Peter C. Tullson

Oxidation of urate in human skeletal muscle during exercise.

The purpose of the present study was to investigate whether high metabolic stress to skeletal muscle, induced by intensive exercise, would lead to an oxidation of urate to allantoin in the exercised muscle. Seven healthy male subjects performed short term (4.39 +/- 0.04 [+/-SE] min) exhaustive cycling exercise. Muscle samples were obtained from m. v. lateralis before and during the first few minutes after the exercise. Venous blood samples were obtained before and up to 45 min after the exercise. The concentration of urate in muscle decreased from a resting level of 0.26 +/- 0.023 to 0.084 +/- 0.016 mumol.g-1 w.w. (p < .05) during the exercise and then rapidly increased during recovery to reach the resting level within 3 min after exercise. The concentration of allantoin in the muscle increased from a resting value of 0.03 +/- 0.007 to 0.10 +/- 0.014 mumol.g-1 w.w. immediately after exercise (p < .05) and then decreased to 0.079 +/- 0.002 mumol.g-1 w.w. during the first 3 min after exercise (p < .05). Plasma urate levels increased slowly from 305 +/- 16 to 426 +/- 20 mumol.liter-1 at 45 min in recovery (p < .05). Plasma allantoin was 11.9 +/- 2.6 mumol.liter-1 at rest and by 5 min the level was more than twofold higher and remained elevated throughout recovery (p < .05). The present results indicate that urate is oxidized to allantoin in the muscle during exercise, probably due to generation of free radicals. Furthermore, the findings support the suggested importance of urate as a free radical scavenger in vivo.

CYTOARCHITECTURAL AND METABOLIC ADAPTATIONS IN MUSCLES WITH MITOCHONDRIAL AND CYTOSOLIC CREATINE KINASE DEFICIENCIES

We have blocked creatine kinase (CK) mediated phosphocreatine (PCr) ⇄ ATP transphosphorylation in mitochondria and cytosol of skeletal muscle by knocking out the genes for the mitochondrial (ScCKmit) and the cytosolic (M-CK) CK isoforms in mice. Animals which carry single or double mutations, if kept and tested under standard laboratory conditions, have surprisingly mild changes in muscle physiology. Strenuous ex vivo conditions were necessary to reveal that MM-CK absence in single and double mutants leads to a partial loss of tetanic force output. Single ScCKmit deficiency has no noticeable effects but in combination the mutations cause slowing of the relaxation rate. Importantly, our studies revealed that there is metabolic and cytoarchitectural adaptation to CK defects in energy metabolism. The effects involve mutation type-dependent alterations in the levels of AMP, IMP, glycogen and phosphomonoesters, changes in activity of metabolic enzymes like AMP-deaminase, alterations in mitochondrial volume and contractile protein (MHC isoform) profiles, and a hyperproliferation of the terminal cysternae of the SR (in tubular aggregates). This suggests that there is a compensatory resiliency of loss-of-function and redirection of flux distributions in the metabolic network for cellular energy in our mutants.

Adenine nucleotide metabolism in contracting skeletal muscle.

During steady-state muscle contractions, ATP production and utilization are well matched. When the rate of ATP hydrolysis exceeds the capacity of a given muscle fiber to phosphorylate ADP, the ADPf and AMPf concentrations rise, first leading to the deamination of adenylates and subsequently to the dephosphorylation of AMP or IMP, or both, to their respective nucleosides and bases. Several proposed roles for the purine nucleotide cycle in skeletal muscle have been reviewed and evaluated. The deaminating limb of the purine nucleotide cycle is most important; it maintains the ATP/ADP ratio and lessens adenine nucleotide degradation. Regulation of glycolytic pathway enzymes by the products of AMP deamination (IMP and NH4+) does not seem likely. During reamination there is a net production of fumarate, with the branch-chain amino acids potentially supplying a significant fraction of the amine; reamination, however, is probably not concurrent with a high rate of deamination. Evidence from some studies of AMP deaminase-deficient persons suggests that an intact purine nucleotide cycle is required for normal muscle function during intense exercise; the issue is clouded, however, by the occurrence of asymptomatic AMP deaminase deficiency. Skeletal muscle is capable of extensive adenine nucleotide degradation during severe, energy-depleting conditions. Purine nucleosides and bases not reincorporated by the salvage pathway must be synthesized de novo. The capacity for de novo synthesis differs among fiber types, being highest in muscle with the highest oxidative capacity.