Oxygenated Species of Cardiolipins as Biomarkers of Mitochondrial Dysfunction

这项应用的目标是鉴定环境诱导的(鱼藤酮--)磷脂生物标志物。 诱导)与帕金森病帕金森病相关的线粒体功能障碍。我们展示了一个 线粒体特异的磷脂，心磷脂(CL)，经历细胞色素c催化的选择性氧化 (Cytc)在神经细胞凋亡的早期。我们的中心假设是接触杀虫剂鱼藤酮， 导致CL的时间和剂量依赖的选择性氧化及其氧化的积累 通过酶促细胞色素C与线粒体功能障碍相关的分子物种 这些反应在细胞凋亡早期就被触发了。CL分子物种的独特轮廓代表了一种新的 鱼藤酮诱导的线粒体功能障碍与帕金森病相关的生物标志物类型。使用氧化法 脂质组学方法我们将首先确定诱导大鼠CL氧化的分子物种的特定模式 原代皮质和中脑神经元以及神经母细胞瘤SH-SY5Y，用鱼藤酮。此外，我们还将 用大鼠鱼藤酮揭示多巴胺能神经元和皮质神经元氧化化学发光物种的特异性 PD的输注模型。最后，我们将确定鱼藤酮特定的CL氧化模式在 鱼藤酮对人外周血淋巴细胞的作用及与正常对照的比较 鱼藤酮输注大鼠模型。为了检验这一假设，制定了以下具体目标：具体目标 1将利用氧化脂质组学来鉴定和表征CL的分子种类以及 CL在原代大鼠皮质神经元和中脑的独特立体特异性氧化产物 在鱼藤酮作用下，神经细胞和SH-SY5Y细胞中也有明显的变化。具体目标2将确立 细胞色素c与CL相互作用参与CL氧化的机制和途径 鱼藤酮对原代培养的大鼠皮质和中脑神经元以及SH-SY5Y细胞的作用。特定的 目标3将确定鱼藤酮诱导的过氧化CL的分子物种的程度 在体外神经元中检测和鉴定蓄积在皮质和中脑线粒体中 大鼠注射鱼藤酮后体内神经元的变化。特定目标4将揭示鱼藤酮特定的CL 人外周血淋巴细胞作为线粒体生物标志物的过氧化模式 与帕金森病相关的功能障碍。

The goal of this application is to identify phospholipid biomarkers of environmentally-induced (rotenone- induced) mitochondrial dysfunction associated with Parkinson's disease PD. We demonstrated that a mitochondria specific phospholipid, cardiolipin (CL), undergoes selective oxidation catalyzed by cytochrome c (cyt c) early during neuronal apoptosis. Our central hypothesis is that exposure to a pesticide, rotenone, causes time- and dose-dependent selective oxidation of CL and accumulation of its oxidized molecular species associated with mitochondrial dysfunction through enzymatic cyt c catalyzed reactions triggered early in apoptosis. The unique profile of CL molecular species represents a new type of biomarkers of rotenone-induced mitochondrial dysfunction associated with PD. Using oxidative lipidomics approach we will first identify specific patterns of CL oxidized molecular species induced in rat primary cortical and midbrain neurons as well as neuroblastoma SH-SY5Y by rotenone. Further, we will reveal the specific profiles of oxidized CL species in dopaminergic and cortical neurons using rat rotenone- infusion model of PD. Finally we will establish the presence of the rotenone-specific CL oxidation patterns in human peripheral blood lymphocytes exposed to rotenone and compare them with those detected in rotenone-infusion rat model. The following Specific Aims were developed to test the hypothesis: Specific Aim 1 will utilize oxidative lipidomics to identify and characterize molecular species of CL as well as unique stereo-specific oxygenated products of CL in primary rat cortical neurons and midbrain neurons as well as in SH-SY5Y cells upon exposure to rotenone. Specific Aim 2 will establish the mechanisms and pathways through which interactions of cyt c with CL are involved in CL oxidation in primary rat cortical and midbrain neurons as well as SH-SY5Y cells exposed to rotenone. Specific Aim 3 will determine the extent to which molecular species of rotenone-induced peroxidized CL detected and identified in neurons in vitro accumulate in mitochondria of cortical and midbrain neurons in vivo after infusion of rotenone to rats. Specific Aim 4 will reveal rotenone-specific CL peroxidation patterns in human peripheral blood lymphocytes as biomarkers of mitochondrial dysfunction associated with PD.