Pharmacological characterization of phyto-cannabinoids and endocannabinoid system

内源性大麻素（eCBs）作为神经递质和神经调节剂普遍存在于生物系统中。与eCBs一样，植物大麻素（pCBs）已被证明可以控制运动活动、认知、感觉知觉以及一些自分泌和内分泌功能。介导这种作用的受体（大麻素和非大麻素）是否相互作用并构成一种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，可能形成同源和异源二聚体，并产生药理学上不同的功能实体。了解内源性大麻素系统（ECS）成分（包括cbr）在正常生理中的作用将有助于更好地理解大麻素介导的药物生物学。因此，我们建议- 1)确定大麻素在CBRs相互作用和异源二聚化中的作用2)表征调节CBRs功能的分子决定因素。利用形态学（免疫组织化学）、生化（Western blot和Co-IP）、生物物理（Pb-FRET分析）和分子（RT-PCR）方法，我们将表征CBR亚型（以及其他ECS成分）在SH-SY5Y细胞、神经元培养和大鼠脑中的表达和功能。一旦大麻素在CBRs二聚化中的作用被确定，我们建议确定效应蛋白（如GRKs）和膜实体（如脂筏）在串扰和功能特性中的作用。此外，我们建议研究CBRs在神经生成和大麻素介导的细胞内蛋白质组/代谢组变化中的作用，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.