Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Islinger M, Grille S, Fahimi HD, Schrader M (2012) The peroxisome: an update on mysteries. Histochem Cell Biol 137:547–574
Schrader M, Thiemann M, Fahimi HD (2003) Peroxisomal motility and interaction with microtubules. Microsc Res Tech 61:171–178
Rucktäschel R, Girzalsky W, Erdmann R (2011) Protein import machineries of peroxisomes. Biochim Biophys Acta 1808:892–900
Trompier D, Savary S (2013) X-adrenoleukodystrophy. Morgan & Claypool Life Science, San Rafael, CA, pp 1–134
Kemp S, Theodoulou FL, Wanders RJ (2011) Mammalian peroxisomal ABC transporters: from endogenous substrates to pathology and clinical significance. Br J Pharmacol 164:1753–1766
Wanders RJ, Waterham HR (2006) Biochemistry of mammalian peroxisomes revisited. Annu Rev Biochem 75:295–332
Wanders RJ (2014) Metabolic functions of peroxisomes in health and disease. Biochimie 98:36–44
Fransen M, Nordgren M, Wang B, Apanasets O (2012) Role of peroxisomes in ROS/RNS-metabolism: implications for human disease. Biochim Biophys Acta 1822:1363–1373
Kassmann CM, Lappe-Siefke C, Baes M, Brugger B, Mildner A, Werner HB, Natt O, Michaelis T, Prinz M, Frahm J, Nave KA (2007) Axonal loss and neuroinflammation caused by peroxisome-deficient oligodendrocytes. Nat Genet 39:969–976
Baes M, Van Veldhoven PP (2012) Mouse models for peroxisome biogenesis defects and β-oxidation enzyme deficiencies. Biochim Biophys Acta 1822:1489–1500
Bottelbergs A, Verheijden S, Van Veldhoven PP, Just W, Devos R, Baes M (2012) Peroxisome deficiency but not the defect in ether lipid synthesis causes activation of the innate immune system and axonal loss in the central nervous system. J Neuroinflammation 9:61
Ferrer I, Aubourg P, Pujol A (2010) General aspects and neuropathology of X-linked adrenoleukodystrophy. Brain Pathol 20:817–830
Weber FD, Wiesinger C, Forss-Petter S, Regelsberger G, Einwich A, Weber WH, Köhler W, Stockinger H, Berger J (2014) X-linked adrenoleukodystrophy: very long-chain fatty acid metabolism is severely impaired in monocytes but not in lymphocytes. Hum Mol Genet 23:2542–2550
Blaylock RL (2013) Immunology primer for neurosurgeons and neurologists part 2: innate brain immunity. Surg Neurol Int 4:118
Bocchini V, Mazzolla R, Barluzzi R, Blasi E, Sick P, Kettenmann H (1992) An immortalized cell line expresses properties of activated microglial cells. J Neurosci Res 31:616–621
Henn A, Lund S, Hedtjärn M, Schrattenholz A, Pörzgen P, Leist M (2009) The suitability of BV2 cells as alternative model system for primary microglia cultures or for animal experiments examining brain inflammation. ALTEX 26:83–94
Fouquet F, Zhou JM, Ralston E, Murray K, Troalen F, Magal E, Robain O, Dubois-Dalcq M, Aubourg P (1997) Expression of the adrenoleukodystrophy protein in the human and mouse central nervous system. Neurobiol Dis 3:271–285
Liu J, Sabeva NS, Bhatnagar S, Li XA, Pujol A, Graf GA (2010) ABCD2 is abundant in adipose tissue and opposes the accumulation of dietary erucic acid (C22:1) in fat. J Lipid Res 51:162–168
Baarine M, Ragot K, Genin EC, El Hajj H, Trompier D, Andreoletti P, Ghandour MS, Menetrier F, Cherkaoui-Malki M, Savary S, Lizard G (2009) Peroxisomal and mitochondrial status of two murine oligodendrocytic cell lines (158N, 158JP): potential models for the study of peroxisomal disorders associated with dysmyelination processes. J Neurochem 111:119–131
Acknowledgment
A part of this work was presented as an oral presentation entitled “Peroxisomal and Oxidative Burst Defects” by Amira Zarrouk, Meriem Yousfi, Hayet Iddir, Thomas Nury, Catherine Gondcaille, Gérard Lizard at the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC)/European Society for Clinical Cell Analysis (ESCCA) Beckman Coulter, Flow Cytometry Course in St Etienne, France (April 11-13, 2013). This work was supported by grants from the INSERM, the Université de Bourgogne, the European Leucodystrophies Association (ELA; project number: 2010-03014), the Conseil Régional de Bourgogne, the Action Intégrée of the Comité Mixte Inter-universitaire Franco-Marocain (CMIFM, AIMA/14/310, EGIDE) from the PHC Volubilis/Toubkal program, Ministère des Affaires Etrangères, the Ministère de l’Enseignement et de la Recherche, and the Projet Sectoriel CNRST.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Debbabi, M. et al. (2017). Flow Cytometric Analysis of the Expression Pattern of Peroxisomal Proteins, Abcd1, Abcd2, and Abcd3 in BV-2 Murine Microglial Cells. In: Schrader, M. (eds) Peroxisomes. Methods in Molecular Biology, vol 1595. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6937-1_25
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6937-1_25
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6935-7
Online ISBN: 978-1-4939-6937-1
eBook Packages: Springer Protocols