Steroidogenic Factor 1: Mediator of Gonadal Function

类固醇生成因子 1：性腺功能调节剂

• 批准号: 7350915
• 负责人: RICHARD J. AUCHUS
• 金额: $ 29.35万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7350915
• 关键词: 5' Flanking Region; Address; Adrenal Glands; Alleles; Candidate Disease Gene; Cell Line; Cell Lineage; Cells; Cholesterol Monooxygenase (Side-Chain-Cleaving); Cloning; Complex; Conserved Sequence; Coupled; DNA Microarray Chip; DNA Microarray format; Deoxyribonuclease I; Deoxyribonucleases; Development; Dominant-Negative Mutation; Elements; Embryo; Endocrine; Event; Exhibits; Fluorescence-Activated Cell Sorting; Gene Expression; Gene Expression Regulation; Gene Targeting; Gene Transfer Techniques; Genes; Gonadal Agenesis; Gonadal structure; Hormones; Insulin; Insulin-Like-Growth Factor I Receptor; Introns; Knock-out; Knockout Mice; Mediating; Mediator of activation protein; Molecular Profiling; Neurons; Nuclear Orphan Receptor; Ovary; Phenotype; Pituitary Gland; Play; Population; Proteins; RNA; Regulation; Reporter Genes; Research Personnel; Role; SF1; Sex Differentiation; Signal Pathway; Site; Staging; Suspension substance; Suspensions; Technology; Testis; Tissues; Transfection; Transgenes; Transgenic Mice; Transgenic Organisms; Urogenital ridge structure; base; cell type; enhanced green fluorescent protein; fetal; gonad function; granulosa cell; in vivo; insight; interest; novel; programs; promoter; protein expression; recombinase; research study; sex; success; transgene expression; ventromedial hypothalamic nucleus

DESCRIPTION (provided by applicant): The orphan nuclear receptor steroidogenic factor 1 (SF-1) plays essential roles in endocrine development and function. SF-1 knockout (KO) mice exhibit adrenal and gonadal agenesis, impaired pituitary gonadotrope function, and marked structural abnormalities of the ventromedial hypothalamic nucleus (VMH). The proposed studies seek to expand our understanding of these pleiotropic roles of SF-1, focusing specifically on its roles in gonadal development. First, we will use the Cre-loxP technology to inactivate SF-1 in specific cell lineages of the testes and ovaries. By using different promoters to direct Cre expression, we envision that we will achieve both cell-type specific inactivation as well as inactivation at different stages of development. We also will use a transgenic enhanced green fluorescent protein (eGFP) reporter gene directed by SF-1 regulatory sequences to purify SF-l-expressing cells from the gonads of wild-type and SF-1 KO mice, allowing us to compare gene expression profiles of the same cell populations in the presence or absence of SF-I. We will use transient transfection experiments with the 5-flanking regions of these candidate genes to determine if they are direct or indirect targets of SF-1. To develop a mechanism to inactivate the expression of genes of interest from early stages of gonadogenesis, we will use BAC transgenesis to target expression of Cre recombinase to the sites where SF-1 is expressed. Finally, we will attempt to define the sequences that regulate SF-1 expression in the gonads, focusing on a conserved sequence in the 6th intron that is associated with a DNase hypersensitive site. These studies will provide novel insights into how SF-1 exerts its multiple actions in gonadal development and function.

描述（由申请人提供）：孤儿核受体类固醇生成因子1（SF-1）在内分泌发育和功能中发挥重要作用。 SF-1 敲除 (KO) 小鼠表现出肾上腺和性腺发育不全、垂体促性腺功能受损以及下丘脑腹内侧核 (VMH) 的明显结构异常。拟议的研究旨在扩大我们对 SF-1 多效性作用的理解，特别关注其在性腺发育中的作用。首先，我们将使用 Cre-loxP 技术灭活睾丸和卵巢特定细胞谱系中的 SF-1。通过使用不同的启动子来指导Cre表达，我们设想我们将实现细胞类型特异性失活以及不同发育阶段的失活。我们还将使用由 SF-1 调控序列指导的转基因增强型绿色荧光蛋白 (eGFP) 报告基因，从野生型和 SF-1 KO 小鼠的性腺中纯化表达 SF-1 的细胞，从而使我们能够比较基因表达在存在或不存在 SF-I 的情况下相同细胞群的概况。我们将对这些候选基因的 5 侧翼区域进行瞬时转染实验，以确定它们是否是 SF-1 的直接或间接靶标。为了开发一种机制，使性腺发生早期阶段感兴趣的基因的表达失活，我们将使用 BAC 转基因将 Cre 重组酶的表达靶向到 SF-1 表达的位点。最后，我们将尝试定义调节性腺中 SF-1 表达的序列，重点关注与 DNase 超敏感位点相关的第 6 个内含子中的保守序列。这些研究将为 SF-1 如何在性腺发育和功能中发挥多种作用提供新的见解。