The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes

The epsilon toxin (Etx) from Clostridium perfringens has been identified as a potential trigger of multiple sclerosis, functioning as a pore-forming toxin that selectively targets cells expressing the plasma membrane (PM) myelin and lymphocyte protein (MAL). Previously, we observed that Etx induces...

Full description

Bibliographic Details
Authors: Dorca Arévalo, Jonatan, Cases Escuté, Mercè, Blanch Lozano, Marta, Rodil, Sergi, Terni, Beatrice, Martín Satué, Mireia, Llobet Berenguer, Artur, 1972-, Blasi Cabús, Joan, Solsona, Carles
Format: article
Status:Published version
Publication Date:2024
Country:España
Institution:Universidad de Barcelona
Repository:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/216569
Online Access:https://hdl.handle.net/2445/216569
Access Level:Open access
Keyword:Toxines bacterianes
Calci
Membranes cel·lulars
Trifosfat d'adenosina
Bacterial toxins
Calcium
Cell membranes
Adenosine triphospahatase
id ES_7f285b79c35b598972ca4d0047f7c577
oai_identifier_str oai:diposit.ub.edu:2445/216569
network_acronym_str ES
network_name_str España
repository_id_str
spelling The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytesDorca Arévalo, JonatanCases Escuté, MercèBlanch Lozano, MartaRodil, SergiTerni, BeatriceMartín Satué, MireiaLlobet Berenguer, Artur, 1972-Blasi Cabús, JoanSolsona, CarlesToxines bacterianesCalciMembranes cel·lularsTrifosfat d'adenosinaBacterial toxinsCalciumCell membranesAdenosine triphospahataseThe epsilon toxin (Etx) from Clostridium perfringens has been identified as a potential trigger of multiple sclerosis, functioning as a pore-forming toxin that selectively targets cells expressing the plasma membrane (PM) myelin and lymphocyte protein (MAL). Previously, we observed that Etx induces the release of intracellular ATP in sensitive cell lines. Here, we aimed to re-examine the mechanism of action of the toxin and investigate the connection between pore formation and ATP release. We examined the impact of Etx on Xenopus laevis oocytes expressing human MAL. Extracellular ATP was assessed using the luciferin-luciferase reaction. Activation of calcium-activated chloride channels (CaCCs) and a decrease in the PM surface were recorded using the two-electrode voltage-clamp technique. To evaluate intracellular Ca2+ levels and scramblase activity, fluorescent dyes were employed. Extracellular vesicles were imaged using light and electron microscopy, while toxin oligomers were identified through western blots. Etx triggered intracellular Ca2+ mobilization in the Xenopus oocytes expressing hMAL, leading to the activation of CaCCs, ATP release, and a reduction in PM capacitance. The toxin induced the activation of scramblase and, thus, translocated phospholipids from the inner to the outer leaflet of the PM, exposing phosphatidylserine outside in Xenopus oocytes and in an Etx-sensitive cell line. Moreover, Etx caused the formation of extracellular vesicles, not derived from apoptotic bodies, through PM fission. These vesicles carried toxin heptamers and doughnut-like structures in the nanometer size range. In conclusion, ATP release was not directly attributed to the formation of pores in the PM, but to scramblase activity and the formation of extracellular vesicles.John Wiley & Sons2024info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/216569Articles publicats en revistes (Patologia i Terapèutica Experimental)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.1002/prp2.70005Pharmacology Research & Perspectives, 2024, vol. 12, num.5https://doi.org/10.1002/prp2.70005cc by-nc (c) Cases, M. et al., 2024https://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/2165692026-05-27T06:46:51Z
dc.title.none.fl_str_mv The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
title The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
spellingShingle The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
Dorca Arévalo, Jonatan
Toxines bacterianes
Calci
Membranes cel·lulars
Trifosfat d'adenosina
Bacterial toxins
Calcium
Cell membranes
Adenosine triphospahatase
title_short The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
title_full The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
title_fullStr The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
title_full_unstemmed The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
title_sort The epsilon toxin from Clostridium perfringens stimulates calcium-activated chloride channels, generating extracellular vesicles in Xenopus oocytes
dc.creator.none.fl_str_mv Dorca Arévalo, Jonatan
Cases Escuté, Mercè
Blanch Lozano, Marta
Rodil, Sergi
Terni, Beatrice
Martín Satué, Mireia
Llobet Berenguer, Artur, 1972-
Blasi Cabús, Joan
Solsona, Carles
author Dorca Arévalo, Jonatan
author_facet Dorca Arévalo, Jonatan
Cases Escuté, Mercè
Blanch Lozano, Marta
Rodil, Sergi
Terni, Beatrice
Martín Satué, Mireia
Llobet Berenguer, Artur, 1972-
Blasi Cabús, Joan
Solsona, Carles
author_role author
author2 Cases Escuté, Mercè
Blanch Lozano, Marta
Rodil, Sergi
Terni, Beatrice
Martín Satué, Mireia
Llobet Berenguer, Artur, 1972-
Blasi Cabús, Joan
Solsona, Carles
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Toxines bacterianes
Calci
Membranes cel·lulars
Trifosfat d'adenosina
Bacterial toxins
Calcium
Cell membranes
Adenosine triphospahatase
topic Toxines bacterianes
Calci
Membranes cel·lulars
Trifosfat d'adenosina
Bacterial toxins
Calcium
Cell membranes
Adenosine triphospahatase
description The epsilon toxin (Etx) from Clostridium perfringens has been identified as a potential trigger of multiple sclerosis, functioning as a pore-forming toxin that selectively targets cells expressing the plasma membrane (PM) myelin and lymphocyte protein (MAL). Previously, we observed that Etx induces the release of intracellular ATP in sensitive cell lines. Here, we aimed to re-examine the mechanism of action of the toxin and investigate the connection between pore formation and ATP release. We examined the impact of Etx on Xenopus laevis oocytes expressing human MAL. Extracellular ATP was assessed using the luciferin-luciferase reaction. Activation of calcium-activated chloride channels (CaCCs) and a decrease in the PM surface were recorded using the two-electrode voltage-clamp technique. To evaluate intracellular Ca2+ levels and scramblase activity, fluorescent dyes were employed. Extracellular vesicles were imaged using light and electron microscopy, while toxin oligomers were identified through western blots. Etx triggered intracellular Ca2+ mobilization in the Xenopus oocytes expressing hMAL, leading to the activation of CaCCs, ATP release, and a reduction in PM capacitance. The toxin induced the activation of scramblase and, thus, translocated phospholipids from the inner to the outer leaflet of the PM, exposing phosphatidylserine outside in Xenopus oocytes and in an Etx-sensitive cell line. Moreover, Etx caused the formation of extracellular vesicles, not derived from apoptotic bodies, through PM fission. These vesicles carried toxin heptamers and doughnut-like structures in the nanometer size range. In conclusion, ATP release was not directly attributed to the formation of pores in the PM, but to scramblase activity and the formation of extracellular vesicles.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/216569
url https://hdl.handle.net/2445/216569
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1002/prp2.70005
Pharmacology Research & Perspectives, 2024, vol. 12, num.5
https://doi.org/10.1002/prp2.70005
dc.rights.none.fl_str_mv cc by-nc (c) Cases, M. et al., 2024
https://creativecommons.org/licenses/by-nc/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc by-nc (c) Cases, M. et al., 2024
https://creativecommons.org/licenses/by-nc/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv John Wiley & Sons
publisher.none.fl_str_mv John Wiley & Sons
dc.source.none.fl_str_mv Articles publicats en revistes (Patologia i Terapèutica Experimental)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869411803667103744
score 15,811543

Similar Items