COBRE: UND: ALPHA-SYNUCLEIN IN BRAIN LIPID METABOLISM

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 阐明α-突触核蛋白的生理功能是重要的，因为这种蛋白质中的突变被认为是家族性帕金森病（PD）的致病因素，并且因为α-突触核蛋白与许多其他神经退行性疾病相关。 本文包含的初步数据表明，α-突触核蛋白显著影响脑和分离的突触体中的稳态脂质质量，表明α-突触核蛋白对脑磷脂代谢具有显著影响。 总的来说，这些变化，沿着其他变化，可以改变膜的基础生物物理状态，以及通过减少脂质介导的信号传导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质代谢的机制以获得关于α-突触核蛋白在脑中的潜在生理作用的更好理解是至关重要的。以下具体目的旨在使用整体动物、细胞培养和体外方法的组合来解决α-突触核蛋白对脑磷脂代谢的影响。 目标1. 使用稳态放射性示踪剂动力学确定α-突触核蛋白对脑磷脂代谢的影响，以评估α-突触核蛋白基因减弱小鼠和对照小鼠中磷脂生物合成和下游加工的动力学。 目标2. 使用稳定同位素和放射性示踪剂测定α-突触核蛋白对离体细胞磷脂代谢的影响，以评估从α-突触核蛋白基因减弱小鼠和对照小鼠分离的神经元和神经胶质中磷脂生物合成和下游加工的动力学。 目标3. 使用稳定同位素和放射性示踪剂比较突变型和野生型α-突触核蛋白对稳定转染的HEK-293细胞中磷脂代谢的影响，以评估磷脂生物合成和下游加工的动力学。 目标4。 使用多种方法的组合测定野生型和突变体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α-突触核蛋白通过直接刺激磷脂酸生物合成来刺激磷脂合成，从而增加可用于特定磷脂下游合成的磷脂酸库的假设。

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.