ROLE OF APOD IN NEURODEGENERATION

APO 在神经退行性变中的作用

• 批准号: 6923686
• 负责人: Shutish C. Patel
• 金额: $ 35.63万
• 依托单位: NEW ENGLAND BIOMEDICAL RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6923686
• 关键词: Niemann Pick disease; apolipoproteins; biological signal transduction; cell line; cholesterol; fibroblasts; fluorescence resonance energy transfer; gene expression; gene targeting; genetically modified animals; glycolipids; green fluorescent proteins; intracellular transport; laboratory mouse; laboratory rat; lipid transport; membrane transport proteins; microarray technology; neural degeneration; neurotoxins; oligodendroglia; protein localization; protein transport; vesicle /vacuole

Apolipoprotein D (apoD), a member of the lipocalin superfamily of transporters has been implicated in neurodegenerative disorders, neural injury and in neural regeneration. For instance, in Alzheimer's disease, there is increased apoD in the entorhinal cortex and in spinal fluid and the increased apoD correlates with inheritance of the apoE4 genotype. In the lysosomal cholesterol storage disorder, Niemann-Pick type C disease (NP-C), there is an induction of the apoD gene and apoD protein levels are increased several-fold in the brain and in plasma. In the regenerating sciatic nerve in the rat, following a crush injury, there is an almost 500-fold increase in apoD levels. ApoD mRNA and protein levels have also been shown to be upregulated following injury to the central nervous system. Finally, apoD gene expression has been shown to be increased in response to atypical neuroleptics suggesting that it may be a modulator of neuronal signal transduction. At a cellular level, we have found that apoD and NPC1, the product of the gene defective in most cases of NP-C, are components of a vesicular trafficking pathway that directs the retroendocytic movement of cholesterol and glycolipids. NPC1 is enriched in astrocytic foot processes around neuronal synapses. ApoD, on the other hand, is present in oligodendrocyte precursor cells, in pericytes, and in perivascular fibroblasts. In cultured peripheral cells and in glia, apoD and NPC1 localize to endocytic vesicles in a cholesterol- and glycolipid-dependent manner. The proposed studies aim to investigate further the role of apoD in neurodegeneration. To enable these studies we will use apoD knockout, apoD transgenic (expressing the human apoD gene) and apoD/NPC1 double knockout mice to investigate the following specific aims: (1) the developmental, behavioral and neurological phenotypes of apoD knockout, apoD transgenic and apod/NPC1 double knockout mice, and their response to neurotoxic and neurodegenerative lesions (2) cholesterol trafficking in apoD knockout and apoD/NPC1 double knockout mice (3) functional interaction(s) of apoD and NPC1 proteins using fluorescence resonance energy transfer (FRET), and (4) apoD and NPC1-mediated regulation of glial and neuronal cholesterol homeostasis.

载脂蛋白D（APOD）是转运蛋白的Lipocalin超家族的成员，与神经退行性疾病，神经损伤和神经再生有关。 例如，在阿尔茨海默氏病中，内嗅皮层和脊髓液中的APOD增加，而增加的APOD与APOE4基因型的遗传相关。 在溶酶体胆固醇储存障碍中，Niemann-pick型C疾病（NP-C），大脑和血浆中APOD基因的诱导和APOD蛋白水平增加了几倍。 在粉碎损伤后，在大鼠的再生坐骨神经中，APOD水平增加了近500倍。 APOD mRNA和蛋白质水平也已显示在中枢神经系统受伤后上调。 最后，已证明APOD基因表达会因非典型神经摄影的响应而增加，这表明它可能是神经元信号转导的调节剂。 在细胞水平上，我们发现APOD和NPC1（在大多数NP-C病例中，基因有缺陷的乘积）是导致胆固醇和糖脂质的逆转录细胞运动的囊泡运输途径的组成部分。 NPC1富含神经元突触周围的星形胶质脚步过程。 另一方面，APOD存在于少突胶质细胞前体细胞，周细胞和血管周成纤维细胞中。 在培养的外围细胞和神经胶质中，ApoD和NPC1以胆固醇和糖脂依赖性方式定位于内吞囊泡。 拟议的研究旨在进一步研究APOD在神经变性中的作用。 To enable these studies we will use apoD knockout, apoD transgenic (expressing the human apoD gene) and apoD/NPC1 double knockout mice to investigate the following specific aims: (1) the developmental, behavioral and neurological phenotypes of apoD knockout, apoD transgenic and apod/NPC1 double knockout mice, and their response to neurotoxic and neurodegenerative lesions (2) cholesterol使用荧光谐振能量转移（FRET）和（4）APOD和NPC1介导的胶质和神经元胆固醇稳态调节调节，使用APOD敲除和APOD/NPC1双基因敲除小鼠（3）功能相互作用。