Abstract
Gap junctional intercellular communication (GJIC) has been described in different cell types including stem cells and has been involved in different biological events. GJIC is required for mouse embryonic stem cell maintenance and proliferation, and various studies suggest that functional GJIC is a common characteristic of human embryonic stem cells (hESC) maintained in different culture conditions. This chapter introduces methods to study gap junctions in hESC, from expression of gap junction proteins to functional study of GJIC in hESC proliferation, apoptosis, colony growth, and pluripotency.
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References
Carpenter M. K., Rosler E. S., Fisk G. J., et al. (2004) Properties of four human embryonic stem cell lines maintained in a feeder-free culture system. Dev Dyn. 229, 243–258.
Wong R. C., Dottori M., Koh K. L., Nguyen L. T., Pera M. F., and Pebay A. (2006) Gap junctions modulate apoptosis and colony growth of human embryonic stem cells maintained in a serum-free system. Biochem Biophys Res Commun. 344, 181–188.
Wong R. C., Pebay A., Nguyen L. T., Koh K. L., and Pera M. F. (2004) Presence of functional gap junctions in human embryonic stem cells. Stem Cells 22, 883–889.
Duval N., Gomes D., Calaora V., Calabrese A., Meda P., and Bruzzone R. (2002) Cell coupling and Cx43 expression in embryonic mouse neural progenitor cells. J Cell Sci. 115, 3241–3251.
Cheng A., Tang H., Cai J., et al. (2004) Gap junctional communication is required to maintain mouse cortical neural progenitor cells in a proliferative state. Dev Biol 272, 203–216.
Potapova I., Plotnikov A., Lu Z., et al. (2004) Human mesenchymal stem cells as a gene delivery system to create cardiac pacemakers. Circ Res. 94, 952–959.
Valiunas V., Doronin S., Valiuniene L., et al. (2004) Human mesenchymal stem cells make cardiac connexins and form functional gap junctions. J Physiol. 555, 617–626.
Todorova M. G., Soria B., and Quesada I. (2008) Gap junctional intercellular communication is required to maintain embryonic stem cells in a non-differentiated and proliferative state. J Cell Physiol. 214, 354–362.
Oyamada Y., Komatsu K., Kimura H., Mori M., and Oyamada M. (1996) Differential regulation of gap junction protein (connexin) genes during cardiomyocytic differentiation of mouse embryonic stem cells in vitro. Exp Cell Res. 229, 318–326.
Worsdorfer P., Maxeiner S., Markopoulos C., et al. (2008) Connexin expression and functional analysis of gap junctional communication in mouse embryonic stem cells. Stem Cells 26, 431–439.
Kumar N. M., and Gilula N. B. (1996) The gap junction communication channel. Cell 84, 381–388.
Revel J. P., and Karnovsky M. J. (1967) Hexagonal array of subunits in intercellular junctions of the mouse heart and liver. J Cell Biol. 33, C7–C12.
Alexander D. B., and Goldberg G. S. (2003) Transfer of biologically important molecules between cells through gap junction channels. Curr Med Chem. 10, 2045–2058.
Krysko D. V., Leybaert L., Vandenabeele P., and D'Herde K. (2005) Gap junctions and the propagation of cell survival and cell death signals. Apoptosis 10, 459–469.
Valiunas V., Bukauskas F. F., and Weingart R. (1997) Conductances and selective permeability of connexin43 gap junction channels examined in neonatal rat heart cells. Circ Res. 80, 708–719.
Neijssen J., Herberts C., Drijfhout J. W., Reits E., Janssen L., and Neefjes J. (2005) Cross-presentation by intercellular peptide transfer through gap junctions. Nature 434, 83–88.
Sheardown S. A., and Hooper M. L. (1992) A relationship between gap junction-mediated intercellular communication and the in vitro developmental capacity of murine embryonic stem cells. Exp Cell Res. 198, 276–282.
De Maio A., Vega V., and Contreras J. (2002) Gap junctions, homeostasis, and injury. J Cell Physiol. 191, 269–282.
Mesnil M., Crespin S., Avanzo J. L., and Zaidan-Dagli M. L. (2005) Defective gap junctional intercellular communication in the carcinogenic process. Biochim Biophys Acta 1719, 125–145.
Trosko J. E., and Chang C. C. (2003) Isolation and characterization of normal adult human epithelial pluripotent stem cells. Oncol Res. 13, 353–357.
Yamasaki H., Krutovskikh V., Mesnil M., Tanaka T., Zaidan-Dagli M. L., and Omori Y. (1999) Role of connexin (gap junction) genes in cell growth control and carcinogenesis. C R Acad Sci III 322, 151–159.
Huettner J. E., Lu A., Qu Y., Wu Y., Kim M., and McDonald J. W. (2006) Gap junctions and connexon hemichannels in human embryonic stem cells. Stem Cells 24, 1654–1667.
Assou S., Lecarrour T., Tondeur S., et al. (2007) A meta-analysis of human embryonic stem cells transcriptome integrated into a web-based expression atlas. Stem Cells 25, 961–973.
Boyer L. A., Lee T. I., Cole M. F., et al. (2005) Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122, 947–956.
El-Fouly M. H., Trosko J. E., and Chang C. C. (1987) Scrape-loading and dye transfer. A rapid and simple technique to study gap junctional intercellular communication. Exp Cell Res. 168, 422–430.
Venance L., Piomelli D., Glowinski J., and Giaume C. (1995) Inhibition by anandamide of gap junctions and intercellular calcium signalling in striatal astrocytes. Nature 376, 590–594.
Lampe P. D., and Lau A. F. (2000) Regulation of gap junctions by phosphorylation of connexins. Arch Biochem Biophys. 384, 205–215.
Lampe P. D., and Lau A. F. (2004) The effects of connexin phosphorylation on gap junctional communication. Int J Biochem Cell Biol. 36, 1171–1186.
Acknowledgements
This study was supported by The University of Melbourne and the National Health and Medical Research Council of Australia (NHMRC 454723). R.C. Wong is currently supported by the California Institute of Regenerative Medicine Grant RC1-00110-1.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC 2006
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Wong, R.C., Pébay, A. (2009). Study of Gap Junctions in Human Embryonic Stem Cells. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 584. Humana Press. https://doi.org/10.1007/978-1-60761-369-5_12
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DOI: https://doi.org/10.1007/978-1-60761-369-5_12
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