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Connexin43 dephosphorylation at serine 282 is associated with connexin43-mediated cardiomyocyte apoptosis

Abstract

Gap junction protein connexin 43 (Cx43) plays an important role in regulating cardiomyocyte survival in addition to regulating electrical coordination. Cx43 dephosphorylation, found in severe cardiac pathologies, is thought to contribute to myocardial injury. However, the mechanisms underlying Cx43 mediation of cell survival and myocardial lesions remain unknown. Here, we found that transfecting an adenovirus carrying a mutant gene of Cx43-serine 282 substituted with alanine (S282A) into neonatal rat ventricular myocytes (NRVMs) induced cell apoptosis and Ca2+ transient desynchronization, whereas using gap junction inhibitor or knocking down Cx43 expression with Cx43-miRNA caused uncoupled Ca2+ signaling without cell death. Similarly, while Cx43-S282A+/+ failed in generation, Cx43-S282A+/− mice exhibited cardiomyocyte apoptosis and ventricular arrhythmias dependent on S282 dephosphorylation. Further, Cx43 dephosphorylation at S282 activated p38 mitogen-activated protein kinase (p38 MAPK), factor-associated suicide and the caspase-8 apoptotic pathway by physically interacting with p38 MAPK. These findings uncovered a specific Cx43 phosphorylation residue involved in regulating cardiomyocyte homeostasis. S282 phosphorylation deficiency acts as a trigger inducing cardiomyocyte apoptosis and cardiac arrhythmias, providing a potential mechanism for Cx43-mediated myocardial injury in severe cardiac diseases.

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Acknowledgements

This work was supported by the National Natural Science Foundation (81370339, 81570206) and Scientific Research Key Program of Beijing Municipal Commission of Education (KZ201710025023).

Author contributions

YY and XY contributed equally to the execution of most of the experiment and data analysis. JX planned and performed all Ca2+, Lucifer Yellow and cell death measurements, MC and ZS planned and performed immunofluorescence staining, TL planned and performed histochemistry, CW isolated and cultured cardiomyocytes and vascular smooth muscle cells, and HY generated and identified model animals. DL and YZ contributed to the design of the experiments, writing and data analysis of this manuscript.

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Conflict of interest

The authors declare that they have no conflict of interest.

Correspondence to Dali Luo.

Supplementary information

  1. Cx43 wt

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