Localisation of wild-type and DeltaF508-CFTR in nasal epithelial cells.
Wild-type and the DeltaF508 mutation of the cystic fibrosis transmembrane conductance regulator (DeltaF508-CFTR) were localised by confocal imaging in DeltaF508/DeltaF508 native airway epithelial cells using a well-characterised CFTR antibody. Surface nasal epithelial cells from three control and three CF individuals were obtained from nasal brushings. Cells were fixed, permeabilised and incubated with first antibody for 18 h at 4 degrees C. Following labelling with second antibody, cells were viewed with the confocal microscope. Wild-type CFTR was localised predominantly apically, whereas DeltaF508-CFTR was located mainly inside the cell in a region close to the nucleus. Incubation of cells with MPB-07 (250 microM) at 37 degrees C for 2 h resulted in pronounced movement of DeltaF508-CFTR to the cell periphery, but did not change the localisation of wild-type CFTR. The results show that DeltaF508-CFTR is mislocalised in native nasal epithelial cells and that its distribution is altered in response to the new CFTR activator, MPB-07. The findings should lead to development of a rational drug treatment for CF patients carrying the DeltaF508 mutation.
Dormer
R L
RL
Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff CF14 4XN, UK. dormer@cardiff.ac.uk
McNeilly
C M
CM
Morris
M R
MR
Pereira
M M
MM
Doull
I J
IJ
Becq
F
F
Mettey
Y
Y
Vierfond
J M
JM
McPherson
M A
MA
D000918:Antibody Specificity; D003550:Cystic Fibrosis / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000032:analysis* / Q000276:immunology; D004847:Epithelial Cells / Q000737:chemistry*; D006801:Humans; D018613:Microscopy, Confocal; D009297:Nasal Mucosa / Q000737:chemistry* / Q000166:cytology
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator