Peroxisomal and mitochondrial status of two murine oligodendrocytic cell lines (158N, 158JP): potential models for the study of peroxisomal disorders associated with dysmyelination processes.
In some neurodegenerative disorders (leukodystrophies) characterized by myelin alterations, the defect of peroxisomal functions on myelin-producing cells (oligodendrocytes) are poorly understood. The development of in vitro models is fundamental to understanding the physiopathogenesis of these diseases. We characterized two immortalized murine oligodendrocyte cell lines: a normal (158N) and a jimpy (158JP) cell line mutated for the proteolipid protein PLP/DM20. Fluorescence microscopy, flow cytometry, and western blotting analysis allow to identify major myelin proteins (PLP colocalizing with mitochondria; myelin basic protein), oligodendrocyte (CNPase and myelin oligodendrocyte glycoprotein), and peroxisomal markers [adrenoleukodystrophy protein, PMP70, acyl-CoA oxidase 1 (ACOX1), l-peroxisomal bifunctional enzyme, and catalase]. Using electron microscopy, peroxisomes were identified in the two cell lines. Gene expression (ATP-binding cassette, Abcd1, Abcd2, Abcd3, and Acox1) involved in peroxisomal transport or beta-oxidation of fatty acids was evaluated using quantitative PCR. 4-phenylbutyrate treatment increases expression of ACOX1, l-peroxisomal bifunctional enzyme, PLP, myelin oligodendrocyte glycoprotein, and CNPase, mainly in 158N cells. In both cell lines, 4-phenylbutyrate-induced ACOX1 and catalase activities while only Abcd2 gene was up-regulated in 158JP. Moreover, the higher mitochondrial activity and content observed in 158JP were associated with higher glutathione content and increased basal production of reactive oxygen species revealing different redox statuses. Altogether, 158N and 158JP cells will permit studying the relationships between peroxisomal defects, mitochondrial activity, and oligodendrocyte functions.
Baarine
Mauhamad
M
Centre de Recherche INSERM, Dijon, France.
Ragot
Kevin
K
Genin
Emmanuelle C
EC
El Hajj
Hammam
H
Trompier
Doriane
D
Andreoletti
Pierre
P
Ghandour
M Said
MS
Menetrier
Franck
F
Cherkaoui-Malki
Mustapha
M
Savary
Stephane
S
Lizard
Gerard
G
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000378:metabolism; D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000378:metabolism; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D042962:Acyl-CoA Oxidase / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism; D002461:Cell Line, Transformed; D004746:Enoyl-CoA Hydratase / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D007535:Isomerases / Q000378:metabolism; D051379:Mice; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D009097:Multienzyme Complexes / Q000378:metabolism; D004676:Myelin Basic Protein / Q000378:metabolism; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein / Q000378:metabolism; D018992:Myelin-Associated Glycoprotein / Q000378:metabolism; D063308:Myelin-Oligodendrocyte Glycoprotein; D009836:Oligodendroglia / Q000000981:diagnostic imaging* / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D063994:Peroxisomal Bifunctional Enzyme; D020675:Peroxisomes / Q000378:metabolism*; D010654:Phenylbutyrates / Q000494:pharmacology; D018709:Statistics, Nonparametric; D014463:Ultrasonography
D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; C571327:Mog protein, mouse; D009097:Multienzyme Complexes; D004676:Myelin Basic Protein; D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D018992:Myelin-Associated Glycoprotein; D063308:Myelin-Oligodendrocyte Glycoprotein; D010654:Phenylbutyrates; C047516:Plp1 protein, mouse; C075773:4-phenylbutyric acid; D015094:3-Hydroxyacyl CoA Dehydrogenases; D002374:Catalase; D042962:Acyl-CoA Oxidase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; C576254:Ehhadh protein, mouse; D004746:Enoyl-CoA Hydratase; D063994:Peroxisomal Bifunctional Enzyme; D007535:Isomerases