Identification of enhancers whose activity defines cortical interneuron types

识别其活性定义皮质中间神经元类型的增强子

基本信息

批准号：
8935930
负责人：
JOHN L. R. RUBENSTEIN
金额：
$ 48.83万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-26 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8935930
关键词：
Adult Agreement Alleles Autistic Disorder Automobile Driving Behavioral Binding Biological Assay Brain Cell Transplants Cells Cerebral cortex ChIP-seq Development Disease Electrophysiology (science)Embryo Enhancers Epilepsy Funding Gene Expression Genes Genetic Enhancer Element Genetic Transcription Genomics Health Histones Human Image Immunohistochemistry In Situ Hybridization Interneuron function Interneurons Knowledge Label Lentivirus Vector Maps Medial Methods Molecular Molecular Genetics Morphology Mus Nature Neonatal Neurons Neurophysiology - biologic function Pattern Phenotype Physiological Physiology Play Positioning Attribute Process Property Proteins RNA Regulatory Element Reporter Research Rhodopsin Role Schizophrenia Series Slice Staging Taxonomy Time Transplantation Viral Whole-Cell Recordings base behavior measurement behavioral study cell type in vivo neurogenetics neuropsychiatry novel optogenetics progenitor ranpirnase research study tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Molecular definitions of neural cell types largely depend on the expression of RNAs or proteins as assessed by in situ hybridization, RNA array and sequencing, and immunohistochemistry. However, recent studies are demonstrating that gene regulatory elements, such as enhancers, can have highly specific spatial and temporal activity patterns in the developing brain. Thus, enhancer activity can be used to define neural cell types, and importantly, also have other broad applications. First, they can be used as tools to drive gene expression in specific cell types, which can then be used to visualize and/or purify the cells (GFP), modify gene expression in the cells (Cre), modify electrical activity (channel rhodopsin), and visualize electrical activity in the cells (GCaMP). Secondly, knowledge about the nature and position of enhancers enables geneticists to identify disease alleles that map in extra-exonic genomic space. Herein, we will focus on identifying enhancers that are active in cortical interneurons, during development and in the mature state. We choose to study cortical GABAergic interneurons because of their central role in cortical function and diseases. We will focus on these GABAergic neurons derived from the medial ganglionic eminence (MGE), which generates the majority of cortical interneurons. Our aim is to identify novel enhancers that are active in cortical interneurons using three assays: 1) histone (H3K27Ac) ChIP-seq (marker of active enhancers); 2) transcription factor (TF) ChIP- seq using TFs that regulate cortical interneurons development and function (Arx, Dlx2 and Lhx6); 3) a newly developed in vivo assay of enhancer function based on viral transduction of the enhancer driving Cre and GFP in immature cortical interneurons. We will then use a series of methods to define the interneuron subtypes that have enhancer activity. Together, our unique approach should define interneuronal cell types (developmental and adult), and simultaneously generate a powerful toolkit that will enable new ways to assess neural function and disease.

描述（由适用提供）：神经元细胞类型的分子定义在很大程度上取决于通过原位杂交，RNA阵列和测序以及免疫组织化学评估的RNA或蛋白质的表达。但是，最近的研究表明，基因调节元素（例如增强子）可以在发育中的大脑中具有高度特异性的空间和临时活动模式。这是可以使用增强剂活性来定义神经元细胞类型，重要的是还具有其他广泛的应用。首先，它们可以用作在特定细胞类型中驱动基因表达的工具，然后可以将其用于可视化和/或纯化细胞（GFP），修饰细胞中的基因表达（CRE），修饰电活动（通道锯状通道），并可视化细胞中的电活动（GCAMP）。其次，了解增强剂的性质和位置使遗传学家能够鉴定出在外部基因组空间中绘制的疾病等位基因。在此，我们将专注于确定在皮质中间神经元，发育和成熟状态中活跃的增强子。由于它们在皮质功能和疾病中的核心作用，我们将重点关注这些GABA能中间神经元。我们将重点关注这些源自内侧神经节杰出（MGE）的GABA能神经元，该神经元产生了大多数皮层中间神经元。我们的目的是使用三个测定法确定在皮质中间神经元中活跃的新型增强剂：1）组蛋白（H3K27AC）芯片seq（活性增强剂的标记）； 2）使用调节皮质间神经元发育和功能的TFS转录因子（TF）CHIP-SEQ（ARX，DLX2和LHX6）； 3）基于未成熟皮质中间神经元中增强子驱动CRE和GFP的病毒转导增强子功能的新开发的增强子功能。然后，我们将使用一系列方法来定义具有增强剂活性的间神经元亚型。我们独特的方法应该共同定义神经元细胞类型（发育和成人），并简单地产生一个强大的工具包，该工具包将启用评估神经功能和疾病的新方法。