Cargando…

AQP4-independent TRPV4 modulation of plasma membrane water permeability

Despite of the major role of aquaporin (AQP) water channels in controlling transmembrane water fluxes, alternative ways for modulating water permeation have been proposed. In the Central Nervous System (CNS), Aquaporin-4 (AQP4) is reported to be functionally coupled with the calcium-channel Transien...

Descripción completa

Detalles Bibliográficos
Autores principales: Barile, Barbara, Mola, Maria Grazia, Formaggio, Francesco, Saracino, Emanuela, Cibelli, Antonio, Gargano, Concetta Domenica, Mogni, Guido, Frigeri, Antonio, Caprini, Marco, Benfenati, Valentina, Nicchia, Grazia Paola
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10500071/
https://www.ncbi.nlm.nih.gov/pubmed/37720545
http://dx.doi.org/10.3389/fncel.2023.1247761
_version_ 1785105846144860160
author Barile, Barbara
Mola, Maria Grazia
Formaggio, Francesco
Saracino, Emanuela
Cibelli, Antonio
Gargano, Concetta Domenica
Mogni, Guido
Frigeri, Antonio
Caprini, Marco
Benfenati, Valentina
Nicchia, Grazia Paola
author_facet Barile, Barbara
Mola, Maria Grazia
Formaggio, Francesco
Saracino, Emanuela
Cibelli, Antonio
Gargano, Concetta Domenica
Mogni, Guido
Frigeri, Antonio
Caprini, Marco
Benfenati, Valentina
Nicchia, Grazia Paola
author_sort Barile, Barbara
collection PubMed
description Despite of the major role of aquaporin (AQP) water channels in controlling transmembrane water fluxes, alternative ways for modulating water permeation have been proposed. In the Central Nervous System (CNS), Aquaporin-4 (AQP4) is reported to be functionally coupled with the calcium-channel Transient-Receptor Potential Vanilloid member-4 (TRPV4), which is controversially involved in cell volume regulation mechanisms and water transport dynamics. The present work aims to investigate the selective role of TRPV4 in regulating plasma membrane water permeability in an AQP4-independent way. Fluorescence-quenching water transport experiments in Aqp4(–/–) astrocytes revealed that cell swelling rate is significantly increased upon TRPV4 activation and in the absence of AQP4. The biophysical properties of TRPV4-dependent water transport were therefore assessed using the HEK-293 cell model. Calcein quenching experiments showed that chemical and thermal activation of TRPV4 overexpressed in HEK-293 cells leads to faster swelling kinetics. Stopped-flow light scattering water transport assay was used to measure the osmotic permeability coefficient (Pf, cm/s) and activation energy (Ea, kcal/mol) conferred by TRPV4. Results provided evidence that although the Pf measured upon TRPV4 activation is lower than the one obtained in AQP4-overexpressing cells (Pf of AQP4 = 0.01667 ± 0.0007; Pf of TRPV4 = 0.002261 ± 0.0004; Pf of TRPV4 + 4αPDD = 0.007985 ± 0.0006; Pf of WT = 0.002249 ± 0.0002), along with activation energy values (Ea of AQP4 = 0.86 ± 0.0006; Ea of TRPV4 + 4αPDD = 2.73 ± 1.9; Ea of WT = 8.532 ± 0.4), these parameters were compatible with a facilitated pathway for water movement rather than simple diffusion. The possibility to tune plasma membrane water permeability more finely through TRPV4 might represent a protective mechanism in cells constantly facing severe osmotic challenges to avoid the potential deleterious effects of the rapid cell swelling occurring via AQP channels.
format Online
Article
Text
id pubmed-10500071
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-105000712023-09-15 AQP4-independent TRPV4 modulation of plasma membrane water permeability Barile, Barbara Mola, Maria Grazia Formaggio, Francesco Saracino, Emanuela Cibelli, Antonio Gargano, Concetta Domenica Mogni, Guido Frigeri, Antonio Caprini, Marco Benfenati, Valentina Nicchia, Grazia Paola Front Cell Neurosci Neuroscience Despite of the major role of aquaporin (AQP) water channels in controlling transmembrane water fluxes, alternative ways for modulating water permeation have been proposed. In the Central Nervous System (CNS), Aquaporin-4 (AQP4) is reported to be functionally coupled with the calcium-channel Transient-Receptor Potential Vanilloid member-4 (TRPV4), which is controversially involved in cell volume regulation mechanisms and water transport dynamics. The present work aims to investigate the selective role of TRPV4 in regulating plasma membrane water permeability in an AQP4-independent way. Fluorescence-quenching water transport experiments in Aqp4(–/–) astrocytes revealed that cell swelling rate is significantly increased upon TRPV4 activation and in the absence of AQP4. The biophysical properties of TRPV4-dependent water transport were therefore assessed using the HEK-293 cell model. Calcein quenching experiments showed that chemical and thermal activation of TRPV4 overexpressed in HEK-293 cells leads to faster swelling kinetics. Stopped-flow light scattering water transport assay was used to measure the osmotic permeability coefficient (Pf, cm/s) and activation energy (Ea, kcal/mol) conferred by TRPV4. Results provided evidence that although the Pf measured upon TRPV4 activation is lower than the one obtained in AQP4-overexpressing cells (Pf of AQP4 = 0.01667 ± 0.0007; Pf of TRPV4 = 0.002261 ± 0.0004; Pf of TRPV4 + 4αPDD = 0.007985 ± 0.0006; Pf of WT = 0.002249 ± 0.0002), along with activation energy values (Ea of AQP4 = 0.86 ± 0.0006; Ea of TRPV4 + 4αPDD = 2.73 ± 1.9; Ea of WT = 8.532 ± 0.4), these parameters were compatible with a facilitated pathway for water movement rather than simple diffusion. The possibility to tune plasma membrane water permeability more finely through TRPV4 might represent a protective mechanism in cells constantly facing severe osmotic challenges to avoid the potential deleterious effects of the rapid cell swelling occurring via AQP channels. Frontiers Media S.A. 2023-08-31 /pmc/articles/PMC10500071/ /pubmed/37720545 http://dx.doi.org/10.3389/fncel.2023.1247761 Text en Copyright © 2023 Barile, Mola, Formaggio, Saracino, Cibelli, Gargano, Mogni, Frigeri, Caprini, Benfenati and Nicchia. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Barile, Barbara
Mola, Maria Grazia
Formaggio, Francesco
Saracino, Emanuela
Cibelli, Antonio
Gargano, Concetta Domenica
Mogni, Guido
Frigeri, Antonio
Caprini, Marco
Benfenati, Valentina
Nicchia, Grazia Paola
AQP4-independent TRPV4 modulation of plasma membrane water permeability
title AQP4-independent TRPV4 modulation of plasma membrane water permeability
title_full AQP4-independent TRPV4 modulation of plasma membrane water permeability
title_fullStr AQP4-independent TRPV4 modulation of plasma membrane water permeability
title_full_unstemmed AQP4-independent TRPV4 modulation of plasma membrane water permeability
title_short AQP4-independent TRPV4 modulation of plasma membrane water permeability
title_sort aqp4-independent trpv4 modulation of plasma membrane water permeability
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10500071/
https://www.ncbi.nlm.nih.gov/pubmed/37720545
http://dx.doi.org/10.3389/fncel.2023.1247761
work_keys_str_mv AT barilebarbara aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT molamariagrazia aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT formaggiofrancesco aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT saracinoemanuela aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT cibelliantonio aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT garganoconcettadomenica aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT mogniguido aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT frigeriantonio aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT caprinimarco aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT benfenativalentina aqp4independenttrpv4modulationofplasmamembranewaterpermeability
AT nicchiagraziapaola aqp4independenttrpv4modulationofplasmamembranewaterpermeability