Pharmacological characterization of phyto-cannabinoids and endocannabinoid system

点击翻译按钮获取中文摘要

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.