Estrogen receptor regulation of brain sexual differentiation

雌激素受体对大脑性别分化的调节

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要在有性生殖的物种中，雄性和雌性表现出不同的社会和生殖行为，例如这些行为通常不需要训练，表明它们是发育性的。在许多哺乳动物物种中，包括灵长类动物和啮齿类动物，性别分化都在大脑中进行了编程。大脑受核受体转录因子 (TF) 的调节，该转录因子与性腺类固醇激素结合，例如如睾酮和雌激素，在小鼠中，围产期睾酮的激增使关键大脑永久男性化。控制典型性别行为的区域，称为围产期终纹床核（BNST）。 BNST，睾酮转化为雌二醇，激活雌激素受体α（ERα）。敲除和药理学研究表明围产期 ERα 信号传导对于 BNST 电路和行为的男性化具体而言，ERα 激活导致 BNST 细胞的男性偏好。出生后的生存和轴突引导，特别是前腹侧室周核（AVPV）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。最近我们小组发现雌二醇调节BNST的表达。成人 BNST ERα+ 细胞中的 Netrin 受体 Unc5b 在新生儿 BNST 和细胞中强烈表达。先前已被证明可以调节神经元存活和轴突引导，该项目研究的目标 1。使用缺乏 Unc5b 的小鼠，雌二醇对 Unc5b 的调节是否有助于 BNST 性别分化 ERα+ 细胞中的表达 (Esr1cre/+;Unc5blx/lx) 目标 1 中的实验检验雌二醇的假设。 Unc5b 的调节有助于雄性偏向的 BNST 细胞存活和/或 AVPV 神经支配。通过多种生物学途径使 BNST 男性化，该项目的目标 2 旨在确定完整的开发中的 BNST 中的 ERα 基因组结合位点和靶基因的全部内容之前，我发现了使用最近发布的低输入 TF 分析方法，称为成人大脑中的第一个 ERα 基因组结合位点 CUT&RUN（目标下切割并核酸酶下释放）这种方法揭示了大脑特异性 ERα。该应用的目标 2 使用的结合位点在参与神经发育过程的基因附近富集。 CUT&RUN 还可测量围产期 BNST/下丘脑中雌二醇调节的 ERα 结合位点，目标 2。使用 CRISPR 介导的方法测量 P4 BNST ERα+ 细胞中围产期雌二醇调节的基因表达。初级神经元中的 CRISPRa 激活（CRISPRa）系统，远端 ERα 结合位点将与总体而言，该项目将揭示核受体如何调节。参与大脑性别分化的基因，将为分子基础提供新的见解性别偏见的心理健康状况和神经发育疾病。