G. Vidal et al., CYTOSOLIC PH REGULATION IN PERFUSED-RAT-LIVER - ROLE OF INTRACELLULARBICARBONATE PRODUCTION, Biochimica et biophysica acta (G). General subjects, 1425(1), 1998, pp. 224-234
The contribution of metabolic bicarbonate to cytosolic pH (pH(cyto)) r
egulation was studied on isolated perfused rat liver using phosphorus-
31 NMR spectroscopy. Removal of external HCO3- decreased proton efflux
from 18.6 +/- 5.0 to 1.64 +/- 0.29 mu mol/min per g liver wet weight
(w.w.) and pH(cyto) from 7.17 +/- 0.06 to 6.87 +/- 0.06. In the nomina
l absence of bicarbonate, inhibition of carbonic anhydrase by acetazol
amide induced a further decrease of proton efflux of 0.69 +/- 0.26 mu
mol/min per g liver w.w, reflecting a reduction in metabolic CO2 hydra
tion, and hence a decrease of H+ and HCO3- supplies. Even though 27% o
f the proton efflux was amiloride-sensitive under bicarbonate-free con
ditions, amiloride did not change pH(cyto), revealing the contribution
of additional regulatory processes. Indeed, pH regulation was affecte
d by the combined use of acetanlido-4'-isothiocyanostilbene-2,2'-disul
fonic acid (SITS) and amiloride since pH(cyto) decreased by 0.16 +/- 0
.05 and proton efflux by 0.60 +/- 0.14 mu mol/min per g liver w.w. The
data suggest that amiloride-sensitive or SITS-sensitive transport act
ivities could achieve, by themselves, pH(cyto) regulation. The involve
ment of two mechanisms, most likely Na+/H+ antiport and Na+:HCO3- symp
ort, was confirmed in the whole organ under intracellular and extracel
lular acidosis, The evidence of Na-dependent transport of HCO3- in the
absence of exogenous bicarbonate implies that the amount of metabolic
bicarbonate is sufficient to effectively participate to pH(cyto) regu
lation. (C) 1998 Elsevier Science B.V. All rights reserved.