Optogenetic Targeting of mGluR5 Receptor Signaling

mGluR5 受体信号转导的光遗传学靶向

基本信息

批准号：
8724139
负责人：
M. FOSTER OLIVE
金额：
$ 19.29万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
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：
2014-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8724139
关键词：
Acute Agonist Alzheimer&apos s Disease Animal Model Behavior Behavioral Behavioral Paradigm Biochemical Brain Brain region C-terminal Calmodulin Carybdea Cattle Cerebral cortex Chemosensitization Chimeric Proteins Cognitive Cubozoa Development Disease Drug Addiction Extracellular Domain Fragile X Syndrome Genetic Glutamates Goals Health Image Imagery Impaired cognition Intake Investigation Laboratories Lead Legal Lentivirus Vector Light Mediating Mediator of activation protein Medical Methamphetamine N-Methyl-D-Aspartate Receptors Neurons Opsin Optics Pharmaceutical Preparations Phosphotransferases Photobleaching Play Property Protein Isoforms Proteins Psychological reinforcement Rattus Receptor Activation Receptor Signaling Relapse Reporter Research Resistance Retinal Retinaldehyde Reversal Learning Rhodopsin Rodent Model Role Schizophrenia Self Administration Signal Transduction Societies Specificity Subfamily lentivirinae Substance Use Disorder Time Transmembrane Domain Viral addiction cell type cost drug reward improved in vivo insight learning extinction memory recognition metabotropic glutamate receptor 5 neuropsychiatry novel novel therapeutics object recognition optogenetics promoter receptor receptor function selective expression socioeconomics spatiotemporal tool vector

项目摘要

DESCRIPTION (provided by applicant): The type 5 metabotropic glutamate receptor (mGluR5) plays a critical role in various aspects of drug addiction. Pharmacological or genetic inactivation of mGluR5 receptors reduces drug reward, reinforcement, and relapse- like behaviors. On the other hand, pharmacological potentiation of mGluR5 function via positive allosteric modulators (PAMs) produces pro-cognitive effects such as facilitated extinction learning and reversal of drug- induced cognitive impairments. However, various challenges and limitations to pharmacological activation of mGluR5 receptors significantly limit its utility as a research tool for detailed investigation of mGluR5 function in drug addiction. Such limitations include poor spatiotemporal control and lack of cell-type specificity of receptor activation, non-selectivity of orthosteric mGluR5 agonists, and suboptimal physiochemical properties of mGluR5 PAMs. These technical limitations can be surmounted by the successful development of optogenetic tools that allow for precise spatiotemporal and cell-type specific activation of mGluR5 receptor signaling. We have recently developed and performed a preliminary characterization of a lentiviral vector that expresses light- activated mGluR5 receptor (OptoXR-mGluR5). The expression of OptoXR-mGluR5 is under the control of Ca2+/calmodulin-dependent kinase IIα (CaMKIIα) promoter, allowing for selective expression in cortical glutamatergic neurons, and the vector also encodes enhanced yellow fluorescent protein (eYFP) reporter protein for visualization of expression. Our preliminary characterization of this lentivirus in the rat cerebral cortex in vivo demonstrates its cell-type specificity and ability to acutely activate mGluR5 signaling. However, the effects of OptoXR-mGluR5 activation in animal models of addiction-related behaviors and cognitive dysfunction have not yet been assessed. In addition, the ability of repeated stimulation of OptoXR-mGluR5 to consistently activate mGluR5 signaling, which would be necessary for behavioral studies conducted over the course of days or weeks, has not yet been explored. Therefore, the overarching goals of the studies proposed in this application are to examine the behavioral effects of acute OptoXR-mGluR5 activation in a rodent model of drug-induced cognitive dysfunction, and to develop novel tools for optogenetic activation of mGluR5 signaling that are amenable to behavioral paradigms involving repeated stimulation. These goals will be achieved under two independent Specific Aims. In Specific Aim 1, we will assess the ability of acute activation of OptoXR-mGluR5 to reverse drug-induced cognitive dysfunction. In Specific Aim 2, we will develop novel optogenetic tools for repeated activation of mGluR5 signaling. Successful development of optogenetic tools for mGluR5 activation will allow for an unprecedented level of investigation into the role of these receptors n various aspects of drug addiction. Optogenetic tools for mGluR5 activation may also provide insight into pathophysiological mechanisms and novel therapeutic avenues for other neuropsychiatric disorders in which mGluR5 receptors are implicated, including Alzheimer's disease, Fragile X syndrome, and schizophrenia.

描述（由适用提供）：5型代谢型谷氨酸受体（MGLUR5）在药物成瘾的各个方面起着至关重要的作用。 MGLUR5受体的药理或遗传灭活可减少药物奖励，增强和救济行为。另一方面，MGLUR5功能通过阳性变构调节剂（PAM）的药物潜力产生了认知效应，例如制备的扩展学习和药物诱导的认知障碍的反转。但是，MGLUR5受体的药物激活的各种挑战和局限性大大限制了其作为一种研究工具，以详细研究MGLUR5在药物成瘾中的功能。这种局限性包括不良的空间时间控制和缺乏受体激活的细胞类型特异性，直角MGLUR5激动剂的非选择性以及MGLUR5 PAM的次优理化特性。可以通过成功开发光遗传学工具来克服这些技术局限性，这些工具可以使MGLUR5受体信号的精确空间时间和细胞类型的特异性激活。我们最近对表达光活化的MGLUR5受体（OptOXR-MGLUR5）的慢病毒载体进行了初步表征。 OptOXR-MGLUR5的表达在CA2+/钙调蛋白依赖性激酶IIα（CAMKIIα）启动子的控制之下，允许在皮质谷氨酸能神经元中选择性表达，并且载体还编码增强的黄色荧光蛋白（EYFP）再生蛋白，以实现表达的可视化。我们在体内大鼠脑皮质中这种慢病毒的初步表征证明了其细胞类型的特异性和能力急性激活mglur5信号传导。然而，尚未评估OptOXR-MGLUR5激活在成瘾相关行为和认知功能障碍的动物模型中的影响。此外，尚未探索OptOXR-MGLUR5的重复模拟始终激活MGLUR5信号传导的能力，这对于在几天或几周内进行的行为研究是必不可少的。因此，本应用中提出的研究的总体目标是检查在药物诱导的认知功能障碍的啮齿动物模型中急性OptOXR-MGLUR5激活的行为效应，并开发出新的工具，以使MGLUR5信号的光遗传学激活可反复刺激到行为范式中，这些信号与行为范式相关。这些目标将在两个独立的特定目标下实现。在特定目标1中，我们将评估OptOXR-MGLUR5急性激活逆转药物诱导的认知功能障碍的能力。在特定的目标2中，我们将开发新型的光遗传学工具，以重复激活MGLUR5信号。成功开发用于MGLUR5激活的光遗传学工具将使对这些受体的作用进行前所未有的研究水平，而药物成瘾的各个方面。用于MGLUR5激活的光遗传学工具还可以提供有关其他神经精神疾病的病理生理机制和新型治疗途径，其中隐含了MGLUR5受体，包括阿尔茨海默氏病，脆弱的X综合征和精神分裂症。