AGOLGA3, A PROTEIN ESSENTIAL FOR SPERMATOGENESIS

AGOLGA3，精子发生必需的蛋白质

• 批准号: 8359752
• 负责人: CAROL C LINDER
• 金额: $ 11.22万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359752
• 关键词: Affect; Apoptosis; Biomedical Research; Cells; Defect; Development; Exhibits; Exons; Funding; Genes; Germ Cells; Goals; Golgi Apparatus; Grant; Human Identifications; Induced Mutation; Infertility; Male Infertility; Meiosis; Mus; Mutant Strains Mice; Mutation; National Center for Research Resources; New Mexico; Nonsense Codon; Pathway interactions; Point Mutation; Principal Investigator; Proteins; Research; Research Infrastructure; Resources; Role; Source; Sperm Head; Sperm Tail; Spermatogenesis; Stem cell transplant; Testis; United States National Institutes of Health; Western Blotting; cell type; cost; human male; male; member; protein expression; protein transport; research study; sertoli cell; sperm cell

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The overall goal of my research is to identify mechanisms responsible for sperm differentiation using mice to understand normal development and aid in the treatment of human male infertility. Mice with the repro27 chemically-induced mutation display male-specific infertility characterized by cell dealth in late meiosis and abnormal differentiatiion leading to abnormal sperm heads and tails. repro27 mice have a point mutation in the golgin subfamily A member 3 (Golga3) gene that inserts a premature stop codon in exon 18. Protein expression analysis using Western blotting suggests that GOLGA3 protein is degraded in mutant mice. GOLGA3 is a Golgi complex protein implicated in protein trafficking of certain proteins, apoptosis, and spermatogenesis. GOLGA3 is expressed in both somatic Sertoli cells and germ cells but its function is not known. The objective of this proposal is to understand GOLGA3's role in spermatogenesis by utilizing repro27 mutant mice known to carry a mutation in this gene. The central hypothesis is that repro27 mutant mice exhibit multiple spermatogenesis defects resulting from disrupted GOLGA3-dependent pathways. Although Golga3 mutations have not been identified in humans, identification of affected genes and pathways may be conserved. Thus, repro27 mice will be used to 1) conduct spermatogonial stem cell transplant experiments to determine if GOLGA3 function is germ cell specific; and 2) study the role of GOLGA3 in the testis at transcriptional and post-translational levels. Spermatogenesis in reciprocal stem cell transplant experiments will be used to determine cell type requirements. Validated protein targets and demonstrated protein interactions will dissect GOLGA3 pathways. Results will facilitate our understanding of spermatogenesis and may provide new clues and treatment options for human male infertility.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 我研究的总体目标是利用小鼠来确定负责精子分化的机制，以了解正常发育并帮助治疗人类男性不育症。携带repro27化学诱导突变的小鼠表现出雄性特有的不育症，其特征是减数分裂晚期细胞死亡和异常分化，导致精子头部和尾部异常。 repro27 小鼠的 golgin 亚家族 A 成员 3 (Golga3) 基因存在点突变，在外显子 18 中插入提前终止密码子。使用蛋白质印迹法进行的蛋白质表达分析表明，GOLGA3 蛋白质在突变小鼠中被降解。 GOLGA3 是一种高尔基复合体蛋白，参与某些蛋白质的蛋白质运输、细胞凋亡和精子发生。 GOLGA3 在体细胞支持细胞和生殖细胞中表达，但其功能尚不清楚。该提案的目的是通过利用已知携带 GOLGA3 基因突变的 repro27 突变小鼠来了解 GOLGA3 在精子发生中的作用。中心假设是 repro27 突变小鼠表现出多种精子发生缺陷，这是由于 GOLGA3 依赖性途径被破坏所致。尽管尚未在人类中鉴定出 Golga3 突变，但受影响基因和通路的鉴定可能是保守的。因此，repro27小鼠将用于1）进行精原干细胞移植实验以确定GOLGA3功能是否具有生殖细胞特异性； 2) 研究 GOLGA3 在转录和翻译后水平上在睾丸中的作用。相互干细胞移植实验中的精子发生将用于确定细胞类型要求。经验证的蛋白质靶标和已证明的蛋白质相互作用将剖析 GOLGA3 通路。结果将有助于我们对精子发生的理解，并可能为人类男性不育症提供新的线索和治疗选择。