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secretion but not
Na+ reabsorption in A6
cells
Departments of 1 Physiology and Biophysics, 2 Cell Biology, and 3 Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35294
The effects of microtubule disruption on arginine
vasotocin (AVT)-stimulated Na+ and
Cl
transport were studied
in A6 cells by measuring short-circuit currents
(Isc) across
cell layers grown in tissue culture on permeable supports. Microtubule
disruption inhibited an AVT-stimulated secretory Cl current but did not
prevent activation of amiloride-sensitive Na+ transport. This AVT-stimulated
secretory Cl current was
significantly inhibited by glibenclamide, an inhibitor of
the cystic fibrosis transmembrane conductance regulator (CFTR). Reverse
transcription of RNA isolated from A6 cells followed by polymerase
chain reaction (PCR) using primers designed to amplify a portion of the
R-domain of CFTR cloned from Xenopus
laevis skin and immunocytochemistry demonstrated the
presence of CFTR in A6 cells and an apparent recruitment of cytoplasmic
CFTR to the apical cell surface after AVT stimulation. In contrast,
indirect immunofluorescent labeling of
Na+ channels using a polyclonal
antibody raised against a biochemically isolated
Na+ channel complex from bovine
renal medulla labeled the apical plasma membrane but failed to
demonstrate intracellular labeling of
Na+ channels (except in
subconfluent cells) or recruitment of
Na+ channels to the apical
membrane region after AVT stimulation.
trafficking; cystic fibrosis transmembrane conductance regulator; cytoskeleton; amiloride-sensitive sodium channel; sodium transport; chloride transport
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