Time course of active Na transport and oxidative metabolism following transepithelial potential perturbation in toad...

The use of an Ussing chamber with well-defined mixing characteristics coupled to a mass spectrometer permits the concurrent evaluation of transepithelial current and oxidative metabolism with improved temporal resolution. The time-course of the amiloride-sensitive current Ia and the rate of suprabasal CO2 production JsbCO2 were observed in 10 toad urinary bladders at short-circuit and after clamping delta psi at 100 mV, serosa positive. Following perturbation of delta psi (0 leads to 100 mV), Ia declined sharply within 1/2 min, remaining near constant approximately 15 min, and then increased slightly. JsbCO2 declined more gradually, remained near constant at approximately 4-7 min, and then declined further. Detailed analysis revealed an early quasi-steady state with near constancy of JsbCO2 starting at 2.9 +/- 1.1 (SD) min and lasting 4.7 +/- 1.8 (SD) min, followed by relaxation to a later steady state at about 15 min. During the early quasi-steady state, Ia was also nearly constant. Considering that in steady states Ia/F approximately or equal to JaNa, the rate of transepithelial active Na transport, during the early quasi-steady state mean values +/- SE of JaNa, JsbCO2 and (JaNa/JsbCO2) were, respectively, 29.9 +/- 1.7%, 59.4 +/- 3.2%, and 56.4 +/- 5.7% of values at short-circuit. Corresponding values during the late steady state were 41.4 +/- 6.0%, 38.2 +/- 6.1%, and 111.3 +/- 8.6%. Thus the flow ratio JaNa/JsbCO2 was depressed significantly during the early quasi-steady state, but returned later to the original value. The results of measurements of Ia and JsbO2 in three hemibladders were qualitatively similar. In terms of a phenomenological "black-box" treatment the findings are consistent with earlier studies indicating incomplete coupling between transport and metabolism. Further studies will be required to clarify the molecular basis for these observations.