Activation of Mitogen‐Activated Protein Kinases in Oligodendrocytes

The proliferation and differentiation of oligodendrocyte progenitors are stringently controlled by an interacting network of growth and differentiation factors. Not much is known, however, about the intracellular signaling pathways activated in oligodendrocytes. In this study, we have examined the activation of m itogen‐ a ctivated p rotein (MAP) kinase [also called e xtracellular s ignal‐ r egulated protein k inases (ERKs)] in primary cultures of developing oligodendrocytes and in a primary oligodendrocyte cell line, CG4, in response to platelet‐derived growth factor (PDGF) and basic fibroblast growth factor. MAP kinase activation was determined by an in‐gel protein kinase renaturation assay using myelin basic protein (MBP) as the substrate. The specificity of MAP kinase activation was further confirmed by an immune complex kinase assay using anti‐MAP kinase antibodies. Stimulation of oligodendrocyte progenitors with the growth factors PDGF and basic fibroblast growth factor and a protein kinase C‐activating tumor promoter, phorbol 12‐myristate 13‐acetate, resulted in a rapid activation of p42 mapk (ERK2) and, to a lesser extent, p44 mapk (ERK1). Immunoblot analysis with anti‐phosphotyrosine antibodies revealed an increased Tyr phosphorylation of a 42‐kDa phosphoprotein band cross‐reacting with anti‐MAP kinase antibodies. The phosphorylation of p42 mapk in PDGF‐treated oligodendrocyte progenitors was preceded by a robust autophosphorylation of the growth factor receptor. Immunoblot analysis with anti‐pan‐ERK antibodies indicated the presence of ERK‐immunoreactive species other than p42 mapk and p44 mapk in oligodendrocytes. The presence of some of the same pan‐ERK‐immunoreactive species and certain renaturable MBP kinase activities was also demonstrable in myelin preparations from rat brain, suggesting that MAP kinases (and other MBP kinases) may function not only during oligodendrogenesis but also in myelinogenesis.