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MBC in Press, published online ahead of print September 6, 2006
Mol. Biol. Cell 10.1091/mbc.E06-03-0243

A more recent version of this article appeared on November 1, 2006
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Submitted on March 28, 2006
Accepted on August 28, 2006

Sphingosine 1-phosphate Induces Myoblast Differentiation through Cx43 Protein Expression: Role for a Gap Junction-dependent and -independent Function

R. Squecco,* C. Sassoli,{dagger} F. Nuti,{ddagger} M. Martinesi,{ddagger} F. Chellini,{dagger} D. Nosi,{dagger} S. Zecchi-Orlandini,{dagger} F. Francini,* L. Formigli,{dagger} and E. Meacci{ddagger}

Departments of {ddagger}Biochemical Sciences, {dagger}Anatomy, Histology and Forensic Medicine, and *Physiological Sciences, University of Florence, Florence I-50134, Italy, and Interuniversity Institute of Myology, Italy

Monitoring Editor: Asma Nusrat

Although sphingosine 1-phosphate (S1P) has been considered a potent regulator of skeletal muscle biology, acting as a physiological anti-mitogenic and prodifferentiating agent, its downstream effectors are poorly known. In the present study, we provide experimental evidence for a novel mechanism by which S1P regulates skeletal muscle differentiation through the regulation of gap junctional protein connexin (Cx)43. Indeed, the treatment with S1P greatly enhanced Cx43 expression and gap junctional intercellular communication during the early phases of myoblast differentiation while the down-regulation of Cx43 by transfection with short interfering RNA blocked myogenesis elicited by S1P. Moreover, calcium and p38 MAPK-dependent pathways were required for S1P-induced increase in Cx43 expression. Interestingly, enforced expression of mutated Cx43{Delta}130-136 reduced gap junction communication and totally inhibited S1P-induced expression of the myogenic markers, myogenin, myosin heavy chain, caveolin-3, and myotube formation. Notably, in S1P-stimulated myoblasts, endogenous or wild type Cx43 protein, but not the mutated form, coimmunoprecipitated and colocalized with F-actin and cortactin in a p38 MAPK-dependent manner. These data, together with the known role of actin remodelling in cell differentiation, strongly support the important contribution of gap junctional communication, Cx43 expression and Cx43/cytoskeleton interaction in skeletal myogenesis elicited by S1P. Keywords: sphingosine 1-phosphate, connexin 43, C2C12 cells, myogenesis, cytoskeleton

Address correspondence to: E. Meacci (elisabetta.meacci{at}unifi.it)




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