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Investigating Cardiac Metabolism in the Isolated Perfused Mouse Heart with Hyperpolarized [1-13C]Pyruvate and 13C/31P NMR Spectroscopy
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Biologia
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JoVE Journal Biologia
Investigating Cardiac Metabolism in the Isolated Perfused Mouse Heart with Hyperpolarized [1-13C]Pyruvate and 13C/31P NMR Spectroscopy

Investigating Cardiac Metabolism in the Isolated Perfused Mouse Heart with Hyperpolarized [1-13C]Pyruvate and 13C/31P NMR Spectroscopy

DOI:
10.3791/63188-v

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14:56 min

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April 21, 2023

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, , , , ,
1Department of Radiology, Hadassah Medical Organization and Faculty of Medicine,Hebrew University of Jerusalem, 2The Wohl Institute for Translational Medicine

Capítulos

  • 00:03Introduction
  • 01:17Krebs-Henseleit Buffer Preparation
  • 01:45Perfusion System Preparation
  • 02:29Calibration and Preparation of the NMR Spectrometer for Acquisition
  • 03:22Animal Preparation, Surgical Procedure, and Perfusion of the Heart in the NMR Tube
  • 09:23Acquiring Data for Cardiac Energetics and pH
  • 10:09DNP Spin Polarization and Dissolution
  • 11:25Hyperpolarized 13C Spectroscopy
  • 12:07Representative Results
  • 14:40Conclusion

Summary

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We describe an experimental setup for administrating hyperpolarized 13C-labeled metabolites in continuous perfusion mode to an isolated perfused mouse heart. A dedicated 13C-NMR acquisition approach enabled the quantification of metabolic enzyme activity in real-time, and a multiparametric 31P-NMR analysis enabled the determination of the tissue ATP content and pH.

Tags

Hyperpolarized MRICardiac MetabolismIsolated Perfused Mouse HeartPyruvate Metabolism13C/31P NMR SpectroscopyKrebs-Henseleit BufferAerobic And Anaerobic MetabolismCardiac EnergeticsPHLactateInorganic PhosphatePerfusion SystemTemperature Control

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