Estradiol signaling pathways mediating sex differences in striatal synaptic plasticity

雌二醇信号通路介导纹状体突触可塑性的性别差异

• 批准号: 10607187
• 负责人: Kim L Blackwell
• 金额: $ 65.24万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2023-08-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607187
• 关键词: Acceleration; Affect; Affinity Chromatography; Animals; Behavior; Behavioral; Brain; Cells; Cocaine; Computer Models; Corpus striatum structure; Data; Disparity; Dopamine; Dorsal; Electrophysiology (science); Estradiol; Estradiol Receptors; Estrogen Receptor alpha; Estrus; Extracellular Signal Regulated Kinases; Feedback; Female; Foundations; Future; Gene Expression; Gene Expression Profiling; Genetic Complementation Test; Goals; Gonadal Steroid Hormones; Impairment; Investigation; Learning; Link; Long-Term Potentiation; Mediating; Metabotropic Glutamate Receptors; Methods; Modeling; Molecular; Mus; Neurons; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Play; Population; Proteins; Protocols documentation; Publishing; Reduce health disparities; Research; Rewards; Ribosomes; Rodent; Role; Saline; Self Administration; Sex Differences; Signal Pathway; Signal Transduction; Slice; Specificity; Substance Use Disorder; Synaptic plasticity; Techniques; Technology; Testing; Time; Transgenic Mice; Transgenic Organisms; Translating; Whole-Cell Recordings; Work; addiction; cell type; cocaine related behaviors; cocaine self-administration; cocaine use; data-driven model; experimental study; extracellular; female sex hormone; gender difference; gene interaction; habit learning; health disparity; improved; innovation; male; novel; sensor; single molecule; single nucleus RNA-sequencing; stem; substance use; transcriptome sequencing; transcriptomics; Acceleration; Affect; Affinity Chromatography; Animals; Behavior; Behavioral; Brain; Cells; Cocaine; Computer Models; Corpus striatum structure; Data; Disparity; Dopamine; Dorsal; Electrophysiology (science); Estradiol; Estradiol Receptors; Estrogen Receptor alpha; Estrus; Extracellular Signal Regulated Kinases; Feedback; Female; Foundations; Future; Gene Expression; Gene Expression Profiling; Genetic Complementation Test; Goals; Gonadal Steroid Hormones; Impairment; Investigation; Learning; Link; Long-Term Potentiation; Mediating; Metabotropic Glutamate Receptors; Methods; Modeling; Molecular; Mus; Neurons; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Play; Population; Proteins; Protocols documentation; Publishing; Reduce health disparities; Research; Rewards; Ribosomes; Rodent; Role; Saline; Self Administration; Sex Differences; Signal Pathway; Signal Transduction; Slice; Specificity; Substance Use Disorder; Synaptic plasticity; Techniques; Technology; Testing; Time; Transgenic Mice; Transgenic Organisms; Translating; Whole-Cell Recordings; Work; addiction; cell type; cocaine related behaviors; cocaine self-administration; cocaine use; data-driven model; experimental study; extracellular; female sex hormone; gender difference; gene interaction; habit learning; health disparity; improved; innovation; male; novel; sensor; single molecule; single nucleus RNA-sequencing; stem; substance use; transcriptome sequencing; transcriptomics

Substance use disorders affect ~15% of the population, with gender differences in all stages of substance use. Female sex-steroid hormones explain some of the disparity, such as accelerated transition from casual use to addiction, as estradiol produces sex-specific differences in rodent learning and cocaine self-administration. Prolonged cocaine use is known to engage dorsal striatal circuits. Synaptic plasticity in such circuits is critical for a variety of types of reward learning, highlighting the potential role such plasticity could play in substance use disorders. Thus, understanding sex differences in cocaine use requires determining how estradiol impacts molecular signaling and synaptic plasticity in the dorsal striatum. Our preliminary results show that estradiol, acting at estradiol receptor type α, impairs long term potentiation (LTP) in dorsomedial striatum (DMS) in estrous females; however, the cell type in which estradiol acts has yet to be identified. Striatal spiny projection neurons (SPNs) are either direct pathway SPNs, which promote action, or indirect pathway SPNs, which inhibit action, and changes in these SPNs are critical for various behavioral consequences. Thus, it is essential to understand LTP deficits in both direct and indirect pathway SPNs. In this proposal, we will test the hypothesis that estradiol impairs LTP in indirect pathway SPNs in the DMS. LTP in the dorsal striatum critically depends on activation of extracellular regulated kinase (ERK), which also is modified by both cocaine and estradiol. Estradiol enhances cocaine-mediated dopamine release and interacts with metabotropic glutamate receptors to modify ERK activation, but an unbiased approach is needed to determine whether estradiol impacts other signaling pathways. Our preliminary results have identified several signaling pathways that are modified by estradiol; however, a critical question is how cocaine interacts with estradiol to modulate these signaling pathways. We propose cutting-edge molecular and transgenic approaches combined with novel computational modeling to test the hypothesis that estradiol-mediated changes in gene expression impair LTP and to determine how cocaine further modifies the signaling pathways underlying synaptic plasticity. In Specific Aim 1, we perform electrophysiology in transgenic mice to determine whether LTP is impaired by estradiol in one or both SPNs. In Specific Aim 2, we use innovative techniques of single nuclei RNA sequencing, translating ribosome affinity purification followed by RNA sequencing and spatial transcriptomics to identify signaling pathways modified by estradiol and cocaine self-administration in a cell-type specific and spatial manner. In Specific Aim 3, we use innovative, data-driven modeling of signaling pathways followed by model-driven experiments to causally test which interactions between critical signaling pathways produce estradiol-mediated deficits in LTP. Successful completion of the proposed research will delineate how estradiol influences synaptic plasticity in dorsal striatum, including in conjunction with cocaine, and provide a foundation for future work to understand sex differences in reward learning and the consequences of cocaine use.

药物使用障碍影响约15％的人口，在底物使用的所有阶段都存在性别差异。 女性性甾体激素解释了一些差异，例如从随意使用到加速过渡到 成瘾，因为雌二醇在啮齿动物学习和可卡因自我给药方面产生了特定的性别差异。 已知可卡因的长时间使用可以接合背纹状体回路。此类电路中的突触可塑性至关重要 对于各种类型的奖励学习，强调这种可塑性在底物中的潜在作用 使用疾病。那就是了解可卡因使用中的性别差异需要确定雌二醇的影响 背纹状体中的分子信号传导和突触可塑性。我们的初步结果表明雌二醇， 作用于雌二醇受体型α型，会损害长期遗传（LTP）在背侧纹状体（DMS）中 发情的女性；但是，雌二醇作用的细胞类型尚未鉴定。纹状体刺突 神经元（SPNS）是直接途径SPN，它促进作用或间接途径SPNS，它抑制 动作以及这些SPN的变化对于各种行为后果至关重要。那是至关重要的 了解LTP在直接和间接途径SPN中定义。在此提案中，我们将检验假设 雌二醇会损害DMS中间接途径SPN中的LTP。背侧纹状体中的LTP严重取决于 细胞外调节激酶（ERK）的激活，可卡因和雌二醇也会修饰。 雌二醇增强可卡因介导的多巴胺释放，并与代谢型谷氨酸接收器相互作用 要修改ERK激活，但是需要一种公正的方法来确定雌二醇是否影响其他 信号通路。我们的初步结果已经确定了几种通过 雌二醇；但是，一个关键的问题是可卡因如何与雌二醇相互作用以调节这些信号 途径。我们提出了结合新的计算的尖端分子和转基因方法 建模以检验雌二醇介导的基因表达变化损害LTP和TO的假设 确定可卡因如何进一步修改突触可塑性的信号通路。在特定目标中 1，我们在转基因小鼠中进行电生理学，以确定雌二醇在一个或 两个SPN。在特定目标2中，我们使用单核RNA测序的创新技术，翻译 核糖体亲和力纯化，然后进行RNA测序和空间转录组学以识别信号传导 雌二醇和可卡因自我给药以细胞类型的特异性和空间方式修饰的途径。在 特定目标3，我们使用创新的，数据驱动的信号通路建模，然后是模型驱动的 实验以偶尔测试关键信号通路之间的哪些相互作用产生雌二醇介导的 LTP中的缺陷。成功完成拟议的研究将描述雌二醇如何影响突触 背纹状体的可塑性，包括与可卡因结合使用，并为将来的工作提供基础 了解奖励学习中的性别差异和可卡因使用的后果。