Quantitative study of the effects of long-term denervation on the extensor digitorum longus muscle of the rat.

BACKGROUND: In order to understand the cellular basis underlying the progressively poorer restorative capacity of long-term denervated muscle, we determined the effects of long-term denervation on the muscle fibers and satellite cell population of the rat extensor digitorum longus (EDL) muscle. METHODS: In 36 male rats, the right hind legs were denervated, and EDL muscles were removed 2, 4, 7, 12, and 18 months later. Muscles were either fixed for electron microscopic analysis or were dissociated into individual muscle fibers for direct fiber counting or for confocal microscopic analysis. RESULTS: The percentage of satellite cells rose from the 2.8% control value to 9.1% at 2 months of denervation; thereafter the percentage decreased to 1.1% at 18 months of denervation. The number of myonuclei per muscle fiber steadily declined from 410 in 4 month control muscle to 158 in 7 month denervated muscle. Up to 7 months of denervation, the total number of muscle fibers per muscle remained relatively constant at somewhat over 5,000. The calculated total satellite cell population in 4 month denervated EDL muscle was the same as that of controls at 65,000, but by 7 months of denervation it had declined to 21,000. With increasing time of denervation, the number of cross-sectional profiles of muscle fibers not containing nuclei rose from 14% in control muscle to 49% in 12 month denervated muscle. This was correlated with a pronounced regular clumping of the nuclei, with pronounced nonnucleated segments between nuclear clumps. CONCLUSIONS: Increasing times of denervation are accompanied by a pronounced decline in the number of myonuclei per muscle fiber and an initial rise and subsequent fall in satellite cell number. These changes are correlated with a decreasing restorative ability of these muscles over the same periods of denervation. Further work on the proliferative capacity of the remaining satellite cells is necessary before firm quantitative conclusions can be made.

Carbohydrate dependence during marathon running.

To test the hypothesis that marathon running is dependent on lipid oxidation, 12 post-absorptive males (31.9 +/- 2.1 yr) ran a treadmill marathon and substrate utilization was assessed. Subjects were placed into a fast (F < or = 2 hr, 45 min; 73.3% VO2max), or a slow (S < or = 3 hr, 45 min; 64.5% VO2max) marathon group. The day before testing subjects rested, but ate their normal diet. Subjects were tested in the morning after an overnight fast, and only tap water, at a rate of 1 l.h, was ingested during exercise. Blood glucose concentration rose at exercise onset, peaked at approximately an hour, but then decreased over time remaining at or above resting levels. Free fatty acids and glycerol rose continuously. No significant differences in plasma FFA, glycerol, or blood glucose concentrations were observed between F or S groups during the marathon. Mean blood lactate concentration was significantly higher (P < 0.05) in the F (2.1 +/- 0.3 mM) group than the S (1.2 +/- 0.2 mM) during exercise. Mean plasma epinephrine was significantly higher in the F (0.9 +/- 0.2 ng.ml-1) than the S (0.6 +/- 0.2 ng.ml-1) group; norepinephrine was also higher in F (3.9 +/- 1.4 ng.ml-1) than the S (2.5 +/- 0.9 ng.ml-1, P < or = 0.05). Blood lactate and epinephrine concentrations correlated significantly (4r = 0.76 and 0.78 in F and S groups, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Antioxidant status and indexes of oxidative stress during consecutive days of exercise.

We tested whether consecutive days of prolonged submaximal exercise would result in oxidant stress sufficient to alter blood antioxidant profiles, progressively change and exhaust blood and plasma antioxidants, and damage RNA. Eleven moderately trained males (24.3 +/- 1.1 yr) exercised 90 min at 65% peak O2 uptake on a cycle ergometer for 3 consecutive days. During day 1 exercise, blood reduced glutathione (GSH) declined 55 +/- 10% and oxidized glutathione (GSSG) increased 28 +/- 7% within 15 min. Total blood glutathione did not significantly change during exercise. GSH levels returned to baseline after 15 min of recovery. On day 3, preexercise GSH and GSSG levels were not significantly different from day 1 preexercise values; essentially similar results were obtained during exercise and recovery. During day 1 exercise, plasma total ascorbate (ascorbate + dehydroascorbate) increased from 53.8 +/- 9.3 to 59.0 +/- 11.3 microM, and percent reduced ascorbate increased from 77.6 +/- 9.3 to 87.3 +/- 9.7%. During day 3 exercise, plasma ascorbate changes were similar to those on day 1. Plasma vitamin E did not change due to exercise on either day 1 or 3. RNA adducts, urinary 8-hydroxyguanosine, did not change significantly due to exercise. Observed increases in GSH oxidation indicate the presence of oxidant stress during prolonged submaximal exercise. Similar redox changes on consecutive days of exercise, with recovery to preexercise values within 15 min, indicate no evidence of persistent or cumulative exercise effects on blood glutathione redox status.(ABSTRACT TRUNCATED AT 250 WORDS)

Exercise, oxidative damage and effects of antioxidant manipulation.

Exercise induces free radical formation in muscle and liver, and oxidative damage, such as lipid peroxidation. The amount of damage depends on exercise intensity, training state and the tissue examined and can be reduced through dietary supplementation of antioxidants such as vitamin E and possibly coenzyme Q10. Supplementation with antioxidants does not increase maximal aerobic capacity or maximal exercise capacity; effects on endurance capacity are unclear. Deficiency of vitamin E or vitamin C greatly reduces endurance capacity, whereas selenium deficiency has no effect on endurance capacity. In studies by the authors, urinary output of the oxidatively damaged RNA base 8-hydroxyguanosine was not affected by several submaximal exercise bouts nor by supplementation with vitamins E and C and beta-carotene in moderately trained humans. In rats, endurance training caused an increase in oxidative damage, as measured by the protein carbonyl concentration of muscle, but not liver. Muscle protein carbonyl concentration returned to normal on detraining. These results indicate that the search for oxidative damage due to exercise and the effects of antioxidant manipulation on such damage should ideally involve examination of several indices of oxidative damage in various tissues after exercise and training.