Modulation of L-type calcium channel expression during retinoic acid-induced differentiation of H9C2 cardiac cells

The molecular mechanisms underlying the developmental regulation of L-type voltage-dependent Ca2+ channels (VDCCs) are still unknown, In this study, w e have characterized the expression patterns of skeletal (alpha(1S)) and ca rdiac (alpha(1C)) L-type VDCCs during cardiogenic differentiation in H9C2 c ells that derived from embry onic rat heart. We report that chronic treatme nt of H9C2 cells with 10 an 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 prev ented 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 mar kers indicated that RA, by preventing myogenic transdifferentiation, preser ves cardiac differentiation of this cell line, Altogether, we provide evide nce that cardiac and skeletal subtype-specific L-type Ca2+ channels are rel evant functional markers of differentiated cardiac and skeletal myocytes, r espectively, In conclusion our data demonstrate that in vitro RA stimulates cardiac (alpha(1C)) L-type Ca2+ channel expression, therefore supporting t he hypothesis that the RA pathway might be involved in the tissue specific expression of Ca2+ channels in mature cardiac cells.