Enzymatic adaptation to physical training under beta-blockade in the rat. Evidence of a beta 2-adrenergic mechanism in skeletal muscle.

Nonselective and beta 1-selective adrenergic antagonists were tested for their effects on enzymatic adaptation to exercise training in rats as follows: trained + placebo (TC); trained + propranolol (TP); trained + atenolol (TA); and corresponding sedentary groups, SC and SP. Trained rats ran 1 h/d at 26.8 m/min, 15% grade, 5 d/wk, 10 wk. Both beta-antagonists were given at doses that decreased exercise heart rates by 25%. Training increased skeletal muscle citrate synthase, cytochrome c oxidase (Cyt-Ox), carnitine palmitoyltransferase (CPT), beta-hydroxyacyl coenzyme A dehydrogenase, mitochondrial malate dehydrogenase (MDH), and alanine aminotransferase (ALT) activities significantly in the TC group, but not in TP. These enzyme activities, except Cyt-Ox and CPT, were also significantly increased in TA. Hepatic phosphoenolpyruvate carboxykinase activity did not alter with training or beta-blockade. Fructose 1,6-bisphosphatase activity was lower in TC than in SC, but unchanged in TP or TA. Hepatic mitochondrial MDH and ALT activities increased with training only in TC. It is concluded that beta 2-adrenergic mechanisms play an essential role in the training-induced enzymatic adaptation in skeletal muscle.

Glycogen synthase in diabetic rat skeletal muscle: activation by insulin.

Glycogen synthase in skeletal muscle of 3-day alloxan-diabetic rats was found to be in a less active state than in normal muscle. Both the activity ratio (activity without G6P divided by activity with 7.2 mM G6P at 4.4 mM UDPG, pH 7.8) and fractional velocity (activity with 0.25 mM G6P divided by activity with 10 mM G6P at 0.03 mM UDPG, pH 6.9) were significantly lower in the diabetic tissue. Correspondingly, the S0.5 for UDPG and A0.5 for G6P were significantly higher in diabetic tissue, suggesting decreased affinity for substrate and activator, respectively. The kinetic changes in the diabetic synthase were identical whether the alloxan-treated animals were maintained on insulin for 7 days prior to withdrawal for 3 days, or studied 3 days immediately after alloxan treatment. The diabetes-induced changes in synthase could be reversed by injecting the diabetic rat with insulin 10 min prior to sacrifice. After insulin treatment, the S0.5 for UDPG and A0.5 for G6P decreased to control levels or lower and the activity ratios and fractional velocities increased to control levels or higher. The activity of glycogen synthase phosphatase was not decreased in diabetic skeletal muscle. This observation, coupled with the rapid response of the diabetic synthase to in vivo insulin treatment, suggests that, unlike the phosphatase in cardiac muscle and liver, the glycogen synthase phosphatase in skeletal muscle is not altered by the diabetic state.

Modified assays to detect activation of glycogen synthase following exercise.

Glycogen levels and glycogen synthase activity were measured in red vastus lateralis muscle of male Sprague-Dawley rats killed at rest, immediately after swimming to exhaustion, or 4 h postexhaustion. Glycogen levels were very low immediately after the exercise but returned to preexercise levels after 4 h of recovery. Activation of glycogen synthase by glucose 6-phosphate (G6P) was determined in the standard assay of Thomas et al. (Anal. Biochem. 25: 486-499, 1968) using 4.4 mM uridine diphosphate glucose (UDPG) (pH 7.8) or in a modified assay using 0.03 mM UDPG (pH 6.9) in the absence or presence of inhibitor (inorganic phosphate or uridine 5'-diphosphate). Activation of glycogen synthase was determined by measuring activity ratio (activity in the absence of G6P divided by activity in the presence of G6P), A0.5 for G6P (concentration of G6P producing half-maximal activation), and fractional velocity (activity with low G6P divided by activity with high G6P). All three measurements indicated glycogen synthase was significantly activated immediately after exhaustion. Activation after 4 h of recovery was not detected using activity ratio but was readily apparent when fractional velocity of A0.5 for G6P were measured.

Glycogen synthase activation in human skeletal muscle: effects of diet and exercise.

We investigated the role of glycogen synthase in supranormal resynthesis (supercompensation) of skeletal muscle glycogen after exhaustive exercise. Six healthy men exercised 60 min by cycling with one leg at 75% VO2max, recovered 3 days on a low-carbohydrate diet, exercised again, and recovered 4 days on high-carbohydrate diet. Glycogen and glycogen synthase activities at several glucose-6-phosphate (G6P) concentrations were measured in biopsy samples of m. vastus lateralis. Dietary alterations alone did not affect glycogen, whereas exercise depleted glycogen stores. After the second exercise bout, glycogen returned to normal within 24 h and reached supercompensated levels by 48 h of recovery. Glycogen synthase activation state strikingly increased after exercise in exercised muscle and remained somewhat elevated for the first 48 h of recovery in both muscles. We suggest that 1) forms of glycogen synthase intermediate to I (G6P-independent) and D (G6P-dependent) forms are present in vivo, and 2) glycogen supercompensation can in part be explained by the formation of intermediate forms of glycogen synthase that exhibit relatively low activity ratios, but an increased sensitivity to activation by G6P.

Postexercise glycogen replenishment in untrained animals: denervation, tenotomy effects.

To test the hypothesis that an intact nerve supply is essential to normal replenishment of muscle glycogen after exhaustive exercise, glycogen concentration in normal gastrocnemius muscles from guinea pigs exhausted 48 hr previously was compared to glycogen in contralateral muscles that had been denervated at the muscle or high in the thigh immediately after exercise. Each animal of another group underwent tenotomy of one Achilles tendon after exercise so that any effect of disuse could be distinguished from effects of denervation. All muscles were able to replenish glycogen to normal control values 48 hr after exercise and surgery; there were no significant (greater than .05) effects of any of the surgical interventions. Therefore, it appears that glycogen concentration in skeletal muscle 48 hr after exhaustive exercise is independent of both neural phenomena and muscle tension.