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1 Department of Clinical Genetics and 3 Department of Otolaryngology, Head and Neck Surgery, Rigshospitalet, University of Copenhagen, DK-2100 Copenhagen; and 2 Department of Medical Physiology and 4 Department of Medical Anatomy, The Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark
Airway
epithelium explants from cystic fibrosis (CF) patients and non-CF
subjects formed monolayered spheres, with the apical ciliated cell
membrane facing the bath and the basolateral cell membrane pointing
toward a fluid-filled lumen. With the use of two microelectrodes,
transepithelial potential difference and changes in potential
difference in response to passage of current pulses were recorded, and
epithelial resistance and the equivalent short-circuit current were
calculated. Non-CF control potential difference and short-circuit
current values were significantly lower than the CF values, and
amiloride inhibited both values. Fluid transport rates were calculated
from repeated measurements of spheroid diameters. The results showed
that 1) non-CF and CF spheroids
absorbed fluid at identical rates (4.4 µl · cm
2 · h
1),
2) amiloride inhibited fluid
absorption to a lower residual level in non-CF than in CF spheroids,
3)
Cl
-channel inhibitors
increased fluid absorption in amiloride-treated non-CF spheroids to a
level equal to that of amiloride-treated CF spheroids,
4) hydrochlorothiazide reduced
the amiloride-insensitive fluid absorption in both non-CF and CF
spheroids, and 5) osmotic water
permeabilities were equal in non-CF and CF spheroids (~27 × 10
7
cm · s
1 · atm
1).
amiloride; hydrochlorothiazide; osmotic permeability; current-voltage relationship; cystic fibrosis
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