CFTR-dependent Cl- secretion in Xenopus laevis lung epithelium.
In our present study we used preparations from Xenopus laevis lungs to perform electrophysiological Ussing chamber measurements, unidirectional flux measurements, and employed molecular approaches to elucidate the presence and function of a cystic fibrosis transmembrane conductance regulator (CFTR) homolog in this tissue. Application of different CFTR blockers (NPPB (5-nitro-2-(3-phenylpropylamino)benzoic acid), niflumic acid (NFA), glibenclamide, lonidamine, CFTR(inh)-172) to the apical side of the tissues was able to significantly decrease the measured short circuit current (I(SC)) indicating a Cl(-) secretion due to luminal located CFTR channels. This was further supported by a net (36)Cl(-) secretion determined by radioactive tracer flux experiments. Further, Xenopus pulmonary epithelia responded to apical chlorzoxazone exposure - a CFTR activator - and this activated current was inhibited by CFTR(inh)-172. We performed reverse transcription-PCR (RT-PCR) and Western blot analysis and with both approaches we found characteristic signals indicating the presence of a CFTR homolog in Xenopus lung. In addition, we were able to detect CFTR in apical membranes of Xenopus lung slices with immunohistological techniques. We conclude that Xenopus lung epithelium exhibits functional CFTR channels and that this tissue represents a valuable model for the investigation of ion transport properties in pulmonary epithelia.
Sommer
Dagmar
D
Institute of Animal Physiology, Justus-Liebig University of Giessen, Wartweg 95, 35392 Giessen, Germany.
Bogdan
Roman
R
Berger
Jens
J
Peters
Dorothea M
DM
Morty
Rory E
RE
Clauss
Wolfgang G
WG
Fronius
Martin
M
D000818:Animals; D002712:Chlorides / Q000378:metabolism*; D002753:Chlorzoxazone / Q000494:pharmacology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000494:pharmacology*; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D004594:Electrophysiology; D005260:Female; D007700:Kinetics; D008168:Lung / Q000187:drug effects / Q000502:physiology*; D009125:Muscle Relaxants, Central / Q000494:pharmacology; D009579:Nitrobenzoates / Q000494:pharmacology; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014982:Xenopus laevis
D002712:Chlorides; D017931:DNA Primers; D009125:Muscle Relaxants, Central; D009579:Nitrobenzoates; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D012313:RNA; D004247:DNA; D002753:Chlorzoxazone