Bicarbonate and fast-twitch muscle: evidence for a major role in pH regulation

Internal pH (pHi) was analyzed in rat extensor digitorum longus (Edl) muscle at 30 degrees C with single-barrel liquid ion-selective electrodes. Average pHi in 284 cells was 7.197 +/- 0.006. Increases in CO2 from nominally 0 to 5% produced an acidification from which recovery took place. In different groups of cells, recovery from the 5% CO2 acidification was significantly inhibited by 100 microM 4,4' diisothiocyanatostilbene 2,2' disulfonic acid (DIDS), Cl removal, Na removal and 2 mM amiloride. Prepulsing with 20 mM NH4 in the presence of CO2/HCO3 typically reduced pHi to only about neutral, whereas 50 mM reduced pHi to 6.7-6.8. In the nominal absence of CO2/HCO3, 20 mM NH4 reduced pHi to about 6.7 from which recovery took place at about 58% of the rate seen in different cells in the presence of CO2/HCO3. In the presence of CO2/HCO3, cells prepulsed with 50 mM NH4 had fully recovered to an average pHi of 7.22 +/- 0.04 about 90 min after removal of NH4. However, 90 min after removal of 20 mM NH4 in the absence of CO2/HCO3, average pHi was significantly less (7.05 +/- 0.03). Intrinsic buffering capacity (beta i) was obtained during pulses of CO2, acetic acid or after an NH4 pulse. beta i was significantly reduced in the absence of HCO3, Cl or Na and HCO3. The data provide significant support for an important role of HCO3 in the control of pHi in fast-twitch muscle.