Switching of actin isoforms in skeletal muscle differentiation using mouse ES cells

Among six actin isoforms, alpha-skeletal and alpha-cardiac actins have similar amino acid components and are highly conserved. Although skeletal muscles essentially express alpha-skeletal actins in the adult tissue, alpha-cardiac isoform actin is prominent in the embryonic muscle tissue. Switching of actin isoforms from alpha-cardiac to alpha-skeletal actin occurs during skeletal muscle differentiation. The cardiac type alpha-actin is expressed in the regeneration and patho-physiological states of the skeletal muscles as well. In the present study, we demonstrate the morphological switching of alpha-type actin isoforms from alpha-cardiac to alpha-skeletal actin in vitro using mouse ES cells for the first time. Immunofluorescent double staining with two specific antibodies revealed that alpha-cardiac actin appeared first in myoblasts. After cell fusion to form myotubes, the cardiac type actin decreased and alpha-skeletal actin conversely increased. Finally, the alpha-skeletal isoform remained as a main actin component in the fully mature skeletal muscle fibers. The exchange of isoforms is not directly linked to the sarcomere formation. As a result, ES cells provide a useful in vitro system for exploring skeletal muscle differentiation.