Endothelin-1 and neuregulin-1 convert embryonic cardiomyocytes into cells of the conduction system in the mouse

Abstract

The cells that form the cardiac conduction system (CCS) are recruited from embryonic cardiomyocytes. Endothelin-1 (ET-1) and Neuregulin-1 (NRG-1) have been associated with this transition in the avian and murine systems, respectively. We established murine embryonic cardiomyocyte cultures induced or not with ET-1 and/or NRG-1 to compare the expression of cardiomyocyte and CCS-specific genes. Semiquantitative reverse transcription-polymerase chain reaction analysis showed that cardiomyogenesis and CCS-specific markers, such as Nkx2.5, GATA4, Irx4, Connexin 40, Connesin 45, HF-1b, and MinK, were up-regulated in the presence of either growth factor. Additionally, immimofluorescence analysis demonstrated that ET-1 or NRG-1 increased the number of cells expressing the Purkinje fiber-specific marker Connexin 40 in induced cultures but did not selectively increase their proliferation rate. Interestingly, additive effects were not observed in ET-1 and NEG-1 combination treatments. Among other possibilities, this observation suggests that these factors may interact to promote the differentiation of the murine CCS. © 2005 Wiley-Liss, Inc.