Increased spontaneous Ca2+ activity in Cardiac Purkinje cells after myocardial infarction; A consequence of a dramatic shift of SERCA isoforms as potential adaptation to acute ischemia?

Stuyvers, B. D.; Boyden, P. A.; Amin, R.; Wiede, L. L.; Dore, J.; ter Keurs, H. E.; Dun, W.; Guo, Y.; Haissaguerre, M.; Hocini, M.; Brette, F.; Bernus, O.; Chaigne, S.; Loyer, V.; Naulin, J.; Quesson, B.; Alafiatayo, A.; Benoukraf, T.

doi:10.1101/2023.10.23.563678

bioRxiv · 10.1101/2023.10.23.563678

Increased spontaneous Ca2+ activity in Cardiac Purkinje cells after myocardial infarction; A consequence of a dramatic shift of SERCA isoforms as potential adaptation to acute ischemia?

Stuyvers, B. D.·Boyden, P. A.·Amin, R.·Wiede, L. L.·Dore, J.·ter Keurs, H. E.·Dun, W.·Guo, Y.·Haissaguerre, M.·Hocini, M.·Brette, F.·Bernus, O.·Chaigne, S.·Loyer, V.·Naulin, J.·Quesson, B.·Alafiatayo, A.·Benoukraf, T.

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Abstract

BackgroundStudies of Purkinje cells (Pcells) from canine hearts have suggested an increase of Ca2+-release by the sarcoplasmic reticulum (SR) but also reported a potential augmentation of SR-Ca2+-uptake after MI. Abnormal increase of SR-Ca2+-uptake in heart cells is novel and contrasts with the reduction of this function in cells of failing heart. Our study examined the origin of this increased SR-Ca2+-uptake by considering a change in SR-Ca2+ pump (SERCA2) expression in Purkinje fibers (PFs) post MI. MethodsPcells were isolated from canine hearts 48Hrs post MI. Intracellular Ca2+-activity was captured by confocal microscopy. Purkinje-typical Ca2+ events were analyzed to probe the regional Ca2+-dynamics within Pcells. A Purkinje-specific numerical model assisted in the interpretation of Ca2+-anomalies detected in Pcells Ca2+-transients. SR-Ca2+-uptake system was studied by immunofluorescence in Pcells from canine, ovine and human hearts post MI. SERCA protein and gene expressions in PFs and myocardium were measured by Western Blots and RT-qPCR in a classical porcine model of MI. Results48Hrs after MI, Pcells showed 60% increase in spark-rate and 37% acceleration of Ca2+ wave decay. In the model of normal wave, 35% increase of Ca2+-uptake rate reproduced the actual post-MI wave alterations. In apparent contrast with increased Ca2+-uptake rate, SERCA2 protein expression was reduced in canine, sheep, and human Pcells after MI. In pig MI model, the protein level of cardiac-specific SERCA2-splicing variant SERCA2a was reduced by 52% in the whole infarcted ventricle whereas the "non-cardiac" SERCA2b level was increased by 120%. In the infarcted regions, PFs showed 30% downregulation of SERCA2a gene expression and 630% upregulation of SERCA2b. ConclusionOur results confirm that elevated spontaneous Ca2+-activity in post-MI PFs is due to increased SR-Ca2+-uptake within Pcells. Data suggest that a replacement of "cardiac" SERCA2a by the "non-cardiac" SERCA2b sub-isoform in cardiac cells in response to ischemia is implicated in this alteration.

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Stuyvers, B. D., Boyden, P. A., Amin, R., Wiede, L. L., Dore, J., ter Keurs, H. E., Dun, W., Guo, Y., Haissaguerre, M., Hocini, M., Brette, F., Bernus, O., Chaigne, S., Loyer, V., Naulin, J., Quesson, B., Alafiatayo, A., Benoukraf, T.. 2023-10-26. Increased spontaneous Ca2+ activity in Cardiac Purkinje cells after myocardial infarction; A consequence of a dramatic shift of SERCA isoforms as potential adaptation to acute ischemia?. https://doi.org/10.1101/2023.10.23.563678

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