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Myocardial subcellular glycogen distribution and sarcoplasmic reticulum Ca2+ handling: effects of ischaemia, reperfusion and ischaemic preconditioning

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Abstract

Ischaemic preconditioning (IPC) protects against myocardial ischaemia–reperfusion injury. The metabolic and ionic effects of IPC remain to be clarified in detail. We aimed to investigate the effect of IPC (2 times 5 min ischaemia) on the subcellular distribution of glycogen and Ca2+-uptake and leakiness by the sarcoplasmic reticulum (SR) in response to ischaemia–reperfusion in cardiomyocytes of isolated perfused rat hearts (Wistar rats, 335 ± 25 g). As estimated by quantitative transmission electron microscopy, the pre-ischaemic contribution [%, mean (95% CI)] of three sub-fractions of glycogen relative to total glycogen was 50 (39:61) as subsarcolemmal, 41 (31:50) as intermyofibrillar, and 9 (5:13) as intramyofibrillar glycogen. After 25 min of ischaemia, the relative contribution (%) of subsarcolemmal glycogen decreased to 39 (32:47) in control hearts (Con) and to 38 (31:45) in IPC. After 15 min reperfusion the contribution of subsarcolemmal glycogen was restored to pre-ischaemic levels in IPC hearts, but not in Con hearts. IPC increased the left ventricular developed pressure following ischaemia–reperfusion compared with Con. In saponin-skinned cardiomyocyte bundles, ischaemia reduced the SR Ca2+-uptake rate, with no effect of IPC. However, IPC reduced a SR Ca2+-leakage at pre-ischaemia, after ischaemia and during reperfusion. In conclusion, subsarcolemmal glycogen was preferentially utilised during sustained myocardial ischaemia. IPC improved left ventricular function reflecting reduced ischaemia–reperfusion injury, mediated a re-distribution of glycogen towards a preferential storage within the subsarcolemmal space during reperfusion, and lowered SR Ca2+-leakage. Under the present conditions, we found no temporal associations between alterations in glycogen localisation and SR Ca2+ kinetics.

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Acknowledgements

The authors thank Karin Trampedach, Susan Bøgebjerg, and Sandra Holm Riggelsen for technical assistance.

Funding

This work was supported by the Danish Council for Independent Research [DFF – 1333-00,144 to J.N.], the Danish Heart Association [72981 to J.N], the Lundbeck Foundation [R108-A10616 to J.N.], and The Danish Council for Strategic Research (11-1115818 to H.E.B.].

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Authors and Affiliations

  1. Department of Sports Science and Clinical Biomechanics, SDU Muscle Research Cluster (SMRC), University of Southern Denmark, 5230, Odense M, Denmark

    Joachim Nielsen & Niels Ørtenblad

  2. Department of Cardiology, Aarhus University Hospital, 8200, Aarhus N, Denmark

    Jacob Johnsen, Kasper Pryds & Hans Erik Bøtker

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  1. Joachim Nielsen
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Contributions

The experiments were performed in the laboratory of the Department of Cardiology, Aarhus University Hospital. The TEM was performed at the Department of Pathology, Odense University Hospital, Denmark. The TEM image analyses were performed at the Department of Sports Science and Clinical Biomechanics, University of Southern Denmark. JN, JJ, NØ and HEB were involved in the design of this study. All authors were involved in acquisition, analysis or interpretation of data for the work. JN drafted the work and all authors revised it critically for important intellectual content. All authors approved the final, submitted version of the manuscript. All authors agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. All persons designated as authors qualify for authorship, and all those who qualify for authorship are listed.

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Correspondence to Joachim Nielsen.

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Nielsen, J., Johnsen, J., Pryds, K. et al. Myocardial subcellular glycogen distribution and sarcoplasmic reticulum Ca2+ handling: effects of ischaemia, reperfusion and ischaemic preconditioning. J Muscle Res Cell Motil 42, 17–31 (2021). https://doi.org/10.1007/s10974-019-09557-3

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