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。

• 批准号: 8798678
• 负责人: TONY M PLANT
• 金额: $ 37.55万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-06 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8798678
• 关键词: Address; Adult; Animal Feed; Biology; Cell physiology; Cells; Cellular biology; 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; 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; transcriptome sequencing

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.

项目描述（由申请人提供）：本项目的长期目标是了解具有代表性的高等灵长类动物恒河猴精原细胞分化的细胞和分子机制。这一建议背后的策略部分是基于这样的发现：虽然精原细胞的增殖发生在没有促性腺激素刺激的情况下，但灵长类动物精原细胞的分化只发生在促性腺激素驱动的情况下。这两个过程中内分泌需求的差异将被用来询问睾丸基因、基因网络和任何决定精原细胞更新或分化的新转录本。主要有三个问题。首先，精原细胞中哪些基因和途径决定了细胞分化而不是自我更新。其次，什么是促性腺激素依赖的支持细胞旁分泌信号，指示未分化精原细胞继续分化的道路。第三，精原细胞分化在多大程度上依赖于维生素A。为了开始回答这些问题，RNA-seq将用于评估生殖细胞和支持细胞基因表达和转录组谱的全球变化，这些变化与将未分化的A型精原细胞放在促性腺激素依赖分化途径的初始步骤有关。在qRTPCR确认后，将使用免疫组织化学（IHC）和/或原位杂交（ISH）来确定qRTPCR显示的与精原细胞分化相关的基因和相关转录物的细胞特异性表达。维生素A缺乏（VAD）对精原细胞增殖的影响将通过饲喂缺乏维生素A的饲料来确定。将讨论两个具体目标；1 .鉴定与未分化型Ap精原细胞走向分化途径相关的生殖细胞和支持细胞基因、基因网络和新的转录本；为了确定猴子未分化的Ap型精原细胞是否依赖于维生素a，这项工作将在一个互动的智力环境中进行，该环境由一组对精原生物学和支持细胞功能相关方面感兴趣的研究人员创建。预计所提出的工作将提供对精原细胞分化的基本见解：精子发生的一个关键方面，将与人类男性不育的治疗和新型男性避孕药的开发有关。