title: Adenoviral transduction of FRET-based biosensors for cAMP in primary adult mouse cardiomyocytes.
abstract: Genetically encoded biosensors that make use of fluorescence resonance energy transfer (FRET) are important tools for the study of compartmentalized cyclic nucleotide signaling in living cells. With the advent of germ line and tissue-specific transgenic technologies, the adult mouse represents a useful tool for the study of cardiovascular pathophysiology. The use of FRET-based genetically encoded biosensors coupled with this animal model represents a powerful combination for the study of cAMP signaling in live primary cardiomyocytes. In this chapter, we describe the steps required during the isolation, viral transduction, and culture of cardiomyocytes from an adult mouse to obtain reliable expression of genetically encoded FRET biosensors for the study of cAMP signaling in living cells.
authors.affiliation: Department of Cardiovascular Medicine, John Radcliffe Hospital, University of Oxford, Oxford, OX3 9DU, UK.
authors.forename: Oliver
authors.initials: O
authors.lastname: Lomas
authors.forename: Marcella
authors.initials: M
authors.lastname: Brescia
authors.forename: Ricardo
authors.initials: R
authors.lastname: Carnicer
authors.forename: Stefania
authors.initials: S
authors.lastname: Monterisi
authors.forename: Nicoletta C
authors.initials: NC
authors.lastname: Surdo
authors.forename: Manuela
authors.initials: M
authors.lastname: Zaccolo
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods; D057791:Cardiac Imaging Techniques / Q000379:methods; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000821:virology; D015398:Signal Transduction; D014161:Transduction, Genetic
chemical_list: D000242:Cyclic AMP