Article (Scientific journals)
Segmental and subcellular distribution of CFTR in the kidney.
JOURET, François; Courtoy, Pierre J.; Devuyst, Olivier
2011In Methods in Molecular Biology, 741, p. 285-99
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Keywords :
Cystic Fibrosis Transmembrane Conductance Regulator/isolation & purification/metabolism; Endocytosis; Epithelial Cells/cytology/metabolism; Immunoblotting; Immunohistochemistry/methods; Intracellular Space/metabolism; Kidney/cytology; Kidney Tubules, Proximal/cytology/metabolism; Mice; Molecular Imaging; Paraffin Embedding; Peroxidase/metabolism
Abstract :
[en] Besides its location at the plasma membrane, CFTR is present in intracellular vesicles along both the exocytic and the endocytic pathways. Immunostaining and subcellular fractionation studies of mouse kidney demonstrate that CFTR is located in endosomes of the cells lining the terminal part of the proximal tubule (PT). The PT cells efficiently reabsorb the ultrafiltered low molecular weight (LMW) proteins by apical endocytosis involving the multiligand receptors megalin and cubilin. The progression from early endosomes to lysosomes depends on the integrity of the cytoskeleton, as well as on vesicular acidification. The latter is mediated by the vacuolar H+-ATPase (V-ATPase) and requires an anionic conductance to dissipate the transmembrane potential gradient. CFTR might ensure such chloride conductance, thereby participating to endosomal acidification and protein uptake by PT cells. Immunostaining with well-characterized antibodies shows that CFTR is located in the terminal segment of PT, where it co-distributes with megalin and cubilin. Subcellular fractionation of total mouse kidneys through Percoll gradients demonstrates the co-localization of CFTR with the V-ATPase and early endosome markers including the Cl-/H+ exchanger, ClC-5, and the small GTPase, Rab5a. Deglycosylation studies and immunoblotting show a distinct glycosylation pattern for CFTR in mouse kidney and lung. The segmental and subcellular distribution of CFTR in mouse kidney supports a role for CFTR in PT receptor-mediated endocytosis of ultrafiltered LMW proteins.
Disciplines :
Urology & nephrology
Author, co-author :
JOURET, François  ;  Centre Hospitalier Universitaire de Liège - CHU > Néphrologie
Courtoy, Pierre J.
Devuyst, Olivier
Language :
English
Title :
Segmental and subcellular distribution of CFTR in the kidney.
Publication date :
2011
Journal title :
Methods in Molecular Biology
ISSN :
1064-3745
eISSN :
1940-6029
Publisher :
Humana Press, Clifton, United States - New Jersey
Volume :
741
Pages :
285-99
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 20 November 2012

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