Microarray analysis of global changes in gene expression during cardiac myocyte differentiation.
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 dimethyl sulfoxide (DMSO) treatment, differentiate efficiently over 10 days 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 expressed sequence tags (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.
Peng
Chang-Fu
CF
Department of Medicine (Molecular Cardiology), Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Wei
Yi
Y
Levsky
Jeffrey M
JM
McDonald
Thomas V
TV
Childs
Geoffrey
G
Kitsis
Richard N
RN
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D016000:Cluster Analysis; D004268:DNA-Binding Proteins / Q000235:genetics; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D050991:GATA4 Transcription Factor; D020869:Gene Expression Profiling / Q000379:methods / Q000706:statistics & numerical data; D005786:Gene Expression Regulation / Q000235:genetics*; D064326:MEF2 Transcription Factors; D051379:Mice; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D018006:Myogenic Regulatory Factors / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000706:statistics & numerical data; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D050991:GATA4 Transcription Factor; D064326:MEF2 Transcription Factors; C578041:Mef2c protein, mouse; D018006:Myogenic Regulatory Factors; D014157:Transcription Factors; D004121:Dimethyl Sulfoxide