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Deciphering the GABA neuron alterations in schizophrenia

破译精神分裂症中 GABA 神经元的改变

基本信息

批准号：
10545717
负责人：
KENNETH N FISH
金额：
$ 48.58万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-15 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10545717
关键词：
Affect Amygdaloid structure Autopsy Behavior Biological Models Bipolar Disorder Brain Cells Cholecystokinin Cognitive deficits Complex Confocal Microscopy Data Development Diagnosis Disease Environmental Risk Factor Enzymes Etiology Fluorescent in Situ Hybridization Foundations Functional disorder Future Gene Expression Genes Genetic Transcription HDAC1 gene Human Immunohistochemistry Impaired cognition Individual Label Lasers Learning Major Depressive Disorder Messenger RNA Methylation Microdissection Mus Myoepithelial cell Neurons Parvalbumins Pathway interactions Pattern Prefrontal Cortex Presynaptic Terminals Primates Process Proteins Reporting Schizophrenia Signal Pathway Signal Transduction Specificity Testing Transcript calbindin calretinin cell type cohort differential expression extracellular gamma-Aminobutyric Acid gene product genetic variant hippocampal pyramidal neuron insight neural circuit neuronal cell body neuronal circuitry neurotransmission novel programs promoter sensor therapeutic target transcriptome transcriptome sequencing

项目摘要

PROJECT SUMMARY Schizophrenia (SZ) results from the complex interplay of genetic variants affecting liability and environmental risk factors that alter developmental trajectories of neural circuits. Despite this etiological diversity, alterations in GABA neurons are a highly conserved feature of SZ and substantial data suggest these alterations contribute to cognitive dysfunction in the disorder. GABA neurotransmission is heavily dependent on GAD67. Many GABA neurons in the PFC of SZ subjects express normal levels of GAD67 mRNA, but this transcript is not detectable in 30-50% of PFC GABA neurons in the disorder. The identities of the affected GABA neurons are largely unknown. Transcriptionally-unique subtypes (TUS) of GABA neurons uniquely target distinct pyramidal neuron (PN) ensembles, allowing them to powerfully modulate certain brain-wide networks and behaviors. Thus, knowing the identity of the affected GABA TUS in SZ and the mechanisms underlying GAD67 mRNA deficits occurring in only some GABA TUS is essential for moving towards a mechanistic understanding of the cortical circuitry alterations in the illness. To meet these needs, the following Aims are proposed: Aim 1. Quantify GAD67 mRNA and protein levels in the soma and axon terminals, respectively, of GABA TUS in SZ. Information from profiling studies of GABA neurons in mice, was used to identify mRNA markers that distinguish 10 GABA TUS in human PFC, which capture >95% of all GABA neurons. Multiplex fluorescence in situ hybridization (FISH) is used to quantify GAD67 mRNA levels within these TUS in the PFC of matched SZ and control (CON) subjects (Cohort 1). In the same subjects, multi-label immunohistochemistry is used to quantify GAD67 protein levels in the axon terminals of the different GABA TUS. The GAD67 mRNA deficit in SZ is predict to be restricted to specific GABA TUS, which have lower terminal GAD67 protein levels. Aim 2. Determine the pattern of altered TUS that is specific to the disease process of SZ. Some alterations in cortical GABA neurons, including lower levels of GAD67, seem to be shared across SZ, bipolar disorder (BP), and major depressive disorder (MDD), whereas those of others are not. The studies of Aim 1 are conducted in an all new second cohort of matched SZ, BP, MDD and CON subjects. SZ, BP, and MDD are predicted to each have a unique constellation of affected GABA neuron subtypes relative to each other. Aim 3. Perform transcriptome sequencing of GABA TUS in SZ and CON subjects. In Cohort 1 subjects, individual neurons of each GABA TUS are identified by FISH, collected using laser microdissection, pooled and subjected to transcriptome sequencing. In SZ, the affected GABA TUS are predicted to share a pattern of differentially-expressed genes that are components of pathways known to regulate GAD67 expression. By identifying the SZ-specific constellation of affected GABA TUS and assessing their transcriptome profile, these studies will provide essential information for future mechanistic tests of TUS-specific GABA neuron circuit dysfunction in model systems and developing therapeutic targets of GABA TUS.

项目总结精神分裂症(SZ)是遗传变异影响易感性和环境的复杂相互作用的结果改变神经回路发育轨迹的风险因素。尽管病原学多样化，但变化在GABA中，神经元是SZ的一个高度保守的特征，大量数据表明这些变化会导致认知功能障碍。GABA的神经传递严重依赖于GAD67。 SZ受试者PFC中的许多GABA神经元表达正常水平的GAD67 mRNA，但该转录本是在疾病中30-50%的PFC GABA神经元中未检测到。受影响的GABA神经元的身份在很大程度上是未知的。转录独特亚型(TU)的GABA神经元具有独特的靶向性锥体神经元(PN)集合，使它们能够强大地调节某些全脑网络和行为。因此，知道深圳受影响的GABA TUS的身份和GAD67的机制仅在某些GABA TU中出现的mRNA缺陷对于迈向机械论理解是必不可少的疾病中大脑皮层回路的改变。为满足这些需求，我们提出了以下目标：目的1.定量检测GAD67在胞体和轴突终末的表达水平。在深圳的GABA TUS。来自小鼠GABA神经元的图谱研究的信息被用来鉴定mRNA 区分人类前额叶中10个GABA TU的标记，这些标记捕获了所有GABA神经元的95%。多路传输荧光原位杂交(FISH)被用来定量PFC中这些TU中GAD67mRNA的水平匹配的SZ和对照组(CON)受试者(队列1)。在同一受试者中，多标记免疫组织化学用于量化不同GABA TU轴突终末的GAD67蛋白水平。GAD67基因的mRNA 据预测，SZ的缺陷仅限于特定的GABA TU，它们的末端GAD67蛋白水平较低。目的2.确定SZ疾病过程中所特有的TUS改变模式。一些人皮质GABA神经元的改变，包括GAD67水平降低，似乎在SZ、双相情感障碍中都存在精神障碍(BP)和严重抑郁障碍(MDD)，而其他人则不是。AIM-1的研究研究对象为SZ、BP、MDD和CON的全新第二队列受试者。SZ、BP和MDD是据预测，每一种都有相对于彼此的受影响的GABA神经元亚型的独特星座。目的3.对SZ和CON受试者的GABATUS进行转录组测序。在第一组受试者中，每个GABA TU的单个神经元通过FISH鉴定，使用激光显微解剖收集，合并并进行转录组测序。在深圳，受影响的GABA TU被预测为共享一个模式差异表达的基因是已知的调节GAD67表达的途径的组成部分。通过鉴定受影响的GABA TU的SZ特异性星座并评估它们的转录组图谱，这些研究将为未来TUS特异性GABA神经元的机械测试提供必要的信息模型系统中的回路功能障碍与GABA TUS治疗靶点的开发。