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Measurement of myofilament calcium in living cardiomyocytes using a targeted genetically encoded indicator

Paul Robinson, Alexander J Sparrow, Connor N Broyles, Kolja Sievert, Yu-Fen Chang, Frances A Brook, Xiaoyu Zhang, Hugh Watkins, Yama A Abassi, Michael A Geeves, Charles Redwood, View ORCID ProfileMatthew J Daniels
doi: https://doi.org/10.1101/268003
Paul Robinson
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
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Alexander J Sparrow
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
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Connor N Broyles
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
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Kolja Sievert
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
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Yu-Fen Chang
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
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Frances A Brook
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
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Xiaoyu Zhang
4ACEA Biosciences, Inc., San Diego, CA 92121, USA
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Hugh Watkins
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
3Department of Cardiology, Oxford University NHS Hospitals Trust, Oxford, UK
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Yama A Abassi
4ACEA Biosciences, Inc., San Diego, CA 92121, USA
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Michael A Geeves
5Department of Biosciences, University of Kent, Canterbury, UK
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Charles Redwood
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
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Matthew J Daniels
1Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, UK
2BHF Centre of Research Excellence, University of Oxford, Oxford, UK
3Department of Cardiology, Oxford University NHS Hospitals Trust, Oxford, UK
6BHF Centre of Regenerative Medicine, University of Oxford, Oxford, UK
7Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan
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  • ORCID record for Matthew J Daniels
  • For correspondence: matthew.daniels@cardiov.ox.ac.uk
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Abstract

Visualising when and where calcium appears and disappears in cardiomyocytes is a major goal of cardiovascular research. Surprisingly we find that the chemical dyes widely used for this purpose disrupt cell contractility, due at least in part due to direct inhibition of the acto-myosin ATPase required to generate force. In order to improve calcium detection methods, we have developed a genetically encoded indicator that sits within the myofilament to directly visualise the changes occurring at the sarcomere. This tool improves on established chemical dyes and untargeted genetically encoded indicators for analysing small molecule modulators of myofilament-based calcium signalling. Importantly this is achieved without any measurable change in contractile function.

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Posted April 16, 2018.
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Measurement of myofilament calcium in living cardiomyocytes using a targeted genetically encoded indicator
Paul Robinson, Alexander J Sparrow, Connor N Broyles, Kolja Sievert, Yu-Fen Chang, Frances A Brook, Xiaoyu Zhang, Hugh Watkins, Yama A Abassi, Michael A Geeves, Charles Redwood, Matthew J Daniels
bioRxiv 268003; doi: https://doi.org/10.1101/268003
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Measurement of myofilament calcium in living cardiomyocytes using a targeted genetically encoded indicator
Paul Robinson, Alexander J Sparrow, Connor N Broyles, Kolja Sievert, Yu-Fen Chang, Frances A Brook, Xiaoyu Zhang, Hugh Watkins, Yama A Abassi, Michael A Geeves, Charles Redwood, Matthew J Daniels
bioRxiv 268003; doi: https://doi.org/10.1101/268003

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