Gamete Interactions in Caenorhabditis elegans

描述：（申请人提供） 受精是一种细胞生物学过程，具有重要的医学、社会和医学意义。 经济影响。配子的基本细胞生物学功能 相互作用在所有细胞中是保守的，无论它们的体细胞或 生殖系起源尽管有大量的研究，精子和卵子的相互作用仍然是 在分子水平上知之甚少。以前关于受精的大多数工作 依赖于生物化学和免疫学方法， 一直缺乏。 我们正在开创性地使用线虫秀丽隐杆线虫， 解决精子-卵子相互作用的分子机制。强大的 针对蠕虫开发的经典和分子遗传学工具并不 传统上在其他生物体中是可用的或很难利用的 用来研究施肥。对C. elegans 有助于识别影响精子的突变，而不是其他突变。 细胞这些突变为定义精子成分提供了独特的机会 精子和卵子相互作用所必需的。两个相互作用的基因突变的蠕虫 基因SPE-9和SPE-13产生具有野生型形态的精子， 即使在配子之间接触后也不能使卵母细胞受精的运动性。 因此，SPE-9或SPE-13功能的破坏会影响任一配子 识别、粘附、信号传导和/或融合。SPE-9基因编码一个精子 一种跨膜蛋白，其胞外结构域包含10个表皮 EGF（EGF）的作用蛋白质的一个共同特征包括 EGF样基序是它们参与细胞外功能， 粘附和配体-受体相互作用。这些结果是一致 SPE-9在专门的细胞间相互作用中发挥作用的假设 需要受精。 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其细胞区域 对配子相互作用很重要。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。此信息将 有助于制定关于SPE-13在 受精和可能的相互作用与SPE-9。我们将启动 几个新的基因的特征，当突变出现表型复制， SPE 9和SPE 13突变。这样的基因有望编码额外的精子 受精所需的成分。最后，我们将进行基因筛查， 为了鉴定SPE-9的卵母细胞受体和其他基因， 精卵相互作用这项工作将提供新的见解细胞-细胞 相互作用，概念和补充研究的受精在其他 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.