Abstract
1. Intracellular pH (pHi) was recorded in isolated guinea-pig ventricular myocytes using the pH-sensitive fluoroprobe, carboxy-SNARF-1 (carboxy-seminaphthorhodafluor). 2. Addition and removal of 10 mM NH4Cl was used to induce an intracellular acid load in a myocyte perfused with HCO3(-)-buffered solution containing amiloride. Under these conditions, subsequent pHi recovery is known to rely upon Na(+)-HCO3- co-transport into the cell. The application of 0.5-5 microM adrenaline resulted in an inhibition of this pHi recovery. 3. In HEPES-buffered solution, where acid extrusion is mediated primarily by Na(+)-H+ antiport, pHi recovery from an acid load was stimulated by the application of adrenaline. 4. In HCO3-/CO2-buffered solution (no amiloride), when both acid-aquivalent extruders are activated by an intracellular acidification, adrenaline was found to slow pHi recovery. 5. When both carriers were inhibited in Na(+)-free, HCO3(-)-buffered medium, adrenaline had no effect on pHi, ruling out any effect of the catecholamine on background acid loading. 6. The voltage clamp technique was used to test if the inhibitory effect of adrenaline on amiloride-resistant, HCO3(-)-dependent pHi recovery was due to an efflux of HCO3- ions through catecholamine-activated anion channels. During pHi recovery, membrane depolarization, sufficient to reverse the electrochemical driving force acting on HCO3-, had no effect upon pHi recovery rate. 7. The above results show that adrenaline has direct but opposite effects on Na(+)-HCO3- co-transport and Na(+)-H+ antiport. In the presence of this agonist, the pHi dependence of Na(+)-HCO3- symport was shifted to the left along the pHi axis by 0.13 +/- 0.03 units (n = 4) whereas that for Na(+)-H+ antiport was shifted in the opposite direction by only 0.07 +/- 0.01 units (n = 3). Following an acid load, the net effect of adrenaline under physiological conditions was, therefore, a slowing of pHi recovery. 8. The application of extracellular ATP (ATPo, 10-50 microM) mimicked the effects of adrenaline on both Na(+)-H+ exchange and Na(+)-HCO3- symport. 9. Adenosine (50 microM) and ADP (50 microM) did not induce any inhibition of Na(+)-HCO3- symport, suggesting that the inhibition induced by ATP was not mediated through P1 or P2-purinergic receptors. 10. We conclude that Na(+)-H+ antiport and Na(+)-HCO3- symport are both coupled to adrenaline and ATPo receptors. Activation of these receptors switches acid-equivalent extrusion from a situation dependent on both HCO3- and H+ ions to one nearly exclusively dependent upon H+.