Lipidomic profile of endocannabinoids from neuronal cells

描述(由申请人提供)：这项提案的广泛、长期目标是开发新的代谢组学方法，以确定与内源性大麻素信号分子的代谢和生物合成有关的途径，如双胺和2-花生四烯基甘油(2-AG)。最终，这些脂肪组学策略可以用于体内或体外系统，以更好地了解在各种刺激下产生的内源性大麻素的概况。内源性大麻素是一类脂质信号分子，通常是来自长链脂肪酸的酰胺或酯。这些化合物具有神经调节活性，似乎在多种生理过程中发挥作用。大脑中发现的两种主要的内源性大麻素是花生四烯基乙醇胺(N-花生四烯基乙醇胺)和2-氨基酚。拟议的研究旨在改进用于研究阿南达胺和其他推定的内源性大麻素的生产和再循环的方法。将探索与某些钙通道以及神经递质受体激活相关的特定代谢和生物合成途径。该项目的具体目标是：1)开发和应用特定代谢物指纹和图谱方法，以确定内源性大麻素的脂肪组学特征，以及2)确定内源性大麻素释放与刺激相关的差异。这些研究将使用分子、生化、分析(薄层色谱、质谱学)和脂组/代谢组学方法。这些创新的比较脂组学研究将详细检查从代谢的山梨酸胺中提取的回收产品。更好地了解内源性大麻素的产生和释放机制将为大麻所作用的系统提供新的见解。公共相关性：这个项目与人类健康直接相关，因为内源性大麻素系统与记忆、情绪、认知、疼痛、发烧和免疫系统的调节有关。因此，对这一系统的药理操作在抽搐、青光眼、运动障碍、焦虑、肥胖和多发性硬化症等情况下具有潜在的治疗效果。了解控制内源性大麻素生产的分子基础对于未来针对这一系统的药物发现工作至关重要。

DESCRIPTION (provided by applicant): The broad, long-term objectives of this proposal are to develop new metabolomic methodologies to identify pathways involved with metabolism and biosynthesis of endogenous cannabinoid signaling molecules such as anandamide and 2-arachidonylglycerol (2-AG). Ultimately, these lipidomic strategies can be employed in in vivo or in vitro systems to better understand the profile of endogenous cannabinoids produced under a variety of stimuli. The endogenous cannabinoids are a family of lipid signaling molecules that are typically amides or esters derived from long-chain fatty acids. These compounds have neuromodulatory activity and appear to play a role in multiple physiological processes. The two major endocannabinoids identified in the brain are anandamide (N-arachidonyl- ethanolamide) and 2-AG. The proposed studies are designed to refine the approaches used to study production and recycling of anandamide as well as other putative endogenous cannabinoids. The specific metabolic and biosynthetic pathways linked to certain calcium channels as well as neurotransmitter receptor activation will be explored. The Specific Aims of this project are: 1) To develop and apply specific metabolite fingerprinting and profiling methodology for identifying the lipidomic profile for endogenous cannabinoids, and 2) To determine stimulus-dependent differences in the release of endogenous cannabinoids. The studies will use molecular, biochemical, analytical (thin layer chromatography, mass spectrometry), and lipidomic/ metabolomic approaches. These innovative comparative lipidomic studies will examine in detail the recycling products derived from metabolized anandamide. A better understanding of the mechanisms of endogenous cannabinoid production and release will provide new insight into the system that is acted upon by marijuana. The Public Relevance: This project has direct relevance to human health because the endocannabinoid system is implicated in modulation of memory, mood, cognition, pain, fever, and the immune system. As such, pharmacological manipulation of this system has potentially therapeutic effects in such conditions as convulsions, glaucoma, movement disorders, anxiety, obesity, and multiple sclerosis. Understanding the molecular basis for controlling the production of the endogenous cannabinoids is critical for future drug discovery efforts targeting this system.