The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes

Abstract The mechanoelectrical feedback (MEF) mechanism in the heart that plays a significant role in the occurrence of arrhythmias, involves cation flux through cation nonselective stretch‐activated channels (SACs). It is well known that nitric oxide (NO) can act as a regulator of MEF. Here we addr...

Full description

Bibliographic Details
Main Authors: Andre G. Kamkin, Olga V. Kamkina, Andrey L. Shim, Andrey Bilichenko, Vadim M. Mitrokhin, Viktor E. Kazansky, Tatiana S. Filatova, Denis V. Abramochkin, Mitko I. Mladenov
Format: Article
Language:English
Published: Wiley 2022-04-01
Series:Physiological Reports
Subjects:
Online Access:https://doi.org/10.14814/phy2.15246
_version_ 1797387902232035328
author Andre G. Kamkin
Olga V. Kamkina
Andrey L. Shim
Andrey Bilichenko
Vadim M. Mitrokhin
Viktor E. Kazansky
Tatiana S. Filatova
Denis V. Abramochkin
Mitko I. Mladenov
author_facet Andre G. Kamkin
Olga V. Kamkina
Andrey L. Shim
Andrey Bilichenko
Vadim M. Mitrokhin
Viktor E. Kazansky
Tatiana S. Filatova
Denis V. Abramochkin
Mitko I. Mladenov
author_sort Andre G. Kamkin
collection DOAJ
description Abstract The mechanoelectrical feedback (MEF) mechanism in the heart that plays a significant role in the occurrence of arrhythmias, involves cation flux through cation nonselective stretch‐activated channels (SACs). It is well known that nitric oxide (NO) can act as a regulator of MEF. Here we addressed the possibility of SAC’s regulation along NO‐dependent and NO‐independent pathways, as well as the possibility of S‐nitrosylation of SACs. In freshly isolated rat ventricular cardiomyocytes, using the patch‐clamp method in whole‐cell configuration, inward nonselective stretch‐activated cation current ISAC was recorded through SACs, which occurs during dosed cell stretching. NO donor SNAP, α1‐subunit of sGC activator BAY41‐2272, sGC blocker ODQ, PKG blocker KT5823, PKG activator 8Br‐cGMP, and S‐nitrosylation blocker ascorbic acid, were employed. We concluded that the physiological concentration of NO in the cell is a necessary condition for the functioning of SACs. An increase in NO due to SNAP in an unstretched cell causes the appearance of a Gd3+‐sensitive nonselective cation current, an analog of ISAC, while in a stretched cell it eliminates ISAC. The NO‐independent pathway of sGC activation of α subunit, triggered by BAY41‐2272, is also important for the regulation of SACs. Since S‐nitrosylation inhibitor completely abolishes ISAC, this mechanism occurs. The application of BAY41‐2272 cannot induce ISAC in a nonstretched cell; however, the addition of SNAP on its background activates SACs, rather due to S‐nitrosylation. ODQ eliminates ISAC, but SNAP added on the background of stretch increases ISAC in addition to ODQ. This may be a result of the lack of NO as a result of inhibition of NOS by metabolically modified ODQ. KT5823 reduces PKG activity and reduces SACs phosphorylation, leading to an increase in ISAC. 8Br‐cGMP reduces ISAC by activating PKG and its phosphorylation. These results demonstrate a significant contribution of S‐nitrosylation to the regulation of SACs.
first_indexed 2024-03-08T22:32:55Z
format Article
id doaj.art-f1d5b65b034e455caa819d1fd31c50a2
institution Directory Open Access Journal
issn 2051-817X
language English
last_indexed 2024-03-08T22:32:55Z
publishDate 2022-04-01
publisher Wiley
record_format Article
series Physiological Reports
spelling doaj.art-f1d5b65b034e455caa819d1fd31c50a22023-12-18T02:20:40ZengWileyPhysiological Reports2051-817X2022-04-01107n/an/a10.14814/phy2.15246The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytesAndre G. Kamkin0Olga V. Kamkina1Andrey L. Shim2Andrey Bilichenko3Vadim M. Mitrokhin4Viktor E. Kazansky5Tatiana S. Filatova6Denis V. Abramochkin7Mitko I. Mladenov8Department of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaDepartment of Physiology Pirogov Russian National Research Medical University Moscow RussiaAbstract The mechanoelectrical feedback (MEF) mechanism in the heart that plays a significant role in the occurrence of arrhythmias, involves cation flux through cation nonselective stretch‐activated channels (SACs). It is well known that nitric oxide (NO) can act as a regulator of MEF. Here we addressed the possibility of SAC’s regulation along NO‐dependent and NO‐independent pathways, as well as the possibility of S‐nitrosylation of SACs. In freshly isolated rat ventricular cardiomyocytes, using the patch‐clamp method in whole‐cell configuration, inward nonselective stretch‐activated cation current ISAC was recorded through SACs, which occurs during dosed cell stretching. NO donor SNAP, α1‐subunit of sGC activator BAY41‐2272, sGC blocker ODQ, PKG blocker KT5823, PKG activator 8Br‐cGMP, and S‐nitrosylation blocker ascorbic acid, were employed. We concluded that the physiological concentration of NO in the cell is a necessary condition for the functioning of SACs. An increase in NO due to SNAP in an unstretched cell causes the appearance of a Gd3+‐sensitive nonselective cation current, an analog of ISAC, while in a stretched cell it eliminates ISAC. The NO‐independent pathway of sGC activation of α subunit, triggered by BAY41‐2272, is also important for the regulation of SACs. Since S‐nitrosylation inhibitor completely abolishes ISAC, this mechanism occurs. The application of BAY41‐2272 cannot induce ISAC in a nonstretched cell; however, the addition of SNAP on its background activates SACs, rather due to S‐nitrosylation. ODQ eliminates ISAC, but SNAP added on the background of stretch increases ISAC in addition to ODQ. This may be a result of the lack of NO as a result of inhibition of NOS by metabolically modified ODQ. KT5823 reduces PKG activity and reduces SACs phosphorylation, leading to an increase in ISAC. 8Br‐cGMP reduces ISAC by activating PKG and its phosphorylation. These results demonstrate a significant contribution of S‐nitrosylation to the regulation of SACs.https://doi.org/10.14814/phy2.152468Br‐cGMPascorbic acidBAY41‐2272KT5823L‐Argininenitric oxide
spellingShingle Andre G. Kamkin
Olga V. Kamkina
Andrey L. Shim
Andrey Bilichenko
Vadim M. Mitrokhin
Viktor E. Kazansky
Tatiana S. Filatova
Denis V. Abramochkin
Mitko I. Mladenov
The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes
Physiological Reports
8Br‐cGMP
ascorbic acid
BAY41‐2272
KT5823
L‐Arginine
nitric oxide
title The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes
title_full The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes
title_fullStr The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes
title_full_unstemmed The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes
title_short The role of activation of two different sGC binding sites by NO‐dependent and NO‐independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes
title_sort role of activation of two different sgc binding sites by no dependent and no independent mechanisms in the regulation of sacs in rat ventricular cardiomyocytes
topic 8Br‐cGMP
ascorbic acid
BAY41‐2272
KT5823
L‐Arginine
nitric oxide
url https://doi.org/10.14814/phy2.15246
work_keys_str_mv AT andregkamkin theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT olgavkamkina theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT andreylshim theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT andreybilichenko theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT vadimmmitrokhin theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT viktorekazansky theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT tatianasfilatova theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT denisvabramochkin theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT mitkoimladenov theroleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT andregkamkin roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT olgavkamkina roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT andreylshim roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT andreybilichenko roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT vadimmmitrokhin roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT viktorekazansky roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT tatianasfilatova roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT denisvabramochkin roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes
AT mitkoimladenov roleofactivationoftwodifferentsgcbindingsitesbynodependentandnoindependentmechanismsintheregulationofsacsinratventricularcardiomyocytes