Differentiation of nonbeating embryonic stem cells into beating cardiomyocytes is dependent on downregulation of PKCβ and ζ in concert with upregulation of PKCε

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Abstract

Cardiomyocyte differentiation overall has been analyzed in vivo and in vitro at the molecular level by homologous recombination, gene mutation studies, and by transgenics; however, the roles of many signal transduction mechanisms that drive this differentiation process are still not fully understood. One set of signal transduction components that has been studied in detail in mature, differentiated cardiomyocytes is the PKC isotype superfamily. However, while the function of each isotype is slowly being uncovered in adult cardiomyocytes, limited information persists concerning their function in the differentiation process of cardiomyocytes. To begin analyzing the function of specific PKC isotypes in the differentiation process, we employed an established model for differentiating ES cells into cardiomyocyte-positive embryoid bodies (EBs) in vitro. RT-PCR, Western analyses, and confocal microscopy all showed that the expression of specific PKC isotypes was significantly changed as ES cells differentiated into cardiomyocytes. More importantly, by using antagonists specific for each isotype we found that this change was a final step in the differentiation process. PKCβ and ζ downregulation served to promote differentiation (beating), while upregulation of PKCε appeared to amplify differentiation (beating). Finally, melding classical tools (i.e., ionic exchange glass beads) with recently developed methods for differentiating ES cells creates a possible novel technique for investigating differentiation of ES cells into cardiomyocytes as well as other cell types. © 2003 Elsevier Science (USA). All rights reserved.

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Zhou, X., Quann, E., & Gallicano, G. I. (2003). Differentiation of nonbeating embryonic stem cells into beating cardiomyocytes is dependent on downregulation of PKCβ and ζ in concert with upregulation of PKCε. Developmental Biology, 255(2), 407–422. https://doi.org/10.1016/S0012-1606(02)00080-5

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