Low Intake of Choline Is Associated with Diminished Strength and Lean Mass Gains in Older Adults.

OBJECTIVES: Choline is an essential micronutrient for many physiological processes related to exercise training including biosynthesis of acetylcholine. Though dietary choline intake has been studied in relation to endurance training and performance, none have studied it during resistance exercise training (RET) in older adults. The objective of the study was to examine the relationship between choline intake and muscle responses to RET in older adults. METHODS: Forty-six, 60-69-year-old individuals (M=19, F=27) underwent 12 weeks of RET (3x/week, 3 sets, 8-12 reps, 75% of maximum strength [1RM], 8 exercises). Body composition (DEXA) and 1RM tests were performed before and after training. After analyzing 1,656 diet logs (3x/week, 46 participants, 12 weeks), participants' mean choline intakes were categorized into three groups: Low (2.9-5.5 mg/kg lean/d), Med-Low (5.6-8.0 mg/kg lean/d), or Adequate (8.1-10.6 mg/kg lean/d). These correspond to <50%, ~63%, and ~85% of Adequate Intake (AI) for choline, respectively. RESULTS: Gains in composite strength (leg press + chest press 1RM) were significantly lower in the Low group compared with the other groups (Low: 30.9 ± 15.1%, Med-Low: 70.3 ± 48.5%, Adequate: 81.9 ± 68.4%; p=0.004). ANCOVA with cholesterol, protein, or other nutrients did not alter this result. Reduced gains in lean mass were also observed in the Low group, compared with higher choline intake of 5.6-10.6 mg/kg lean/d (1.3 ± 0.6% vs. 3.2 ± 0.6%, p<0.05). CONCLUSION: These data suggest that this population of older adults does not consume adequate choline and lower choline intake is negatively and independently associated with muscle responses to RET.

PGC-1α4 gene expression is suppressed by the IL-6-MEK-ERK 1/2 MAPK signalling axis and altered by resistance exercise, obesity and muscle injury.

AIM: PGC-1α4 is a novel regulator of muscle hypertrophy; however, there is limited understanding of the regulation of its expression and role in many (patho)physiological conditions. Therefore, our purpose was to elicit signalling mechanisms regulating gene expression of Pgc1α4 and examine its response to (patho)physiological stimuli associated with altered muscle mass. METHODS: IL-6 knockout mice and pharmacological experiments in C2C12 myocytes were used to identify regulation of Pgc1α4 transcription. To examine Pgc1α4 gene expression in (patho)physiological conditions, obese and lean Zucker rats with/without resistance exercise (RE), ageing mice and muscle regeneration from injury were examined. RESULTS: In IL-6 knockout mice, Pgc1α4mRNA was ~sevenfold greater than wild type. In C2C12 cells, Pgc1α4mRNA was suppressed ~70% by IL-6. Suppression of Pgc1α4 by IL-6 was prevented by MEK-ERK-MAPK inhibition. RE led to ~260% greater Pgc1α4mRNA content in lean rats. However, obese Zucker rats exhibited ~270% greater Pgc1α4mRNA than lean, sedentary with no further augmentation by RE. No difference was seen in IL-6mRNA or ERK-MAPK phosphorylation in Zucker rats. Aged mice demonstrated ~50% lower Pgc1α4mRNA and ~fivefold greater ERK-MAPK phosphorylation than young despite unchanged Il-6mRNA. During muscle regeneration, Pgc1α4 content is ~30% and IL-6mRNA >threefold of uninjured controls 3 days following injury; at 5 days, Pgc1α4 was >twofold greater in injured mice with no difference in IL-6mRNA. CONCLUSION: Our findings reveal a novel mechanism suppressing Pgc1α4 gene expression via IL-6-ERK-MAPK and suggest this signalling axis may inhibit Pgc1α4 in some, but not all, (patho)physiological conditions.

Cumulative responses of muscle protein synthesis are augmented with chronic resistance exercise training.

AIM: The purpose of this study was to determine the anabolic response of a single bout of high intensity resistance exercise (RE) following 5 weeks of RE training. METHODS: To complete these studies, Sprague-Dawley rats were assigned by body mass to RE, exercise control (EC), or sedentary cage control (CC) groups and studied over 36 h after 5 weeks of RE (squat-like) training. Cumulative (final 36 h) fractional rates of muscle protein synthesis (FSR) were determined by ²H2O, and acute (16 h post-RE) rates of muscle protein synthesis (RPS) were determined by flooding with l-[2,3,4,5,6-³H]phenylalanine. Regulators of peptide-chain initiation, 4E-BP1, eIF4E and the association of the two were determined by Western blotting and immunoprecipitation respectively. RESULTS: No differences were observed with acute measures of RPS obtained 16 h following the final exercise bout in the plantaris or soleus muscles (P > 0.05). Consistent with this observation, 4E-BP1 was similarly phosphorylated and bound to eIF4E among all groups. However, upon determination of the cumulative response, FSR was significantly increased in the plantaris of RE vs. EC and CC (0.929±0.094, 0.384±0.039, 0.300±0.022% h(-1) respectively; P<0.001), but not the soleus. CONCLUSION: With the advantage of determining cumulative FSR, the present study demonstrates that anabolic responses to RE are still evident after chronic RE training, primarily in muscle composed of fast-twitch fibres.

Increased training loads do not magnify cancellous bone gains with rodent jump resistance exercise.

This study sought to elucidate the effects of a low- and high-load jump resistance exercise (RE) training protocol on cancellous bone of the proximal tibia metaphysis (PTM) and femoral neck (FN). Sprague-Dawley rats (male, 6 mo old) were randomly assigned to high-load RE (HRE; n = 16), low-load RE (LRE; n = 15), or sedentary cage control (CC; n = 11) groups. Animals in the HRE and LRE groups performed 15 sessions of jump RE during 5 wk of training. PTM cancellous volumetric bone mineral density (vBMD), assessed by in vivo peripheral quantitative computed tomography scans, significantly increased in both exercise groups (+9%; P < 0.001), resulting in part from 130% (HRE; P = 0.003) and 213% (LRE; P < 0.0001) greater bone formation (measured by standard histomorphometry) vs. CC. Additionally, mineralizing surface (%MS/BS) and mineral apposition rate were higher (50-90%) in HRE and LRE animals compared with controls. PTM bone microarchitecture was enhanced with LRE, resulting in greater trabecular thickness (P = 0.03) and bone volume fraction (BV/TV; P = 0.04) vs. CC. Resorption surface was reduced by nearly 50% in both exercise paradigms. Increased PTM bone mass in the LRE group translated into a 161% greater elastic modulus (P = 0.04) vs. CC. LRE and HRE increased FN vBMD (10%; P < 0.0001) and bone mineral content (∼ 20%; P < 0.0001) and resulted in significantly greater FN strength vs. CC. For the vast majority of variables, there was no difference in the cancellous bone response between the two exercise groups, although LRE resulted in significantly greater body mass accrual and bone formation response. These results suggest that jumping at minimal resistance provides a similar anabolic stimulus to cancellous bone as jumping at loads exceeding body mass.

A rat resistance exercise regimen attenuates losses of musculoskeletal mass during hindlimb suspension.

Exposure to microgravity and/or spaceflight causes dramatic losses in both muscle and bone mass. In normal gravity, resistance exercise has been effectively used to increase muscle and bone mass. We tested a novel form of resistance exercise training using flywheel technology as a countermeasure to offset the loss of musculoskeletal mass during 4 weeks of adult rat hindlimb suspension (HS), an unloading model of microgravity. Male, Sprague-Dawley rats (6-month old) were operantly conditioned to perform resistance exercise, and then randomly assigned to groups of sedentary control (CON), HS, and HS with resistance exercise training (HSRT; 2 sets of approximately 21 repetitions, 3 days week(-1) for 4 weeks during suspension). In soleus, HS resulted in lower (P < 0.05) muscle mass to body mass ratio (approximately 50% of controls) and rates of protein synthesis. HSRT significantly attenuated the loss of muscle mass in soleus muscle, and rates of protein synthesis for soleus were similar for HSRT and controls. There were no differences among groups for mass or rates of protein synthesis in extensor digitorum longus. In cancellous regions of the distal femur, HS resulted in significant reductions of bone mineral density (BMD), but this was restored to control levels with HSRT. Cortical regions of the femur were not different among HS, HSRT or control groups. Together, these data suggest that resistance training using flywheel technology may be a promising tool to attenuate losses of the musculoskeletal system during periods of hindlimb unloading.

Skeletal muscle PGF(2)(alpha) and PGE(2) in response to eccentric resistance exercise: influence of ibuprofen acetaminophen.

PGs have been shown to modulate skeletal muscle protein metabolism as well as inflammation and pain. In nonskeletal muscle tissues, the over the counter analgesic drugs ibuprofen and acetaminophen function through suppression of PG synthesis. We previously reported that ibuprofen and acetaminophen inhibit the normal increase in skeletal muscle protein synthesis after high intensity eccentric resistance exercise. The current study examined skeletal muscle PG levels in the same subjects to further investigate the mechanisms of action of these drugs in exercised skeletal muscle. Twenty-four males (25 +/- 3 yr) were assigned to 3 groups that received the maximal over the counter dose of ibuprofen (1200 mg/d), acetaminophen (4000 mg/d), or a placebo after 10-14 sets of 10 eccentric repetitions at 120% of concentric 1 repetition maximum using the knee extensors. Preexercise and 24 h postexercise biopsies of the vastus lateralis revealed that the exercise-induced change in PGF(2alpha) in the placebo group (77%) was significantly different (P < 0.05) from those in the ibuprofen (-1%) and acetaminophen (-14%) groups. However, the exercise-induced change in PGE(2) in the placebo group (64%) was only significantly different (P < 0.05) from that in the acetaminophen group (-16%). The exercise-induced changes in PGF(2alpha) and PGE(2) were not different between the ibuprofen and acetaminophen groups. These results suggest that ibuprofen and acetaminophen have a comparable effect on suppressing the normal increase in PGF(2alpha) in human skeletal muscle after eccentric resistance exercise, which may profoundly influence the anabolic response of muscle to this form of exercise.

Insulin action on rates of muscle protein synthesis following eccentric, muscle-damaging contractions.

The purpose of this study was to determine whether eccentric, muscle-damaging contractions affect insulin action on muscle protein synthesis. Male Wistar rats (n = 28) were anaesthetized either once or twice separated by 7 days' rest, and one limb was electrically stimulated to contract eccentrically, while the contralateral limb served as a non-stimulated control. Twenty-four and 48 h after contractions, rates of protein synthesis were assessed in soleus and red or white gastrocnemius muscles during a hindlimb perfusion with or without insulin (20 000 microU mL(-1)). Rates of protein synthesis were not different in non-stimulated muscle, with or without insulin (P > 0.05). In red or white gastrocnemius without insulin, rates of protein synthesis were significantly reduced (P < 0.05) 24 and 48 h after a single session and 48 h after a double session of muscle contractions. However, protein synthesis was normalized with insulin 24 and 48 h after contractions in red, and 48 h after contractions in white gastrocnemius. In soleus muscle, protein synthesis was impaired only 48 h after the second session, but partially restored by insulin (P < 0.05). These results indicate that muscle becomes more sensitive to insulin action on rates of protein synthesis after muscle-damaging contractions.

Effect of exercise training on in vivo lipolysis in intra-abdominal adipose tissue in rats.

Intra-abdominal obesity is associated with cardiovascular disease and non-insulin-dependent diabetes mellitus, and physical training has been suggested to alleviate these conditions. We compared epinephrine-stimulated lipolysis in vivo in three intra-abdominal adipose tissues (ATs: retroperitoneal, parametrial, and mesenteric) and in subcutaneous AT, and we also studied the effect of physical training. Moreover, we studied the effect of physical training on epinephrine-stimulated lipolysis in muscle in vivo. Female rats were either swim trained (15 wk, n = 8) or sedentary (n = 7). Under anesthesia, a two-stage intravenous epinephrine infusion (60 min of 80 and 200 ng. kg(-1). min(-1), respectively) was carried out, and local interstitial glycerol concentration was measured by the microdialysis technique. Blood flow was measured by microspheres. Training increased blood flow in all ATs [on average: 73 +/- 12 (trained) vs. 14 +/- 4 (sedentary) ml. 100 g(-1). min(-1), P < 0. 05]; nevertheless, epinephrine-stimulated interstitial glycerol concentrations were increased or unchanged. Interstitial glycerol concentration was higher in intra-abdominal than in subcutaneous AT in both trained and sedentary rats. In skeletal muscle, interstitial glycerol concentration and blood flow did not differ between trained and sedentary rats. In conclusion, in vivo lipolysis is higher both in the basal state and during epinephrine-stimulation in intra-abdominal than in subcutaneous AT, and training may be beneficial in alleviating intra-abdominal obesity by enhancing lipolysis in intra-abdominal fat depots.

Insulin stimulation of muscle protein synthesis in obese Zucker rats is not via a rapamycin-sensitive pathway.

The obese Zucker rat is resistant to insulin for glucose disposal, but it is unknown whether this insulin resistance is accompanied by alterations of insulin-mediated muscle protein synthesis. We examined rates of muscle protein synthesis either with or without insulin in lean and obese Zucker rats with the use of a bilateral hindlimb preparation. Additional experiments examined insulin's effect on protein synthesis with or without rapamycin, an inhibitor of protein synthesis. Protein synthesis in red and white gastrocnemius was stimulated by insulin compared with control (no insulin) in obese (n = 10, P<0.05) but not in lean (n = 10, P>0.05) Zucker rats. In white gastrocnemius, rapamycin significantly reduced rates of protein synthesis compared with control in lean (n = 6) and obese (n = 6) rats; however, in red gastrocnemius, the attenuating effect of rapamycin occurred only in obese rats. The addition of insulin to rapamycin resulted in rates of synthesis that were similar to those for rapamycin alone for lean rats and to those for insulin alone (augmented) for obese rats in both tissues. Our results demonstrate that insulin enhances protein synthesis in muscle that is otherwise characterized as insulin resistant. Furthermore, rapamycin inhibits protein synthesis in muscle of obese Zucker rats; however, stimulation of protein synthesis by insulin is not via a rapamycin-sensitive pathway.

Fluid snacks to help persons with type 1 diabetes avoid late onset postexercise hypoglycemia.

PURPOSE: The present study assessed whether whole milk, skim milk, or two commercially available sports drinks are effective in preventing late onset postexercise hypoglycemia (LOPEH) in persons with type 1 diabetes mellitus. METHODS: Subjects ingested water, whole milk, skim milk, sport drink A (carbohydrate and electrolytes), or sport drink B (carbohydrate, fat, and protein) before, during, and after 1 h of bicycle exercise at 60% VO2max in the late afternoon. Drinks were isocaloric (470 +/- 150 kcal) and the number of calories consumed was based on individual energy expenditure. No adjustment in insulinization was allowed in anticipation of exercise. RESULTS: During water trials all subjects became hypoglycemic. Most drinks lead to a moderate hyperglycemia (range of mean values = 200-280 mg x dL(-1)) during the period between the end of exercise and dinner, but this was not the case for whole milk (range 80-120 mg x dL(-1)). Glycemia peaked about 1.5 h after dinner and declined over the next 90 min. Persistent hyperglycemia (range of means = 200-310 mg x dL(-1)) from after exercise to about 4 h postexercise was observed with sports drink B. A decline in glycemia in the evening was greatest during the skim milk trial and required subjects to ingest more carbohydrate as a late evening snack. The least decline during this period occurred during the whole milk trial. Subjects experienced pre-bed and early morning (0300 h) hypoglycemia in 7 of the 28 trials. CONCLUSIONS: These data show that whole milk and sports drinks that are designed for both quick (sport drink A) and long lasting (sport drink B) nutrient replenishment can be used by persons with type 1 diabetes in an effort to avoid LOPEH.

Effect of exercise training on in vivo insulin-stimulated glucose uptake in intra-abdominal adipose tissue in rats.

Intra-abdominal obesity may be crucial in the pathogenesis of the insulin-resistance syndrome, and training may alleviate this condition. We compared insulin-mediated glucose uptake in vivo in three intra-abdominal adipose tissues (ATs; retroperitoneal, parametrial, and mesenteric) and in subcutaneous AT and also studied the effect of training. Rats were either swim trained (15 wk, n = 9) or sedentary (n = 16). While the rats were under anesthesia, a hyperinsulinemic ( approximately 900 pM), euglycemic clamp was carried out and local glucose uptake was measured by both the 2-deoxy-D-[(3)H]glucose and microdialysis techniques. Blood flow was measured by microspheres. Upon insulin stimulation, blood flow generally decreased in AT. Flow was higher in mesenteric tissue than in other ATs, whereas insulin-mediated glucose uptake did not differ between ATs. Training doubled the glucose infusion rate during hyperinsulinemia, in part, reflecting an effect in muscle. During hyperinsulinemia, interstitial glucose concentrations were lower, glucose uptake per 100 g of tissue was higher in AT in trained compared with sedentary rats, and training influenced glucose uptake identically in all ATs. In conclusion, differences between ATs in insulin sensitivity with respect to glucose uptake do not explain that insulin resistance is associated with intra-abdominal rather than subcutaneous obesity. Furthermore, training may be beneficial by enhancing insulin sensitivity in intra-abdominal fat depots.

Attenuated insulin action on glucose uptake and transport in muscle following resistance exercise in rats.

A previous study reported elevations of insulin-mediated muscle protein synthesis following four days of resistance exercise in rats (Fluckey et al. 1996. Am J Physiol 270, E313-E319). The purpose of this study was to determine if insulin-stimulated muscle glucose uptake (a-v diff.) and 2-deoxyglucose (2-DG) transport were altered under similar conditions. The protocol consisted of squat-like exercises during four sessions with progressively increased weight (70-190 g). Each session consisted of 50 repetitions and sessions were separated by 48 h. Sixteen hours after the last exercise session, basal glucose uptake in perfused hindlimbs was not different (P > 0.05) between exercised (n=6) and non-exercised controls (n=6). However, there was a significant (P < 0.05) attenuation of insulin-stimulated (20 000 microU mL-1) glucose uptake in exercised vs. non-exercised rats (491 +/- 31 vs. 664 +/- 58 micromol glucose-1 g-1 [15-min insulin period]-1, respectively). Following resistance exercise, insulin-stimulated 2-DG transport, measured during the last 10 min of the perfusion period, was significantly reduced (P < 0.05) in the soleus, white gastrocnemius and extensor digitorum longus muscles. Additionally, GLUT-4 glucose transporter protein content was significantly reduced (P < 0.05) in white gastrocnemius and extensor digitorum longus muscles. These results demonstrate that insulin-stimulated glucose uptake and transport are reduced after resistance exercise. Furthermore, the applied resistance exercise protocol causes directionally opposite changes of insulin action in two major metabolic pathways, i.e. glucose transport and protein synthesis.

Mechanisms associated with hypoxia- and contraction-mediated glucose transport in muscle are fibre-dependent.

The purpose of this study was to examine the effects of hypoxia and muscle contractions on rates of 2-deoxyglucose (2-DG) transport in red and white portions of the gastrocnemius muscle of the rat. 2-DG transport was measured during the last 10 min of a 60-min hindlimb perfusion in male Wistar rats ( approximately 300 g), with or without muscle contractions of one limb. The medium was gassed with either 95% oxygen and 5% carbon dioxide or 95% nitrogen and 5% carbon dioxide to achieve normal or hypoxic conditions, respectively. Muscle contractions began after 30 min of perfusion and consisted of isometric muscle actions (200-ms trains, 100 Hz; one train per second) for two sets of 5 min, with 1-min rest between sets. 2-DG transport in white gastrocnemius was higher (P < 0.05) than basal during hypoxia (4.8-fold) and following contractions using oxygenated or hypoxic medium (4.6-fold and 5.4-fold, respectively; n=6 for each group). 2-DG transport was not different (P > 0.05) between these stimulated conditions. Similarly, 2-DG transport in red gastrocnemius was 5.1- and 4.8-fold higher (P < 0.05) than basal during hypoxia and following contractions in oxygenated medium, respectively. However, 2-DG transport following contractions during hypoxic conditions in red gastrocnemius was, unlike white gastrocnemius, higher (8.9-fold over basal; P < 0.05) than in all other conditions. These results suggest that mechanisms associated with hypoxia- and muscle contraction-mediated glucose transport are fibre type-dependent, with additive effects of the two stimuli in fast-twitch, oxidative fibres.

Hypertrophy of skeletal muscle in diabetic rats in response to chronic resistance exercise.

This study had the following objectives: 1) to determine whether diabetic rats could increase muscle mass due to a physiological manipulation (chronic resistance exercise), 2) to determine whether exercise training status modifies the effect of the last bout of exercise on elevations in rates of protein synthesis, and 3) to determine whether chronic resistance exercise alters basal glycemia. Groups consisted of diabetic or nondiabetic rats that performed progressive resistance exercise for 8 wk, performed acute resistance exercise, or remained sedentary. Arterial plasma insulin in diabetic groups was reduced by about one-half (P < 0.05) compared with nondiabetic groups. Soleus and gastrocnemius-plantaris complex muscle wet weights were lower because of diabetes, but in response to chronic exercise these muscles hypertrophied in diabetic (0.028 +/- 0.003 vs. 0.032 +/- 0.0015 g/cm for sedentary vs. exercised soleus and 0.42 +/- 0.068 vs. 0.53 +/- 0.041 g/cm for sedentary vs. exercised gastrocnemius-plantaris, both P < 0.05) but not in nondiabetic (0.041 +/- 0.0026 vs. 0.042 +/- 0.003 g/cm for sedentary vs. exercised soleus and 0.72 +/- 0.015 vs. 0.69 +/- 0.013 g/cm for sedentary vs. exercised gastrocnemius-plantaris) rats when muscle weight was expressed relative to tibial length or body weight (data not shown). Another group of diabetic rats that lifted heavier weights showed muscle hypertrophy. Rates of protein synthesis were higher in red gastrocnemius in chronically exercised than in sedentary rats: 155 +/- 11 and 170 +/- 7 nmol phenylalanine incorporated x g muscle(-1) x h(-1) in exercised diabetic and nondiabetic rats vs. 110 +/- 14 and 143 +/- 7 nmol phenylalanine incorporated x g muscle(-1) x h(-1) in sedentary diabetic and nondiabetic rats. These elevations, however, were lower than in acutely exercised (but untrained) rats: 176 +/- 15 and 193 +/- 8 nmol phenylalanine incorporated x g muscle(-1) x h(-1) in diabetic and nondiabetic rats. Finally, chronic exercise training in diabetic rats was associated with reductions in basal glycemia, and such reductions did not occur in sedentary diabetic groups. These data demonstrate that, despite lower circulating insulin concentrations, diabetic rats can increase muscle mass in response to a physiological stimulus.

Estimation of rat muscle blood flow by microdialysis probes perfused with ethanol, [14C]ethanol, and 3H2O.

We used the perfused rat hindquarter to evaluate whether the microdialysis ethanol technique can be used to qualitatively estimate nutritive skeletal muscle blood flow. Four microdialysis probes were inserted in different hindlimb muscles in each of 16 rats. Hindquarters were perfused at blood flow rates ranging from 0 to 21 ml. 100 g-1. min-1. The microdialysis probes were perfused at 2 microliter/min with perfusate containing ethanol, [14C]ethanol, and 3H2O. Within and between experiments outflow-to-inflow ratios (o/i) generally varied inversely with blood flow. When a low flow or no flow was maintained in hindquarters, o/i ratios first increased with time (for at least 60 min) and then leveled off. The long time constant impaired detection of rapid oscillations in blood flow, especially at low blood flow rates. Contractions per se apparently decreased o/i ratios independent of blood flow. Ethanol and [14C]ethanol o/i ratios did not differ. 3H2O o/i paralleled ethanol and [14C]ethanol o/i ratios but it was significantly lower. In conclusion, differences in skeletal muscle blood flow can be detected by the microdialysis technique. However, the slow changes in o/i, in particular at low blood flow rates, limit the usefulness of the technique for measuring dynamic changes in blood flow; caution must also be exerted during muscle contractions. 3H2O and [14C]ethanol are good alternatives to ethanol in the determination of blood flow by microdialysis.

Regulation of skeletal muscle UCP-2 and UCP-3 gene expression by exercise and denervation.

The factors that regulate gene expression of uncoupling proteins 2 and 3 (UCP-2 and UCP-3) in skeletal muscle are poorly understood, but both genes are clearly responsive to the metabolic state of the organism. Therefore, we tested the hypothesis that denervation and acute and/or chronic exercise (factors that profoundly affect metabolism) would alter UCP-2 and UCP-3 gene expression. For the denervation studies, the sciatic nerve of rat and mouse hindlimb was sectioned in one leg while the contralateral limb served as control. Northern blot analysis revealed that denervation was associated with a 331% increase (P < 0.001) in UCP-3 mRNA and a 200% increase (P < 0. 01) in UCP-2 mRNA levels in rat mixed gastrocnemius (MG) muscle. In contrast, denervation caused a 53% decrease (P < 0.001) in UCP-3 and a 63% increase (P < 0.01) in UCP-2 mRNA levels in mouse MG. After acute exercise (2-h treadmill running), rat UCP-3 mRNA levels were elevated (vs. sedentary control) 252% (P < 0.0001) in white gastrocnemius and 63% (P < 0.05) in red gastrocnemius muscles, whereas UCP-2 levels were unaffected. To a lesser extent, elevations in UCP-3 mRNA (22%; P < 0.01) and UCP-2 mRNA (55%; P < 0.01) levels were observed after acute exercise in the mouse MG. There were no changes in either UCP-2 or UCP-3 mRNA levels after chronic exercise (9 wk of wheel running). These results indicate that acute exercise and denervation regulate gene expression of skeletal muscle UCPs.

Effect of resistance exercise training on cortical and cancellous bone in mature male rats.

The effect of resistance training on tibial cancellous and cortical bone was evaluated in rats by using static histomorphometry and Northern analysis. Five-month-old male Sprague-Dawley rats were randomly assigned to exercise (Ex; n = 8) or control (Con; n = 4) groups. Animals were operantly conditioned to press two levers, facilitating full extension and flexion of the hindlimbs ("squats"), while wearing an unweighted vest. After an 8-wk familiarization period, Ex animals performed 3 sessions/wk for 17-19 sessions with progressively increased amounts of weight applied to the vest. Con rats completed the same exercise protocol without applied resistance. No difference in cross-sectional, medullary, or cortical bone area was observed between Ex and Con rats in the tibial diaphysis. In contrast, the cancellous bone area in the proximal tibial metaphysis was significantly larger in trained rats. Trabecular number, trabecular thickness, and the percentage of cancellous bone covered by osteoid were significantly greater in the Ex animals compared with Con animals. In addition, steady-state mRNA levels for osteocalcin for the Ex group were 456% those expressed in the Con group. The data demonstrate that resistance training increases cancellous bone area in sexually mature male rats and suggest that it does so, in part, by stimulating bone formation.

Influence of muscle mass and work on post-exercise glucose and insulin responses in young untrained subjects.

The 18 h post-exercise glucose and insulin responses of six male and six female subjects were measured following one- or two-leg cycling to determine the influence of muscle mass involvement and work. Each subject performed three exercise trials on a Cybex Met 100 cycle ergometer: (1) two-leg exercise for 30 min at 60% of the two-leg VO2 max; (2) one-leg exercise for 30 min at 60% of one-leg VO2 max; and (3) one-leg exercise (one-leg TW) at 60% of the one-leg VO2 max with the total work performed equal to that of the two-leg trial (duration approximately 50 min). These trials were preceded by 2 days of inactivity and followed by an 18 h post-exercise 75 g oral glucose tolerance test (OGTT). The glucose response during the baseline OGTT demonstrated that the subjects had normal glucose tolerance with fasting serum glucose levels of 5.1 mM, and 1 and 2 h serum glucose less than 7.8 mM, respectively. The 18 h post-exercise glucose responses were significantly lower following the two-leg trial (P < 0.05), with the area under the curve values being 129.9 mM h-1 less than the resting control level. The 18 h post-exercise insulin AUC response of the two-leg trial was significantly lower than either of the one-leg responses (14.7 pM below the one-leg and 5.0 pM below the one-leg TW) but was not associated with a change in C-peptide. The 18 h post-exercise insulin levels of the one-leg and one-leg TW trials were above or near the resting control values, but were not accompanied by a significant change in C-peptide. In conclusion, the data presented here show that the amount of muscle tissue utilized during an exercise bout can influence both the glucose and insulin responses, whereas the amount of total work employed during the exercise had no effect on either of these parameters.

Insulin stimulation of protein synthesis in rat skeletal muscle following resistance exercise is maintained with advancing age.

This study examined whether or not insulin elevates rates of protein synthesis in muscle following four days of resistance exercise in young (4-mo), middle-aged (12-mo), and old (32-mo) rats. Thirty-six male Fischer 344/BN F1 rats (n = 12 in each group) performed an operantly conditioned activity which required full extension of the hindlimbs with weights over the scapula (ACUTE; n = 6 for each age group) or with no additional weight (nonexercised; NONEX; n = 6 for each age group). Acutely exercised animals engaged in four distinct exercise sessions with each session separated by 48 h. Rates of protein synthesis were assessed in soleus, gastrocnemius (GAST), and extensor digitorum longus (EDL) muscles 16 h after the last exercise bout using a bilateral hindlimb perfusion to measure the incorporation of tritiated phenylalanine (F) into muscle protein. One limb of the bilateral hindlimb preparation received a medium that contained rat insulin at a physiological concentration (6.25 ng.ml-1), while the other limb did not. Rates of protein synthesis in soleus with insulin supplementation were significantly higher within all age groups following resistance exercise vs ACUTE without insulin and NONEX with or without insulin (p < .05). Rates of protein synthesis in soleus were not different within age groups for NONEX with or without insulin (p < .05), but rates of protein synthesis for young NONEX were significantly higher (p < .05) than middle-aged or old NONEX (204 +/- 9 vs 149 +/- 6 or 141 +/- 9 nmol F incorporated.g-1.h-1, respectively; means +/- SE). Rates of protein synthesis in GAST with insulin were also significantly higher within all age groups following resistance exercise than ACUTE without insulin or NONEX with or without insulin (p < .05). Unlike soleus, rates of protein synthesis in GAST were significantly higher for old NONEX vs young NONEX (68 +/- 6 vs 45 +/- 5 nmol F incorporated.g-1.h-1, respectively; P < .05), but not middle-aged NONEX (51 +/- 3 nmol F incorporated.g-1.h-1). Translational efficiency (rates of protein synthesis.unit of RNA-1.h-1) for GAST supplemented with insulin was significantly greater in ACUTE with insulin than ACUTE without insulin or NONEX with or without insulin (p < .05). There were no effect of age, insulin, or exercise on rates of protein synthesis in EDL (p > .05). These data suggest that following resistance exercise, insulin increased rates of protein synthesis in both soleus and GAST regardless of age, and it appeared that this insulin-mediated elevation may have occurred at the level of translation.

Augmented insulin action on rates of protein synthesis after resistance exercise in rats.

This study investigated whether insulin has a modulatory effect on protein synthesis rates in skeletal muscle after four sessions of resistance exercise. Male rats engaged in resistance exercise (Acute) that required full extension of the hindlimbs with weights over the scapula or performed the standing movement with no additional weight (Nonex). Two separate studies were conducted. Rates of protein synthesis for study 1 (Acute, n = 6; Nonex, n = 6) were assessed 16 h postexercise by incorporation of [3H]phenylalanine ([3H]F) into muscle protein by use of an in vivo flooding dose protocol. Rates of protein synthesis in soleus of Acute (100 +/- 9 nmol F.g-1.h-1) were significantly higher than in Nonex (72 +/- 9 nmol F.g-1.h-1, P < 0.05). Rates of protein synthesis were significantly higher in gastrocnemius of Acute vs. Nonex (48 +/- 7 vs. 25 +/- 2 nmol F.g-1.h-1) but not in extensor digitorum longus (EDL). Assessment of protein synthesis rates for study 2 was conducted 16 h after resistance exercise with use of [3H]F incorporation into muscle protein during in situ bilateral hindlimb perfusion, with each leg perfused simultaneously but separately. Perfusion medium for one leg, but not the other, contained insulin (6.25 ng/ml). Soleus and gastrocnemius of Acute had higher protein synthesis rates than Nonex only in the leg that received insulin. For gastrocnemius, rates of protein synthesis in Acute without insulin were significantly lower than in Nonex with or without insulin. Insulin had no effect on protein synthesis rates for any muscle in Nonex rats. Neither exercise nor insulin affected protein synthesis rates in EDL. We conclude that insulin is a necessary component in elevated protein synthesis rates after resistance exercise in muscles composed of primarily slow-or fast-twitch fibers, and that a physiological perturbation (resistance exercise in this study) is required to observe such modulation, because rates of protein synthesis in Nonex muscles were not influenced by insulin.