COBRE: UND: ALPHA-SYNUCLEIN IN BRAIN LIPID METABOLISM

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。阐明α-突触核蛋白的生理功能很重要，因为该蛋白的突变被认为是家族性帕金森病(PD)的致病因素，并且与其他一些神经退行性疾病有关。初步数据显示，α-突触核蛋白显著影响大脑和分离的突触体内的稳态脂质质量，这表明α-突触核蛋白对脑磷脂代谢有显著影响。总的来说，这些变化和其他变化一起，可以改变膜的基本生物物理状态，并通过减少脂质介导的信号来减少细胞内第二信使的产生。因此，解决α-突触核蛋白影响脑脂代谢的机制(S)，以更好地了解α-突触核蛋白在脑中的潜在生理作用是至关重要的。以下具体目标旨在结合整体动物、细胞培养和体外方法来研究α-突触核蛋白对大脑磷脂代谢的影响。目的1.用稳态放射性示踪动力学方法研究α-突触核蛋白对小鼠脑内磷脂代谢的影响，以评价α-突触核蛋白基因修饰小鼠和对照小鼠脑磷脂生物合成和下游加工的动力学。目的2.用稳定同位素和放射性示踪剂检测α-突触核蛋白对细胞磷脂代谢的影响，以评价α-突触核蛋白基因抑制小鼠和对照小鼠神经元和胶质细胞磷脂生物合成和下游加工的动力学。目的3.利用稳定同位素和放射性示踪剂比较突变型和野生型α-突触核蛋白对稳定表达的HEK-293细胞磷脂代谢的影响，以评价磷脂生物合成和下游加工的动力学。目的4.结合多种方法测定野生型和突变型α-突触核蛋白在体外直接改变磷脂生物合成的能力。我们将检验这一假设，即α-突触核蛋白通过直接刺激磷脂酸的生物合成来刺激磷脂的合成，从而增加可用于下游合成特定磷脂的磷脂酸库。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Elucidating the physiological function of alpha-synuclein is important because mutations in this protein are implicated as a causative factor in familial Parkinson disease (PD) and because alpha-synuclein is associated with a number of other neurodegenerative diseases. Preliminary data contained herein indicates that alpha-synuclein dramatically affects steady-state lipid mass in brain and in isolated synaptosomes, suggesting that alpha-synuclein has a significant impact on brain phospholipid metabolism. Collectively, these changes, along with others, could alter the basal biophysical state of the membrane as well as decrease intracellular second messenger generation via a reduction in lipid-mediated signaling. Thus, it is critical to address the mechanism(s) by which alpha-synuclein affects brain lipid metabolism to gain a greater understanding regarding the potential physiological role for alpha-synuclein in the brain. The following specific aims are designed to address the impact that alpha-synuclein has on brain phospholipid metabolism using a combination of whole animal, cell culture, and in vitro approaches. Aim 1. Determine the effect of alpha-synuclein on brain phospholipid metabolism in vivo using steady-state radiotracer kinetics to assess the kinetics of phospholipid biosynthesis and downstream processing in alpha-synuclein gene-abated and control mice. Aim 2. Determine the effect of alpha-synuclein on cellular phospholipid metabolism ex vivo using stable isotopes and radiotracers to assess the kinetics of phospholipid biosynthesis and downstream processing in neurons and glia isolated from alpha-synuclein gene-abated and control mice. Aim 3. Compare the effects of mutant and wild-type alpha-synuclein on phospholipid metabolism in stably transfected HEK-293 cells using stable isotopes and radiotracers to assess the kinetics of phospholipid biosynthesis and downstream processing. Aim 4. Determine the ability of wild-type and mutant alpha-synuclein to directly alter phospholipid biosynthesis in vitro using a combination of approaches. We will test the hypothesis that alpha-synuclein stimulates phospholipid synthesis through a direct stimulation of phosphatidic acid biosynthesis, thereby increasing the phosphatidic acid pool available for use in downstream synthesis of specific phospholipids.