Abstract
Primary adult murine cardiomyocytes are an important tool for the investigation of cardiovascular physiology, especially with the development of transgenic mouse models. Fluorescence Resonance Energy Transfer (FRET) based probes have contributed greatly to the understanding of subcellular cAMP signalling. However, the transfer of genes encoding such probes into adult murine cardiomyocytes has proved difficult. Here we present a method for the effective adenovirus-mediated transduction of FRET biosensors that allow the investigation of cAMP signalling in adult murine cardiomyocytes in under 24 hours in culture. Furthermore we show that the contractility of adult murine cardiomyocytes in culture is not impaired by adenoviral gene transfer of FRET biosensors. Combined with a microscopy chamber perfusion system, that allows the removal of pharmacological agents such as specific inhibitors of phosphodiesterases (PDEs), this method allows the cost-effective, reproducible and rapid investigation of sub-cellular cAMP signalling and hence PDE activity in adult murine cardiomyocytes.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
