Herpesvirus-mediated delivery of a genetically encoded fluorescent Ca(2+) sensor to canine cardiomyocytes

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. In...

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Bibliographic Details
Main Authors: Prorok János
Kovács Péter Pál
Kristóf Attila
Nagy Norbert
Tombácz Dóra
Tóth Judit S.
Ördög Balázs
Jost Norbert László
Virág László
Papp Gyula
Varró András
Tóth András
Boldogkői Zsolt
Format: Article
Published: 2009
Series:Journal of biomedicine & biotechnology 2009
doi:10.1155/2009/361795

mtmt:1239395
Online Access:http://publicatio.bibl.u-szeged.hu/9049
Description
Summary: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.
Physical Description:Terjedelem: 12 p.-Azonosító: 361795
ISSN:1110-7251