Flow Cytometric Analysis of the Expression Pattern of Peroxisomal Proteins, Abcd1, Abcd2, and Abcd3 in BV-2 Murine Microglial Cells

Autor: Amira Zarrouk, Sébastien Terreau, Imen Helali, Flore Geillon, Maryem Bezine, Thomas Nury, Amina Najid, El Mostafa Karym, Anne Vejux, Mustapha Cherkaoui-Malki, Pierre Andreoletti, Meryam Debbabi, Doriane Trompier, Catherine Gondcaille, Gérard Lizard, Stéphane Savary
Rok vydání: 2017
Předmět:
Zdroj: Methods in Molecular Biology ISBN: 9781493969357
DOI: 10.1007/978-1-4939-6937-1_25
Popis: 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.
Databáze: OpenAIRE