Pharmacological characterization of phyto-cannabinoids and endocannabinoid system

内源性大麻素（eCB）在生物系统中无处不在，并作为神经递质和神经调质发挥作用。与eCB一样，植物大麻素（pCB）已被证明可以控制运动活动，认知，感官知觉以及几种自分泌和内分泌功能。介导这种效应的受体（大麻素和非大麻素）是否相互作用并构成新的功能复合物尚不清楚。因此，我们假设大麻素受体（CBRs）在被大麻素激活后，可能会形成同源和异源二聚体，并产生一个独特的功能实体。了解内源性大麻素系统（ECS）组分（包括CBR）在正常生理学中的作用将有助于更好地了解大麻素介导的药理学。我们建议：1）确定大麻素对CBR相互作用和异源二聚化的作用; 2）表征调节CBR功能的分子决定因素; 3）表征大麻素在轴突发生和蛋白质组变化中的作用;利用形态学（免疫组织化学）、生物化学（Western印迹和Co-IP）、生物物理学（Pb-FRET分析）和分子（RT-PCR）方法，我们将表征CBR亚型（和其他ECS组分）在SH-SY 5 Y细胞、神经元培养物和大鼠脑中的表达和功能。一旦确定大麻素对CBR二聚化的作用，我们建议确定效应蛋白如GRKs和膜实体如脂筏对串扰和功能特性的作用。此外，我们建议研究CBRs对轴突发生和大麻素介导的细胞内蛋白质组/代谢组变化的作用，以实现广泛的生物学功能。

Endocannabinoids (eCBs) are omnipresent in biological systems and function as neurotransmitters and neuromodulators. Like eCBs, phytocannabinoids (pCBs) have been shown to control motor activity, cognition, sensory perception, as well as several autocrine and endocrine functions. Whether the receptors (cannabinoid and non-cannabinoid) that mediate such effect interact and constitute a novel functional complex is unknown. Therefore, we hypothesize that cannabinoid receptors (CBRs), upon activation with cannabinoids, might form homo-and heterodimers and create a pharmacologically distinct functional entity. Understanding the role of endocannabinoid system (ECS) components, including CBRs, in normal physiology will allow for a better understanding of cannabinoids mediated pharmacobiology.Therefore, we propose to- 1) determine the role of cannabinoids on CBRs interaction and heterodimerization2) characterize the molecular determinants that regulate CBRs function.3) characterize the role of cannabinoids in neuritogenesis and changes in proteome/metabolomeUsing morphological (immunohistochemistry), biochemical (Western blot and Co-IP), biophysical (Pb-FRET analysis), and molecular (RT-PCR) approaches, we will characterize CBR subtypes (and other ECS components) expression and functions in SH-SY5Y cells, neuronal culture and rat brain. Once the role of cannabinoids on CBRs dimerization is identified, we propose to determine the role of effector proteins such as GRKs and membrane entities such as lipid rafts on cross-talk and functional properties. In addition, we propose to study the role of CBRs on neuritogenesis and cannabinoid-mediated changes in intracellular proteome/metabolome to fulfil a wide array of biological functions.