Abstract
Materials and methods We generated a transgenic mouse model with cardiomyocyte-specific expression of the highly sensitive fluorescence resonance energy transfer (FRET)-based cGMP biosensor red cGES-DE5. This sensor mouse allows us to performed FRET measurements of cGMP in freshly isolated adult mouse ventricular myocytes, to visulaize realtime dynamics of cGMP. To analyze cAMP and cGMP interactions (cGMP/cAMP signaling crosstalk), FRET experiments were performed in cardiomyocytes isolated from mice, transgenically expressing the FRET-based cAMP sensor Epac1-camps.
Highlights
CGMP is an important second messenger which is involved in the regulation of cardiac contractility and pathological hypertrophy
PDE3 is the major cGMP-PDE in adult mouse ventricular cardiomyocytes
Stimulation of the soluble guanylyl cyclase with NO-donors such as SNAP had no effect. Constitutive activity of this cyclase is involved in basal cGMP production, since stimulating cardiomyocytes with the sGC inhibitor ODQ showed a decrease of basal cGMP levels
Summary
From 6th International Conference on cGMP: Generators, Effectors and Therapeutic Implications Erfurt, Germany. 28-30 June 2013. From 6th International Conference on cGMP: Generators, Effectors and Therapeutic Implications Erfurt, Germany. Background cGMP is an important second messenger which is involved in the regulation of cardiac contractility and pathological hypertrophy. Signaling by cGMP is considered cardioprotective, but little is known about the spatio-temporal dynamics of cGMP and its regulation in adult cardiomyocytes
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