Cargando…

Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293

Microdomains formed by proteins of endoplasmic reticulum and plasma membrane play a key role in store-operated Ca(2+) entry (SOCE). Ca(2+) release through inositol 1,4,5-trisphosphate receptor (IP(3)R) and subsequent Ca(2+) store depletion activate STIM (stromal interaction molecules) proteins, sens...

Descripción completa

Detalles Bibliográficos
Autores principales: Kolesnikov, Dmitrii, Perevoznikova, Anastasiia, Gusev, Konstantin, Glushankova, Lyubov, Kaznacheyeva, Elena, Shalygin, Alexey
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8124839/
https://www.ncbi.nlm.nih.gov/pubmed/33946319
http://dx.doi.org/10.3390/ijms22094767
_version_ 1783693325951303680
author Kolesnikov, Dmitrii
Perevoznikova, Anastasiia
Gusev, Konstantin
Glushankova, Lyubov
Kaznacheyeva, Elena
Shalygin, Alexey
author_facet Kolesnikov, Dmitrii
Perevoznikova, Anastasiia
Gusev, Konstantin
Glushankova, Lyubov
Kaznacheyeva, Elena
Shalygin, Alexey
author_sort Kolesnikov, Dmitrii
collection PubMed
description Microdomains formed by proteins of endoplasmic reticulum and plasma membrane play a key role in store-operated Ca(2+) entry (SOCE). Ca(2+) release through inositol 1,4,5-trisphosphate receptor (IP(3)R) and subsequent Ca(2+) store depletion activate STIM (stromal interaction molecules) proteins, sensors of intraluminal Ca(2+), which, in turn, open the Orai channels in plasma membrane. Downstream to this process could be activated TRPC (transient receptor potential-canonical) calcium permeable channels. Using single channel patch-clamp technique we found that a local Ca(2+) entry through TRPC1 channels activated endogenous Ca(2+)-activated chloride channels (CaCCs) with properties similar to Anoctamin6 (TMEM16F). Our data suggest that their outward rectification is based on the dependence from membrane potential of both the channel conductance and the channel activity: (1) The conductance of active CaCCs highly depends on the transmembrane potential (from 3 pS at negative potentials till 60 pS at positive potentials); (2) their activity (NPo) is enhanced with increasing Ca(2+) concentration and/or transmembrane potential, conversely lowering of intracellular Ca(2+) concentration reduced the open state dwell time; (3) CaCC amplitude is only slightly increased by intracellular Ca(2+) concentration. Experiments with Ca(2+) buffering by EGTA or BAPTA suggest close local arrangement of functional CaCCs and TRPC1 channels. It is supposed that Ca(2+)-activated chloride channels are involved in Ca(2+) entry microdomains.
format Online
Article
Text
id pubmed-8124839
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-81248392021-05-17 Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293 Kolesnikov, Dmitrii Perevoznikova, Anastasiia Gusev, Konstantin Glushankova, Lyubov Kaznacheyeva, Elena Shalygin, Alexey Int J Mol Sci Article Microdomains formed by proteins of endoplasmic reticulum and plasma membrane play a key role in store-operated Ca(2+) entry (SOCE). Ca(2+) release through inositol 1,4,5-trisphosphate receptor (IP(3)R) and subsequent Ca(2+) store depletion activate STIM (stromal interaction molecules) proteins, sensors of intraluminal Ca(2+), which, in turn, open the Orai channels in plasma membrane. Downstream to this process could be activated TRPC (transient receptor potential-canonical) calcium permeable channels. Using single channel patch-clamp technique we found that a local Ca(2+) entry through TRPC1 channels activated endogenous Ca(2+)-activated chloride channels (CaCCs) with properties similar to Anoctamin6 (TMEM16F). Our data suggest that their outward rectification is based on the dependence from membrane potential of both the channel conductance and the channel activity: (1) The conductance of active CaCCs highly depends on the transmembrane potential (from 3 pS at negative potentials till 60 pS at positive potentials); (2) their activity (NPo) is enhanced with increasing Ca(2+) concentration and/or transmembrane potential, conversely lowering of intracellular Ca(2+) concentration reduced the open state dwell time; (3) CaCC amplitude is only slightly increased by intracellular Ca(2+) concentration. Experiments with Ca(2+) buffering by EGTA or BAPTA suggest close local arrangement of functional CaCCs and TRPC1 channels. It is supposed that Ca(2+)-activated chloride channels are involved in Ca(2+) entry microdomains. MDPI 2021-04-30 /pmc/articles/PMC8124839/ /pubmed/33946319 http://dx.doi.org/10.3390/ijms22094767 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kolesnikov, Dmitrii
Perevoznikova, Anastasiia
Gusev, Konstantin
Glushankova, Lyubov
Kaznacheyeva, Elena
Shalygin, Alexey
Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293
title Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293
title_full Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293
title_fullStr Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293
title_full_unstemmed Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293
title_short Electrophysiological Properties of Endogenous Single Ca(2+) Activated Cl(−) Channels Induced by Local Ca(2+) Entry in HEK293
title_sort electrophysiological properties of endogenous single ca(2+) activated cl(−) channels induced by local ca(2+) entry in hek293
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8124839/
https://www.ncbi.nlm.nih.gov/pubmed/33946319
http://dx.doi.org/10.3390/ijms22094767
work_keys_str_mv AT kolesnikovdmitrii electrophysiologicalpropertiesofendogenoussingleca2activatedclchannelsinducedbylocalca2entryinhek293
AT perevoznikovaanastasiia electrophysiologicalpropertiesofendogenoussingleca2activatedclchannelsinducedbylocalca2entryinhek293
AT gusevkonstantin electrophysiologicalpropertiesofendogenoussingleca2activatedclchannelsinducedbylocalca2entryinhek293
AT glushankovalyubov electrophysiologicalpropertiesofendogenoussingleca2activatedclchannelsinducedbylocalca2entryinhek293
AT kaznacheyevaelena electrophysiologicalpropertiesofendogenoussingleca2activatedclchannelsinducedbylocalca2entryinhek293
AT shalyginalexey electrophysiologicalpropertiesofendogenoussingleca2activatedclchannelsinducedbylocalca2entryinhek293