Na+-H+ antiport and monensin effects on cytosolic pH and iodide transport in FRTL-5 rat thyroid cells.
Na+-H+ exchange may proceed via an endogenous antiporter or by exposure to the Na+ ionophore monensin. We investigated the characteristics of Na+-H+ exchange induced by antiporter stimulation and by monensin in FRTL-5 rat thyroid cells. We also examined the effects of intracellular pH (pHi) changes on iodide uptake and efflux. pHi was determined using 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein. The resting pHi was 7.33 +/- 0.02 units; this level correlated directly with extracellular pH. In acid-loaded cells, Km for external Na+ activation of the antiporter was 7.1 mM and maximum velocity was 0.3801 delta pH units/min. Dimethylamiloride was 42 times more potent than amiloride in inhibiting sodium-dependent recovery in acidified cells. Metabolic inhibition reduced the initial alkalinization rate. Monensin increased pHi, and this response was dependent on extracellular Na+ and HCO3- but not on antiporter function. Low-dose monensin (1 microM) and 1 mM NH4Cl enhanced 125I uptake. High-dose monensin (100 microM), but not NH4Cl, reduced iodide uptake. Neither NH4Cl nor monensin altered 125I efflux. Thus FRTL-5 cells possess an amiloride-sensitive Na+-H+ exchanger, which is not essential for maintaining basal pHi but is affected by ATP depletion. Monensin also alkalinizes these cells but independently of the antiporter. Iodide uptake, but not efflux, is affected by changes in intracellular Na+ and H+ levels.
Smallridge
R C
RC
Department of Clinical Physiology, Walter Reed Army Institute of Research, Washington, District of Columbia 20307-5100.
Gist
I D
ID
Kiang
J G
JG
D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007454:Iodides / Q000378:metabolism*; D007700:Kinetics; D008985:Monensin / Q000494:pharmacology*; D010042:Ouabain / Q000494:pharmacology; D051381:Rats; D012964:Sodium / Q000378:metabolism*; D017923:Sodium-Hydrogen Exchangers; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism*
D002352:Carrier Proteins; D007454:Iodides; D017923:Sodium-Hydrogen Exchangers; D000643:Ammonium Chloride; C039465:5-dimethylamiloride; D010042:Ouabain; D000584:Amiloride; D008985:Monensin; D012964:Sodium