It is well established that abnormalities in [Ca2+] regulation occur in heart diseases. Actually, independent studies demonstrated that Orai1/2/3 and TRPC protein related with store-operated calcium channels (SOCC) have a role in cardiac pathologies. Ischemia/reperfusion (I/R) stimulates transcription factor activation that modifies the expression of genes implicated in the pathogenesis of this process. Previous results described an increase in the expression of Orai1 and TRPC5 in cardiomyocytes after I/R, although the molecular mechanisms that mediate this regulation are still unknown. The aim of this study is to examine the molecular mechanisms implicated in the regulation of SOCC in cardiomyocytes after I/R focusing on the handling of intracellular [Ca2+]. Experiments were performed in a rat model of myocardial I/R, in adult (ARVM) and neonatal rat ventricular myocytes (NRVM), and in ventricular samples of heart-failure patients. Immunofluorescence was used to investigate CREB activation, and the protein expression was analyzed by Western blot. Calcium diastolic studies were realized using microfluorimetric technic with FURA-2AM. To evoke intracellular Ca2+ transients, ARVMs were field stimulated at 0.5 Hz and NRVMs at 1 Hz. An activation of CREB after I/R was observed in adult and neonatal rat cardiomyocytes. Furthermore, it was demonstrated that this activation was mediated by PKA, but not for EPAC2 or ERK. I/R induced an CREB-dependent ORAI protein expression increase and also an increase in the diastolic calcium in NRVM and ARVM from I/R animal models. Additionally, it was observed that ORAI1 inhibition with SYNTA-66 or GSK reduced the calcium diastolic increase induced by I/R. We demonstrated, for the first time, the activation of the transcription factor CREB in cardiomyocytes after I/R. This activation induces an up-regulation of ORAI1, suggesting that this channel plays a role in the I/R induced calcium diastolic increase.
Meeting Abstract|
E–C Coupling Meeting 2021|
November 12 2021
Implication of CREB in the calcium regulation by ischemia/reperfusion in cardiomyocytes: Calcium Signaling and Excitation–Contraction in Cardiac, Skeletal and Smooth Muscle
Débora Falcón,
Débora Falcón
1Departamento de Fisiología Médica y Biofísica, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
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Isabel Mayoral,
Isabel Mayoral
1Departamento de Fisiología Médica y Biofísica, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
2Departamento Cirugía, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
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Antonio Ordóñez,
Antonio Ordóñez
2Departamento Cirugía, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
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Tarik Smani
Tarik Smani
1Departamento de Fisiología Médica y Biofísica, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
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Débora Falcón
1Departamento de Fisiología Médica y Biofísica, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
Isabel Mayoral
1Departamento de Fisiología Médica y Biofísica, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
2Departamento Cirugía, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
Antonio Ordóñez
2Departamento Cirugía, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
Tarik Smani
1Departamento de Fisiología Médica y Biofísica, Instituto de Biomedicina de Sevilla, Universidad de Sevilla, Seville, Spain
Online ISSN: 1540-7748
Print ISSN: 0022-1295
© 2021 Falcón et al.
2021
This article is available under a Creative Commons License (Attribution–Noncommercial–Share
Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Gen Physiol (2022) 154 (9): e2021ecc7.
Citation
Débora Falcón, Isabel Mayoral, Antonio Ordóñez, Tarik Smani; Implication of CREB in the calcium regulation by ischemia/reperfusion in cardiomyocytes: Calcium Signaling and Excitation–Contraction in Cardiac, Skeletal and Smooth Muscle. J Gen Physiol 5 September 2022; 154 (9): e2021ecc7. doi: https://doi.org/10.1085/jgp.2021ecc7
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