Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells
Thanks to the crosstalk between Na<sup>+</sup> and Ca<sup>2+</sup> channels, Na<sup>+</sup> and Ca<sup>2+</sup> homeostasis interplay in so-called excitable cells enables the generation of action potential in response to electrical stimulation. Here, w...
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| Format: | Article |
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MDPI AG
2022-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/2/827 |
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| author | Léa Réthoré Joohee Park Jérôme Montnach Sébastien Nicolas Joseph Khoury Elodie Le Seac’h Kamel Mabrouk Harold De Pomyers Hélène Tricoire-Leignel César Mattei Daniel Henrion Ziad Fajloun Michel De Waard Claire Legendre Christian Legros |
| author_facet | Léa Réthoré Joohee Park Jérôme Montnach Sébastien Nicolas Joseph Khoury Elodie Le Seac’h Kamel Mabrouk Harold De Pomyers Hélène Tricoire-Leignel César Mattei Daniel Henrion Ziad Fajloun Michel De Waard Claire Legendre Christian Legros |
| author_sort | Léa Réthoré |
| collection | DOAJ |
| description | Thanks to the crosstalk between Na<sup>+</sup> and Ca<sup>2+</sup> channels, Na<sup>+</sup> and Ca<sup>2+</sup> homeostasis interplay in so-called excitable cells enables the generation of action potential in response to electrical stimulation. Here, we investigated the impact of persistent activation of voltage-gated Na<sup>+</sup> (Na<sub>V</sub>) channels by neurotoxins, such as veratridine (VTD), on intracellular Ca<sup>2+</sup> concentration ([Ca<sup>2+</sup>]<sub>i</sub>) in a model of excitable cells, the rat pituitary GH3b6 cells, in order to identify the molecular actors involved in Na<sup>+</sup>-Ca<sup>2+</sup> homeostasis crosstalk. By combining RT-qPCR, immunoblotting, immunocytochemistry, and patch-clamp techniques, we showed that GH3b6 cells predominantly express the Na<sub>V</sub>1.3 channel subtype, which likely endorses their voltage-activated Na<sup>+</sup> currents. Notably, these Na<sup>+</sup> currents were blocked by ICA-121431 and activated by the β-scorpion toxin Tf2, two selective Na<sub>V</sub>1.3 channel ligands. Using Fura-2, we showed that VTD induced a [Ca<sup>2+</sup>]<sub>i</sub> increase. This effect was suppressed by the selective Na<sub>V</sub> channel blocker tetrodotoxin, as well by the selective L-type Ca<sub>V</sub> channel (LTCC) blocker nifedipine. We also evidenced that crobenetine, a Na<sub>V</sub> channel blocker, abolished VTD-induced [Ca<sup>2+</sup>]<sub>i</sub> elevation, while it had no effects on LTCC. Altogether, our findings highlight a crosstalk between Na<sub>V</sub> and LTCC in GH3b6 cells, providing a new insight into the mode of action of neurotoxins. |
| first_indexed | 2024-03-10T01:18:00Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T01:18:00Z |
| publishDate | 2022-01-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-57ada7469988481e8614e44ef136cdde2023-11-23T14:05:01ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-01-0123282710.3390/ijms23020827Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 CellsLéa Réthoré0Joohee Park1Jérôme Montnach2Sébastien Nicolas3Joseph Khoury4Elodie Le Seac’h5Kamel Mabrouk6Harold De Pomyers7Hélène Tricoire-Leignel8César Mattei9Daniel Henrion10Ziad Fajloun11Michel De Waard12Claire Legendre13Christian Legros14INSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceInstitut du Thorax, Université de Nantes INSERM UMR 1087-CNRS UMR 6291, 44007 Nantes, FranceInstitut du Thorax, Université de Nantes INSERM UMR 1087-CNRS UMR 6291, 44007 Nantes, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceInstitut de Chimie Radicalaire, UMR CNRS 7273, Aix-Marseille Université, 13013 Marseille, FranceLatoxan Laboratory, 26800 Portes lès Valence, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceDepartment of Biology, Faculty of Science III, Michel Slayman Tripoli Campus, Lebanese University, Tripoli 1352, LebanonInstitut du Thorax, Université de Nantes INSERM UMR 1087-CNRS UMR 6291, 44007 Nantes, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceINSERM, CNRS, MITOVASC, Equipe CarME, SFR ICAT, University of Angers, 49000 Angers, FranceThanks to the crosstalk between Na<sup>+</sup> and Ca<sup>2+</sup> channels, Na<sup>+</sup> and Ca<sup>2+</sup> homeostasis interplay in so-called excitable cells enables the generation of action potential in response to electrical stimulation. Here, we investigated the impact of persistent activation of voltage-gated Na<sup>+</sup> (Na<sub>V</sub>) channels by neurotoxins, such as veratridine (VTD), on intracellular Ca<sup>2+</sup> concentration ([Ca<sup>2+</sup>]<sub>i</sub>) in a model of excitable cells, the rat pituitary GH3b6 cells, in order to identify the molecular actors involved in Na<sup>+</sup>-Ca<sup>2+</sup> homeostasis crosstalk. By combining RT-qPCR, immunoblotting, immunocytochemistry, and patch-clamp techniques, we showed that GH3b6 cells predominantly express the Na<sub>V</sub>1.3 channel subtype, which likely endorses their voltage-activated Na<sup>+</sup> currents. Notably, these Na<sup>+</sup> currents were blocked by ICA-121431 and activated by the β-scorpion toxin Tf2, two selective Na<sub>V</sub>1.3 channel ligands. Using Fura-2, we showed that VTD induced a [Ca<sup>2+</sup>]<sub>i</sub> increase. This effect was suppressed by the selective Na<sub>V</sub> channel blocker tetrodotoxin, as well by the selective L-type Ca<sub>V</sub> channel (LTCC) blocker nifedipine. We also evidenced that crobenetine, a Na<sub>V</sub> channel blocker, abolished VTD-induced [Ca<sup>2+</sup>]<sub>i</sub> elevation, while it had no effects on LTCC. Altogether, our findings highlight a crosstalk between Na<sub>V</sub> and LTCC in GH3b6 cells, providing a new insight into the mode of action of neurotoxins.https://www.mdpi.com/1422-0067/23/2/827GH3b6 cellsvoltage-gated sodium channelL-type voltage-gated calcium channelveratridinepatch-clampFura-2 |
| spellingShingle | Léa Réthoré Joohee Park Jérôme Montnach Sébastien Nicolas Joseph Khoury Elodie Le Seac’h Kamel Mabrouk Harold De Pomyers Hélène Tricoire-Leignel César Mattei Daniel Henrion Ziad Fajloun Michel De Waard Claire Legendre Christian Legros Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells International Journal of Molecular Sciences GH3b6 cells voltage-gated sodium channel L-type voltage-gated calcium channel veratridine patch-clamp Fura-2 |
| title | Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells |
| title_full | Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells |
| title_fullStr | Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells |
| title_full_unstemmed | Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells |
| title_short | Pharmacological Dissection of the Crosstalk between Na<sub>V</sub> and Ca<sub>V</sub> Channels in GH3b6 Cells |
| title_sort | pharmacological dissection of the crosstalk between na sub v sub and ca sub v sub channels in gh3b6 cells |
| topic | GH3b6 cells voltage-gated sodium channel L-type voltage-gated calcium channel veratridine patch-clamp Fura-2 |
| url | https://www.mdpi.com/1422-0067/23/2/827 |
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