Fluid and ion transport in corneal endothelium: insensitivity to modulators of Na(+)-K(+)-2Cl- cotransport.
The role of Na(+)-K(+)-2Cl- cotransport in ion and fluid transport of the corneal endothelium was examined by measuring changes in corneal hydration and uptake of 86Rb by the endothelial cell layer. Isolated, intact rabbit corneas maintain normal hydration when they are superfused at the endothelial surface with bicarbonate (HCO3-)-Ringer solutions as a result of equilibrium between active ion and fluid transport out of the stromal tissue and leak of fluid into stromal tissue from the aqueous humor. Furosemide and bumetanide did not alter this equilibrium when they were added to the superfusion medium. Uptake of 86Rb by the endothelium of the incubated cornea was increased 25% by bumetanide, but uptake in the presence of ouabain (70% less than that of controls) was not changed by bumetanide. In Na(+)-free medium, uptake of 86Rb was reduced by 58%, but it was unchanged in Cl(-)-free medium. Calyculin A, a protein phosphatase inhibitor and activator of Na(+)-K(+)-Cl- cotransport, was without effect on 86Rb uptake. Hypertonicity (345 mosmol/kg) increased uptake slightly, whereas hypotonicity (226 mosmol/kg) caused a 33% decrease. Neither of these changes was significantly different when bumetanide was present in the media. It is concluded that Na(+)-K(+)-2Cl- cotransporter activity is not exhibited by the in situ corneal endothelium and does not play a role in the ion and fluid transport of this cell layer. Its presence in cultured endothelial cells may reflect the reported importance of this protein in growth, proliferation, and differentiation.
Riley
M V
MV
Eye Research Institute, Oakland University, Rochester, Michigan 48309-4401, USA.
Winkler
B S
BS
Starnes
C A
CA
Peters
M I
MI
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002034:Bumetanide / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D003315:Cornea / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D004728:Endothelium, Corneal / Q000166:cytology / Q000502:physiology*; D005665:Furosemide / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008353:Mannitol / Q000493:pharmacokinetics; D010477:Perfusion; D011817:Rabbits; D012413:Rubidium / Q000493:pharmacokinetics; D012414:Rubidium Radioisotopes; D012964:Sodium / Q000378:metabolism; D028021:Sodium-Potassium-Chloride Symporters; D017154:Stromal Cells / Q000502:physiology; D014508:Urea / Q000378:metabolism
D002352:Carrier Proteins; D002712:Chlorides; D012414:Rubidium Radioisotopes; D028021:Sodium-Potassium-Chloride Symporters; D002034:Bumetanide; D005576:Colforsin; D008353:Mannitol; D005665:Furosemide; D014508:Urea; D012964:Sodium; D012413:Rubidium