Herpesvirus-mediated delivery of a genetically encoded fluorescent ca 2+ sensor to canine cardiomyocytes

János Prorok, Péter P. Kovács, Attila A. Kristf, Norbert Nagy, Dra Tombácz, Judit S. Tth, Balázs Ördg, Norbert Jost, Lászl Virág, Julius G. Papp, András Varr, András Tth, Zsolt Boldogki

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Abstract

We report the development and application of a pseudorabies virus-based system for delivery of troponeon, a fluorescent Ca 2 + sensor to adult canine cardiomyocytes. The efficacy of transduction was assessed by calculating the ratio of fluorescently labelled and nonlabelled cells in cell culture. Interaction of the virus vector with electrophysiological properties of cardiomyocytes was evaluated by the analysis of transient outward current (I to), kinetics of the intracellular Ca 2 + transients, and cell shortening. Functionality of transferred troponeon was verified by FRET analysis. We demonstrated that the transfer efficiency of troponeon to cultured adult cardiac myocytes was virtually 100. We showed that even after four days neither the amplitude nor the kinetics of the I to current was significantly changed and no major shifts occurred in parameters of [ Ca 2 + ] i transients. Furthermore, we demonstrated that infection of cardiomyocytes with the virus did not affect the morphology, viability, and physiological attributes of cells.

Original languageEnglish
Article number361795
JournalJournal of Biomedicine and Biotechnology
Volume2009
DOIs
Publication statusPublished - Aug 24 2009

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ASJC Scopus subject areas

  • Biotechnology
  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

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