Expression of myelin genes: Comparative analysis of Oli‐neu and N20.1 oligodendroglial cell lines

Abstract

The use of immortalized cells has been instrumental as a tool in which to study gene regulation. However it is crucial to understand the status of a given cell line, especially when investigating the regulation of genes whose expression is developmentally regulated. Several immortalized cell lines have been derived from primary cultures of mouse oligodendrocytes. Two such cell lines – N20.1 and Oli-neu – were characterized here in terms of their relative expression of myelin genes at both the mRNA and protein level. Analysis of the splice isoforms expressed by the myelin proteolipid protein (Plp1), myelin basic protein (Mbp), and 2’,3’-cyclic nucleotide 3’-phosphodiesterase (Cnp) genes, along with the relative amount of protein expressed by these genes suggest that the cell lines are representative of immature oligodendrocytes, although Oli-neu cells appear to be further along the differentiation pathway compared to N20.1 cells. Previous studies have shown that the developmental increase in Plp1 gene expression that occurs during the active myelination period is governed by transcription regulatory elements present within the first intron. The responsiveness of one of these elements – the so-called antisilencer/enhancer (ASE) – was investigated in both cell lines. Results presented here suggest that the ASE has a much more potent effect in Oli-neu cells. Thus, the two cell lines appear to be at different stages and will be useful as a means to study transcription regulatory elements whose influence changes during development.

Oncostatin M protects against demyelination by inducing a protective microglial phenotype

“…C). To reveal whether this was a direct protective effect of OSM on the oligodendrocytes, we induced TNF‐α and IFN‐γ mediated cell death in the oligodendrocyte cell line, OliNeu (Pereira et al,). While cell viability decreased 72 h after addition of TNF‐α and IFN‐γ, pretreatment with different concentrations of OSM was unable to prevent this (Fig.…”

Drug discovery for remyelination and treatment of MS

“…Immortalized cell lines have the advantage of being relatively homogenous and rapidly expandable to quantities required for high‐throughput screening, but how closely they reflect the biology of the oligodendrocyte lineage in vivo is more questionable. For example, the Oli‐neu line (used by Joubert et al) does not mature sufficiently to robustly express markers of myelination (see review by (Pereira, Dobretsova, Hamdan, & Wight, ). Therefore, Joubert et al quantified total actin‐positive process length as a proxy of differentiation (Joubert et al, ).…”

NF‐Y‐dependent regulation of glutamate receptor 4 expression and cell survival in cells of the oligodendrocyte lineage

“…Oli‐neu cells are an established model for OPC (Jung et al, 1995; Pereira, Dobretsova, Hamdan, & Wight, 2011) previously used to study transcriptional regulation in oligodendroglia (Fratangeli et al, 2013). AMPAR expression has not been reported in Oli‐neu cells so we used qPCR and immunocytochemistry to detect AMPAR subunits GluA1–4.…”

“…Oli-neu cells are an established model for OPC (Jung et al, 1995;Pereira, Dobretsova, Hamdan, & Wight, 2011) previously used to study transcriptional regulation in oligodendroglia (Fratangeli et al, 2013).…”