Gonadal expression of FSH receptor

FSH 受体的性腺表达

• 批准号: 7602939
• 负责人: LESLIE L. HECKERT
• 金额: $ 30.61万
• 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/7602939
• 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; Genomics; Gonadotropins; Human; Hypersensitivity; Information Resources; Investigation; Knockout Mice; Maps; Modeling; Mutation; Nucleic Acid Regulatory Sequences; Ovary; Process; Property; Proteins; Rattus; Regulation; Regulatory Element; Reproductive Health; Reproductive system; Research; Role; Site; Testing; Testis; Tissues; Transcriptional Regulation; Transfection; Transgenic Mice; Transgenic Organisms; chromatin immunoprecipitation; comparative; 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是控制性腺功能，配子发生和生育能力的内分泌轴的关键组成部分。调节FSHR转录的机制的表征为揭示了Sertoli和Granulosa细胞中选择性基因表达涉及的蛋白质的机会，并扩展了我们对调节生殖健康的因素的理解。迄今为止，FSHR的转录研究仅集中在启动子和5'侧翼序列上。尽管这些分析对于我们对FSHR的理解至关重要，但似乎该区域以外的调节序列对于FSHR转录调节很重要。幸运的是，由于基因组研究的广泛进展，我们识别此类监管序列的能力已大大提高，这提供了大量的新信息和资源。提出的研究结合了计算分析，转录测定和转基因方法，以进一步了解控制FSHR转录的过程。在目标I中，建议研究使用小鼠基因敲除模型和染色质免疫沉淀来评估与FSHR调节有关的转录因子的体内作用，并扩展我们对通过近端启动子调节转录的机制的理解。在AIM II中，比较基因组学，DNase I超敏反应和瞬时转染分析将用于识别FSHR基因座内的远端调节区域。 DNA/蛋白质结合研究将用于鉴定与重要调节元件相关的蛋白质。 AIM III建议使用转基因小鼠在体内表征FSHR调节元件。含FSHR的酵母人工染色体（YAC）将用于产生转基因小鼠，该小鼠将在各种组织中表达。 YAC中提出的调节元件内的突变将通过同源重组产生，并在转基因小鼠中进行了正确的表达，以评估其在体内的作用。拟议的研究将揭示FSHR表达所需的调节元素和蛋白质，不仅可以更好地理解其转录控制，而且导致了有关性腺功能，生育能力和广义基因转录的基本原理。通过促进我们在这些领域的理解，我们增强了评估人类生殖健康问题并制定有助于生育问题和管理的策略的能力。