的精子和 即使在配子之间接触后也不能使卵母细胞受精的能动性。 因此，Spe-9或Spe-13功能的中断会影响配子 识别、黏附、信号和/或融合。Spe-9基因编码精子。 具有胞外结构域的跨膜蛋白，含有10个表皮 生长因子(EGF)样重复序列。蛋白质的一个共同特征包括 类EGF基序参与细胞外功能，如 粘附剂和配体-受体的相互作用。这些结果与 SPE-9在专门化细胞间相互作用中的作用假说 受精所需的。 为了更准确地确定SPE-9在受精过程中的作用，我们 将确定其在精子中的定位，并揭示细胞区域 对配子相互作用很重要。此外，我们将调查如何 蛋白质的不同结构域对其生物学功能有贡献。我们会 克隆spe-13基因并对其基因产物进行分析。这一信息将是 有助于建立关于SPE-13在体内作用的分子模型 受精和可能与spe-9的相互作用。我们将启动 几个新基因的特征，当突变时，这些基因似乎具有表型复制 Spe-9和Spe-13突变。这些基因有望编码更多的精子。 受精所需的成分。最后，我们将进行基因筛查 确定SPE-9的卵母细胞受体和其他必需的基因 精子和卵子的相互作用。这项工作将对细胞-细胞提供新的见解 其他受精的互作、受精概念及互补研究 有机体。

DESCRIPTION: (provided by applicant) Fertilization is a cell biological process with important medical, social and economic implications. The underlying cell biological functions of gamete interactions are conserved in all cells regardless of their somatic or germ-line origins. Despite intense study, sperm-egg interactions are still poorly understood at the molecular level. Most previous work on fertilization has relied on biochemical and immunological approaches while a genetic approach has been lacking. We are pioneering the use of the nematode worm Caenorhabditis elegans for addressing the molecular mechanisms of sperm-egg interactions. The powerful tools of classical and molecular genetics developed for the worm are not available or are very difficult to utilize in the other organisms traditionally used for studying fertilization. The reproductive biology of C. elegans facilitates the identification of mutations that affect sperm and no other cells. These mutations provide a unique opportunity to define sperm components required for sperm-egg interactions. Worms with mutations in two interacting genes, spe-9 and spe-13, produce spermatozoa with wild-type morphology and motility that cannot fertilize oocytes even after contact between gametes. Therefore, disruption of spe-9 or spe-13 function affects either gamete recognition, adhesion, signaling and/or fusion. The spe-9 gene encodes a sperm transmembrane protein with an extracellular domain that contains ten epidermal growth factor (EGF)-like repeats. A common feature of proteins that include EGF-like motifs is their involvement in extracellular functions such as adhesive and ligand-receptor interactions. These results are consistent with the hypothesis that SPE-9 functions in the specialized cell-cell interactions required for fertilization. In order to more precisely define the role of SPE-9 during fertilization, we will determine its localization in sperm and reveal the cellular regions important for gamete interactions. Furthermore, we will investigate how different domains of the protein contribute to its biological function. We will clone the spe-13 gene and analyze its gene product. This information will be useful in formulating a molecular model concerning the role of SPE-13 during fertilization and possible interactions with SPE-9. We will initiate the characterization of several new genes that when mutated appear to phenocopy the spe-9 and spe-13 mutations. Such genes are expected to encode additional sperm components required for fertilization. Finally, we will conduct genetic screens to identify the oocyte receptor for SPE-9 and other genes required for sperm-egg interactions. This work will provide new insight into cell-cell interactions, conception and complement studies of fertilization in other organisms.