Estradiol-mediated Gene Signatures in Hippocampal Memory

海马记忆中雌二醇介导的基因特征

基本信息

批准号：
10619592
负责人：
Jennifer J Tuscher
金额：
$ 10.46万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-09 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10619592
关键词：
ATAC-seq Adult Aging Anabolism Aromatase Astrocytes Binding Biological Assay Brain Brain region CREB1 gene CRISPR interference CRISPR-mediated transcriptional activation CRISPR/Cas technology Categories Cell Nucleus Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Cognition DNA Data Development Dimerization Dorsal Electrophysiology (science)Enzymes Epigenetic Process Estradiol Estrogen Receptor alpha Estrogen Receptors Estrogens Female Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Glutamates Gonadal Steroid Hormones Hippocampus Hormonal Hour Individual Interneurons Intervention Learning Longevity Mediating Membrane Memory Mental Health Mental disorders Molecular Morphology National Institute of Mental Health Neurobiology Neuroendocrinology Neuronal Plasticity Neurons Oligodendroglia Outcome Pathway interactions Physiology Play Population Process Production Property Public Health Recreation Regulation Regulator Genes Research Resolution Response Elements Rodent Role Signal Transduction Testing Therapeutic Intervention Transcriptional Regulation Transposase Work cell type cognitive function estrogenic experimental study genetic signature genomic locus improved insight long term memory male neuronal excitability neuropsychiatric disorder neuroregulation novel pharmacologic prevent programs receptor response sex single nucleus RNA-sequencing therapeutic target tool transcription factor transcriptomics treatment strategy

ATAC-seq Adult Aging Anabolism Aromatase Astrocytes Binding Biological Assay Brain Brain region CREB1 gene CRISPR interference CRISPR-mediated transcriptional activation CRISPR/Cas technology Categories Cell Nucleus Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Cognition DNA Data Development Dimerization Dorsal Electrophysiology (science)Enzymes Epigenetic Process Estradiol Estrogen Receptor alpha Estrogen Receptors Estrogens Female Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Glutamates Gonadal Steroid Hormones Hippocampus Hormonal Hour Individual Interneurons Intervention Learning Longevity Mediating Membrane Memory Mental Health Mental disorders Molecular Morphology National Institute of Mental Health Neurobiology Neuroendocrinology Neuronal Plasticity Neurons Oligodendroglia Outcome Pathway interactions Physiology Play Population Process Production Property Public Health Recreation Regulation Regulator Genes Research Resolution Response Elements Rodent Role Signal Transduction Testing Therapeutic Intervention Transcriptional Regulation Transposase Work cell type cognitive function estrogenic experimental study genetic signature genomic locus improved insight long term memory male neuronal excitability neuropsychiatric disorder neuroregulation novel pharmacologic prevent programs receptor response sex single nucleus RNA-sequencing therapeutic target tool transcription factor transcriptomics treatment strategy

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项目摘要

Project Summary The sex-steroid hormone 17-estradiol (E2) has a well-established role in mediating neuronal physiology, altering gene expression, and facilitating memory formation. Despite decades of research on E2’s neuromodulatory effects, much remains unknown regarding which specific gene targets regulated by E2 contribute to its effects on neuronal function and memory. This is in part due to a lack of experimental feasibility, as the tools necessary for examining cell-type specific transcriptional regulation by E2, and targeted manipulation of specific E2-sensitive genes, were not previously available. Here, I propose to overcome these hurdles by using single-nuclei RNA- and ATAC-sequencing, which will allow for comprehensive and unbiased identification of gene targets regulated by E2 in both sexes at the single-cell level (Aim 1). This approach is advantageous to the few existing studies examining transcriptomic regulation of sex-steroid hormones, which use brain homogenates containing a variety of cell types for sequencing experiments. Although such work has been informative, the receptors through which E2 orchestrates transcriptional control are expressed in a wide variety of distinct cell types. As such, higher resolution approaches such 10X snRNA-seq and snATAC-seq at the individual cell level will be necessary to understand the full myriad of estrogenic regulation of gene expression. Another limitation of previous studies has been the inability to directly manipulate E2-sensitive gene targets to empirically test the requirement of such changes in expression on memory formation. This proposal will overcome such limitations by using cutting-edge CRISPR/dCas9-based epigenetic editing approaches to test the requirement of E2 action at specific gene loci in mediating its beneficial mnemonic effects (Aim 2). Finally, although circulating levels of E2 have been the focus of much neuroendocrinology research, E2 biosynthesis also occurs locally in the adult brain of both sexes, via a process regulated by the enzyme aromatase (Cyp19a1). Aim 3 will use a CRISPR/dCas9 approach to bidirectionally regulate aromatase expression specifically in neurons, to test the role of neuron-specific E2 synthesis in memory formation. Collectively, these experiments will provide much needed insight into how E2 acts as a key modulator of neuronal function and memory in both sexes.

项目摘要性类替代的马酮17-雌二醇（E2）在介导神经元生理学中具有良好的作用，改变基因表达并支持记忆形成。尽管对E2的研究进行了数十年的研究神经调节作用，关于哪些特定基因靶标由E2调节，尚不清楚有助于其对神经元功能和记忆的影响。这部分是由于缺乏实验性可行性，作为通过E2检查细胞类型的特定转录调控所需的工具，并针对操纵以前没有特定的E2敏感基因。在这里，我建议通过使用单核RNA和ATAC序列，这将允许对基因靶标在单细胞水平的两个性别中受E2调节的靶标（AIM 1）。这种方法对很少有研究使用脑匀浆的性生物激素转录组调节的研究包含多种用于测序实验的细胞类型。尽管这样的工作很有帮助，但 E2策划转录控制的受体在各种不同的细胞中表达类型。因此，更高的分辨率接近单个细胞水平的10倍SNRNA-SEQ和SNATAC-SEQ 了解基因表达的全部雌激素调节是必要的。另一个限制先前的研究一直无法直接操纵E2敏感基因靶标，以经验测试对记忆形成的表达变化的要求。该建议将克服此类限制通过使用基于尖端的CRISPR/DCAS9的表观遗传编辑方法来测试E2动作的需求在介导其有益助记符效应的特定基因基因座（AIM 2）。最后，尽管循环水平 E2一直是许多神经内分泌研究的重点，E2生物合成也发生在成年人两性的大脑，通过酶芳香酶调节的过程（CYP19A1）。 AIM 3将使用 CRISPR/DCAS9双向调节芳香酶表达在神经元中的表达，以测试神经特异性E2合成在记忆形成中的作用。总的来说，这些实验将提供很多需要深入了解E2如何充当两个性别中神经元功能和记忆的关键调节剂。