The expression of PLP/DM-20 mRNA is restricted to the oligodendrocyte-lineage cells in the adult rat spinal cord

Kazuya Takeda, Mari Dezawa, Masaaki Kitada

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Proteolipid protein (PLP) is the major component of myelin; its gene encodes two major splicing variants: PLP and DM-20. Compared with PLP, DM-20 lacks the amino acids encoded by exon IIIb. The expression of PLP/DM-20 in cells outside the oligodendrocyte-lineage is unclear. To address this issue, we analyzed the detailed expression pattern of PLP/DM-20 mRNA in the adult rat spinal cord by in situ hybridization (ISH) with a cRNA probe complementary to DM-20 mRNA, which has been used to detect both PLP and DM-20 both mRNA. ISH did not label the cells expressing NeuN nor glial fibrillary acidic protein but detected those expressing Olig2, indicating that PLP/DM-20 mRNA are expressed only in oligodendrocyte-lineage cells. This cell population was expected to contain NG2-expressing oligodendrocyte precursor cells (OPCs), because some exhibited the expression of glutathione S-transferase pi isoform in the nucleus. A recent publication showed that OPCs express PLP but not DM-20 mRNA. However, no OPCs were detected. We performed ISH with a cRNA probe that specifically recognizes PLP mRNA to successfully detect some OPCs. Additionally, OPCs were detected by ISH with a cRNA probe complementary to DM-20 mRNA that was digested via alkaline hydrolysis prior to ISH. These findings collectively demonstrate that PLP and DM-20 mRNA expression is restricted to oligodendrocyte-lineage cells, and imply that the undigested cRNA probe complementary to the full-length DM-20 mRNA sequence only recognizes DM-20 mRNA and not the PLP counterpart when applied to ISH without denaturation/digestion methods.

Original languageEnglish
Pages (from-to)147-161
Number of pages15
JournalHistochemistry and Cell Biology
Volume145
Issue number2
DOIs
Publication statusPublished - 2016 Feb 1

Keywords

  • DM-20
  • Immunohistochemistry
  • In situ hybridization
  • Oligodendrocyte differentiation
  • Oligodendrocyte precursor cells
  • Proteolipid protein

ASJC Scopus subject areas

  • Histology
  • Molecular Biology
  • Medical Laboratory Technology
  • Cell Biology

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