Role of DAX1 in Testis Determination and Function

DAX1 在睾丸测定和功能中的作用

基本信息

批准号：
7285191
负责人：
James Larry JAMESON
金额：
$ 30万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
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/7285191
关键词：
Adult Animal Model Backcrossings Binding Binding Sites Biological Assay Candidate Disease Gene Complex DNA Development Doctor of Philosophy Embryo Exhibits Female Gene Dosage Gene Duplication Gene Expression Gene Targeting Genes Genetic Genetic Transcription Goals Gonadal structure Grant Histologic Immunoprecipitation Knock-out Libraries Link Male Infertility Mammals Mediating Modeling Molecular Mus Mutagenesis Mutation Nuclear Orphan Receptor Other Genetics Ovarian Pathway interactions Patients Phenotype Play Proteins Regulation Repression Role SF1 Structure Study models Testis Transcription Repressor/Corepressor Two-Hybrid System Techniques Yeasts base cell type chromatin immunoprecipitation design gene repression in vivo insight male mouse model novel promoter protein protein interaction sex sex determination success transcription factor yeast two hybrid system

项目摘要

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.

在过去的 8 年里，我们识别并描述了具有天然突变的患者孤儿核受体 DAX1 和 SF1，并证明 DAX1 部分通过抑制 SF1 起作用介导的转录。通过定向诱变，我们开发了小鼠 Daxl 敲除 (KO) 模型并确定了 Daxl 在性腺决定、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，这些研究为性腺机制提供了意想不到的见解发展。我们提出了 3 个相互关联的目标，重点关注 Daxl 作为转录的结构和功能阻遏物，识别 Daxl 调节的遗传途径，并开发动物模型来揭示相互作用 Daxl 与其他参与睾丸发育和功能的基因。具体来说，在目标 1 中，我们将确定介导DAX1转录抑制的分子伙伴。自然发生的 DAX1 突变将被鉴定和表征，以阐明 DAX1 体内功能所需的关键结构域。在胚胎性腺文库的酵母双杂交筛选中鉴定出的候选蛋白将进一步特点。目标 2 的目标是使用微阵列识别 Daxl 调控的遗传途径在 12 dpc（一个时间点）时对野生型与 Daxl 缺陷胚胎性腺中表达的基因进行分析当 Daxl 表达在男性和女性中存在差异时。目标 3 旨在探索 Daxl 与体内其他遗传途径。使用缺乏 Daxl 的小鼠，我们将探索性腺在各种细胞类型中选择性拯救 Daxl 的小鼠表型。此外，缺乏 Daxl 的小鼠会与其他菌株杂交，遗传途径发生改变，建议与 Daxl 相交（例如，Sfl、抱歉，PTC）。这些目标的成功应该提供 DAX1 和转录抑制之间的关键联系连接参与性腺发育的各种其他转录因子的途径。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Hypogonadotropic hypogonadism in subjects with DAX1 mutations.

DOI：
10.1016/j.mce.2011.04.017
发表时间：
2011-10-22
期刊：
MOLECULAR AND CELLULAR ENDOCRINOLOGY
影响因子：
4.1
作者：
Jadhav, Unmesh;Harris, Rebecca M.;Jameson, J. Larry
通讯作者：
Jameson, J. Larry

X-linked sex-determining region Y box 3 (SOX3) gene mutations are uncommon in men with idiopathic oligoazoospermic infertility.

X 连锁性别决定区 Y 框 3 (SOX3) 基因突变在特发性少精症不育症男性中并不常见。

DOI：
10.1210/jc.2004-0191
发表时间：
2004
期刊：
The Journal of clinical endocrinology and metabolism.
影响因子：
0
作者：
Raverot,Gerald;Lejeune,Herve;Kotlar,Tom;Pugeat,Michel;Jameson,JLarry
通讯作者：
Jameson,JLarry

Analysis of DAX1 (NR0B1) and steroidogenic factor-1 (NR5A1) in children and adults with primary adrenal failure: ten years' experience.

DOI：
10.1210/jc.2006-0603
发表时间：
2006-08
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
0
作者：
Lin L;Gu WX;Ozisik G;To WS;Owen CJ;Jameson JL;Achermann JC
通讯作者：
Achermann JC

Heterozygous missense mutations in steroidogenic factor 1 (SF1/Ad4BP, NR5A1) are associated with 46,XY disorders of sex development with normal adrenal function.

DOI：
10.1210/jc.2006-1672
发表时间：
2007-03
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
0
作者：
Lin L;Philibert P;Ferraz-de-Souza B;Kelberman D;Homfray T;Albanese A;Molini V;Sebire NJ;Einaudi S;Conway GS;Hughes IA;Jameson JL;Sultan C;Dattani MT;Achermann JC
通讯作者：
Achermann JC