TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response

Abstract Despite successful clinical application of non-equilibrium atmospheric pressure plasma (APP), the details of the molecular mechanisms underlying APP-inducible biological responses remain ill-defined. We previously reported that exposure of 3T3L1 cells to APP-irradiated buffer raised the cyt...

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Autores principales:	Masayoshi Kawase, Weijian Chen, Kota Kawaguchi, Mazvita R. Nyasha, Shota Sasaki, Hiroyasu Hatakeyama, Toshiro Kaneko, Makoto Kanzaki
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6b5df954d6514b66b79681c89b1a47dd
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spelling	oai:doaj.org-article:6b5df954d6514b66b79681c89b1a47dd2021-12-02T17:41:07ZTRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response10.1038/s41598-020-66510-y2045-2322https://doaj.org/article/6b5df954d6514b66b79681c89b1a47dd2020-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-66510-yhttps://doaj.org/toc/2045-2322Abstract Despite successful clinical application of non-equilibrium atmospheric pressure plasma (APP), the details of the molecular mechanisms underlying APP-inducible biological responses remain ill-defined. We previously reported that exposure of 3T3L1 cells to APP-irradiated buffer raised the cytoplasmic free Ca2+ ([Ca2+]i) concentration by eliciting Ca2+ influx in a manner sensitive to transient receptor potential (TRP) channel inhibitors. However, the precise identity of the APP-responsive channel molecule(s) remains unclear. In the present study, we aimed to clarify channel molecule(s) responsible for indirect APP-responsive [Ca2+]i rises. siRNA-mediated silencing experiments revealed that TRPA1 and TRPV1 serve as the major APP-responsive Ca2+ channels in 3T3L1 cells. Conversely, ectopic expression of either TRPA1 or TRPV1 in APP-unresponsive C2C12 cells actually triggered [Ca2+]i elevation in response to indirect APP exposure. Desensitization experiments using 3T3L1 cells revealed APP responsiveness to be markedly suppressed after pretreatment with allyl isothiocyanate or capsaicin, TRPA1 and TRPV1 agonists, respectively. APP exposure also desensitized the cells to these chemical agonists, indicating the existence of a bi-directional heterologous desensitization property of APP-responsive [Ca2+]i transients mediated through these TRP channels. Mutational analyses of key cysteine residues in TRPA1 (Cys421, Cys621, Cys641, and Cys665) and in TRPV1 (Cys258, Cys363, and Cys742) have suggested that multiple reactive oxygen and nitrogen species are intricately involved in activation of the channels via a broad range of modifications involving these cysteine residues. Taken together, these observations allow us to conclude that both TRPA1 and TRPV1 channels play a pivotal role in evoking indirect APP-dependent [Ca2+]i responses.Masayoshi KawaseWeijian ChenKota KawaguchiMazvita R. NyashaShota SasakiHiroyasu HatakeyamaToshiro KanekoMakoto KanzakiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Masayoshi Kawase Weijian Chen Kota Kawaguchi Mazvita R. Nyasha Shota Sasaki Hiroyasu Hatakeyama Toshiro Kaneko Makoto Kanzaki TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response
description	Abstract Despite successful clinical application of non-equilibrium atmospheric pressure plasma (APP), the details of the molecular mechanisms underlying APP-inducible biological responses remain ill-defined. We previously reported that exposure of 3T3L1 cells to APP-irradiated buffer raised the cytoplasmic free Ca2+ ([Ca2+]i) concentration by eliciting Ca2+ influx in a manner sensitive to transient receptor potential (TRP) channel inhibitors. However, the precise identity of the APP-responsive channel molecule(s) remains unclear. In the present study, we aimed to clarify channel molecule(s) responsible for indirect APP-responsive [Ca2+]i rises. siRNA-mediated silencing experiments revealed that TRPA1 and TRPV1 serve as the major APP-responsive Ca2+ channels in 3T3L1 cells. Conversely, ectopic expression of either TRPA1 or TRPV1 in APP-unresponsive C2C12 cells actually triggered [Ca2+]i elevation in response to indirect APP exposure. Desensitization experiments using 3T3L1 cells revealed APP responsiveness to be markedly suppressed after pretreatment with allyl isothiocyanate or capsaicin, TRPA1 and TRPV1 agonists, respectively. APP exposure also desensitized the cells to these chemical agonists, indicating the existence of a bi-directional heterologous desensitization property of APP-responsive [Ca2+]i transients mediated through these TRP channels. Mutational analyses of key cysteine residues in TRPA1 (Cys421, Cys621, Cys641, and Cys665) and in TRPV1 (Cys258, Cys363, and Cys742) have suggested that multiple reactive oxygen and nitrogen species are intricately involved in activation of the channels via a broad range of modifications involving these cysteine residues. Taken together, these observations allow us to conclude that both TRPA1 and TRPV1 channels play a pivotal role in evoking indirect APP-dependent [Ca2+]i responses.
format	article
author	Masayoshi Kawase Weijian Chen Kota Kawaguchi Mazvita R. Nyasha Shota Sasaki Hiroyasu Hatakeyama Toshiro Kaneko Makoto Kanzaki
author_facet	Masayoshi Kawase Weijian Chen Kota Kawaguchi Mazvita R. Nyasha Shota Sasaki Hiroyasu Hatakeyama Toshiro Kaneko Makoto Kanzaki
author_sort	Masayoshi Kawase
title	TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response
title_short	TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response
title_full	TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response
title_fullStr	TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response
title_full_unstemmed	TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response
title_sort	trpa1 and trpv1 channels participate in atmospheric-pressure plasma-induced [ca2+]i response
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/6b5df954d6514b66b79681c89b1a47dd
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TRPA1 and TRPV1 channels participate in atmospheric-pressure plasma-induced [Ca2+]i response

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