title: Redox Imaging Using Cardiac Myocyte-Specific Transgenic Biosensor Mice.
abstract: BACKGROUND
  Changes in redox potentials of cardiac myocytes are linked to several cardiovascular diseases. Redox alterations are currently mostly described qualitatively using chemical sensors, which however do not allow quantifying redox potentials, lack specificity, and the possibility to analyze subcellular domains. Recent advances to quantitatively describe defined redox changes include the application of genetically encoded redox biosensors.
  
  
  OBJECTIVE
  Establishment of mouse models, which allow the quantification of the glutathione redox potential (EGSH) in the cytoplasm and the mitochondrial matrix of isolated cardiac myocytes and in Langendorff-perfused hearts based on the use of the redox-sensitive green fluorescent protein 2, coupled to the glutaredoxin 1 (Grx1-roGFP2).
  
  
  RESULTS
  We generated transgenic mice with cardiac myocyte-restricted expression of Grx1-roGFP2 targeted either to the mitochondrial matrix or to the cytoplasm. The response of the roGFP2 toward H2O2, diamide, and dithiothreitol was titrated and used to determine the EGSHin isolated cardiac myocytes and in Langendorff-perfused hearts. Distinct EGSHwere observed in the cytoplasm and the mitochondrial matrix. Stimulation of the cardiac myocytes with isoprenaline, angiotensin II, or exposure to hypoxia/reoxygenation additionally underscored that these compartments responded independently. A compartment-specific response was also observed 3 to 14 days after myocardial infarction.
  
  
  CONCLUSIONS
  We introduce redox biosensor mice as a new tool, which allows quantification of defined alterations of EGSHin the cytoplasm and the mitochondrial matrix in cardiac myocytes and can be exploited to answer questions in basic and translational cardiovascular research.
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Lija
authors.initials: L
authors.lastname: Swain
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Andrea
authors.initials: A
authors.lastname: Kesemeyer
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Stefanie
authors.initials: S
authors.lastname: Meyer-Roxlau
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Christiane
authors.initials: C
authors.lastname: Vettel
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Anke
authors.initials: A
authors.lastname: Zieseniss
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Annemarie
authors.initials: A
authors.lastname: Güntsch
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Aline
authors.initials: A
authors.lastname: Jatho
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Andreas
authors.initials: A
authors.lastname: Becker
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Maithily S
authors.initials: MS
authors.lastname: Nanadikar
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Bruce
authors.initials: B
authors.lastname: Morgan
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Sven
authors.initials: S
authors.lastname: Dennerlein
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Ajay M
authors.initials: AM
authors.lastname: Shah
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Ali
authors.initials: A
authors.lastname: El-Armouche
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.).
authors.forename: Viacheslav O
authors.initials: VO
authors.lastname: Nikolaev
authors.affiliation: From the Institute of Cardiovascular Physiology, Georg August University Göttingen, Germany (L.S., A.K., A.Z., A.G., A.J., A.B., M.S.N., D.M.K.); Institute of Pharmacology, Technical University Dresden, Germany (S.M.-R., A.E.-A.); Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany (C.V.); Cellular Biochemistry, Department of Biology, University of Kaiserslautern, Germany (B.M.); Department of Cellular Biochemistry, University Medical Center Göttingen, Germany (S.D.); Cardiovascular Division, King's College London, British Heart Foundation Centre, United Kingdom (A.M.S.); and German Center for Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany (V.O.N.); and Institute of Experimental Cardiovascular Research, Hamburg, Germany (V.O.N.). doerthe.katschinski@med.uni-goettingen.de.
authors.forename: Dörthe M
authors.initials: DM
authors.lastname: Katschinski
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism*; D006321:Heart / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D032383:Myocytes, Cardiac / Q000378:metabolism*; D009924:Organ Culture Techniques; D010084:Oxidation-Reduction; D010101:Oxygen Consumption / Q000502:physiology
chemical_list: D049452:Green Fluorescent Proteins
keywords: angiotensin II; cytoplasm; diamide; ischemia; reactive oxygen species