Estrogen receptor regulation of brain sexual differentiation

雌激素受体对大脑性别分化的调节

• 批准号: 10251067
• 负责人: Jordan Bruno Gegenhuber
• 金额: $ 2.77万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-07-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10251067
• 关键词: Adult; Aggressive behavior; Animals; Anxiety; Apoptosis; Axon; Behavior; Binding; Binding Sites; Biological; Biological Process; Brain; Brain region; Caring; Cell Count; Cell Nucleus; Cell Survival; Cells; ChIP-seq; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Nucleotidylexotransferase; Data; Development; Disease; Distal; Enhancers; Estradiol; Estrogen Nuclear Receptor; Estrogen Receptor alpha; Estrogen Receptors; Estrogens; Female; Feminization; Fluorescent in Situ Hybridization; Functional disorder; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genomics; Goals; Gonadal Steroid Hormones; Hormones; Hypothalamic structure; Impairment; In Situ Hybridization; In Vitro; Injections; Knock-out; Label; Limbic System; Link; Masculine; Measures; Mediating; Mental Health; Methods; Molecular; Mus; Mutant Strains Mice; Neocortex; Neonatal; Neurodevelopmental Disorder; Neurons; Nuclear Receptors; Partner in relationship; Pathway interactions; Perinatal; Pharmacology Study; Physiological; Play; Presynaptic Terminals; Primates; Process; Publishing; RNA; Regulation; Reproductive Behavior; Rodent; Role; Running; Sex Bias; Sex Differences; Sex Differentiation; Signal Transduction; Social Behavior; Specific qualifier value; Structure of terminal stria nuclei of preoptic region; System; Systems Development; Testing; Testosterone; Tissues; Training; autism spectrum disorder; axon guidance; brain behavior; crosslink; experimental study; improved; insight; male; mutant; nerve supply; netrin receptor; neural circuit; novel; nuclease; postnatal; pup; sex; sexual dimorphism; steroid hormone; tool; transcription factor; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT In sexually reproducing species, males and females display different social and reproductive behaviors, such as mating and aggression. These behaviors typically require no training, indicating they are developmentally programmed in the brain. In many mammalian species, including primates and rodents, sexual differentiation of the brain is regulated by nuclear receptor transcription factors (TFs), which bind gonadal steroid hormones, such as testosterone and estrogens. In mice, a perinatal surge of testosterone permanently masculinizes a key brain region controlling sex-typical behaviors, called the bed nucleus of the stria terminalis (BNST). Within the perinatal BNST, testosterone is converted to estradiol, which activates estrogen receptor α (ERα). Previous genetic knockout and pharmacological studies demonstrate perinatal ERα signaling is necessary and sufficient for masculinization of BNST circuitry and behavior. Specifically, ERα activation leads to a male-bias in BNST cell survival and axon guidance, particularly to the anteroventral periventricular nucleus (AVPV), between postnatal day 4 (P4) and P10. This project seeks to identify and characterize ERα genomic binding sites and target genes involved in BNST sexual differentiation. Recently our group discovered that estradiol regulates the expression of the netrin receptor, Unc5b, in adult BNST ERα+ cells. Unc5b is robustly expressed in the neonatal BNST and has previously been shown to regulate neuron survival and axon guidance. Aim 1 of this project investigates whether estradiol regulation of Unc5b contributes to BNST sexual differentiation, using mice lacking Unc5b expression in ERα+ cells (Esr1cre/+;Unc5blx/lx). The experiments in Aim 1 test the hypothesis that estradiol regulation of Unc5b contributes to male-biased BNST cell survival and/or AVPV innervation. Because ERα likely masculinizes the BNST through multiple biological pathways, Aim 2 of this project seeks to identify the complete repertoire of ERα genomic binding sites and target genes in the developing BNST. Previously, I discovered the first ERα genomic binding sites in the adult brain, using a recently published low-input TF profiling method, called CUT&RUN (Cleavage Under Targets & Release Under Nuclease). This approach revealed brain-specific ERα binding sites are enriched near genes involved in neurodevelopmental processes. Aim 2 of this application uses CUT&RUN to measure estradiol-regulated ERα binding sites in the perinatal BNST/hypothalamus. Aim 2 also measures perinatal estradiol-regulated gene expression in P4 BNST ERα+ cells. Using a CRISPR-mediated activation (CRISPRa) system in primary neurons, distal ERα binding sites will be causally linked to the expression of perinatal estradiol-regulated genes. Overall, the project will reveal how nuclear receptors regulate genes involved in brain sexual differentiation and, in doing so, will provide novel insight into the molecular basis of sex-biased mental health conditions and neurodevelopmental diseases.

项目摘要/摘要 在性繁殖物种中，雄性和女性表现出不同的社会和生殖行为，例如 交配和积极进取。这些行为通常不需要培训，表明它们正在发展 在大脑中编程。在许多哺乳动物物种中，包括主要和啮齿动物， 大脑受核接收器转录因子（TFS）的调节，该因子结合性腺类固醇激素，这样 作为睾丸激素和雌激素。在小鼠中，睾丸激素的围产期激增永久性男性大脑 控制性典型行为的区域，称为质末端的床核（BNST）。在围产期内 BNST，将睾丸激素转化为雌二醇，该雌二醇激活雌激素受体α（ERα）。以前的遗传 敲除和药物研究表明，围产期ERα信号是必要的，足以满足 BNST电路和行为的男性化。具体而言，ERα激活导致BNST细胞的雄性偏见 生存和轴突引导，特别是对腹膜前核（AVPV）的生存和指导。 第4天（P4）和P10。该项目旨在识别和表征ERα基因组结合位点和靶基因 参与第一个性别差异。最近，我们的小组发现雌二醇调节表达 在成年BNSTERα+细胞中的Netrin受体UNC5B。 UNC5B在新生儿BNST中强烈表达 以前已显示出调节神经元存活和轴突指导。该项目的目标1调查 使用缺乏UNC5B的小鼠 ERα+细胞（ESR1CRE/+; UNC5BLX/LX）中的表达。 AIM 1中的实验检验了雌二醇的假设 UNC5B的调节有助于男性偏见的BNST细胞存活和/或AVPV神经。因为ERα可能 通过多种生物学途径使BNST男性化，该项目的目标2旨在确定完整 ERα基因组结合位点和靶基因的曲目在发育中的BNST中。以前，我发现了 使用最近发表的低输入TF分析方法，成人大脑中的第一个ERα基因组结合位点称为 切割和运行（在目标下进行切割和核酸酶下释放）。这种方法揭示了大脑特异性ERα 结合位点富含与神经发育过程有关的基因。该应用程序使用的目标2 切割和运行以测量围产期BNST/下丘脑中雌二醇调节的ERα结合位点。 AIM 2也是 测量P4 BNSTERα+细胞中雌性雌二醇调节的基因表达。使用CRISPR介导的 原发性神经元中的激活（CRISPRA）系统，远端ERα结合位点将与该系统联系在一起 围产期雌二醇调节基因的表达。总体而言，该项目将揭示核接收器如何规范 与大脑性别分化有关的基因，并这样做将提供对分子基础的新见解 性偏见的心理健康状况和神经发育疾病。