Abstract
Diabetes is commonly associated with an elevated level of reactive carbonyl species due to alteration of glucose and fatty acid metabolism. These metabolic changes cause an abnormality in cardiac Ca2+ regulation that can lead to cardiomyopathies. In this study, we explored how the reactive α-dicarbonyl methylglyoxal (MGO) affects Ca2+ regulation in mouse ventricular myocytes. Analysis of intracellular Ca2+ dynamics revealed that MGO (200 μM) increases action potential (AP)-induced Ca2+ transients and sarcoplasmic reticulum (SR) Ca2+ load, with a limited effect on L-type Ca2+ channel-mediated Ca2+ transients and SERCA-mediated Ca2+ uptake. At the same time, MGO significantly slowed down cytosolic Ca2+ extrusion by Na+/Ca2+ exchanger (NCX). MGO also increased the frequency of Ca2+ waves during rest and these Ca2+ release events were abolished by an external solution with zero [Na+] and [Ca2+]. Adrenergic receptor activation with isoproterenol (10 nM) increased Ca2+ transients and SR Ca2+ load, but it also triggered spontaneous Ca2+ waves in 27% of studied cells. Pretreatment of myocytes with MGO increased the fraction of cells with Ca2+ waves during adrenergic receptor stimulation by 163%. Measurements of intracellular [Na+] revealed that MGO increases cytosolic [Na+] by 57% from the maximal effect produced by the Na+-K+ ATPase inhibitor ouabain (20 μM). This increase in cytosolic [Na+] was a result of activation of a tetrodotoxin-sensitive Na+ influx, but not an inhibition of Na+-K+ ATPase. An increase in cytosolic [Na+] after treating cells with ouabain produced similar effects on Ca2+ regulation as MGO. These results suggest that protein carbonylation can affect cardiac Ca2+ regulation by increasing cytosolic [Na+] via a tetrodotoxin-sensitive pathway. This, in turn, reduces Ca2+ extrusion by NCX, causing SR Ca2+ overload and spontaneous Ca2+ waves.
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Acknowledgements
The authors would like to thank Dr. L. Michel Espinoza-Fonseca (University of Michigan, USA) for assisting with preparing purified NKA.
Funding
This work was supported by the National Institutes of Health Grants R01HL151990 (to A.V.Z.) and R01 HL092321 (to S.L.R.).
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E.B., S.L.R. and A.V.Z. contributed to the conception and design of the study. E.B., J.S., S.L.R., and A.V.Z. performed the experimental work and analysis of results. E.B., J.S., S.L.R. and A.V.Z. contributed to the writing of the manuscript.
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All animal experiments were carried out in accordance with the National Institutes of Health guide for the care and use of Laboratory animals [21]. Male and female C57Bl6/J mice (Jackson Laboratories, US) were housed according to approved IACUC protocol IACUC#23–027 (Loyola University Chicago).
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Bovo, E., Seflova, J., Robia, S.L. et al. Protein carbonylation causes sarcoplasmic reticulum Ca2+ overload by increasing intracellular Na+ level in ventricular myocytes. Pflugers Arch - Eur J Physiol 476, 1077–1086 (2024). https://doi.org/10.1007/s00424-024-02972-7
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DOI: https://doi.org/10.1007/s00424-024-02972-7