Coordinated regulation of TRPV5-mediated Ca²⁺ transport in primary distal convolution cultures.

Détails

ID Serval
serval:BIB_80D3958A3D52
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Coordinated regulation of TRPV5-mediated Ca²⁺ transport in primary distal convolution cultures.
Périodique
Pflügers Archiv : European Journal of Physiology
Auteur(s)
van der Hagen E.A., Lavrijsen M., van Zeeland F., Praetorius J., Bonny O., Bindels R.J., Hoenderop J.G.
ISSN
1432-2013 (Electronic)
ISSN-L
0031-6768
Statut éditorial
Publié
Date de publication
11/2014
Peer-reviewed
Oui
Volume
466
Numéro
11
Pages
2077-2087
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov' tPublication Status: ppublish
Résumé
Fine-tuning of renal calcium ion (Ca(2+)) reabsorption takes place in the distal convoluted and connecting tubules (distal convolution) of the kidney via transcellular Ca(2+) transport, a process controlled by the epithelial Ca(2+) channel Transient Receptor Potential Vanilloid 5 (TRPV5). Studies to delineate the molecular mechanism of transcellular Ca(2+) transport are seriously hampered by the lack of a suitable cell model. The present study describes the establishment and validation of a primary murine cell model of the distal convolution. Viable kidney tubules were isolated from mice expressing enhanced Green Fluorescent Protein (eGFP) under the control of a TRPV5 promoter (pTRPV5-eGFP), using Complex Object Parametric Analyser and Sorting (COPAS) technology. Tubules were grown into tight monolayers on semi-permeable supports. Radioactive (45)Ca(2+) assays showed apical-to-basolateral transport rates of 13.5 ± 1.2 nmol/h/cm(2), which were enhanced by the calciotropic hormones parathyroid hormone and 1,25-dihydroxy vitamin D3. Cell cultures lacking TRPV5, generated by crossbreeding pTRPV5-eGFP with TRPV5 knockout mice (TRPV5(-/-)), showed significantly reduced transepithelial Ca(2+) transport (26 % of control), for the first time directly confirming the key role of TRPV5. Most importantly, using this cell model, a novel molecular player in transepithelial Ca(2+) transport was identified: mRNA analysis revealed that ATP-dependent Ca(2+)-ATPase 4 (PMCA4) instead of PMCA1 was enriched in isolated tubules and downregulated in TRPV5(-/-) material. Immunohistochemical stainings confirmed co-localization of PMCA4 with TRPV5 in the distal convolution. In conclusion, a novel primary cell model with TRPV5-dependent Ca(2+) transport characteristics was successfully established, enabling comprehensive studies of transcellular Ca(2+) transport.
Pubmed
Web of science
Création de la notice
15/01/2015 15:02
Dernière modification de la notice
20/08/2019 14:41
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