In vivo model with targeted cAMP biosensor reveals changes in receptor-microdomain communication in cardiac disease.
3',5'-cyclic adenosine monophosphate (cAMP) is an ubiquitous second messenger that regulates physiological functions by acting in distinct subcellular microdomains. Although several targeted cAMP biosensors are developed and used in single cells, it is unclear whether such biosensors can be successfully applied in vivo, especially in the context of disease. Here, we describe a transgenic mouse model expressing a targeted cAMP sensor and analyse microdomain-specific second messenger dynamics in the vicinity of the sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA). We demonstrate the biocompatibility of this targeted sensor and its potential for real-time monitoring of compartmentalized cAMP signalling in adult cardiomyocytes isolated from a healthy mouse heart and from an in vivo cardiac disease model. In particular, we uncover the existence of a phosphodiesterase-dependent receptor-microdomain communication, which is affected in hypertrophy, resulting in reduced β-adrenergic receptor-cAMP signalling to SERCA.
1] Emmy Noether Group of the DFG, European Heart Research Institute Göttingen, University Medical Center Göttingen, D-37075 Göttingen, Germany [2] Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany [3] Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany.
Julia U
JU
Sprenger
1] Emmy Noether Group of the DFG, European Heart Research Institute Göttingen, University Medical Center Göttingen, D-37075 Göttingen, Germany [2] Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany [3] Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany.
Ruwan K
RK
Perera
Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany.
Julia H
JH
Steinbrecher
1] Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany [2] German Center for Cardiovascular Research (DZHK), D-93053 Regensburg, Germany.
Stephan E
SE
Lehnart
Department of Internal Medicine II, University Hospital Regensburg, D-93053 Regensburg, Germany.
Lars S
LS
Maier
1] Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany [2] German Center for Cardiovascular Research (DZHK), D-93053 Regensburg, Germany.
Gerd
G
Hasenfuss
1] Emmy Noether Group of the DFG, European Heart Research Institute Göttingen, University Medical Center Göttingen, D-37075 Göttingen, Germany [2] Department of Cardiology and Pulmonology, Heart Research Center Göttingen, University Medical Center Göttingen, Georg August University, D-37075 Göttingen, Germany [3] Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, D-20246 Hamburg, Germany [4] German Center for Cardiovascular Research (DZHK), D-93053 Regensburg, Germany.
Viacheslav O
VO
Nikolaev
D000318:Adrenergic beta-Agonists; D000818:Animals; D015374:Biosensing Techniques; D002135:Calcium-Binding Proteins; D006332:Cardiomegaly / Q000209:etiology / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D031541:Fluorescence Resonance Energy Transfer; D020662:Guanine Nucleotide Exchange Factors; D006331:Heart Diseases / Q000209:etiology / Q000378:metabolism*; D007545:Isoproterenol; D008822:Mice, Transgenic; D032383:Myocytes, Cardiac / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D011897:Random Allocation; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*; D000099211:Phospholamban
D000318:Adrenergic beta-Agonists; D002135:Calcium-Binding Proteins; D000242:Cyclic AMP; D020662:Guanine Nucleotide Exchange Factors; D007545:Isoproterenol; D010727:Phosphoric Diester Hydrolases; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000099211:Phospholamban; C482601:Epac protein, mouse