Molecular Bases Committing Primate Spermatogonia to a Pathway of Differentiation.

使灵长类精原细胞走向分化途径的分子基础。

基本信息

批准号：
8271046
负责人：
TONY M PLANT
金额：
$ 40.56万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-06 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8271046
关键词：
Address Adult Animal Feed Biology Cell physiology Cells Cellular biology Commit Cues Development Diet Endocrine Environment Experimental Models Gene Expression Gene Expression Profile Genes Germ Germ Cells Global Change Goals Gonadotropins Human Immunohistochemistry In Situ Hybridization Infertility Interruption Knowledge Lead Macaca mulatta Male Contraceptions Male Contraceptive Agents Microarray Analysis Mitosis Molecular Molecular Biology Monkeys Mus Paracrine Communication Pathway interactions Primates Process Proliferating Quantitative Reverse Transcriptase PCR RNA Research Personnel Rodent Role Signal Pathway Signal Transduction Spermatogenesis Spermatogonia Testis Transcript Translating Tretinoin Undifferentiated Vitamin A Vitamin A Deficiency Work base comparative human male insight interest male nonhuman primate novel self-renewal sertoli cell species difference tool

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to understand the cellular and molecular mechanisms that control spermatogonial differentiation in the rhesus monkey, a representative higher primate. The strategy underlying the proposal is based, in part, on the finding that while spermatogonial proliferation occurs in the absence of gonadotropin stimulation, differentiation of primate spermatogonia occurs only in the presence of a gonadotropin drive. This difference in the endocrine requirements of the two processes will be used to interrogate the testicular genes, gene networks and any novel transcripts that determine the decision of spermatogonia to renew or to differentiate. There are three major questions. First, what genes and pathways within spermatogonia underlie the decision of the cell to differentiate rather than undergo self-renewal. Second, what are the gonadotropin dependent Sertoli cell paracrine signals that instruct undifferentiated spermatogonia to proceed down the path of differentiation. Third, to what extent is spermatogonial differentiation dependent on Vitamin A. To begin to answer these questions, RNA-seq will be used to assess global changes in germ cell and Sertoli cell gene expression and transcriptome profiles associated with the initial step in placing undifferentiated type A spermatogonia on the gonadotropin dependent pathway of differentiation. Following confirmation by qRTPCR, cell specific expression of those genes and relevant transcripts that are shown by qRTPCR to be up- or down-regulated in association with spermatogonial differentiation will then be determined using immunohistochemistry (IHC) and/or in situ hybridization (ISH). The effects of Vitamin A deficiency (VAD) on spermatogonial proliferation will be determined by feeding the animals a Vitamin A deficient diet. Two Specific Aims will be addressed; 1 to identify germ cell and Sertoli cell genes, gene networks and novel transcripts associated with the decision of undifferentiated type Ap spermatogonia to commit to a pathway of differentiation, and 2 To determine whether the decision of undifferentiated Type Ap spermatogonia to commit to the path of differentiation in the monkey is dependent on Vitamin A. This work will be conducted in an interactive intellectual environment that is generated by a group of investigators interested in related aspects of spermatogonial biology and Sertoli cell function. It is anticipated that the proposed work will provide fundamental insight into spermatogonial differentiation: a critical aspect of spermatogenesis that will be relevant to the treatment of infertility in the human male and to the development of novel male contraceptives. PUBLIC HEALTH RELEVANCE: The understanding of the molecular bases underlying the decision of undifferentiated spermatogonia to proceed down the pathway of differentiation or to renew themselves is important because this is a key step in the process of spermatogenesis. To date, this step in the spermatogenic lineage has been investigated primarily to rodents, and studies of non-human primates are urgently needed.

描述（由申请人提供）：该项目的长期目标是了解控制精子分化的细胞和分子机制，该机制是代表性较高的灵长类动物。该提案基础的策略部分基于以下发现：尽管精子增殖发生在没有促性腺激素刺激的情况下，但灵长类动物精子的分化仅在促性腺激素驱动的存在下才会发生。这两个过程的内分泌需求的这种差异将用于询问睾丸基因，基因网络和任何确定精子剂续订或分化的新成绩单。有三个主要问题。首先，精子中哪些基因和途径是细胞决定区分而不是自我更新的决定。其次，促性腺激素依赖性的Sertoli细胞旁分泌信号是什么，这些信号指示未分化的精子进行分化的路径。第三，依赖于维生素A的精子分化在多大程度上是在回答这些问题的情况下，RNA-Seq将用于评估生殖细胞和Sertoli细胞基因表达的全球变化以及与放置与分化依赖性促肿瘤蛋白依赖性路径上A上的A型A精子的初步步骤相关的初始步骤。在QRTPCR确认后，将使用免疫组织化学（IHC）和/或原位杂交（ISH）确定这些基因和相关转录物的细胞特异性表达和相关的转录本与精子分化相关的相关转录物。维生素A缺乏症（VAD）对精子增生的影响将通过为动物喂养维生素A缺乏饮食来确定。将解决两个具体目标； 1要鉴定生殖细胞和塞托利细胞基因，基因网络和与未分化类型AP的决定有关的决定，以实现分化途径的决定，而2确定未分化类型AP精子的决定是否依赖于猴子的互动依赖于维生素的互动。精子生物学和Sertoli细胞功能的各个方面。预计拟议的工作将提供对精子分化的基本见解：精子发生的关键方面，这将与人类男性的不育治疗以及新型男性避孕药的发展有关。公共卫生相关性：对未分化的精子沿分化途径或更新本身的决定的分子碱基的理解很重要，因为这是精子发生过程中的关键步骤。迄今为止，已经主要研究了啮齿动物的精子谱系中的这一步骤，并且迫切需要对非人类灵长类动物的研究。