A phosphorus nuclear magnetic resonance study of metabolites and intracellular pH in rabbit vascular smooth muscle.

1. 31P nuclear magnetic resonance (n.m.r.) spectroscopy was used to investigate metabolites, intracellular pH (pHi) and the effects of pHi on tone in rabbit blood vessels. The vessels were bathed in mammalian Ringer solution and maintained at 20 degrees C while inside the spectrometer. 2. Vascular spectra showed relatively low phosphocreatine (PCr) concentrations compared to skeletal muscle. The [PCr]/[ATP] ratio was only 1.32 +/- 0.09 (n = 7). There was also a prominent phosphomonoester (PME) peak. Similar features have been reported for other smooth muscles examined by 31P n.m.r. 3. The [PCr] was higher and the inorganic phosphate (Pi) concentration lower than values deduced from chemical analysis of arterial extracts. However, the [PCr] value fell within the range obtained for other smooth muscles when studied by 31P n.m.r. 4. Measurement of pHi under control conditions (external pH 7.25) gave a mean value of 7.19 +/- 0.03 at 20 degrees C (n = 5). Metabolic inhibition brought about by 0.5 mM-cyanide and 0.2 mM-fluoride did not significantly alter pHi. At higher inhibitor concentrations (3 and 1 mM respectively) there was a significant acidosis. 5. The effects of NH4Cl upon pH were investigated in metabolically inhibited preparations. During 10 min applications of 30 mM-NH4Cl (isosmotically substituted for NaCl) the pHi rose; during subsequent NH4Cl removal it fell below control values. In the least inhibited tissues the total pHi excursion between NH4Cl applications and removals was 0.5 unit. 6. Rabbit ear vessels have been found to increase vascular tone during manoeuvres which were expected to decrease pHi. From the direct measurement of pHi reported in this study, it is concluded that the vascular tone changes brought about by NH4Cl application and withdrawal may be attributed to the alteration of pHi.