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Hyperpolarized 13C lactate as a substrate for in vivo metabolic studies in skeletal muscle

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Abstract

Resting skeletal muscle has a preference for the oxidation of lipids compared to carbohydrates and a shift towards carbohydrate oxidation is observed with increasing exercise. Lactate is not only an end product in skeletal muscle but also an important metabolic intermediate for mitochondrial oxidation. Recent advances in hyperpolarized MRS allow the measurement of substrate metabolism in vivo in real time. The aim of this study was to investigate the use of hyperpolarized 13C lactate as a substrate for metabolic studies in skeletal muscle in vivo. Carbohydrate metabolism in healthy rat skeletal muscle at rest was studied in different nutritional states using hyperpolarized [1-13C]lactate, a substrate that can be injected at physiological concentrations and leaves other oxidative processes undisturbed. 13C label incorporation from lactate into bicarbonate in fed animals was observed within seconds but was absent after an overnight fast, representing inhibition of the metabolic flux through pyruvate dehydrogenase (PDH). A significant decrease in 13C labeling of alanine was observed comparing the fed and fasted group, and was attributed to a change in cellular alanine concentration and not a decrease in enzymatic flux through alanine transaminase. We conclude that hyperpolarized [1-13C]lactate can be used to study carbohydrate oxidation in resting skeletal muscle at physiological levels. The herein proposed method allows probing simultaneously both PDH activity and variations in alanine tissue concentration, which are associated with metabolic dysfunctions. A simple alteration of the nutritional state demonstrated that the observed pyruvate, alanine, and bicarbonate signals are indeed sensitive markers to probe metabolic changes in vivo.

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Abbreviations

ALT:

Alanine transaminase

LDH:

Lactate dehydrogenase

PDH:

Pyruvate dehydrogenase

TCA:

Tricarboxylic acid cycle

MCT:

Monocarboxylate transporter

CA:

Carbonic anhydrase

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Acknowledgments

We would like to thank Tian Cheng and Andrea Capozzi for their suggestions and improvements on the lactate recipe and Jacquelina Romero, Laure Bardouillet and Mario Lepore for veterinary support.

Conflict of interest

The authors declare no conflicts of interest.

Animal studies

All institutional and national guidelines for the care and use of laboratory animals were followed.

Funding

This work was supported by the Swiss National Science Foundation (Grant PP00P2_133562 and 31003AB_131087), the Centre d’Imagerie BioMédicale (CIBM) of the UNIL, UNIGE, HUG, CHUV, EPFL, and the Leenaards and Jeantet Foundations.

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Authors and Affiliations

  1. Laboratory for Functional and Metabolic Imaging, LIFMET-IPSB-SB, Ecole Polytechnique Fédérale de Lausanne (EPFL), Station 6, 1015, Lausanne, Switzerland

    Jessica A. M. Bastiaansen & Rolf Gruetter

  2. Institute of Physics of Biological Systems, Ecole Polytechnique Fédérale de Lausanne, 1015, Lausanne, Switzerland

    Hikari A. I. Yoshihara, Yuhei Takado & Arnaud Comment

  3. Department of Cardiology, Lausanne University Hospital, 1011, Lausanne, Switzerland

    Hikari A. I. Yoshihara

  4. Laboratory of Biological Geochemistry, Ecole Polytechnique Fédérale de Lausanne, 1015, Lausanne, Switzerland

    Yuhei Takado

  5. Department of Radiology, Geneva University Hospital and Faculty of Medicine, University of Geneva, 1211, Geneva 4, Switzerland

    Rolf Gruetter

  6. Department of Radiology, University of Lausanne, 1015, Lausanne, Switzerland

    Rolf Gruetter

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  1. Jessica A. M. Bastiaansen
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  2. Hikari A. I. Yoshihara
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  3. Yuhei Takado
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  4. Rolf Gruetter
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  5. Arnaud Comment
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Correspondence to Jessica A. M. Bastiaansen.

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Bastiaansen, J.A.M., Yoshihara, H.A.I., Takado, Y. et al. Hyperpolarized 13C lactate as a substrate for in vivo metabolic studies in skeletal muscle. Metabolomics 10, 986–994 (2014). https://doi.org/10.1007/s11306-014-0630-5

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