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Articles in PresS, published online ahead of print April 16, 2002
Physiol Genomics, 10.1152/physiolgenomics.00027.2002
Submitted on March 14, 2002
Accepted on April 14, 2002
1 Medicine (Molecular Cardiology), Albert Einstein College of Medicine, Bronx, New York, USA; Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
2 Molecular Genetics, Albert Einstein College of Medicine, Bronx, New York, USA
3 Anatomy & Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA
4 Medicine (Molecular Cardiology), Albert Einstein College of Medicine, Bronx, New York, USA; Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York, USA
* To whom correspondence should be addressed. E-mail: kitsis{at}aecom.yu.edu.
Significant progress has been made in defining pathways that mediate the formation of the mammalian heart. Little is known, however, about the genetic program that directs the differentiation of cardiac myocytes from their precursor cells. A major hindrance to this kind of investigation has been the absence of an appropriate cell culture model of cardiac myocyte differentiation. Recently, a subline of P19 cells (P19CL6) was derived that, following dimethylsulfoxide (DMSO) treatment, differentiate efficiently over 10 d into spontaneously beating cardiac myocytes. We demonstrate that these cells are indeed cardiac myocytes as they express cell type-specific markers and exhibit electrophysiological properties indicative of cardiac myocytes. The requirement for DMSO stimulation in this paradigm was shown to be limited to the first 4 days, suggesting that critical events in the differentiation process occur over this interval. To uncover relationships among known genes and identify novel genes that mediate cardiac myocyte differentiation, a detailed time course of changes in global gene expression was carried out using cDNA microarrays. In addition to the activation of genes encoding cardiac transcription factors and structural proteins, increases were noted in the expression of multiple known genes and ESTs. Analysis of the former suggested the involvement of a variety of signaling pathways in cardiac myocyte differentiation. The 16 ESTs whose expression was increased during the early, stimulus-dependent phase of cardiac myocyte differentiation, may be novel regulators of this process. Thus, this first report of large-scale changes in gene expression during cardiac myocyte differentiation has delineated relationships among the expression patterns of known genes and identified a number of novel genes that merit further study.
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