TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line
The Ca<sup>2+</sup> release in microdomains formed by intercompartmental contacts, such as mitochondria-associated endoplasmic reticulum membranes (MAMs), encodes a signal that contributes to Ca<sup>2+</sup> homeostasis and cell fate control. However, the composition and func...
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MDPI AG
2023-09-01
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| author | Nolwenn Tessier Mallory Ducrozet Maya Dia Sally Badawi Christophe Chouabe Claire Crola Da Silva Michel Ovize Gabriel Bidaux Fabien Van Coppenolle Sylvie Ducreux |
| author_facet | Nolwenn Tessier Mallory Ducrozet Maya Dia Sally Badawi Christophe Chouabe Claire Crola Da Silva Michel Ovize Gabriel Bidaux Fabien Van Coppenolle Sylvie Ducreux |
| author_sort | Nolwenn Tessier |
| collection | DOAJ |
| description | The Ca<sup>2+</sup> release in microdomains formed by intercompartmental contacts, such as mitochondria-associated endoplasmic reticulum membranes (MAMs), encodes a signal that contributes to Ca<sup>2+</sup> homeostasis and cell fate control. However, the composition and function of MAMs remain to be fully defined. Here, we focused on the transient receptor potential vanilloid 1 (TRPV1), a Ca<sup>2+</sup>-permeable ion channel and a polymodal nociceptor. We found TRPV1 channels in the reticular membrane, including some at MAMs, in a rat cardiomyoblast cell line (SV40-transformed H9c2) by Western blotting, immunostaining, cell fractionation, and proximity ligation assay. We used chemical and genetic probes to perform Ca<sup>2+</sup> imaging in four cellular compartments: the endoplasmic reticulum (ER), cytoplasm, mitochondrial matrix, and mitochondrial surface. Our results showed that the ER Ca<sup>2+</sup> released through TRPV1 channels is detected at the mitochondrial outer membrane and transferred to the mitochondria. Finally, we observed that prolonged TRPV1 modulation for 30 min alters the intracellular Ca<sup>2+</sup> equilibrium and influences the MAM structure or the hypoxia/reoxygenation-induced cell death. Thus, our study provides the first evidence that TRPV1 channels contribute to MAM Ca<sup>2+</sup> exchanges. |
| first_indexed | 2024-03-10T22:55:54Z |
| format | Article |
| id | doaj.art-080129ae2739452aa4fad1a8fe882142 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T22:55:54Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-080129ae2739452aa4fad1a8fe8821422023-11-19T10:00:24ZengMDPI AGCells2073-44092023-09-011218232210.3390/cells12182322TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell LineNolwenn Tessier0Mallory Ducrozet1Maya Dia2Sally Badawi3Christophe Chouabe4Claire Crola Da Silva5Michel Ovize6Gabriel Bidaux7Fabien Van Coppenolle8Sylvie Ducreux9Univ Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceUniv Lyon, CarMeN Laboratory, INSERM, INRA, INSA Lyon, Université Claude Bernard Lyon 1, 69500 Bron, FranceThe Ca<sup>2+</sup> release in microdomains formed by intercompartmental contacts, such as mitochondria-associated endoplasmic reticulum membranes (MAMs), encodes a signal that contributes to Ca<sup>2+</sup> homeostasis and cell fate control. However, the composition and function of MAMs remain to be fully defined. Here, we focused on the transient receptor potential vanilloid 1 (TRPV1), a Ca<sup>2+</sup>-permeable ion channel and a polymodal nociceptor. We found TRPV1 channels in the reticular membrane, including some at MAMs, in a rat cardiomyoblast cell line (SV40-transformed H9c2) by Western blotting, immunostaining, cell fractionation, and proximity ligation assay. We used chemical and genetic probes to perform Ca<sup>2+</sup> imaging in four cellular compartments: the endoplasmic reticulum (ER), cytoplasm, mitochondrial matrix, and mitochondrial surface. Our results showed that the ER Ca<sup>2+</sup> released through TRPV1 channels is detected at the mitochondrial outer membrane and transferred to the mitochondria. Finally, we observed that prolonged TRPV1 modulation for 30 min alters the intracellular Ca<sup>2+</sup> equilibrium and influences the MAM structure or the hypoxia/reoxygenation-induced cell death. Thus, our study provides the first evidence that TRPV1 channels contribute to MAM Ca<sup>2+</sup> exchanges.https://www.mdpi.com/2073-4409/12/18/2322TRPV1TRP channelsCa<sup>2+</sup> homeostasisER–mitochondria contact sitesH9c2hypoxia–reoxygenation |
| spellingShingle | Nolwenn Tessier Mallory Ducrozet Maya Dia Sally Badawi Christophe Chouabe Claire Crola Da Silva Michel Ovize Gabriel Bidaux Fabien Van Coppenolle Sylvie Ducreux TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line Cells TRPV1 TRP channels Ca<sup>2+</sup> homeostasis ER–mitochondria contact sites H9c2 hypoxia–reoxygenation |
| title | TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line |
| title_full | TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line |
| title_fullStr | TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line |
| title_full_unstemmed | TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line |
| title_short | TRPV1 Channels Are New Players in the Reticulum–Mitochondria Ca<sup>2+</sup> Coupling in a Rat Cardiomyoblast Cell Line |
| title_sort | trpv1 channels are new players in the reticulum mitochondria ca sup 2 sup coupling in a rat cardiomyoblast cell line |
| topic | TRPV1 TRP channels Ca<sup>2+</sup> homeostasis ER–mitochondria contact sites H9c2 hypoxia–reoxygenation |
| url | https://www.mdpi.com/2073-4409/12/18/2322 |
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