Redox Imaging Using Cardiac Myocyte-Specific Transgenic Biosensor Mice
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Redox Imaging Using Cardiac Myocyte-Specific Transgenic Biosensor Mice. / Swain, Lija; Kesemeyer, Andrea; Meyer-Roxlau, Stefanie; Vettel, Christiane; Zieseniss, Anke; Güntsch, Annemarie; Jatho, Aline; Becker, Andreas; Nanadikar, Maithily S; Morgan, Bruce; Dennerlein, Sven; Shah, Ajay M; El-Armouche, Ali; Nikolaev, Viacheslav O; Katschinski, Dörthe M.
In: CIRC RES, Vol. 119, No. 9, 14.10.2016, p. 1004-1016.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Redox Imaging Using Cardiac Myocyte-Specific Transgenic Biosensor Mice
AU - Swain, Lija
AU - Kesemeyer, Andrea
AU - Meyer-Roxlau, Stefanie
AU - Vettel, Christiane
AU - Zieseniss, Anke
AU - Güntsch, Annemarie
AU - Jatho, Aline
AU - Becker, Andreas
AU - Nanadikar, Maithily S
AU - Morgan, Bruce
AU - Dennerlein, Sven
AU - Shah, Ajay M
AU - El-Armouche, Ali
AU - Nikolaev, Viacheslav O
AU - Katschinski, Dörthe M
N1 - © 2016 American Heart Association, Inc.
PY - 2016/10/14
Y1 - 2016/10/14
N2 - RATIONALE: 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).METHODS AND 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 EGSH in isolated cardiac myocytes and in Langendorff-perfused hearts. Distinct EGSH were 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 EGSH in the cytoplasm and the mitochondrial matrix in cardiac myocytes and can be exploited to answer questions in basic and translational cardiovascular research.
AB - RATIONALE: 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).METHODS AND 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 EGSH in isolated cardiac myocytes and in Langendorff-perfused hearts. Distinct EGSH were 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 EGSH in the cytoplasm and the mitochondrial matrix in cardiac myocytes and can be exploited to answer questions in basic and translational cardiovascular research.
U2 - 10.1161/CIRCRESAHA.116.309551
DO - 10.1161/CIRCRESAHA.116.309551
M3 - SCORING: Journal article
C2 - 27553648
VL - 119
SP - 1004
EP - 1016
JO - CIRC RES
JF - CIRC RES
SN - 0009-7330
IS - 9
ER -