GENETICS OF CAENORHABIDITIS ELEGANS SPERM MORPHOGENESIS

秀丽隐杆线虫精子形态发生的遗传学

• 批准号: 3467284
• 负责人: STEVEN W. L'HERNAULT
• 金额: $ 9.29万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3467284
• 关键词: Caenorhabditis elegans; alleles; cell differentiation; complementary DNA; cytogenetics; developmental genetics; electron microscopy; endonuclease; gene expression; gene mutation; genetic manipulation; genetic mapping; genetic recombination; genetically modified animals; histogenesis; laboratory rabbit; microscopy; molecular cloning; molecular genetics; morphology; nucleic acid hybridization; nucleic acid sequence; spermatogenesis

The purpose of this project is to identify and characterize genes involved in generating a specialized cell during its differentiation. The cell chosen for study is the amoeboid sperm of the nematode C. elegans because every cellular stage that appears during its differentiation can be genetically dissected, differentiation will occur in vitro and be gentically dissected, differentiation will occur in vitro and sufficient material for biochemical analysis is obtainable. Four of the over 30 genes known to affect spermatogenesis in C. elegans have been selected for further genetic, cytological and molecular analysis. Each of these four genes arrests development at a different cytological stage and will allow analysis of the intracellular events at these stages of spermatogenesis. The specific aims are: (1) to isolate additional alleles of these four spermatogenesis defective (spe) mutants especially rearrangements and transposon insertions that will aid in their molecular analysis; (2) to build on the existing phenotypic data to identify how these 4 genes alter sperm morphogenesis; (3) to search for suppressors of these 4 genes to determine how they interact with other sperm proteins; (4) to clone these 4 spe genes taking advantage of the growing genomic physical map (already 90 percent) of overlapping cosmid clones and/or transposon insertion mutants whenever possible. Transgenic worms will be created to confirm that the sep gene has been cloned. The sequence of the cloned gene might suggest its function and will permit generation of an antibody, allowing additional cytological analysis of these 4 spe genes. This project has general significance because it is likely that the principles of cellular differentiation are similar in all organisms. Genetic methods can be used to define and alter parameters that control cell shape and the assymetric partitioning of cellular components during division in this system, phenomena that are clearly abnormal in cancerous cells but are very difficult to analyze. This research will also be useful for the study of spermatogenesis in mammals where both limited genetics and association of spermatocytes with the Sertoli cell makes it difficult to identify and study sperm specific genes.

该项目的目的是识别和表征基因 在分化过程中参与产生特化细胞。 选择的研究细胞是线虫的变形精子 C.因为每一个细胞阶段，出现在其 分化可以从遗传学上进行解剖，分化将 发生在体外，并进行遗传解剖，分化将 发生在体外，并且有足够的材料用于生化分析， 可获得的。 已知的30多个基因中有4个 精子发生在C.已经选择了秀丽线虫， 遗传学、细胞学和分子分析。 这四 基因在不同的细胞学阶段阻止发育， 将允许分析在这些阶段的细胞内事件， 精子发生 具体目标是：（1）分离出附加的 这四个精子发生缺陷突变体的等位基因 特别是重组和转座子插入， （2）建立在现有的表型基础上， 确定这4个基因如何改变精子形态发生的数据;（3） 寻找这4个基因的抑制因子，以确定它们是如何 与其它精子蛋白相互作用;（4）克隆这4个spe基因 利用不断增长的基因组物理图谱（已经有90 重叠粘粒克隆和/或转座子插入 变种人尽可能 转基因蠕虫将被创造出来， 证实sep基因已被克隆。 的序列 克隆的基因可能会提示它的功能， 的抗体，允许额外的细胞学分析，这4 spe基因 该项目具有普遍意义，因为它是 细胞分化的原理可能与 所有的有机体。 遗传学方法可以用来定义和改变 控制像元形状和不对称划分的参数 在这个系统中细胞成分分裂的过程中， 在癌细胞中明显异常， 来分析。 这项研究也将有助于研究 哺乳动物的精子发生， 精母细胞与支持细胞的结合使其 难以识别和研究精子特异性基因。