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Redox Imaging Using Cardiac Myocyte-Specific Transgenic Biosensor Mice.

Lija Swain¹, Andrea Kesemeyer¹, Stefanie Meyer-Roxlau¹

¹From 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.).

Circulation Research

|August 25, 2016

Summary

New mouse models enable quantitative measurement of glutathione redox potential (E_GSH) in cardiac cells. This tool allows precise analysis of redox changes in cytoplasm and mitochondria, aiding cardiovascular disease research.

Keywords:

angiotensin II cytoplasm diamide ischemia reactive oxygen species

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Area of Science:

Cardiovascular Biology
Redox Biology
Genetically Engineered Models

Background:

Cardiac myocyte redox potential alterations are implicated in cardiovascular diseases.
Current methods for assessing redox status are qualitative, lack specificity, and cannot analyze subcellular domains.
Genetically encoded redox biosensors offer quantitative analysis of redox changes.

Purpose of the Study:

To establish mouse models for quantifying glutathione redox potential (E_GSH) in cardiac myocyte cytoplasm and mitochondrial matrix.
To enable measurements in isolated myocytes and Langendorff-perfused hearts using Grx1-roGFP2 biosensor.

Main Methods:

Generated transgenic mice with cardiac-specific Grx1-roGFP2 expression in mitochondria or cytoplasm.
Titrated roGFP2 response to H₂O₂, diamide, and dithiothreitol for E_GSH determination.
Measured E_GSH in isolated myocytes and perfused hearts under various physiological and pathological conditions.

Main Results:

Demonstrated distinct E_GSH in cytoplasmic and mitochondrial compartments of cardiac myocytes.
Showed independent compartment-specific redox responses to isoprenaline, angiotensin II, and hypoxia/reoxygenation.
Observed compartment-specific redox alterations post-myocardial infarction.

Conclusions:

Introduced novel redox biosensor mice for quantitative E_GSH measurement in cardiac myocytes.
These models allow precise analysis of subcellular redox changes.
The tool is valuable for basic and translational cardiovascular research.