Show simple item record

dc.contributor.authorSalman, Mootaz M.
dc.contributor.authorKitchen, Philip
dc.contributor.authorWoodroofe, M. Nicola
dc.contributor.authorBrown, James E.
dc.contributor.authorBill, Roslyn M.
dc.contributor.authorConner, Alex C.
dc.contributor.authorConner, Matthew T.
dc.date.accessioned2017-10-30T11:58:39Z
dc.date.available2017-10-30T11:58:39Z
dc.date.issued2017-10-13
dc.identifier.citationSalman MM., Kitchen P., Woodroofe MN., Brown JE., Bill RM., Conner AC., Conner MT. (2017) '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', European Journal of Neuroscience, 46(9) pp. 2542-2547. doi: 10.1111/ejn.13723
dc.identifier.issn0953-816X
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.
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).
dc.language.isoen
dc.publisherFederation of European Neuroscience Societies and John Wiley & Sons Ltd
dc.relation.urlhttp://doi.wiley.com/10.1111/ejn.13723
dc.subjectaquaporin 4
dc.subjectastrocyte
dc.subjectcalcium
dc.subjectcalmodulin
dc.subjectmild therapeutic hypothermia
dc.subjectTRPV4
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 mechanism
dc.typeJournal article
dc.identifier.journalEuropean Journal of Neuroscience
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.accepted2017-09-30
rioxxterms.funderUniversity of Wolverhampton
rioxxterms.identifier.projectUoW301017MC
rioxxterms.versionVoR
rioxxterms.licenseref.urihttps://creativecommons.org/CC BY-NC-ND 4.0
rioxxterms.licenseref.startdate2017-10-30
dc.source.volume46
dc.source.issue9
dc.source.beginpage2542
dc.source.endpage2547
refterms.dateFCD2018-10-19T09:24:44Z
refterms.versionFCDVoR
refterms.dateFOA2017-10-30T00:00:00Z
html.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.


Files in this item

Thumbnail
Name:
Hypothermia and AQP4 EJN paper ...
Size:
565.9Kb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record