Hypothermia increases aquaporin 4 (AQP4) plasma membrane abundance in human primary cortical astrocytes via a calcium/transient receptor potential vanilloid 4 (TRPV4)- and calmodulin-mediated mechanism

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Hdl Handle:
http://hdl.handle.net/2436/620810
Title:
Hypothermia increases aquaporin 4 (AQP4) plasma membrane abundance in human primary cortical astrocytes via a calcium/transient receptor potential vanilloid 4 (TRPV4)- and calmodulin-mediated mechanism
Authors:
Salman, Mootaz M. ( 0000-0002-5683-1706 ) ; Kitchen, Philip; Woodroofe, M. Nicola; Brown, James E.; Bill, Roslyn M.; Conner, Alex C.; Conner, Matthew T. ( 0000-0001-5662-9121 )
Abstract:
Human aquaporin 4 (AQP4) is the primary water channel protein in brain astrocytes. Hypothermia is known to cause astrocyte swelling in culture, but the precise role of AQP4 in this process is unknown. Primary human cortical astrocytes were cultured under hypothermic (32 °C) or normothermic (37 °C) conditions. AQP4 transcript, total protein and surface-localized protein were quantified using RT-qPCR, sandwich ELISA with whole cell lysates or cell surface biotinylation, followed by ELISA analysis of the surface-localized protein, respectively. Four-hour mild hypothermic treatment increased the surface localization of AQP4 in human astrocytes to 155 4% of normothermic controls, despite no change in total protein expression levels. The hypothermiamediated increase in AQP4 surface abundance on human astrocytes was blocked using either calmodulin antagonist (trifluoperazine, TFP); TRPV4 antagonist, HC-067047 or calcium chelation using EGTA-AM. The TRPV4 agonist (GSK1016790A) mimicked the effect of hypothermia compared with untreated normothermic astrocytes. Hypothermia led to an increase in surface localization of AQP4 in human astrocytes through a mechanism likely dependent on the TRPV4 calcium channel and calmodulin activation. Understanding the effects of hypothermia on astrocytic AQP4 cell surface expression may help develop new treatments for brain swelling based on an in-depth mechanistic understanding of AQP4 translocation.
Citation:
Hypothermia increases aquaporin 4 (AQP4) plasma membrane abundance in human primary cortical astrocytes via a calcium/transient receptor potential vanilloid 4 (TRPV4)- and calmodulin-mediated mechanism 2017 European Journal of Neuroscience
Publisher:
Federation of European Neuroscience Societies and John Wiley & Sons Ltd
Journal:
European Journal of Neuroscience
Issue Date:
13-Oct-2017
URI:
http://hdl.handle.net/2436/620810
DOI:
10.1111/ejn.13723
Additional Links:
http://doi.wiley.com/10.1111/ejn.13723
Type:
Article
Language:
en
ISSN:
0953-816X
Sponsors:
BMRC Sheffield Hallam University, RIHS University of Wolverhampton, School of Life and Health Sciences Aston University and the HCED/Iraq grant number GD-13-3 (M Salman).
Appears in Collections:
FSE

Full metadata record

DC FieldValue Language
dc.contributor.authorSalman, Mootaz M.en
dc.contributor.authorKitchen, Philipen
dc.contributor.authorWoodroofe, M. Nicolaen
dc.contributor.authorBrown, James E.en
dc.contributor.authorBill, Roslyn M.en
dc.contributor.authorConner, Alex C.en
dc.contributor.authorConner, Matthew T.en
dc.date.accessioned2017-10-30T11:58:39Z-
dc.date.available2017-10-30T11:58:39Z-
dc.date.issued2017-10-13-
dc.identifier.citationHypothermia increases aquaporin 4 (AQP4) plasma membrane abundance in human primary cortical astrocytes via a calcium/transient receptor potential vanilloid 4 (TRPV4)- and calmodulin-mediated mechanism 2017 European Journal of Neuroscienceen
dc.identifier.issn0953-816Xen
dc.identifier.doi10.1111/ejn.13723-
dc.identifier.urihttp://hdl.handle.net/2436/620810-
dc.description.abstractHuman aquaporin 4 (AQP4) is the primary water channel protein in brain astrocytes. Hypothermia is known to cause astrocyte swelling in culture, but the precise role of AQP4 in this process is unknown. Primary human cortical astrocytes were cultured under hypothermic (32 °C) or normothermic (37 °C) conditions. AQP4 transcript, total protein and surface-localized protein were quantified using RT-qPCR, sandwich ELISA with whole cell lysates or cell surface biotinylation, followed by ELISA analysis of the surface-localized protein, respectively. Four-hour mild hypothermic treatment increased the surface localization of AQP4 in human astrocytes to 155 4% of normothermic controls, despite no change in total protein expression levels. The hypothermiamediated increase in AQP4 surface abundance on human astrocytes was blocked using either calmodulin antagonist (trifluoperazine, TFP); TRPV4 antagonist, HC-067047 or calcium chelation using EGTA-AM. The TRPV4 agonist (GSK1016790A) mimicked the effect of hypothermia compared with untreated normothermic astrocytes. Hypothermia led to an increase in surface localization of AQP4 in human astrocytes through a mechanism likely dependent on the TRPV4 calcium channel and calmodulin activation. Understanding the effects of hypothermia on astrocytic AQP4 cell surface expression may help develop new treatments for brain swelling based on an in-depth mechanistic understanding of AQP4 translocation.en
dc.description.sponsorshipBMRC Sheffield Hallam University, RIHS University of Wolverhampton, School of Life and Health Sciences Aston University and the HCED/Iraq grant number GD-13-3 (M Salman).en
dc.language.isoenen
dc.publisherFederation of European Neuroscience Societies and John Wiley & Sons Ltden
dc.relation.urlhttp://doi.wiley.com/10.1111/ejn.13723en
dc.rightsArchived with thanks to European Journal of Neuroscienceen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectaquaporin 4en
dc.subjectastrocyteen
dc.subjectcalciumen
dc.subjectcalmodulinen
dc.subjectmild therapeutic hypothermiaen
dc.subjectTRPV4en
dc.titleHypothermia increases aquaporin 4 (AQP4) plasma membrane abundance in human primary cortical astrocytes via a calcium/transient receptor potential vanilloid 4 (TRPV4)- and calmodulin-mediated mechanismen
dc.typeArticleen
dc.identifier.journalEuropean Journal of Neuroscienceen
dc.contributor.institutionBiomolecular Sciences Research Centre; Sheffield Hallam University; Sheffield UK-
dc.contributor.institutionInstitute of Cancer and Genomic Sciences; University of Birmingham; Birmingham UK-
dc.contributor.institutionBiomolecular Sciences Research Centre; Sheffield Hallam University; Sheffield UK-
dc.contributor.institutionSchool of Life & Health Sciences; Aston University; Aston Triangle Birmingham B4 7ET UK-
dc.contributor.institutionSchool of Life & Health Sciences; Aston University; Aston Triangle Birmingham B4 7ET UK-
dc.contributor.institutionInstitute of Clinical Sciences; University of Birmingham; Edgbaston Birmingham B15 2TT UK-
dc.contributor.institutionResearch Institute of Health Sciences; Wolverhampton School of Sciences; University of Wolverhampton; Wulfruna St Wolverhampton WV1 1LY UK-
dc.date.accepted2017-09-
rioxxterms.funderBMRC Sheffield Hallam University, RIHS University of Wolverhampton, School of Life and Health Sciences AstonUniversity and the HCED/Iraq grant number GD-13-3 (M Salman).en
rioxxterms.identifier.projectUoW301017MCen
rioxxterms.versionVoRen
rioxxterms.licenseref.urihttps://creativecommons.org/CC BY-NC-ND 4.0en
rioxxterms.licenseref.startdate2017-10-30en
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