Characterization and Use of Fluorescent Endocannabinoid Transporter Substrates

描述(申请人提供)：内源性大麻素在大脑中释放，以响应突触活动，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递方面发挥着关键作用。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到介导细胞重新摄取的运输活动。几种假定的再摄取抑制剂已被证明调节内源性大麻素信号，提示转运体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，存在着很大的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型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转运。这将使我们能够1)研究转运的机制和动力学，2)确定其在脑内的时空分布，以及3)通过分子克隆来鉴定它。转运蛋白的鉴定和鉴定将为开发潜在的治疗药物提供新的策略，这些药物可以调节内源性大麻素的水平，用于广泛的应用，包括止痛、止吐、高血压和神经退行性变。与公共健康相关：大脑产生的内源性大麻素与特定的受体相互作用，调节神经元之间的信号传递。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到介导细胞重新摄取的运输活动。由于内源性大麻素转运的分子基础尚不清楚，我们建议使用一种新型的荧光大麻素来1)研究它的功能特性，2)确定它在脑中的位置，3)通过分子克隆来鉴定它。转运蛋白的特性和鉴定将为调节内源性大麻素水平提供潜在的治疗策略，用于广泛的应用，包括止痛、止吐、高血压和神经退行性变。

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.