Examining the Role of EAAT3 in OCD-like Behavior

检查 EAAT3 在强迫症样行为中的作用

• 批准号: 9464749
• 负责人: Jared Michael Kopelman
• 金额: $ 4.9万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-29 至 2020-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9464749
• 关键词: 9p24; Ablation; Acute; Adult; Affect; Agonist; Alleles; Amphetamines; Attenuated; Autistic Disorder; Autopsy; Behavior; Behavioral; Biological Assay; Brain; Brain region; Breeding; Cells; Chromosomes; Chronic; Code; Complex; Compulsive Behavior; Corpus striatum structure; Data; Development; Dopamine D1 Receptor; Etiology; Family Study; Generations; Genes; Genetic; Genetic Polymorphism; Glutamate Transporter; Glutamates; Grooming; Human; Hyperactive behavior; Impairment; Knock-in Mouse; Knock-out; Knockout Mice; Lead; Link; Luciferases; Medical; Mental disorders; Messenger RNA; Modeling; Mus; Neurons; Neurosciences; Obsessive-Compulsive Disorder; Pathogenesis; Patients; Pattern; Pharmacology; Phenotype; Population; Proteins; Reporter; Reversal Learning; Role; Suggestion; Testing; Transgenic Mice; Twin Studies; Up-Regulation; Western Blotting; World Health Organization; anxiety-like behavior; behavioral response; calmodulin-dependent protein kinase II; excitatory amino acid transporter 3; imaging study; improved; in vivo calcium imaging; inhibitor/antagonist; mouse model; overexpression; prepulse inhibition; prevent; protein expression; protein function; repetitive behavior; response; standard of care; stereotypy; tool; uptake

PROJECT SUMMARY/ABSTRACT Obsessive Compulsive Disorder (OCD) is a debilitating psychiatric illness with a complex etiology. Studies in humans have identified hyperactivity of corticostriatal circuits in patients with OCD. Furthermore, twin and family studies show a significant role for genetics in the etiology of OCD, with multiple studies identifying association of polymorphisms in the gene SLC1A1 with OCD. The most common of these OCD- associated polymorphisms increases expression of the encoded protein – the neuronal glutamate transporter, excitatory amino acid transporter-3 (EAAT3). This OCD-linked allele is associated with increased SLC1A1 expression in lymphoblastoid cells, human postmortem brain, a luciferase reporter assay, and transfected HEK cells, where there is also a functional increase in EAAT3 protein activity, as evidenced by increased glutamate uptake. There is also increased EAAT3 protein expression in striatum of Sapap3-knockout (KO) mice, a model of OCD-like behavior. These results suggest that increased SLC1A1 expression may result in increased generation of abnormal repetitive behaviors, and lead to the prediction that reducing EAAT3 may reduce compulsive behavior. Indeed, Slc1a1-STOP knock-in mice that have ablated EAAT3 protein expression and function show blunted responses during pharmacologically-induced repetitive behavior. Specifically, these mice have attenuated increases in stereotypy and hyperlocomotion in response to amphetamine, and attenuated grooming increases in response to a dopamine D1 receptor agonist. In addition, preliminary data indicate that a specific EAAT3 inhibitor (EAAT3i) can reduce compulsive-like behavior. Chronic EAAT3i administration reduced amphetamine-induced hyperlocomotion, and acute administration decreased grooming in a mouse model of autism-like behavior. However, the role of EAAT3 in OCD-like behavior and associated corticostriatal circuit activity has not been studied. I will therefore use advanced neuroscience tools to determine if chronic ablation and/or acute inhibition of EAAT3 normalizes behavior and striatal activity in a mouse model of OCD-like behavior, and directly test whether EAAT3 overexpression causes OCD-like behavior. My overarching hypothesis is that EAAT3 is necessary and sufficient for the development and expression of OCD-like behavior in mice.

项目摘要/摘要 强迫症（OCD）是一种具有复杂病因的精神病患者。 人类的研究确定了强迫症患者的皮质纹状体回路的多动症。此外， 双胞胎和家庭研究表明了遗传学在强迫症病因中的重要作用，并进行了多项研究 鉴定基因SLC1A1中多态性与OCD的缔合。这些强迫症中最常见的 相关的多态性增加了编码蛋白的表达 - 神经谷氨酸转运蛋白， 兴奋性氨基酸转运蛋白3（EAAT3）。该OCD连接等位基因与SLC1A1的增加有关 在淋巴母细胞中的表达，人类后大脑，荧光素酶报告基因测定法和翻译HEK EAAT3蛋白活性也有功能增加的细胞，谷氨酸增加了 吸收。在SAPAP3-KNOCKOUT（KO）小鼠的纹状体中，EAAT3蛋白表达也增加了 强迫症的行为。这些结果表明，SLC1A1表达的增加可能导致增加 产生异常的重复行为，并导致预测减少EAAT3可能会减少 强迫行为。实际上，具有消融的EAAT3蛋白表达和 功能显示在药物诱导的重复行为期间的反应钝。具体来说，这些 小鼠响应苯丙胺的刻板印象和超倒态剂的增加而减弱了，并且 响应多巴胺D1受体激动剂的响应减弱的修饰增加。此外，初步数据 表明特定的EAAT3抑制剂（EAAT3I）可以减少强迫性行为。慢性eaat3i 给药降低了苯丙胺诱导的超替代性，急性给药降低了修饰 在类似自闭症行为的鼠标模型中。但是，EAAT3在类似OCD的行为和相关的作用 皮质纹状体电路活性尚未研究。因此，我将使用先进的神经科学工具来 确定慢性消融和/或急性抑制EAAT3是否使A中的行为和纹状体活性归一化 OCD样行为的鼠标模型，并直接测试EAAT3过表达是否引起类似OCD 行为。我的总体假设是，EAAT3对于开发和 小鼠中强迫症行为的表达。