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

基本信息

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
项目类别：
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

项目摘要

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.

药物滥用障碍影响约 15% 的人口，在药物使用的各个阶段都存在性别差异。女性性类固醇激素解释了一些差异，例如从随意使用到加速转变成瘾，因为雌二醇在啮齿类动物的学习和可卡因自我给药方面产生性别特异性差异。众所周知，长期使用可卡因会影响背侧纹状体回路，突触可塑性至关重要。对于各种类型的奖励学习，强调这种可塑性在实质上可能发挥的潜在作用因此，了解可卡因使用的性别差异需要确定雌二醇的影响。背侧纹状体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α型雌二醇受体，损害背内侧纹状体（DMS）的长时程增强（LTP）发情女性；然而，雌二醇起作用的细胞类型尚未确定。神经元 (SPN) 要么是促进作用的直接途径 SPN，要么是抑制作用的间接途径 SPN 这些 SPN 的行动和变化对于各种行为后果至关重要。直接和间接途径 SPN 中的 LTP 缺陷在本提案中，我们将检验该假设。雌二醇损害 DMS 中 LTP 间接通路 SPN 的 LTP 关键取决于。细胞外调节激酶（ERK）的激活，该激酶也被可卡因和雌二醇修饰。雌二醇增强可卡因介导的多巴胺释放并与代谢型谷氨酸受体相互作用修改 ERK 激活，但需要一种公正的方法来确定雌二醇是否影响其他我们的初步结果已经确定了几种被修饰的信号通路。然而，一个关键问题是可卡因如何与雌二醇相互作用来调节这些信号传导我们提出了与新颖的计算相结合的尖端分子和转基因方法。建立模型来检验雌二醇介导的基因表达变化会损害 LTP 的假设，并确定可卡因如何进一步改变突触可塑性的信号通路。 1，我们对转基因小鼠进行电生理学检查，以确定 LTP 是否受到雌二醇的损害在具体目标 2 中，我们使用单核 RNA 测序、翻译的创新技术。核糖体亲和纯化，然后进行 RNA 测序和空间转录组学以鉴定信号传导通过雌二醇和可卡因以细胞类型特异性和空间方式自我给药来改变途径。具体目标 3，我们使用创新的、数据驱动的信号通路建模，然后使用模型驱动因果实验测试关键信号通路之间的相互作用产生雌二醇介导 LTP 的缺陷。成功完成拟议的研究将描述雌二醇如何影响突触。背侧纹状体的可塑性，包括与可卡因结合，并为未来的工作奠定基础了解奖励学习中的性别差异以及使用可卡因的后果。