Short-term effects of thyroid hormones on the Na/H antiport in L-6 myoblasts:: High molecular specificity for 3,3′,5-triiodo-L-thyronine

The thyroid hormones L-T-3 and L-T-4 were shown to activate the Na/H antiport in L-6 cells from rat skeletal muscle by a rapid, nongenomic mechanism. Under pH equilibrium conditions, a significant rise in the intracellular pH, measured by the fluorescent pH indicator 2',7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein was observed after the addition of physiological concentrations (10(-10) M) of either L-T-3 or L-T-4, but with different time courses. L-T-3 at all concentrations increased the pH after a delay of 2 min, whereas L-T-4 showed a concentration-dependent lag time, going from 11 min at 10(-11) M down to 5 min for a hormone concentration of 10(-6) M. The effect of L-T-4 was blocked in the presence of the 5'-deiodinase inhibitor 6-n-propyl-2-thiouracil, suggesting that the difference in lag time between L-T-3 and L-T-4 was due to the 5'-deiodination process that transforms L-T-4 into the bioactive L-T-3. In short term studies (<5 min), a high molecular specificity for L-T-3 was found, as L-T-4, rT(3), the D-isomer of T-3, and the deaminated analogues were ineffective at physiological concentrations. In analogy with the results found at equilibrium, intracellular pH recovery from an acid load and set-point were increased after 2 min for L-T-3 (10(-9) M) and after 10 min for L-T-4 (10(-9) M). The effect of the hormones on the intracellular pH was completely blocked by the specific antiport inhibitor 5-(ethyl-N-isopropyl)amiloride. These findings suggest that thyroid hormones may play an active role in the recovery from muscular acidosis through direct stimulation of the Na/H antiport.