Protein kinase C activity modulates myelin gene expression in enriched oligodendrocytes

Protein kinase C (PKC) and its potential role in myelin gene expression were investigated in primary cultured rat oligodendrocytes. The major myelin genes were expressed in a developmentally coordinated manner in cultured oligodendrocytes. PKC activity in these cells was similarly regulated with differential expression transiently and was most abundant in 9‐day cells in vitro. PKC‐α and PKC‐β mRNAs were present at low levels throughout development in these cells, and their expression increased in 18–25 day cells. Immunocytochemical colocalization of PKC with oligodendrocyte‐specific markers—O4, galactosyl cerebroside, MBP, and PLP—in enriched oligodendrocyte cultures suggested that the PKC predominantly contributed by oligodendrocytes. PKC inhibition resulting from long‐term exposure to 4β‐phorbol‐12,13‐dibutyrate (4β‐PDB) reduced steady‐state levels of MBP, PLP, MAG, CNP, and PKC‐α mRNAs, as detected by slot blots or in situ hybridization, and downregulated the oligodendrocyte‐specific markers O4, galactosyl cerebroside, and the major consitutent proteins MBP and PLP, as detected by immunocytochemistry. PKC‐mediated downmodulation of myelin gene expression was most profound in normally differentiating oligodendrocytes at or before the onset of myelin protein synthesis. Six‐day oligodendrocytes were most susceptible to such modulation. To elucidate the mechanism of reduction in various myelin gene messages upon modulation of PKC, we analyzed mRNA levels in oligodendrocytes, which were pretreated with either the transcriptional inhibitor actinomycin D or the protein synthesis blocker cycloheximide before exposure to 4β‐PDB. Our results demonstrate that the PKC inhibition‐mediated loss in myelin mRNA levels did not require the transcription of any genes, but appeared to be at least partially dependent on continuous protein synthesis. © 1993 Wiley‐Liss, Inc.