Apparent Km of mitochondria for oxygen computed from Vmax measured in permeabilized muscle fibers is lower in water enriched in oxygen by electrolysis than injection

Joffrey Zoll, Jamal Bouitbir, Pascal Sirvent, Alexis Klein, Antoine Charton, Liliana Jimenez, François R Péronnet, Bernard Geny, Ruddy Richard, Joffrey Zoll, Jamal Bouitbir, Pascal Sirvent, Alexis Klein, Antoine Charton, Liliana Jimenez, François R Péronnet, Bernard Geny, Ruddy Richard

Abstract

Background: It has been suggested that oxygen (O2) diffusion could be favored in water enriched in O2 by a new electrolytic process because of O2 trapping in water superstructures (clathrates), which could reduce the local pressure/content relationships for O2 and facilitate O2 diffusion along PO2 gradients.

Materials and methods: Mitochondrial respiration was compared in situ in saponin-skinned fibers isolated from the soleus muscles of Wistar rats, in solution enriched in O2 by injection or the electrolytic process 1) at an O2 concentration decreasing from 240 µmol/L to 10 µmol/L (132 mmHg to 5 mmHg), with glutamate-malate or N, N, N', N'-tetramethyl-p-phenylenediamine dihydrochloride (TMPD)-ascorbate (with antimycin A) as substrates; and 2) at increasing adenosine diphosphate (ADP) concentration with glutamate-malate as substrate.

Results: As expected, maximal respiration decreased with O2 concentration and, when compared to glutamate-malate, the apparent Km O2 of mitochondria for O2 was significantly lower with TMPD-ascorbate with both waters. However, when compared to the water enriched in O2 by injection, the Km O2 was significantly lower with both electron donors in water enriched in O2 by electrolysis. This was not associated with any increase in the sensitivity of mitochondria to ADP; no significant difference was observed for the Km ADP between the two waters.

Conclusion: In this experiment, a higher affinity of the mitochondria for O2 was observed in water enriched in O2 by electrolysis than by injection. This observation is consistent with the hypothesis that O2 diffusion can be facilitated in water enriched in O2 by the electrolytic process.

Keywords: Km O2; TMPD–ascorbate; glutamate–malate; mitochondrial respiration; saponin-skinned fibers.

Figures

Figure 1
Figure 1
Vmax in solutions enriched in O2 by injection and electrolysis for O2 concentrations, as well as apparent Km of the mitochondria for O2 with the two waters and two electron donors. Notes: (A) Vmax in solutions enriched in O2 by injection and electrolysis for O2 concentrations ranging between 10 µmol/L and 240 µmol/L in rat permeabilized soleus muscle fibers with glutamate–malate and TMPD–ascorbate as substrates. (B) Apparent Km of the mitochondria for O2 with the two waters and the two electron donors. Mean ± standard deviation; n=12; #statistically different from glutamate–malate and *from injection, P<0.05. Abbreviations: dw, dry weight; TMPD, tetramethyl-p-phenylenediamine; n, number.
Figure 2
Figure 2
Vmax in solutions enriched in O2 by injection and electrolysis at increasing ADP concentrations, and apparent Km of the mitochondria for ADP with the two waters. Notes: (A) Vmax in solutions enriched in O2 by injection and electrolysis at increasing ADP concentrations in rat permeabilized soleus muscle fibers with glutamate–malate as substrates. (B) Apparent Km of the mitochondria for ADP with the two waters. Mean ± standard deviation; n=11; no significant difference was observed between the two waters. Abbreviations: dw, dry weight; ADP, adenosine diphosphate; n, number.

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Source: PubMed

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