Characterization and Use of Fluorescent Endocannabinoid Transporter Substrates

描述（由申请人提供）：内源性大麻素在大脑中根据突触活动释放，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转运活动，介导再摄取到细胞中。一些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，这表明该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，存在大量的争议和一些相互矛盾的说法。因此，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)研究转运机制和动力学，2)确定其在大脑中的时空分布，3)通过分子克隆对其进行鉴定。转运体的表征和鉴定将为开发调节内源性大麻素水平的潜在治疗药物提供新的策略，其广泛应用包括镇痛、止吐、高血压和神经退行性变。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转运活动，介导再摄取到细胞中。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荧光大麻素来研究其功能特性，2)确定其在大脑中的位置，3)通过分子克隆对其进行鉴定。转运体的表征和鉴定将为调节内源性大麻素水平提供潜在的治疗策略，其广泛应用包括镇痛、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.