Expression of the cystic fibrosis gene in non-epithelial invertebrate cells produces a regulated anion conductance.
The nature of involvement of the cystic fibrosis gene product (CFTR) in epithelial anion transport is not yet understood. We have expressed CFTR in Sf9 insect cells using the baculovirus expression vector system. Reactivity with antibodies against 12 different epitopes spanning the entire sequence suggested that the complete polypeptide chain was synthesized. Immunogold labeling showed localization to both cell-surface and intracellular membranes. Concomitant with CFTR expression, these cells exhibited a new cAMP-stimulated anion permeability. This conductance, monitored both by radioiodide efflux and patch clamping, strongly resembled that present in several CFTR-expressing human epithelial cells. These findings demonstrate that CFTR can function in heterologous nonepithelial cells and lend support to the possibility that CFTR may itself be a regulated anion channel.
Kartner
N
N
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
Hanrahan
J W
JW
Jensen
T J
TJ
Naismith
A L
AL
Sun
S Z
SZ
Ackerley
C A
CA
Reyes
E F
EF
Tsui
L C
LC
Rommens
J M
JM
Bear
C E
CE
D000818:Animals; D000838:Anions; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D018118:Chloride Channels; D000242:Cyclic AMP / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004553:Electric Conductivity; D005822:Genetic Vectors; D006801:Humans; D007313:Insecta; D007473:Ion Channels / Q000235:genetics* / Q000502:physiology; D007700:Kinetics; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology; D014162:Transfection
D000838:Anions; C505032:CFTR protein, human; D018118:Chloride Channels; D007473:Ion Channels; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000242:Cyclic AMP