The Neurometabolome of a Sensory Neuronal Network

描述(由申请人提供)：了解健康和患病的大脑需要了解构成我们大脑的神经元和支持细胞内发生的生化知识。众所周知，神经系统中的单个细胞有不同的蛋白质补充，但这种异质性对细胞代谢组的影响还不是很清楚。虽然网络中使用的神经递质逐个神经元不同，但哺乳动物神经系统中代谢物的细胞间差异是什么？对于大多数代谢物来说，答案是未知的。利用一套基于小体积采样、毛细管电泳光诱导荧光检测、单细胞质谱仪、毛细管分离与傅立叶变换质谱仪相结合的分析技术，将以无与伦比的化学细节研究背根神经节(DRG)感觉神经元网络的神经代谢组。背根节神经元参与了疼痛的机制--这是许多疾病最具破坏性的副作用之一。神经递质和代谢物在单个神经元和它们的功能神经元隔间，如树突、轴突和细胞体，将被描述。这项工作的第一阶段是汇编关于DRG神经元代谢组的信息，并检查是否有不寻常的分子。接下来，使用DRG及其神经代谢物作为模型，解决了几个具体的问题：(1)不同功能区段的神经代谢物是否不同？(2)电或化学刺激时释放的小分子补体的哪一亚群，从而可能参与细胞间的通讯？(3)炎症诱导的疼痛如何改变DRG神经元的神经代谢物？这些努力符合代谢组学路线图倡议的范围，通过开发创新的方法来研究神经递质和其他小分子对模型感觉神经元网络功能的划分。

DESCRIPTION (provided by applicant): Understanding the healthy and the diseased brain requires knowledge of the biochemistry occurring within the neurons and supporting cells that make up our brain. It is well known that individual cells in the nervous system have distinct protein complements, but the consequences of this heterogeneity on the cellular metabolome are much less well understood. While neurotransmitters used in a network vary neuron-by-neuron, what are the cell-to-cell variations of the metabolome in the mammalian nervous system? For most metabolites, the answer is unknown. Using a suite of analytical technologies based on small-volume sampling, capillary electrophoresis with laser- induced fluorescence detection, single cell mass spectrometry, and capillary separations coupled to Fourier-transform mass spectrometry, the neurometabolome of a network of dorsal root ganglion (DRG) sensory neurons will be studied with unmatched chemical detail. DRG neurons are involved in the mechanisms of pain - one of most devastating side effects of many disorders. The neurotransmitters and metabolites in individual neurons and their functional neuronal compartments such as dendrites, axons, and cell bodies, will be characterized. The first stage of this work is to compile information on the metabolome of the DRG neurons, and examine this for unusual molecules. Next, using the DRG and its neurometabolome as the model, several specific questions are addressed: (1) Does the neurometabolome of functionally distinct neuronal compartments differ? (2) What subset of the small-molecule complement is released upon electrical or chemical stimulation, and thus may participate in cell-to-cell communication? (3) How does inflammation- induced pain change the neurometabolome of the DRG neurons? These efforts fit within the scope of the metabolomics roadmap initiative by developing innovative methodologies to investigate neurotransmitter and other small molecule compartmentalization on the function of a model sensory neuron network.