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Endocrinology Vol. 140, No. 2 683-689
Copyright © 1999 by The Endocrine Society

ARTICLES

Short-Term Effects of Thyroid Hormones on the Na/H Antiport in L-6 Myoblasts: High Molecular Specificity for 3,3',5-Triiodo-L-Thyronine1

Sandra Incerpi, Paolo Luly, Paolo De Vito and Ricardo N. Farias

Department of Biology, University of Rome Tor Vergata, 00133 Rome; and the Department of Biology, University of Rome 3 (S.I.), 00146 Rome, Italy; and the Departamento de Bioquimica de la Nutricion, Instituto Superior de Investigaciones Biologicas, Universidad Nacional de Tucuman (R.N.F.), 4000 Tucuman, Argentina

Address all correspondence and requests for reprints to: Dr. Sandra Incerpi, Department of Biology, University of Rome Tor Vergata, via della Ricerca Scientifica, 00133 Rome, Italy. E-mail: incerpi{at}uniroma3.it

The thyroid hormones L-T3 and L-T4 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-T3 or L-T4, but with different time courses. L-T3 at all concentrations increased the pH after a delay of 2 min, whereas L-T4 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-T4 was blocked in the presence of the 5'-deiodinase inhibitor 6-n-propyl-2-thiouracil, suggesting that the difference in lag time between L-T3 and L-T4 was due to the 5'-deiodination process that transforms L-T4 into the bioactive L-T3. In short term studies (<5 min), a high molecular specificity for L-T3 was found, as L-T4, rT3, the D-isomer of T3, 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-T3 (10-9 M) and after 10 min for L-T4 (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.




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