Related Experiment Videos
Kinetic evidence for Na(+)-glucose co-transport in the pigmented rabbit conjunctiva
1Department of Pharmaceutical Sciences, University of Southern California, Los Angeles, USA.
Purpose:
To obtain kinetic evidence for the existence of a Na(+)-coupled glucose co-transport process on the mucosal (tear) side of the pigmented rabbit conjunctiva.
Methods:
The excised pigmented rabbit conjunctiva was mounted in the modified Ussing chamber for measurement of 22Na and 3H-3-O-methyl-D-glucose (3-O-MG) fluxes.
Results:
In the presence of 5 mM glucose, the conjunctival tissue showed net Na+ absorption in the mucosal-to-serosal direction at an approximate rate of 0.15 microEq/cm2/h. This net Na+ absorption was abolished by serosally added 0.5 mM ouabain, but not affected by mucosally added 0.1 mM or 1 mM amiloride. There was a 40-60% reduction in net Na+ absorption under the glucose-free condition or in the mucosal presence of 0.5 mM phlorizin. Moreover, serosally added ouabain and mucosally added phlorizin (both at 0.5 mM) significantly decreased the 3-O-MG permeability coefficient in the mucosal-to-serosal direction by about 70%, whereas mucosally instilled 0.1-1.0 mM amiloride was without any effect. Three-O-MG absorption in the mucosal-to-serosal direction appeared to be coupled with Na+ transport with a 1:1 stoichiometry. In addition, this process exhibited temperature dependency, saturability, and directionality.
Conclusion:
Our findings are consistent with Na(+)-glucose cotransport as being one of the mechanisms for mucosal Na+ entry into the epithelial cells of the pigmented rabbit conjunctiva.