Role of DAX1 in Testis Determination and Function

DAX1 在睾丸测定和功能中的作用

• 批准号: 7069596
• 负责人: James Larry JAMESON
• 金额: $ 30.73万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-11 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7069596
• 关键词: Role; DAX1; Testis; Determination; Function

DESCRIPTION (provided by applicant): Over the last 8 years, we identified and characterized patients with naturally occurring mutations in the orphan nuclear receptors, DAX1 and SF1, and demonstrated that DAX1 acts in part by inhibiting SF1- mediated transcription. Using targeted mutagenesis, we developed a murine Dax1 knockout (KO) model and identified roles for Dax1 in gonadal determination, testis development, and adult testis function. In this grant, we propose to extend these studies, which have provided unexpected insight into mechanisms of gonadal development. We propose 3 inter-related aims that focus on Dax1 structure and function as a transcriptional repressor, identify genetic pathways regulated by Dax1, and develop animal models to unravel the interplay of Dax1 with other genes involved in testis development and function. Specifically, in Aim 1 we will identify molecular partners that mediate DAX1 transcriptional repression. Naturally occurring DAX1 mutations will be identified and characterized to elucidate key structural domains required for DAX1 function in vivo. Candidate proteins identified in a yeast two-hybrid screen of an embryonic gonadal library will be further characterized. The goal of Aim 2 is to identify the genetic pathways regulated by Dax1 using microarray analyses of genes expressed in wild type versus Dax1-deficient embryonic gonads at 12 dpc, a timepoint when Dax1 expression diverges in males and females. Aim 3 is designed to explore the interaction of Dax1 with other genetic pathways in vivo. Using mice that lack Dax1, we will explore the gonadal phenotypes of mice with selective rescue of Dax1 in various cell types. In addition, Dax1-deficient mice will be crossed to other strains with alterations in genetic pathways proposed to intersect with Dax1 (e.g., Sf1, Sry, Ptc). Success in these aims should provide a critical link between DAX1 and transcriptional repression pathways that link a variety of other transcription factors involved in gonadal development.

描述（由申请人提供）：在过去的8年中，我们确定并表征了孤儿核受体DAX1和SF1中天然突变的患者，并证明DAX1部分通过抑制SF1介导的转录来起作用。使用靶向诱变，我们开发了一种鼠DAX1敲除（KO）模型，并确定了DAX1在性腺测定，睾丸发育和成人睾丸功能中的作用。在这笔赠款中，我们建议扩展这些研究，这些研究为性腺发育机制提供了意外的见解。我们提出了3个相互关联的目的，该目标的重点是DAX1结构和功能作为转录阻遏物，识别由DAX1调节的遗传途径，并开发动物模型以揭示DAX1与睾丸发育和功能涉及的其他基因的相互作用。具体而言，在AIM 1中，我们将确定介导DAX1转录抑制的分子伴侣。自然发生的DAX1突变将被鉴定并表征为阐明体内DAX1功能所需的关键结构域。将进一步表征在胚胎性腺库的酵母两杂交筛选中鉴定出的候选蛋白。 AIM 2的目的是使用DAX1调节的遗传途径，使用在12 dpc的野生型与DAX1缺陷型胚胎性腺中表达的基因的微阵列分析，这是雄性和女性中DAX1表达的时间点。 AIM 3旨在探索DAX1与体内其他遗传途径的相互作用。使用缺乏DAX1的小鼠，我们将在各种细胞类型中选择性拯救DAX1的小鼠探索小鼠的性腺表型。此外，DAX1缺陷型小鼠将跨越其他菌株，并与DAX1相交的遗传途径发生了变化（例如SF1，SRY，PTC）。这些目标的成功应提供DAX1与转录抑制途径之间的关键联系，该途径将涉及性腺发育中涉及的其他转录因子联系起来。