Gonadal expression of FSH receptor

FSH 受体的性腺表达

• 批准号: 7804509
• 负责人: LESLIE L. HECKERT
• 金额: $ 30.31万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7804509
• 关键词: 5' Flanking Region; Area; Binding; Biological Assay; Cells; Chromatin; Chromosomes, Artificial, Yeast; Conserved Sequence; DNA-Binding Proteins; Deoxyribonuclease I; Distal; Endocrine; Enhancers; Environment; Fertility; Follicle Stimulating Hormone; Follicle Stimulating Hormone Receptor; Functional RNA; Gametogenesis; Gene Expression; Genes; Genetic Transcription; Genome; Gonadotropins; Human; Hypersensitivity; Investigation; Knockout Mice; Maps; Modeling; Mutation; Nucleic Acid Regulatory Sequences; Ovary; Process; Property; Proteins; Rattus; Regulation; Regulatory Element; Reproductive Health; Reproductive system; Research; Resources; Role; Site; Testing; Testis; Tissues; Transcriptional Regulation; Transfection; Transgenic Mice; Transgenic Organisms; chromatin immunoprecipitation; comparative genomics; gonad function; granulosa cell; homologous recombination; in vivo; promoter; receptor; response; sertoli cell; transcription factor

DESCRIPTION (provided by applicant): Fshr is a critical component of the endocrine axis that controls gonad function, gametogenesis, and fertility. Characterization of the mechanisms regulating Fshr transcription provides the opportunity to reveal proteins involved in selective gene expression in Sertoli and granulosa cells and expand our understanding of factors regulating reproductive health. To date, transcriptional studies of Fshr have focused only on promoter and 5' flanking sequences. While these analyses are critical to our understanding of Fshr, it appears that regulatory sequences outside this region are important for Fshr transcriptional regulation. Fortunately, our ability to identify such regulatory sequences has advanced significantly due to the extensive progress in genome research, which provides a wealth of new information and resources. The proposed studies combine computation analysis, transcriptional assays, and transgenic approaches to further our understanding of the processes controlling Fshr transcription. In Aim I, studies are proposed to use mouse knockout models and chromatin immunoprecipitation to evaluate the in vivo roles of transcription factors implicated in Fshr regulation and expand our understanding of the mechanisms regulating transcription through the proximal promoter. In Aim II, comparative genomics, DNase I hypersensitivity mapping and transient transfection analysis will be used to identify distal regulatory regions within the Fshr locus. DNA/protein binding studies will be used to identify proteins associated with important regulatory elements. Aim III proposes to use transgenic mice for in vivo characterization of Fshr regulatory elements. An Fshr-containing yeast artificial chromosome (YAC) will be used to generate transgenic mice that will be characterized for expression in various tissues. Mutations within proposed regulatory elements will be generated in the YAC by homologous recombination and tested for correct expression in transgenic mice, to evaluate their role in vivo. The proposed studies will reveal regulatory elements and proteins needed for Fshr expression, leading not only to a better understanding of its transcriptional control but to the basic principles governing gonadal function, fertility, and generalized gene transcription. By advancing our understanding in these areas, we enhance our ability to evaluate matters of human reproductive health and to develop strategies to assist fertility problems and management.

描述（由申请人提供）：Fshr是内分泌轴的重要组成部分，控制性腺功能、配子发生和生育。Fshr转录调控机制的表征为揭示支持细胞和颗粒细胞中参与选择性基因表达的蛋白提供了机会，并扩大了我们对调节生殖健康因素的理解。迄今为止，Fshr的转录研究仅集中在启动子和5'侧链序列上。虽然这些分析对我们理解Fshr至关重要，但似乎该区域外的调控序列对Fshr的转录调控也很重要。幸运的是，由于基因组研究的广泛进展，我们识别这些调控序列的能力已经显著提高，这提供了丰富的新信息和资源。提出的研究结合了计算分析，转录分析和转基因方法，以进一步了解控制Fshr转录的过程。在Aim I中，研究人员提出使用小鼠敲除模型和染色质免疫沉淀来评估涉及Fshr调控的转录因子在体内的作用，并扩大我们对通过近端启动子调控转录的机制的理解。在Aim II中，比较基因组学、dna酶I超敏定位和瞬时转染分析将用于确定Fshr位点内的远端调控区域。DNA/蛋白质结合研究将用于鉴定与重要调控元件相关的蛋白质。目的三提出利用转基因小鼠在体内表征Fshr调控元件。一个含有fshrr的酵母人工染色体（YAC）将被用于产生转基因小鼠，其特征是在各种组织中表达。提出的调控元件的突变将通过同源重组在YAC中产生，并在转基因小鼠中测试其正确表达，以评估其在体内的作用。这些研究将揭示Fshr表达所需的调控元件和蛋白质，不仅有助于更好地了解其转录控制，还有助于了解性腺功能、生育能力和通用基因转录的基本原理。通过增进我们对这些领域的了解，我们提高了我们评价人类生殖健康问题和制定战略以协助生育问题和管理的能力。