Adenoviral Transduction of FRET-Based Biosensors for cAMP in Primary Adult Mouse Cardiomyocytes
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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.
KeywordsPrimary cardiomyocyte Mouse Adenovirus cAMP FRET Live-cell imaging
This study was supported by the British Heart Foundation (PG/10/75/28537 and RG/12/3/29423) and the NSF-NIH CRCNS program (NIH R01 AA18060) to MZ and Wellcome Trust PhD Programme for Clinicians at the University of Oxford to OCL.
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