Effects of electrical fields on cardiomyocyte differentiation of embryonic stem cells

The effects of electromagnetic fields (EMFs) on the differentiation of cardiomyocytes in embryoid bodies derived from pluripotent embryonic stem (ES) cells were investigated. A single direct current (DC) field pulse was applied to 4‐day‐old embryoid bodies. The electrical field induced a hyperpolarization of the anode‐facing side of embryoid bodies and a depolarization at the cathode‐facing side. Significant effects of a single electrical field pulse applied for 90 s on cardiomyocyte differentiation were achieved with field strengths of 250 and 500 V/m, which increased both the number of embryoid bodies differentiating beating foci of cardiomyocytes and the size of the beating foci. The 500‐V/m electrical field increased intracellular reactive oxygen species (ROS), but not [Ca2+]i and activated nuclear factor kappa B (NF‐κB). A comparable increase in the number of beating embryoid bodies was achieved by an incubation for 1 h with H2O2 (1–10 nM), indicating that the electrical field effect was transduced via the intracellular generation of ROS. Because the radical scavengers dehydroascorbate and pyrrolidinedithiocarbamate (APDC) and the NF‐κB antagonist N‐tosyl‐L‐phenylalanine chloromethyl ketone (TPCK) inhibited cardiac differentiation, we assume that ROS and NF‐κB may play a role in early cardiac development. J. Cell. Biochem. 75:710–723, 1999. © 1999 Wiley‐Liss, Inc.

[1]  L. Jaffe,et al.  Neurites grow faster towards the cathode than the anode in a steady field. , 1979, The Journal of experimental zoology.

[2]  L. Jaffe,et al.  Strong electrical currents leave the primitive streak of chick embryos. , 1979, Science.

[3]  I. Cameron,et al.  Environmental magnetic fields: Influences on early embryogenesis , 1993, Journal of cellular biochemistry.

[4]  R. Schwartz,et al.  Competition between negative acting YY1 versus positive acting serum response factor and tinman homologue Nkx-2.5 regulates cardiac alpha-actin promoter activity. , 1997, Molecular endocrinology.

[5]  T. Libermann,et al.  Multiple myeloma cell adhesion-induced interleukin-6 expression in bone marrow stromal cells involves activation of NF-kappa B. , 1996, Blood.

[6]  K. Chien,et al.  Cardiotrophin-1 and the role of gp130-dependent signaling pathways in cardiac growth and development , 1997, Journal of Molecular Medicine.

[7]  K. Chien,et al.  Cardiotrophin-1 displays early expression in the murine heart tube and promotes cardiac myocyte survival. , 1996, Development.

[8]  E. Dejana,et al.  Embryonic stem cells differentiate in vitro to endothelial cells through successive maturation steps. , 1996, Blood.

[9]  K R Robinson,et al.  The responses of cells to electrical fields: a review , 1985, The Journal of cell biology.

[10]  J. Hescheler,et al.  DC electrical field-induced c-fos expression and growth stimulation in multicellular prostate cancer spheroids. , 1997, British Journal of Cancer.

[11]  K. Hotary,et al.  Endogenous electrical currents and the resultant voltage gradients in the chick embryo. , 1990, Developmental biology.

[12]  R. G. Allen,et al.  Oxidative influence on development and differentiation: an overview of a free radical theory of development. , 1989, Free radical biology & medicine.

[13]  S. Gollnick,et al.  Activation of multiple transcription factors and fos and jun gene family expression in cells exposed to a single electric pulse. , 1995, Experimental cell research.

[14]  A. Guimond,et al.  Characterization of 45-kDa/54-kDa HSP27 kinase, a stress-sensitive kinase which may activate the phosphorylation-dependent protective function of mammalian 27-kDa heat-shock protein HSP27. , 1995, European journal of biochemistry.

[15]  P. Colepicolo,et al.  Post-implantation mouse embryos have the capability to generate and release reactive oxygen species. , 1995, Reproduction, fertility, and development.

[16]  A M Wobus,et al.  Cardiomyocytes differentiated in vitro from embryonic stem cells developmentally express cardiac-specific genes and ionic currents. , 1994, Circulation research.

[17]  J Bernhardt,et al.  On the generation of potential differences across the membranes of ellipsoidal cells in an alternating electrical field , 1973, Biophysik.

[18]  M. Wiles.  Embryonic stem cell differentiation in vitro. , 1993, Methods in enzymology.

[19]  A. Bradley,et al.  Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector , 1986, Nature.

[20]  H. Pahl,et al.  Reactive oxygen intermediates as second messengers of a general pathogen response. , 1996, Pathologie-biologie.

[21]  J. Remacle,et al.  Low levels of reactive oxygen species as modulators of cell function. , 1995, Mutation research.

[22]  M. Baggiolini,et al.  Chemiluminescence detection of H2O2 produced by human neutrophils during the respiratory burst. , 1987, Analytical biochemistry.

[23]  P. Mehlen,et al.  hsp27 as a Switch between Differentiation and Apoptosis in Murine Embryonic Stem Cells* , 1997, The Journal of Biological Chemistry.

[24]  J. Hescheler,et al.  Spontaneous calcium oscillations in embryonic stem cell-derived primitive endodermal cells. , 1998, Experimental cell research.

[25]  K. R. Robinson,et al.  Self‐generated electrical currents through Xenopus neurulae. , 1984, The Journal of physiology.

[26]  R. Nuccitelli,et al.  Endogenous ionic currents and DC electric fields in multicellular animal tissues. , 1992, Bioelectromagnetics.

[27]  D. Zechner,et al.  A Role for the p38 Mitogen-activated Protein Kinase Pathway in Myocardial Cell Growth, Sarcomeric Organization, and Cardiac-specific Gene Expression , 1997, The Journal of cell biology.

[28]  J. Hiscott,et al.  Biological and biochemical inhibitors of the NF-κB/Rel proteins and cytokine synthesis , 1996 .

[29]  S. Ghosh,et al.  Rel/NF-?B and I?B proteins: an overview , 1997 .

[30]  K. Hotary,et al.  Endogenous electrical currents and voltage gradients in Xenopus embryos and the consequences of their disruption. , 1994, Developmental biology.

[31]  D. Männel,et al.  Dithiocarbamates as potent inhibitors of nuclear factor kappa B activation in intact cells , 1992, The Journal of experimental medicine.

[32]  J. Hescheler,et al.  Electrical fields enhance growth of cancer spheroids by reactive oxygen species and intracellular Ca2+. , 1997, The American journal of physiology.

[33]  J. Hiscott,et al.  Selective Inhibition of IκBα Phosphorylation and HIV-1 LTR-Directed Gene Expression by Novel Antioxidant Compounds☆ , 1997 .

[34]  R. Nuccitelli,et al.  Electrical controls of development. , 1977, Annual review of biophysics and bioengineering.

[35]  R. Steward,et al.  In vivo self-association of the Drosophila rel-protein dorsal. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[36]  M. Daucher,et al.  Activation of the serum response factor by p65/NF‐kappaB. , 1996, The EMBO journal.

[37]  R. Nuccitelli,et al.  Embryonic fibroblast motility and orientation can be influenced by physiological electric fields , 1984, The Journal of cell biology.

[38]  B. Wiedenmann,et al.  Development of G Protein‐mediated Ca2+ Channel Regulation in Mouse Embryonic Stem Cell‐derived Neurons , 1997, The European journal of neuroscience.

[39]  Y. Sun,et al.  Redox regulation of transcriptional activators. , 1996, Free radical biology & medicine.

[40]  D Kosman,et al.  The dorsal morphogen gradient regulates the mesoderm determinant twist in early Drosophila embryos. , 1991, Genes & development.

[41]  W. Webb,et al.  Optical imaging of cell membrane potential changes induced by applied electric fields. , 1986, Biophysical journal.

[42]  K. Frenkel,et al.  Hydrogen peroxide formation by cells treated with a tumor promoter. , 1991, Free radical research communications.

[43]  W. C. Parkinson,et al.  Effects of steady electric fields on human retinal pigment epithelial cell orientation and migration in culture , 1992, Acta ophthalmologica.

[44]  B. Rovin,et al.  Activation of nuclear factor-κB correlates with MCP-1 expression by human mesangial cells , 1995 .

[45]  J. Hescheler,et al.  The embryoid body as a novel in vitro assay system for antiangiogenic agents. , 1998, Laboratory investigation; a journal of technical methods and pathology.