Multiple na,k-atpase subunits colocalize in the brush border of mouse choroid plexus epithelial cells

Inga Baasch Christensen, Lei Cheng, Jonathan R. Brewer, Udo Bartsch, Robert A. Fenton, Helle H. Damkier, Jeppe Praetorius*

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Abstrakt

The unusual accumulation of Na,K-ATPase complexes in the brush border membrane of choroid plexus epithelial cells have intrigued researchers for decades. However, the full range of the expressed Na,K-ATPase subunits and their relation to the microvillus cytoskeleton remains unknown. (2) Methods: RT-PCR analysis, co-immunoprecipitation, native PAGE, mass spectrometry, and differential centrifugation were combined with high-resolution immunofluores-cence histochemistry, proximity ligase assays, and stimulated emission depletion (STED) microscopy on mouse choroid plexus cells or tissues in order to resolve these issues. (3) Results: The choroid plexus epithelium expresses Na,K-ATPase subunits α1, α2, β1, β2, β3, and phospholemman. The α1, α2, β1, and β2, subunits are all localized to the brush border membrane, where they appear to form a complex. The ATPase complexes may stabilize in the brush border membrane via anchor-ing to microvillar actin indirectly through ankyrin-3 or directly via other co-precipitated proteins. Aquaporin 1 (AQP1) may form part of the proposed multi-protein complexes in contrast to another membrane protein, the Na-K-2Cl cotransporter 1 (NKCC1). NKCC1 expression seems necessary for full brush border membrane accumulation of the Na,K-ATPase in the choroid plexus. (4) Conclusion: A multitude of Na,K-ATPase subunits form molecular complexes in the choroid plexus brush border, which may bind to the cytoskeleton by various alternative actin binding proteins.

OriginalsprogEngelsk
Artikelnummer1569
TidsskriftInternational Journal of Molecular Sciences
Vol/bind22
Udgave nummer4
Sider (fra-til)1-27
Antal sider27
ISSN1661-6596
DOI
StatusUdgivet - feb. 2021

Bibliografisk note

Funding Information:
Funding: This research was funded by the Danish Council for Independent Research (0602-02404B), the Lundbeck Foundation (R54-A5378), the Novo Nordisk Foundation (NNF17OC0028584), AU Ideas—Aarhus University Research Foundation.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

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