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J Biol Chem, Vol. 274, Issue 41, 29063-29070, October 8, 1999

Modulation of L-type Calcium Channel Expression during Retinoic Acid-induced Differentiation of H9C2 Cardiac Cells

Claudine Ménard, Sandrine Pupier, Dominique Mornet, Magali Kitzmann, Joël Nargeot, and Philippe Lory

From the IGH-CNRS UPR 1142, 141 rue de la Cardonille, 34396 Montpellier cedex 05, France and the  Muscles et Pathologies, INSERM, St Eloi, 34000 Montpellier, France

The molecular mechanisms underlying the developmental regulation of L-type voltage-dependent Ca²⁺ channels (VDCCs) are still unknown. In this study, we have characterized the expression patterns of skeletal (_1S) and cardiac (_1C) L-type VDCCs during cardiogenic differentiation in H9C2 cells that derived from embryonic rat heart. We report that chronic treatment of H9C2 cells with 10 nM all-trans-retinoic acid (all-trans-RA) enhanced cardiac Ca²⁺ channel expression, as demonstrated by reverse transcription-polymerase chain reaction, immunoblotting, and indirect immunofluorescence studies, as well as patch-clamp experiments. In addition, RA treatment prevented expression of functional skeletal L-type VDCCs, which were restricted to myotubes that spontaneously appear in control H9C2 cultures undergoing myogenic transdifferentiation. The use of specific skeletal and cardiac markers indicated that RA, by preventing myogenic transdifferentiation, preserves cardiac differentiation of this cell line. Altogether, we provide evidence that cardiac and skeletal subtype-specific L-type Ca²⁺ channels are relevant functional markers of differentiated cardiac and skeletal myocytes, respectively. In conclusion, our data demonstrate that in vitro RA stimulates cardiac (_1C) L-type Ca²⁺ channel expression, therefore supporting the hypothesis that the RA pathway might be involved in the tissue specific expression of Ca²⁺ channels in mature cardiac cells.

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