Swelling and Ca2+-activated anion conductances in C127 epithelial cells expressing WT and delta F508-CFTR.
CFTR is a chloride channel that is required for fluid secretion and salt absorption in many exocrine epithelia. Mutations in CFTR cause cystic fibrosis. CFTR expression influences some ion channels, but the range of channels influenced, the mechanism of the interaction and the significance for cystic fibrosis are not known. Possible interactions between CFTR and other ion channels were studied in C127 mouse mammary epithelial cell lines stably transfected with CFTR, delta F508-CFTR, or vector. Cell lines were compared quantitatively using an 125I efflux assay and qualitatively using whole-cell patch-clamp recording. As expected, 125I efflux was significantly increased by forskolin only in the CFTR line, and forskolin-stimulated whole-cell currents were time- and voltage independent. All three lines responded to hypotonic challenge with large 125I efflux responses of equivalent magnitude, and whole-cell currents were outwardly rectified and inactivated at positive voltages. Unexpectedly, basal 125I efflux was significantly smaller in the delta F508-CFTR cell line than in either the CFTR or control cell lines (P < 0.0001), and the magnitude of the efflux response to ionomycin was largest in the vector cell line and smallest in the cell line expressing delta F508-CFTR (P < 0.01). Whole-cell responses to ionomycin had a linear instantaneous I-V relation and activated at depolarizing voltages. Forskolin responses showed simple summation with responses to ionomycin or hypotonic challenge. Thus, we found no evidence for interactions between CFTR and the channels responsible for swelling-mediated responses. Differences were found in basal and ionomycin-stimulated efflux, but these may arise from variations in the clonally selected cell lines that are unrelated to CFTR expression.
Xia
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Cystic Fibrosis Research Laboratory, Building 420, Jordan Hall, Stanford University, Stanford, CA 94305-2130, USA.
Krouse
M E
ME
Fang
R H
RH
Wine
J J
JJ
D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D052688:Bovine papillomavirus 1 / Q000502:physiology; D002118:Calcium / Q000494:pharmacology / Q000502:physiology*; D002460:Cell Line; D048429:Cell Size / Q000502:physiology*; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology*; D005576:Colforsin / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004553:Electric Conductivity; D005260:Female; D007038:Hypotonic Solutions / Q000494:pharmacology; D066298:In Vitro Techniques; D015759:Ionomycin / Q000494:pharmacology; D051379:Mice
D018118:Chloride Channels; D007038:Hypotonic Solutions; D005576:Colforsin; D015759:Ionomycin; D002118:Calcium