Cultured rat skeletal muscle cells treated with cytochalasin exhibit normal dystrophin expression and intracellular free calcium control

Abstract

Many studies performed to elucidate the molecular and cellular processes involved in muscular dystrophies have led to the working hypothesis of a key role for the cytoskeleton elements linking the extracellular matrix to myofibrils. It was recently suggested that cytochalasin B treatment of mouse soleus muscle promoted cell damage mediated by a cytosolic increase in free calcium concentration. Since intracellular calcium overload may be a primary event resulting from the alteration of cytoskeletal structure, this study was intended to evaluate whether or not the integrity of the F-actin microfilament network is necessary for calcium homeostasis. The developmental establishment of the normal cytoarchitecture was altered by treatment of myoblasts with the actin-disrupting agents cytochalasin B and D, and the effects were compared with those in myoblasts treated with colchicine. These drugs modified the morphogenesis in that they prevented the formation of elongated myotubes by myoblast fusion, but did not prevent the maturation of contractile myogenic cells. The subcellular organisation of actin filaments visualised by confocal fluorescence microscopy was modified by colchicine and cytochalasins, but appearance of contractile apparatus and mechanical activity were not precluded. Sarcolemmal addressing of dystrophin, the subsarcolemmal protein lacking in Duchenne muscular dystrophy, was not prevented by cytochalasin. The evaluation of the basal activity of cytosolic calcium measured with indo-1 suggested that the disruption of actin or microtubules did not prevent developing muscle cells to maintain a low basal calcium activity. We propose that the global integrity of the cytoskeleton network is not crucial for the maintenance of calcium homeostasis in muscle cells developing in vitro. These results are discussed with regard to current theories attempting to understand the functional consequences of an abnormal expression of the dystrophin-glycoprotein complex interacting with the extracellular matrix and the cytoskeleton.