A primary culture system of mouse thick ascending limb cells with preserved function and uromodulin processing.

Détails

ID Serval
serval:BIB_D13D89DDAFF4
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
A primary culture system of mouse thick ascending limb cells with preserved function and uromodulin processing.
Périodique
Pflügers Archiv : European Journal of Physiology
Auteur(s)
Glaudemans B., Terryn S., Gölz N., Brunati M., Cattaneo A., Bachi A., Al-Qusairi L., Ziegler U., Staub O., Rampoldi L., Devuyst O.
ISSN
1432-2013 (Electronic)
ISSN-L
0031-6768
Statut éditorial
Publié
Date de publication
2014
Peer-reviewed
Oui
Volume
466
Numéro
2
Pages
343-356
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Résumé
The epithelial cells lining the thick ascending limb (TAL) of the loop of Henle perform essential transport processes and secrete uromodulin, the most abundant protein in normal urine. The lack of differentiated cell culture systems has hampered studies of TAL functions. Here, we report a method to generate differentiated primary cultures of TAL cells, developed from microdissected tubules obtained in mouse kidneys. The TAL tubules cultured on permeable filters formed polarized confluent monolayers in ∼12 days. The TAL cells remain differentiated and express functional markers such as uromodulin, NKCC2, and ROMK at the apical membrane. Electrophysiological measurements on primary TAL monolayers showed a lumen-positive transepithelial potential (+9.4 ± 0.8 mV/cm(2)) and transepithelial resistance similar to that recorded in vivo. The transepithelial potential is abolished by apical bumetanide and in primary cultures obtained from ROMK knockout mice. The processing, maturation and apical secretion of uromodulin by primary TAL cells is identical to that observed in vivo. The primary TAL cells respond appropriately to hypoxia, hypertonicity, and stimulation by desmopressin, and they can be transfected. The establishment of this primary culture system will allow the investigation of TAL cells obtained from genetically modified mouse models, providing a critical tool for understanding the role of that segment in health and disease.
Mots-clé
Animals, Cells, Cultured, Loop of Henle/cytology, Mice, Mice, Knockout, Potassium Channels, Inwardly Rectifying/biosynthesis, Solute Carrier Family 12, Member 1/biosynthesis, Uromodulin/biosynthesis
Pubmed
Web of science
Création de la notice
10/10/2016 10:25
Dernière modification de la notice
20/08/2019 15:51
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