Effects of 2-deoxy-D-glucose, amiloride, vasopressin, and ouabain on active conductance and ENa in the toad bladder

The effects of various agents on active sodium transport were studied in the toad bladder in terms of the equivalent circuit comprising an active conductance Ka, an electromotive force ENa, and a parallel passive conductance Kp. For agents which affect Ka, but not ENa or Kp, the inverse slope of the plot of total conductance K against short-circuit current IO evaluates ENa, and the intercept represents Kp. Studies employing 5 X 10(-7) M amiloride to depress Ka indicate a changing ENa, invalidating the use of the slope technique with this agent. An alternative suitable technique employs 10(-5) M amiloride, which reduces IO reversibly to near zero without effect on Kp. Despite curvilinearity of the K-IO plot under these conditions, Kp may therefore be estimated fairly precisely from the residual conductance. It then becomes possible to follow the dynamic behavior of Ka and ENa (in the absence of 10(-5) M amiloride) by frequent measurements of K and IO, utilizing the relationships Ka=K-Kp, and ENa=IO/(K-Kp). 2-deoxy-D-glucose (7.5 X 10(-3)M) depressed both Ka and ENa. All of the above effects were noted promptly; Kp was unaffected. The "electromotive force of Na transport" ENa appears not to be a pure energetic parameter, but to relfect kinetic factors as well, in accordance with thermodynamic considerations.