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cotransporter in bovine
corneal endothelium
School of Optometry, Indiana University, Bloomington, Indiana 47401
Functional studies
support the presence of the Na+-HCO3
cotransporter (NBC) in corneal endothelium and possibly corneal
epithelium; however, molecular identification and membrane localization
have not been reported. To test whether NBC is expressed in bovine cornea, Western blotting was performed, which showed a single band at
~130 kDa for freshly isolated and cultured endothelial cells, but no
band for epithelium. Two isoforms of NBC have recently been cloned in
kidney (kNBC) and pancreas (pNBC). RT-PCR was run using cultured and
fresh bovine corneal endothelial and fresh corneal epithelial total RNA
and specific primers for kNBC and pNBC. RT-PCR analysis for pNBC was
positive in endothelium and weak in epithelium. The RT-PCR product was
subcloned and confirmed as pNBC by sequencing. No specific bands for
kNBC were obtained from corneal cells. Indirect immunofluorescence and
confocal microscopy indicated that NBC locates predominantly to the
basolateral membrane in corneal endothelial cells. Furthermore,
Na+-dependent HCO3 fluxes and
HCO3-dependent cotransport with Na+ were
elicited only from the basolateral side of corneal endothelial cells.
Therefore, we conclude that pNBC is present in the basolateral membrane
of both fresh and cultured bovine corneal endothelium and weakly
expressed in the corneal epithelium.
sodium bicarbonate cotransporter; cornea; endothelium; bicarbonate transport
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