Characterization and Use of Fluorescent Endocannabinoid Transporter Substrates

荧光内源性大麻素转运蛋白底物的表征和使用

• 批准号: 7590454
• 负责人: MICHAEL PATRICK KAVANAUGH
• 金额: $ 17.69万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7590454
• 关键词: 2-arachidonylglycerol; AM404; Absence of pain sensation; Accounting; Adsorption; Adult; Biological Assay; Brain; CNR1 gene; Calcium; Cannabinoids; Cell Line; Cells; Cerebellum; Complementary DNA; Conflict (Psychology); Dependence; Detection; Endocannabinoids; Exhibits; Fluorescence; Fluorescence-Activated Cell Sorting; G-Protein-Coupled Receptors; Glioma; Hypertension; Incubated; Kinetics; Laser Scanning Confocal Microscopy; Libraries; Location; Maps; Measures; Mediating; Molecular; Molecular Cloning; Mus; Nature; Nerve Degeneration; Neuraxis; Neurons; Neurotransmitters; Physiological; Plasmids; Play; Presynaptic Receptors; Property; Prosencephalon; Radiolabeled; Relative (related person); Role; Signal Transduction; Slice; Somatic Cell; Spatial Distribution; Specificity; Synapses; Synaptic Transmission; System; Temperature; Therapeutic; Therapeutic Agents; Time; Vomiting; anandamide; base; cDNA Library; density; inhibitor/antagonist; novel; postsynaptic; presynaptic; public health relevance; radiotracer; receptor; response; reuptake; spatiotemporal; tool; uptake; vector

DESCRIPTION (provided by applicant): Endocannabinoids are released in the brain in response to synaptic activity and they interact with presynaptic receptors in a retrograde fashion to inhibit release of transmitter. They play key roles in modulating signaling between neurons. As with most other neurotransmitters, the termination of endocannabinoid action is thought to involve a transport activity that mediates reuptake into cells. Several putative inhibitors of reuptake have been shown to modulate endocannabinoid signaling, suggesting that the transporter may be an important target for gaining an understanding of endocannabinoid actions as well as for potential therapeutic applications. However, there is a great deal of controversy and a number of conflicting accounts concerning the mechanism and molecular basis of endocannabinoid transport. Because of this, we propose to use a novel fluorescent cannabinoid substrate as a selective tool to study endocannabinoid transport. This will allow us to 1) study the mechanism and kinetics of transport, 2) determine its spatiotemporal distribution in brain, and 3) to identify it by molecular cloning. Characterization and identification of the transporter will provide new strategies for developing potential therapeutic agents that modulate endocannabinoid levels for a wide range of applications including analgesia, anti-emesis, hypertension, and neurodegeneration. PUBLIC HEALTH RELEVANCE: Endocannabinoids produced in the brain interact with specific receptors to modulate signaling between neurons. As with most other neurotransmitters, the termination of endocannabinoid action is thought to involve a transport activity that mediates reuptake into cells. Because the molecular basis of endocannabinoid transport is unknown, we propose to use a novel fluorescent cannabinoid to 1) study its functional properties, 2) to determine its locations in the brain, and 3) to identify it by molecular cloning. Characterization and identification of the transporter will provide potential therapeutic strategies to modulate endocannabinoid levels for a wide range of applications including analgesia, anti-emesis, hypertension, and neurodegeneration.

描述（由申请人提供）：内源性大麻素在大脑中响应突触活动而释放，并且它们以逆行方式与突触前受体相互作用以抑制递质的释放。它们在调节神经元之间的信号传导中起着关键作用。与大多数其他神经递质一样，内源性大麻素作用的终止被认为涉及介导细胞再摄取的转运活动。几种假定的再摄取抑制剂已被证明可以调节内源性大麻素信号传导，这表明转运蛋白可能是了解内源性大麻素作用以及潜在治疗应用的重要靶点。然而，关于内源性大麻素转运的机制和分子基础存在大量争议和一些相互矛盾的说法。正因为如此，我们建议使用一种新的荧光大麻素底物作为选择性工具来研究内源性大麻素的运输。这将使我们能够1）研究运输的机制和动力学，2）确定其在脑中的时空分布，3）通过分子克隆来鉴定它。转运蛋白的表征和鉴定将为开发潜在的治疗剂提供新的策略，所述治疗剂调节内源性大麻素水平以用于广泛的应用，包括镇痛、止吐、高血压和神经变性。公共卫生相关性：大脑中产生的内源性大麻素与特定受体相互作用，调节神经元之间的信号传导。与大多数其他神经递质一样，内源性大麻素作用的终止被认为涉及介导细胞再摄取的转运活动。由于内源性大麻素转运的分子基础是未知的，我们建议使用一种新的荧光大麻素1）研究其功能特性，2）确定其在大脑中的位置，3）通过分子克隆来识别它。转运蛋白的表征和鉴定将提供潜在的治疗策略，以调节内源性大麻素水平，用于广泛的应用，包括镇痛，止吐，高血压和神经退行性变。