Role of DAX1 in Testis Determination and Function

DAX1 在睾丸测定和功能中的作用

• 批准号: 6914899
• 负责人: James Larry JAMESON
• 金额: $ 31.27万
• 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/6914899
• 关键词: Role; DAX1; Testis; Determination; Function

EXCEED THE SPACE PROVIDED. Over the last 8 years, we identified and characterized patients with naturally occurring mutations in the orphan nuclear receptors, DAXl and SF1, and demonstrated that DAXl acts in part by inhibiting SF1- mediated transcription. Using targeted mutagenesis, we developed a murine Daxl knockout (KO) model and identified roles for Daxl 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 Daxl structure and function as a transcriptional represser, identify genetic pathways regulated by Daxl, and develop animal models to unravel the interplay of Daxl with other genes involved in testis development and function. Specifically, in Aim 1 we will identify molecular partners that mediate DAXl 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 Daxl using microarray analyses of genes expressed in wild type versus Daxl-deficient embryonic gonads at 12 dpc, a timepoint when Daxl expression diverges in males and females. Aim 3 is designed to explore the interaction of Daxl with other genetic pathways in vivo. Using mice that lack Daxl, we will explore the gonadal phenotypes of mice with selective rescue of Daxl in various cell types. In addition, Daxl-deficient mice will be crossed to other strains with alterations in genetic pathways proposed to intersect with Daxl (e.g., Sfl, 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. PERFORMANCE SITE ========================================Section End===========================================

超过提供的空间。 在过去的8年中，我们确定并表征了孤儿核受体DAXL和SF1中天然突变的患者，并证明DAXL部分通过抑制SF1介导的转录而作用。使用靶向诱变，我们开发了一种鼠DAXL敲除（KO）模型，并确定了DAXL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DAXL结构和功能作为转录阻遏物，识别由DAXL调节的遗传途径，并开发动物模型以揭示DAXL与睾丸发育和功能所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DAXL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DAXL调节的遗传途径，使用在12 dpc的野生型和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DAXL表达的时间点。 AIM 3旨在探索DAXL与体内其他遗传途径的相互作用。使用缺乏DAXL的小鼠，我们将在各种细胞类型中选择性营救DAXL的小鼠探索小鼠的性腺表型。此外，DAXL缺陷小鼠将越过其他菌株，并在提出与DAXL相交的遗传途径发生变化（例如SFL，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。表演网站=================================== ==================================