Accumulation of protein carbonyls within cerebellar astrocytes in murine experimental autoimmune encephalomyelitis.
Accumulation of protein carbonyls within cerebellar astrocytes in murine experimental autoimmune encephalomyelitis.
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DOI：
10.1002/jnr.22488
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发表时间：
2010-11-15
影响因子：
4.2
通讯作者：
Bizzozero, Oscar A.
中科院分区：
文献类型：
--
作者：
Zheng, Jianzheng;Bizzozero, Oscar A.
关键词：
我们实验室最近的研究表明，蛋白质羰基化与多发性硬化症（MS）和实验性自身免疫性脑脊髓炎（EAE）的病理生理有关。本研究旨在通过MOG35-55肽主动免疫C57/BL6小鼠制备EAE动物小脑的靶细胞和修饰蛋白，以确定疾病进展过程中蛋白质羰基化的变化。在该模型中，蛋白质羰基化在疾病高峰期（急性期）达到最大，此后（慢性期）减少。双免疫荧光显微镜显示，在急性期和慢性期，羰基积聚在白质星形胶质细胞中，在小胶质细胞/巨噬细胞中也有较小程度的积聚。令人惊讶的是，T细胞、少突胶质细胞和神经元几乎没有染色。通过2D-oxyblot和质谱分析，确定β-actin、β-tubulin、GFAP和HSC-71是整个疾病过程中羰基化的主要靶点。采用下拉/western blot方法，我们发现羰基化β-肌动蛋白、β-微管蛋白和GFAP的比例在慢性期显著增加，而在急性期无显著增加。这些结果表明，随着疾病从炎症阶段发展到神经退行性阶段，可能会不适当地去除氧化细胞骨架蛋白。此外，在EAE的慢性期，羰基化GFAP的广泛积累可能是造成这一阶段星形胶质细胞形状异常的原因。
Recent work from our laboratory has implicated protein carbonylation in the pathophysiology of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). The present study was designed to determine the changes in protein carbonylation during the disease progression, and to identify the target cells and modified proteins in the cerebellum of EAE animals, prepared by active immunization of C57/BL6 mice with MOG35-55 peptide. In this model, protein carbonylation was maximal at the peak of the disease (acute phase) to decrease thereafter (chronic phase). Double immunofluorescence microscopy of affected cerebella showed that carbonyls accumulate in white matter astrocytes, and to a lesser extent in microglia/macrophages, both in the acute and chronic phase. Surprisingly, T cells, oligodendrocytes and neurons were barely stained. By 2D-oxyblot and mass spectrometry, β-actin, β-tubulin, GFAP and HSC-71 were identified as the major targets of carbonylation throughout disease. Using a pull-down/western blot method we found a significant increase in the proportion of carbonylated β-actin, β-tubulin and GFAP in the chronic phase but not in the acute phase. These results suggest that as disease progresses from the inflammatory to the neurodegenerative phase there may be an inappropriate removal of oxidized cytoskeletal proteins. Additionally, the extensive accumulation of carbonylated GFAP in the chronic phase of EAE may be responsible for the abnormal shape of astrocytes observed at this stage.
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影响因子：
3.7
作者：
Maisonneuve E;Ducret A;Khoueiry P;Lignon S;Longhi S;Talla E;Dukan S
通讯作者：
Dukan S
影响因子：
4.2
作者：
Bizzozero, OA;Ziegler, JL;Bolognani, F
通讯作者：
Bolognani, F
影响因子：
3.3
作者：
Aksenov, MY;Aksenova, MV;Markesbery, WR
通讯作者：
Markesbery, WR
影响因子：
5.7
作者：
MacKenzie-Graham, Allan;Tiwari-Woodruff, Seema K.;Voskuhl, Rhonda R.
通讯作者：
Voskuhl, Rhonda R.
影响因子：
4
作者：
Goldman-Wohl, DS;Ariel, I;Yagel, S
通讯作者：
Yagel, S