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Vol. 16, Issue 8, 3632-3641, August 2005

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Vasopressin-dependent Myogenic Cell Differentiation Is Mediated by Both Ca²⁺/Calmodulin-dependent Kinase and Calcineurin Pathways

Bianca Maria Scicchitano ^*, Lucia Spath ^*, Antonio Musarò ^* , Mario Molinaro ^*, Nadia Rosenthal , Clara Nervi ^*, and Sergio Adamo ^* 

^* Department of Histology and Medical Embryology, University of Rome "La Sapienza," 00161 Roma, Italy; Interuniversity Institute of Myology, University of Rome "La Sapienza," 00161 Roma, Italy; and Mouse Biology Programme, European Molecular Biology Laboratory, 00016 Monterotondo, Italy

Submitted January 21, 2005; Revised May 11, 2005; Accepted May 24, 2005
Monitoring Editor: Marianne Bronner-Fraser

Arg⁸-vasopressin (AVP) promotes the differentiation of myogenic cell lines and mouse primary satellite cells by mechanisms involving the transcriptional activation of myogenic bHLH regulatory factors and myocyte enhancer factor 2 (MEF2). We here report that AVP treatment of L6 cells results in the activation of calcineurin-dependent differentiation, increased expression of MEF2 and GATA2, and nuclear translocation of the calcineurin target NFATc1. Interaction of these three factors occurs at MEF2 sites of muscle specific genes. The different kinetics of AVP-dependent expression of early (myogenin) and late (MCK) muscle-specific genes correlate with different acetylation levels of histones at their MEF2 sites. The cooperative role of calcineurin and Ca²⁺/calmodulin-dependent kinase (CaMK) in AVP-dependent differentiation is demonstrated by the effect of inhibitors of the two pathways. We show here, for the first time, that AVP, a "novel" myogenesis promoting factor, activates both the calcineurin and the CaMK pathways, whose combined activation leads to the formation of multifactor complexes and is required for the full expression of the differentiated phenotype. Although MEF2–NFATc1 complexes appear to regulate the expression of an early muscle-specific gene product (myogenin), the activation of late muscle-specific gene expression (MCK) involves the formation of complexes including GATA2.

Abbreviations used: AVP, arg⁸-vasopressin; CaMK, calcium/calmodulin-dependent kinase; CSA, cyclosporin A; HDAC, histone deacetylase; MCK, muscle creatine kinase; MEF2, myocyte enhancer factor 2; MHC, myosin heavy chain; NFAT, nuclear factor of activated T-cells.

Address correspondence to: Sergio Adamo (sergio.adamo{at}uniroma1.it).

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