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. 1992;451:365–385. doi: 10.1113/jphysiol.1992.sp019169

Na(+)-HCO3- symport in the sheep cardiac Purkinje fibre.

C Dart 1, R D Vaughan-Jones 1
PMCID: PMC1176166  PMID: 1403816

Abstract

1. Intracellular pH (pHi) was recorded in isolated sheep cardiac Purkinje fibres using liquid sensor ion-selective microelectrodes in conjunction with conventional (3 M-KCl) microelectrodes (to record membrane potential). 2. In HEPES-buffered solution (pH0 7.4), pHi recovery from an intracellular acid load (20 mM-NH4Cl removal) was blocked by 1 mM-amiloride, consistent with the inhibition of Na(+)-H+ exchange. Replacement of the HEPES buffer with CO2-HCO3- caused a transient acidosis followed by an amiloride-resistant recovery of pHi to more alkaline levels (n = 43). This implies the presence of a HCO3(-)-dependent pHi regulatory mechanism. 3. Comparison of the membrane potential with the equilibrium potential for HCO3- ions (EHCO3) estimated during amiloride-resistant pHi recovery, showed that for polarized fibres (membrane potential Em approximately -80 mV), there was a net outward electrochemical driving force for HCO3- ions. Hence the amiloride-resistant pHi recovery cannot be explained in terms of passive HCO3- influx through membrane channels. 4. Removal of external Na+ (Na0+ replaced by N-methyl-D-glucamine) inhibited HCO3(-)-dependent pHi recovery, whereas removal of external Cl- (leading to depletion of internal Cl-; Cl0- replaced by glucuronate) or short-term removal of extracellular K+ had no inhibitory effect. We suggest that a Na(+)-HCO3- co-influx causes the recovery. Replacement of external Na+ with Li+ greatly reduced HCO3(-)-dependent pHi recovery indicating that Li0+ cannot readily substitute for Na0+ on the co-transport. 5. The stilbene drug DIDS (4,4-diisothiocyano-stilbene-disulphonic acid, 500 microM) slowed HCO3(-)-dependent pHi recovery. 6. Depolarization of the membrane potential in high K0+ (44.5 mM) solution or with 5 mM-BaCl2 had no effect upon the rate of HCO3(-)-sensitive pHi recovery. This observation, when coupled with the fact that activation of HCO3(-)-dependent pHi recovery was associated with no consistent change of membrane potential, suggests that the Na(+)-HCO3- co-influx is electroneutral and voltage insensitive. 7. HCO3(-)-dependent pHi recovery was unaffected by the Na(+)-K(+)-2Cl- co-transport inhibitor, bumetanide (150 microM). 8. The contribution of Na(+)-H+ exchange and Na(+)-HCO3- co-transport to net acid efflux was assessed. At a pHi of 6.6, we estimate that the co-transport should account for 20% of total acid equivalent efflux.(ABSTRACT TRUNCATED AT 400 WORDS)

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Selected References

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