BV-2小鼠小胶质细胞中过氧化物酶体蛋白Abcd1、Abcd2和Abcd3表达模式的流式细胞术分析
Flow Cytometric Analysis of the Expression Pattern of Peroxisomal Proteins, Abcd1, Abcd2, and Abcd3 in BV-2 Murine Microglial Cells.
作者信息
Debbabi Meryam, Nury Thomas, Helali Imen, Karym El Mostafa, Geillon Flore, Gondcaille Catherine, Trompier Doriane, Najid Amina, Terreau Sébastien, Bezine Maryem, Zarrouk Amira, Vejux Anne, Andreoletti Pierre, Cherkaoui-Malki Mustapha, Savary Stéphane, Lizard Gérard
机构信息
Laboratoire 'Biochimie du peroxysome, inflammation et métabolisme lipidique', EA7270/INSERM, Faculté des Sciences Gabriel, Université de Bourgogne Franche Comté, 6 Bd Gabriel, 21000, Dijon, France.
Faculté de Médecine, Laboratoire de Nutrition-Aliments Fonctionnels et Santé Vasculaire (LR12ES05), Monastir & Faculté de Médecine, Université de Monastir, Sousse, Tunisia.
出版信息
Methods Mol Biol. 2017;1595:257-265. doi: 10.1007/978-1-4939-6937-1_25.
Microglial cells play important roles in neurodegenerative diseases including peroxisomal leukodystrophies. The BV-2 murine immortalized cells are widely used in the context of neurodegenerative researches. It is therefore important to establish the expression pattern of peroxisomal proteins by flow cytometry in these cells. So, the expression pattern of various peroxisomal transporters (Abcd1, Abcd2, Abcd3) contributing to peroxisomal β-oxidation was evaluated on BV-2 cells by flow cytometry and complementary methods (fluorescence microscopy, and RT-qPCR). By flow cytometry a strong expression of peroxisomal proteins (Abcd1, Abcd2, Abcd3) was observed. These data were in agreement with those obtained by fluorescence microscopy (presence of numerous fluorescent dots in the cytoplasm characteristic of a peroxisomal staining pattern) and RT-qPCR (high levels of Abcd1, Abcd2, and Abcd3 mRNAs). Thus, the peroxisomal proteins (Abcd1, Abcd2, Abcd3) are expressed in BV-2 cells, and can be analyzed by flow cytometry.
小胶质细胞在包括过氧化物酶体脑白质营养不良在内的神经退行性疾病中发挥着重要作用。BV-2小鼠永生化细胞在神经退行性研究中被广泛使用。因此，通过流式细胞术确定这些细胞中过氧化物酶体蛋白的表达模式很重要。所以，通过流式细胞术以及补充方法（荧光显微镜和RT-qPCR）评估了参与过氧化物酶体β-氧化的各种过氧化物酶体转运蛋白（Abcd1、Abcd2、Abcd3）在BV-2细胞上的表达模式。通过流式细胞术观察到过氧化物酶体蛋白（Abcd1、Abcd2、Abcd3）有强烈表达。这些数据与通过荧光显微镜（细胞质中存在许多具有过氧化物酶体染色模式特征的荧光点）和RT-qPCR（Abcd1、Abcd2和Abcd3 mRNA的高水平）获得的数据一致。因此，过氧化物酶体蛋白（Abcd1、Abcd2、Abcd3）在BV-2细胞中表达，并且可以通过流式细胞术进行分析。
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