ERK MAP kinase activation is required for acetylcholine receptor inducing activity-induced increase in all five acetylcholine receptor subunit mRNAs as well as synapse-specific expression of acetylcholine receptor ε- transgene

The AChR is a pentamer of four different subunits in a stoichiometry of α₂βγδ in embryonic and α₂βεδ in adult animals. Transcription of AChR subunit genes is most active in synaptic nuclei in adult skeletal muscle cells, and is regulated by neural factors such as ARIA. We report here that ARIA up-regulated specifically the expression of all five AChR subunits in C2C12 cells. The mRNA level of erbB2, erbB3, rapsyn, MUSK, SHP-2 and β- actin remained unchanged in response to ARIA stimulation in C2C12 cells. The ARIA-induced increase in AChR subunit expression in C2C12 cells was inhibited by the erbB kinase inhibitor tyrphostin AG1478 and the MEK inhibitor PD98059, but not by the PI3 kinase inhibitor wortmannin, suggesting an important role of the erbB protein tyrosine kinases and MAP kinase in the regulation of the expression of the five different AChR subunits. To determine the signaling pathways in vivo, we studied the expression of reporter genes driven by the ε-promoter in injected muscles. The in vivo expression of the ε-transgene was inhibited by co-expression of dominant negative mutants of key components in the MAP kinase pathway including ras, raf and MEK, but not the dominant negative mutant of PI3 kinase. These results suggest that ERK MAP kinase activation is required for ARIA-induced increase in all five AChR subunit mRNAs as well as synapse-specific expression of AChR ε-transgene.