REGULATION OF INTRACELLULAR PH IN GUINEA-PIG CEREBRAL-CORTEX EX-VIVO STUDIED BY P-31 AND H-1 NUCLEAR-MAGNETIC-RESONANCE SPECTROSCOPY - ROLEOF EXTRACELLULAR BICARBONATE AND CHLORIDE
Trm. Pirttila et Ra. Kauppinen, REGULATION OF INTRACELLULAR PH IN GUINEA-PIG CEREBRAL-CORTEX EX-VIVO STUDIED BY P-31 AND H-1 NUCLEAR-MAGNETIC-RESONANCE SPECTROSCOPY - ROLEOF EXTRACELLULAR BICARBONATE AND CHLORIDE, Journal of neurochemistry, 62(2), 1994, pp. 656-664
The role of transmembrane processes that are dependent on external ani
ons in the regulation of cerebral intracellular pH (pH(i)), high-energ
y metabolites, and lactate was investigated using P-31 and H-1 NMR spe
ctroscopy in an ex vivo brain slice preparation. During oxygenated sup
erfusion, removal of external HCO3-/CO2 in the presence of Na+ led to
a sustained split of the inorganic phosphate (P-i) peak so that the pH
(l) indicated by one part of the peak was 0.38 pH units more alkaline
and by the other part 0.10 pH units more acidic at 5 min than in the p
resence of HCO3-. The pH in the compartment with a higher pH(l) value
returned to 7.29 +/- 0.04 by 10.5 min of superfusion in a HCO3--free m
edium, whereas the pH(l) in an acidic compartment was reduced to 7.02.
In the presence of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid
or the absence of external Cl-, removal of HCO3- caused alkalinizatio
n without split of the P-l peak. Both treatments reduced the rate of p
H(i) normalization following alkalinization. Simultaneous omission of
external HCO3- and Na+ did not inhibit alkalinization of the pH(i) fol
lowing CO2 exit. All these data show that the acid loading mechanism a
t neutral pH(l) is mediated by an Na+-independent anion transport. Dur
ing severe hypoxia, pH(l) dropped from 7.29 +/- 0.05 to 6.13 +/- 0.16
and from 7.33 +/- 0.03 to 6.67 +/- 0.05 in the absence and presence of
HCO3-, respectively, in Na+-containing medium. Lactate accumulated to
18.7 +/- 2.8 and 19.6 +/- 1.5 mmol/kg under the respective conditions
. In the HCO3--free medium supplemented with 1 mM amiloride, the pH(l)
fell only to 6.94 +/- 0.08 despite the lactate concentration of 18.9
+/- 2.4 mmol/kg. Acidification caused by hypoxia was also small in the
slice preparations superfused in the absence of both HCO3- and Cl-, a
s the pH(i) was 7.01 +/- 0.12 at a lactate concentration of 24.5 +/- 2
.4 mmol/kg. These data indicate that apart from anaerobic glucose meta
bolism, separate acidifying mechanisms are functioning during hypoxia
under these conditions. Recovery of phosphocreatine levels following r
eoxygenation was >75% relative to the prehypoxic level in the slice pr
eparations superfused in the absence of HCO3- but <47% in those prepar
ations superfused without HCO3- and Cl-. This indicates that either ne
utral pH(i) or absence of Cl- during hypoxia was deleterious to the en
ergy metabolism. The present data indicate that Cl-/HCO3-( )exchange m
echanisms have distinct roles in cerebral H+ homeostasis depending on
the level of pH(i) and energy state.