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Endothelin induces differentiation of ANP‐EGFP expressing embryonic stem cells towards a pacemaker phenotype
Author(s) -
Gassanov Natig,
Er Fikret,
Zagidullin Naufal,
Hoppe Uta C.
Publication year - 2004
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.04-1619fje
Subject(s) - immunostaining , embryonic stem cell , connexin , green fluorescent protein , microbiology and biotechnology , biology , atrial natriuretic peptide , phenotype , medicine , transfection , cellular differentiation , embryonic heart , stem cell , endothelin 1 , cell culture , endocrinology , gap junction , immunology , intracellular , immunohistochemistry , receptor , gene , biochemistry , genetics
Currently, only limited insight into mechanisms promoting the differentiation and specification of the mammalian cardiac conduction system is available. Therefore, we established a murine embryonic stem (ES) cell line stably expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of the human atrial natriuretic peptide (ANP) promoter to further characterize the development of very early stages of the mammalian cardiac conduction tissue. The cardiac nature of ANP‐EGFP positive cells was confirmed by immunostaining. In ANP‐EGFP expressing ES cell‐derived cardiomyocytes, a distinct sublineage of pacemaker cells could be identified. Pacemaker cells displayed a spindle shape and exhibited a higher spontaneous beating rate, faster I f current activation and larger I f current densities compared with triangular atrial‐like cardiocytes. Exposure to endothelin‐1 significantly increased the percentage of pacemaker‐like cells without affecting their electrophysiological properties. These findings were corroborated by immunostaining with antibodies against connexin 40 and connexin 45, known markers for cardiac conduction tissue. Conversely, treatment of ANP‐EGFP expressing ES cells with neuregulin‐1 exhibited no effect on differentiation. These results indicate that ANP‐EGFP expression enables the identification of ES cell‐derived pacemaker cells by their fluorescence and morphology and that endothelin‐1 promotes the development of ANP‐EGFP positive cardiomyocytes to a pacemaker‐like phenotype.