Intracellular pH and distribution of weak acids across cell membranes. A study of D- and L-lactate and of DMO in rat diaphragm.

Autor: Roos, A
Sprache: Englisch
Veröffentlicht: 1975
Quelle: PubMed Central (PMC)
Online Zugang: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1309555
http://www.ncbi.nlm.nih.gov/pubmed/239228
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1309555
Erfassungsnummer: ftpubmed:oai:pubmedcentral.nih.gov:1309555

Zusammenfassung

1. The steady-state distribution ratios of D- and L-lactate between fibre water and external fluid were measured in 'intact' rat hemidiaphragm preparations exposed for 2-5 hr to a variety of solutions of normal ionic strength and osmolarity. The studies were designed to minimize the effects, on these distributions, of conversion of lactate and of generation of lactic acid by the muscle. 2. At D-lactate concentrations between 2.3 and 118 mM, at normal pH and PCO2, the D-lactate distribution ratio, obtained from the distribution of [2-(3)H]D-lactate was independent of concentration; it averaged 0.349. As the concentration of D-lactate was reduced below 2.3 mM, its distribution ratio progressively fell to less than 0.1. 3. Radiochromatograms of extracts of incubated muscle showed that the tritium label was not attached to substances other than lactate. 4. At L-lactate concentrations of 59 and 108 mM, at normal pH and PCO2, the average L-lactate distribution ratios, obtained by enzymatic analysis, were respectively 0.395 and 0392. 5. At 19-89 mM D-lactate, depolarizing the muscle fibres by high K(49-127 mM), at normal pH, PCO2, and [K]0[Cl]0 product, only slightly affected the D-lactate distribution ratio which averaged 0.405. 6. The D-lactate distribution ratio and intracellular pH (pHi), obtained with the DMO method (5,5-dimethyl-2,4-oxazolidinedione), were measured in thirty sets of studies after exposure of the muscle to solutions buffered to pH values ranging between 5.99 and 8.13, and containing 18.5-118 mM D-lactate and 6-129 mM-K. 7. The relation between the distribution ratios of D-lactate ([TL]i/[TL]O) and of H ions ([Ho/[H]i) in these studies could be expressed by [TL]i/[T]O = 0.646 [H]o/[H]i+0.056. 8. It was concluded that it is predominantly the undissociated lactic acid molecules, rather than the much more numerous lactate ions, which permeate the fibre membrane; and that the steady-state lactate distribution ratio is determined by the transmembrane pH gradient, and not by membrane potential. 9. The ...