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：
8811416
负责人：
M. FOSTER OLIVE
金额：
$ 19.02万
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 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8811416
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 behavioral study 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 II¿ (CaMKII¿) 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（由申请人提供）：5 型代谢型谷氨酸受体 (mGluR5) 在药物成瘾的各个方面发挥着关键作用，另一方面，mGluR5 受体的药理学或基因失活会减少药物奖励、强化和复发样行为。，通过正变构调节剂 (PAM) 药理学增强 mGluR5 功能可产生促认知效应，例如促进消退学习和逆转然而，mGluR5 受体的逻辑药物激活的各种挑战和限制极大地限制了其作为详细研究药物成瘾中 mGluR5 功能的研究工具的实用性，这些限制包括时空控制差和缺乏细胞类型特异性。受体激活的影响、正位 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克服。开发光遗传学工具，可实现 mGluR5 受体信号传导的精确时空和细胞类型特异性激活。我们最近开发了表达光激活 mGluR5 受体（OptoXR-mGluR5）的慢病毒载体，并对其进行了初步表征。 -mGluR5 受 Ca2+/钙调蛋白依赖性激酶 II 的控制¿ (CaMKII¿) 启动子，允许在皮质谷氨酸能神经元中选择性表达，并且该载体还编码增强型黄色荧光蛋白 (eYFP) 报告蛋白，用于大鼠大脑皮层中表达可视化的初步表征，证明了其细胞。 -类型特异性和能力然而，OptoXR-mGluR5 激活对成瘾相关行为和认知功能障碍的动物模型的影响尚未得到评估。此外，重复刺激 OptoXR-mGluR5 持续激活 mGluR5 信号的能力。对于在数天或数周内进行的行为研究是否必要，尚未进行探索，因此，本申请中提出的研究的总体目标是检查急性行为的影响。 OptoXR-mGluR5 在药物引起的认知功能障碍的啮齿动物模型中的激活，以及开发适用于涉及重复刺激的行为范式的 mGluR5 信号光遗传学激活的新工具，这些目标将在两个独立的特定目标下实现。 1，我们将评估 OptoXR-mGluR5 的急性激活逆转药物引起的认知功能障碍的能力。在具体目标 2 中，我们将开发用于重复激活的新型光遗传学工具。 mGluR5 信号传导的光遗传学工具的成功开发将使对这些受体在药物成瘾的各个方面的作用的研究达到前所未有的水平，也可以为药物成瘾的病理生理机制和新的治疗途径提供深入的了解。其他与 mGluR5 受体有关的神经精神疾病，包括阿尔茨海默病、脆性 X 综合征和精神分裂症。