Exercise training, Glut-4 protein abundance and glutamine in skeletal muscle of mature and very old horses

Abstract

Two groups of unfit Standardbred mares (adult: 9-14 years, 540 kg, n=7) and old (20-25 years, 530 kg, n=5) were used to test two hypotheses, first, that aging and training would alter plasma and muscle glutamine [Gln] and glutamate [Glu] and second, that aging and training would alter Glut-4 expression in skeletal muscle. All animals were housed on pasture with free access to grass and all received hay and supplementation with a commercially prepared supplement (15% crude protein and 3.00 Mcal/kg dry matter) in individual stalls. Mares were fed to meet or exceed NRC (2007) nutrient recommendations for moderate to heavy exercise. The mares were exercise trained in a free-stall motorised circular exercise machine for 30 min/d, 5 d/week, for 8 weeks. Work intensity during training was set at a relative intensity of ~60% of the maximum heart rate, previously determined during an incremental exercise test (GXT). Blood samples and muscle biopsies (gluteus) obtained before and after 8 weeks of training were used for measurement of [Gln], [Glu] and Glut-4 abundance. Samples were collected before the initiation of training and at 24 h after cessation of last bout of exercise in the training period. All samples were immediately frozen in liquid nitrogen and stored at -80 °C until enzymatic analysis for [Gln], [Glu] and Western Blot analysis for Glut-4 protein abundance. Data were analysed by one-way or two-way ANOVA for repeated measures and the Pearson correlation method. Post-hoc differences were identified with the Tukey test. Significance was set at P<0.05. There were no differences (P>0.05) in muscle [Glu] due to aging. Training decreased (P<0.05) muscle [Glu] from 7,561±701 nmol/g of tissue (mean ± standard error) in pre-training samples to 4,491±701 nmol/g of tissue post-training. Plasma [Gln] decreased (P<0.05) with training (368±14 nmol/ml vs 317±14 nmol/ml). There was a trend (P=0.063) towards an effect of aging. There were significant interactions between age and training for plasma [Gln]. Old mares had lower (P<0.05) post-exercise plasma [Gln] (224±21 nmol/ml) when compared with pre-exercise plasma [Gln] (372±21 nmol/ml). Post-training, plasma [Gln] was lower (P<0.05) in the old mares compared to adult mares [Gln] (224±21 nmol/ml vs 410±18 nmol/ml). There was an effect (P<0.05) of age on muscle [Gln] (old = 6,126 ±870 nmol/g of tissue; adult = 3,176±735 nmol/g of tissue); however, there were no changes (P>0.05) due to training. Glut-4 abundance analysis did not differ (P>0.05) between the young adult and old horses; however, there was a trend (P=0.063) towards an effect of training when samples from both groups were pooled. It was concluded that training and aging produce changes in plasma and muscle [Gln], which may affect immune function in athletic horses, but not in Glut-4.