Gamete Interactions in Caenorhabditis elegans

秀丽隐杆线虫配子相互作用

• 批准号: 6636648
• 负责人: ANDREW W. SINGSON
• 金额: $ 22.7万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6636648
• 关键词: Caenorhabditis elegans; alleles; cell cell interaction; egg /ovum; epidermal growth factor; fertilization; gene mutation; genetic screening; germ cells; green fluorescent proteins; immunofluorescence technique; ligands; membrane proteins; membrane transport proteins; molecular cloning; molecular dynamics; molecular genetics; mutant; phenotype; protein structure function; sperm

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.

描述：（申请人提供） 受精是一个重要的医学，社会和 经济影响。配子的基本细胞生物学功能 不论其体细胞或 种系起源。尽管进行了深入研究，但精子 - 蛋的相互作用仍在 在分子水平上了解不足。以前关于受精的大多数工作 依赖生化和免疫学方法，而遗传方法 缺乏。 我们正在开创秀丽隐杆线虫的线虫蠕虫的使用 解决精子相互作用的分子机制。强大的 为蠕虫开发的古典和分子遗传学工具不是 传统上可以使用或在其他生物中使用非常困难 用于研究施肥。秀丽隐杆线虫的生殖生物学 促进影响精子的突变的识别，没有其他 细胞。这些突变为定义精子组件提供了独特的机会 精子互动所需。两种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 即使配子之间接触后也无法施肥卵母细胞的运动。 因此，SPE-9或SPE-13功能的破坏会影响任何配子 识别，粘附，信号传导和/或融合。 SPE-9基因编码精子 跨膜蛋白具有一个含有十个表皮的细胞外域 生长因子（EGF）类似重复。包括蛋白质的共同特征 EGF样基序是它们参与细胞外功能，例如 粘合剂和配体 - 受体相互作用。这些结果与 SPE-9在专门的细胞 - 细胞相互作用中起作用的假设 受精所必需的。 为了更精确地定义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 对于配子互动很重要。此外，我们将研究如何 蛋白质的不同结构域有助于其生物学功能。我们将 克隆SPE-13基因并分析其基因产物。这些信息将是 在制定有关SPE-13作用的分子模型中有用 受精和与SPE-9的可能相互作用。我们将启动 几个新基因的表征，当突变时似乎是表观的 SPE-9和SPE-13突变。这些基因有望编码其他精子 受精所需的成分。最后，我们将进行遗传筛选 确定SPE-9和其他基因所需的卵母细胞受体 精子 - 蛋的相互作用。这项工作将为细胞细胞提供新的见解 其他其他人的相互作用，概念和补充研究 有机体。