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Severe diabetes prohibits elevations in muscle protein synthesis after acute resistance exercise in rats

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This study determined whether rates of protein synthesis increase after acute resistance exercise in skeletal muscle from severely diabetic rats. Previous studies consistently show that postexercise rates of protein synthesis are elevated in nondiabetic and moderately diabetic rats. Severely diabetic rats performed acute resistance exercise (n = 8) or remained sedentary (n = 8). A group of nondiabetic age-matched rats served as controls (n = 9). Rates of protein synthesis were measured 16 h after exercise. Plasma glucose concentrations were >500 mg/dl in the diabetic rats. Rates of protein synthesis (nmol phenylalanine incorporated. g muscle(-1). h(-1), means +/- SE) were not different between exercised (117 +/- 7) and sedentary (106 +/- 9) diabetic rats but were significantly (P < 0.05) lower than in sedentary nondiabetic rats (162 +/- 9) and in exercised nondiabetic rats (197 +/- 7). Circulating insulin concentrations were 442 +/- 65 pM in nondiabetic rats and 53 +/- 11 and 72 +/- 19 pM in sedentary and exercised diabetic rats, respectively. Plasma insulin-like growth factor I concentrations were reduced by 33% in diabetic rats compared with nondiabetic rats, and there was no difference between exercised and sedentary diabetic rats. Muscle insulin-like growth factor I was not affected by resistance exercise in diabetic rats. The results show that there is a critical concentration of insulin below which rates of protein synthesis begin to decline in vivo. In contrast to previous studies using less diabetic rats, severely diabetic rats cannot increase rates of protein synthesis after acute resistance exercise.

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... In nondiabetic and moderately diabetic rats, acute moderate-intensity resistance exercise causes increased rates of protein synthesis when the phenylalanine flooding dose is used to assess such rates (13). Severe insulin deficiency retards protein synthesis, and a previous study has shown that severely diabetic rats cannot increase rates of protein synthesis after moderate-intensity resistance exercise (15). ...
... In vivo rates of protein synthesis. To date, our report (15) has been the only one to make the observation that severely diabetic rats cannot increase rates of protein synthesis after resistance exercise. Therefore, we repeated that study using five or six additional rats per group. ...
... Rates of protein synthesis for gastrocnemius muscle in response to exercise are provided in Fig. 1. These responses are similar (not significantly different from data in Fig. 1 of Ref. 15) to those previously reported, except the basal rates of protein synthesis are somewhat lower than those previously reported. Rates of protein synthesis were higher in the exercised nondiabetic group, but no elevation was apparent in the severely diabetic group. ...
Article
Rates of protein synthesis are reduced in severely diabetic rats. A potential mechanism through which insulin can stimulate protein synthesis is modulation of the activity of eukaryotic initiation factor 2B (eIF2B). The activity of this factor is elevated after exercise in nondiabetic rats but is markedly lower in skeletal muscle from nonexercised severely diabetic rats. We tested the hypothesis that a failure to increase eIF2B activity after exercise is one potential reason for a failure of severely diabetic rats to increase rates of protein synthesis after resistance exercise. Diabetic (partial pancreatectomy, plasma glucose >475 mg/dl) and nondiabetic male Sprague-Dawley rats (approximately 300 g) performed acute moderate-intensity resistance exercise or remained sedentary. Rates of protein synthesis were higher in nondiabetic rats and increased significantly with exercise, while no elevation was found in severely diabetic rats. The activity of eIF2B was higher (P < 0.05) in exercised nondiabetic than in sedentary nondiabetic rats (0.096 +/- 0.016 and 0.064 +/- 0.02 pmol GDP exchanged/min, respectively), but no difference was observed between sedentary and exercised diabetic rats (0.037 +/- 0.001 and 0.044 +/- 0.008 pmol GDP exchanged/min, respectively), and these activities were lower (P < 0.05) than in nondiabetic animals. These data suggest that severe hypoinsulinemia is associated with an inability to increase eIF2B activity in response to exercise.
... It has been reported that exercise increases both insulin sensitivity [14] and protein balance [15] in muscles from diabetic (D) subjects. That has also been observed in response to acute exercise bout [16][17][18], revealing an effect of the contractile activity per se. This protein balance seems to be related to some specific atrogenes and miR-NAs, which have been shown to be modulated in response to diabetes [19,20] and/or to an acute exercise bout [21]. ...
... Muscle contraction regulates the metabolic homeostasis of muscle cells, and that includes increased muscle glucose disposal [18,39] and a positive protein balance [4,16,17]. After a bout of resistance exercise, both the protein synthesis and degradation increase from 3 to 24 h in muscle of ND human, the synthesis increasing more than the degradation, thus achieving a positive protein balance [16]. ...
... After a bout of resistance exercise, both the protein synthesis and degradation increase from 3 to 24 h in muscle of ND human, the synthesis increasing more than the degradation, thus achieving a positive protein balance [16]. However, in D rats, there is a critical concentration of insulin below which, the rate of protein synthesis declines [17]. In this study, in response to acute contraction, we observed an increase in Trim63 mRNA and TRIM63 protein expression, detectable only in muscles from untreated D rats. ...
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Diabetes mellitus (DM) induces a variable degree of muscle sarcopenia, which may be related to protein degradation and to the expression of both E3 ubiquitin ligases and some specific microRNAs (miRNAs). The present study investigated the effect of diabetes and acute muscle contraction upon the TRIM63 and FBXO32 expression as well as the potential involvement of some miRNAs. Diabetes was induced by streptozotocin and studied after 30 days. Soleus muscles were harvested, stimulated to contract in vitro for twitch tension analysis (0.5 Hz), 30 min later for tetanic analysis (100 Hz), and 30 min later were frozen. TRIM63 and FBXO32 proteins were quantified by western blotting; Trim63 mRNA, Fbxo32 mRNA, miR-1-3p, miR-29a-3p, miR-29b-3p, miR-133a-3p, and miR-133b-3p were quantified by qPCR. Diabetes induced sarcopenia by decreasing (P < 0.05) muscle weight/tibia length index, maximum tetanic contraction and relaxation rates, and absolute twitch and tetanic forces (P < 0.05). Diabetes decreased (P < 0.05) the Trim63 and Fbxo32 mRNAs (30%) and respective proteins (60%), and increased (P < 0.01) the miR-29b-3p (2.5-fold). In muscle from diabetic rats, acute contractile stimulus increased TRIM63 protein, miR-1-3p, miR-29a-3p, and miR-133a/b-3p, but decreased miR-29b-3p (P < 0.05). Independent of the metabolic condition, after muscle contraction, both TRIM63 and FBXO32 proteins correlated significantly with miR-1-3p, miR-29a/b-3p, and miR-133a/b-3p. All diabetes-induced regulations were reversed by insulin treatment. Concluding, the results depict that muscle wasting in long-term insulinopenic condition may not be accompanied by increased proteolysis, pointing out the protein synthesis as an important modulator of muscle sarcopenia in DM.
... Moderately diabetic rats also maintain the ability to elicit an anabolic response to resistance exercise (14)(15)(16). However, the elevations in rates of protein synthesis in diabetic rats are contingent upon the exercise intensity not being too severe (15) as well as the degree of hypoinsulinemia (17). Severely diabetic rats (plasma insulin less than Х80 pM) are not able to increase rates of protein synthesis after moderate-intensity resistance exercise, suggesting that there is a low but critical concentration of circulating insulin below which appropriate anabolic responses are inhibited (17). ...
... However, the elevations in rates of protein synthesis in diabetic rats are contingent upon the exercise intensity not being too severe (15) as well as the degree of hypoinsulinemia (17). Severely diabetic rats (plasma insulin less than Х80 pM) are not able to increase rates of protein synthesis after moderate-intensity resistance exercise, suggesting that there is a low but critical concentration of circulating insulin below which appropriate anabolic responses are inhibited (17). ...
... We have previously reported that the plasma concentration of IGFBP-1 is increased, whereas the concentration of IGFBP-3 is decreased, in diabetic rats compared with nondiabetic control animals (17). These data are consistent with the large majority of both the clinical and preclinical literature. ...
Article
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These studies examined whether passive immunization against insulin-like growth factor I (IGF-I) would prevent increases in rates of protein synthesis in skeletal muscle of diabetic rats after resistance exercise. Male Sprague-Dawley rats were pancreatectomized and randomly assigned to either an exercise or a sedentary group. Animals in each of these groups received either an IGF-I antibody or a nonspecific IgG from a subcutaneous osmotic pump. Exercise did not change plasma or gastrocnemius IGF-I concentrations in nondiabetic rats. However, plasma and muscle IGF-I concentrations were higher in IgG-treated diabetic rats that exercised compared with respective sedentary groups (P < 0.05). Passively immunized diabetic rats did not exhibit the same exercise-induced increase in IGF-I concentrations. In nondiabetic rats, protein synthesis rates were higher after exercise in both control and immunized groups. In diabetic rats, exercise increased protein synthesis in the IgG-treated animals but not in those treated with IGF-I antibody. There was also a significant positive correlation between both plasma and gastrocnemius IGF-I concentrations and rates of protein synthesis in diabetic (P < 0.01), but not nondiabetic, rats. These results suggest that IGF-I is compensatory for insulin in hypoinsulinemic rats by facilitating an anabolic response after acute resistance exercise.
... On the other hand, non-diabetic rats that were subjected to highintensity intermittent training showed higher expression of GLUT-4 protein in skeletal muscle [27]. Fedele et al. [28] showed that in contrast to non-diabetic animals, partial pancreatectomy diabetic animals did not show increased protein synthesis after acute resistance training . Therefore, it seems that both continuous and intermittent training may play important roles in the prevention and treatment of diabetes, but further studies are needed to characterize the intensity and frequency that are best suited to each of these protocols. ...
... In an attempt to elucidate the possible mechanisms involved in cellular growth alterations in skeletal muscle that are induced by exercise training, protein synthesis and degradation rates were evaluated in the soleus muscle , but no differences were observed among the groups. Fedele et al. [28] also found no change in muscle protein synthesis in diabetic rats after acute resistance training. On the other hand, Farrell et al. [51] showed that diabetic rats presented a higher muscle protein synthesis after moderate endurance training. ...
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This study aimed to examine the effects of intermittent and continuous swimming training on muscle protein metabolism in neonatal alloxan-administered rats. Wistar rats were used and divided into six groups: sedentary alloxan (SA), sedentary control (SC), continuous trained alloxan (CA), intermittent trained alloxan (IA), continuous trained control (CC) and intermittent trained control (IC). Alloxan (250 mg/kg body weight) was injected into newborn rats at 6 days of age. The continuous training protocol consisted of 12 weeks of swimming training in individual cylinder tanks while supporting a load that was 5% of body weight; uninterrupted swimming for 1 h/day, five days a week. The intermittent training protocol consisted of 12 weeks of swimming training in individual cylinder tanks while supporting a load that was 15% of body weight; 30 s of activity interrupted by 30 s of rest for a total of 20 min/day, five days a week. At 28 days, the alloxan animals displayed higher glycemia after glucose overload than the control animals. No differences in insulinemia among the groups were detected. At 120 days, no differences in serum albumin and total protein among the groups were observed. Compared to the other groups, DNA concentrations were higher in the alloxan animals that were subjected to continuous training, whereas the DNA/protein ratio was higher in the alloxan animals that were subjected to intermittent training. It was concluded that continuous and intermittent training sessions were effective in altering muscle growth by hyperplasia and hypertrophy, respectively, in alloxan-administered animals.
... Figure 2 shows that there is a delay of at least 6 h after exercise before elevations in protein synthesis occurred in gastrocnemius muscle; the delay was even longer in soleus muscle. The percent increases in gastrocnemius muscle (data not shown) for synthesis at 12 h (Fig. 2) postexercise were similar to those reported in prior studies (14,15). The increases at 24 h, however, exceed (increase of 42%) those that we normally observe at 16 h postexercise(ϳ20-30%). ...
... This was an expected finding, since the rats were fed identically before exercise and this type of exercise does not alter circulating insulin concentrations (14). These data agree with our previous work showing that elevations in protein synthesis after similar exercise are independent of insulin concentrations as long as a low but critical amount of insulin is available in vivo (15). Such a concept is compatible with many studies (16,31) that show that severe diabetes or the absence of insulin in an in situ hindlimb perfusion model (30) markedly reduces basal rates of protein synthesis. ...
Article
The temporal pattern for changes in rates of protein synthesis and glucose uptake after resistance exercise, especially relative to each other, is not known. Male Sprague-Dawley rats performed acute resistance exercise (n = 7) or remained sedentary (n = 7 per group), and the following were assessed in vivo 1, 3, 6, 12 and 24 h later: rates of protein synthesis, rates of glucose uptake, phosphatidylinositol 3-kinase (PI3-kinase) activity, and p70(S6k) activity. Rates of protein synthesis in mixed gastrocnemius muscle did not increase until 12 h after exercise (e.g., at 12 h, sedentary = 138 +/- 4 vs. exercised = 178 +/- 6 nmol phenylalanine incorporated x g muscle(-1) x h(-1), mean +/- SE, P < 0.05), whereas at 6 h after exercise rates of glucose uptake were significantly elevated (sedentary = 0.18 +/- 0.020 vs. exercised = 0.38 +/- 0.024 micromol glucose 6-phosphate incorporated x kg muscle(-1) x min(-1), P < 0.05). At 24 h after exercise, rates of protein synthesis were still elevated, whereas glucose uptake had returned to basal levels. Arterial insulin concentrations were not different between groups at any time. Non-insulin-stimulated activities of PI3-kinase and p70(S6k) were higher at 6, 12, and 24 h after exercise (P < 0.05), and, generally, these occurred when rates of protein synthesis (12 and 24 h) and glucose uptake were elevated (6 and 12 but not 24 h) by exercise. These data suggest that regulators of protein synthesis and glucose uptake may respond to the same contraction-generated signals with different kinetics or that they respond to different intra- or extracellular signals that are generated by exercise.
... Much of the available information on the topic of the response of muscle-protein metabolism to insulin and exercise is from studies in rats, from both in vivo and in situ muscle preparations. Farrell and colleagues have performed many excellent studies examining the influence of insulin on muscle-protein synthesis following resistance exercise (Farrell, Fedele, Vary, Kimball, & Jefferson, 1998;Farrell et al., 1999Farrell et al., , 2000Fedele et al., 2000;Fluckey, Vary, Jefferson, & Farrell, 1996;Kostyak, Kimball, Jefferson, & Farrell, 2001). However, the model used in these studies provides information that is focused on a different aspect of the interaction of insulin and exercise on muscle-protein metabolism. ...
... These authors examined the response of muscle-protein synthesis following resistance exercise in rats that were hypoinsulinemic. In this rat model, severe, but not moderate, hyposinsulinemia inhibited the increase in muscle-protein synthesis due to resistance exercise (Farrell et al., 1998(Farrell et al., , 1999Fedele et al., 2000;Fluckey et al., 1996;Kostyak et al., 2001). Interestingly, the response to hyposinsulinemia was dependent on the intensity of the exercise. ...
Article
In recent years, a body of literature examining the response of muscle-protein metabolism to exercise and nutrition has arisen. Developments in methods, especially stable isotopic tracer methodology, have allowed much information to be gathered in vivo in humans. The metabolic mechanism behind increased muscle mass requires that muscle-protein synthesis exceeds breakdown, i.e., net muscle-protein synthesis. Increased net muscle-protein balance may occur due to exercise, but net synthesis may occur only with the addition of nutrients, particularly a source of amino acids. The major impact of increased amino acid availability on net muscle-protein balance is due to stimulation of muscle-protein synthesis and less to inhibition of muscle-protein breakdown. Amino acids seem to stimulate muscle-protein synthesis, not only by mass action, i.e., providing substrate, but also as signals for initiation of protein synthesis. Stimulation of muscle-protein synthesis by amino acid ingestion may be linked to increased intracellular amino acid levels and/or to changing amino acid levels in the blood. Carbohydrate ingestion, most likely through the action of insulin, also may play a role in the response of muscle to exercise and nutrition. There is very little research in humans in vivo on the intracellular signaling that is linked to muscle-protein synthesis. It is clear that intracellular signaling responds to both insulin and amino acids, but the interactions with exercise are not well known; however, the details of the pathways have only just begun to be investigated, especially in humans. Delineation of these pathways is complicated, and there is little doubt that multiple intracellular signaling pathways with several levels of communication are involved in the hypertrophy process in response to nutrition and exercise. A systematic investigation of the relationship of the signaling to insulin and amino acids combined with exercise will provide important information, especially for populations vulnerable to muscle loss.
... First of all, the effect of postexercise hyperinsulinaemia has been shown to decrease mixed muscle protein degradation whereas synthesis is unaffected (Biolo et al. 1999) and, presumably, the effect is primarily on lysosomal degradation and not myofibrillar breakdown, which follows on the ubiquitin-proteasome pathway. Second, a recent study on postabsorptive exercise in diabetic/nondiabetic rats observed that insulin only played a permissive role at low concentrations in stimulating protein synthesis (Fedele et al. 2000). Thus, it was concluded that the effect of insulin on protein synthesis was only apparent in the low range of plasma insulin, whereas a further increase in insulin did not enhance net protein synthesis additionally (Fedele et al. 2000). ...
... Second, a recent study on postabsorptive exercise in diabetic/nondiabetic rats observed that insulin only played a permissive role at low concentrations in stimulating protein synthesis (Fedele et al. 2000). Thus, it was concluded that the effect of insulin on protein synthesis was only apparent in the low range of plasma insulin, whereas a further increase in insulin did not enhance net protein synthesis additionally (Fedele et al. 2000). ...
Article
J Physiol 2001 August 15: 535(1): 301–11(1) Age-associated loss of skeletal muscle mass and strength can partly be counteracted by resistance training, causing a net synthesis of muscular proteins. Protein synthesis is influenced synergistically by post-exercise amino acid supplementation, but the importance of the timing of protein intake remains unresolved. (2) The study investigated the importance of immediate (P0) or delayed (P2) intake of an oral protein supplement upon muscle hypertrophy and strength over a period of resistance training in elderly males. (3) Thirteen men (age 74 ± 1 years; body mass index (BMI), 25 ± 1 kg m- 2 (means ± SEM)) completed a 12-week resistance training program (three times per week) receiving oral protein in liquid form (10 g protein, 7 g carbohydrate, 3 g fat) immediately after (P0) or 2 h after (P2) each training session. Muscle hypertrophy was evaluated by magnetic resonance imaging (MRI) and from muscle biopsies and muscle strength was determined using dynamic and isokinetic strength measurements. Body composition was determined from dual-energy X-ray absorptiometry (DEXA) and food records were obtained over 4 days. The plasma insulin response to protein supplementation was also determined. (4) In response to training, the cross-sectional area of m. quadriceps femoris (54.6 ± 0.5–58.3 ± 0.5 cm2) and mean fiber area (4047 ± 320–5019 ± 615 μ m2) increased in the P0 group, whereas no significant increase was observed in P2. For P0 both dynamic and isokinetic strength increased, by 46 and 15%, respectively (P P
... Refeeding is capable of reversing these changes. These starvation-induced changes in IGF-I, IGF-I receptor, and binding protein concentrations are parallel to the changes in these factors stemming from diabetes (3,4,14). Circulating levels of IGFBPs were not measured in the present study. ...
... Along with marked hypoinsulinemia in the pancreatectomized rats, there was also a concomitant decrease in both total plasma (Fig. 1) and gastrocnemius IGF-I peptide content (Fig. 2), a finding that our laboratory (13,14) and others (28) have previously observed. In the present study, a 48-h fast lowered plasma and gastrocnemius IGF-I concentrations in both nondiabetic and diabetic rats. ...
Article
The purpose of this study was to examine whether immune neutralization of muscle-produced insulin-like growth factor I (IGF-I) would prevent an appropriate anabolic response to refeeding in diabetic rats. Male Sprague-Dawley rats were made diabetic by partial pancreatectomy and were randomly assigned to be either control-fed, fasted, or fasted-refed (n = 7-8 per group). Diabetes decreased rates of protein synthesis and increased rates of protein degradation in incubated epitrochlearis muscles (P < 0.05). In both groups of rats, fasting lowered protein synthesis and increased proteolysis and subsequent refeeding returned both parameters to near basal values (P < 0.05). Neutralization of muscle IGF-I by the addition of IGF-I antibody to the incubation medium reduced protein synthesis an average of 22% for all groups (P < 0.05). However, rates of protein degradation were not affected. In nondiabetic rats, refeeding increased protein synthesis in both control and antibody-treated muscles (P < 0.05). Refeeding also increased protein synthesis in the control muscles from diabetic rats (P < 0.01). In contrast, muscles from diabetic rats that were incubated with anti-IGF-I did not increase protein synthesis in response to refeeding. These data suggest that immune neutralization of muscle IGF-I in hypoinsulinemic rats negated the ability of endogenous IGF-I to promote protein synthesis and thereby prevented an appropriate anabolic response.
... Thus, the impairments in muscle growth and relative muscle wasting in poorly controlled diabetic rodent models may be diminished with increased physical activity. However, severely diabetic Px rat models do not display increased rates of muscle protein synthesis after acute resistance exercise [16,17], perhaps because a permissive amount of insulin may need to be present [18]. In contrast, it may be that regular aerobic type exercise can increase muscle protein synthesis via insulin/ IGF-1 independent mechanisms [19]. ...
... Indeed, much of what we know about diabetic myopathy has been from studies published on rodents living in sedentary cage conditions [3,[8][9][10]. Studies employing resistance type exercise in Px rats [16,17] or via non-physiological ablation of the gastrocnemius to overload the soleus and plantaris muscles in rat models of diabetes [30] show that resistance exercise fails to increase protein synthesis during severe hypoinsulinaemia/hyperglycaemia. In this study, we used a volitional wheel running model to increase levels of aerobic type physical activity in Px rats and examined various markers of muscle protein synthesis in response to nutrient feeding (oral gavage of leucine). ...
Article
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Poorly controlled type 1 diabetes mellitus can cause reduced skeletal muscle mass and weakness during adolescence, which may affect long-term management of the disease. The aim of this study was to determine whether regular voluntary physical activity and leucine feeding restore rates of protein synthesis and deficits in skeletal muscle mass in a young, hypoinsulinaemic/hyperglycaemic rat model of diabetes. Four-week-old male Sprague-Dawley rats were partially pancreatectomised (Px) to induce hypoinsulinaemia/hyperglycaemia and housed with/without access to running wheels for 3 weeks (n = 12-14/group). Sham surgery rats (shams) served as sedentary controls (n = 18). Protein synthesis and markers of protein anabolism were assessed in the fasted state and following leucine gavage. Fibre type and cross-sectional areas of the gastrocnemius muscle were measured using a metachromatic ATPase stain. Compared with sedentary behaviour, regular activity lowered fasting glycaemia and reduced fed hyperglycaemia in Px rats. Active-Px rats, which ran 2.2  ±  0.71 km/night, displayed greater muscle mass and fibre areas similar to shams, while sedentary-Px rats displayed a 20-30% loss in muscle fibre areas. Muscle protein synthesis (basal and in response to leucine gavage) was impaired in sedentary-Px (by ~65%), but not in active-Px rats, when compared with shams. Following leucine gavage, the phosphorylation status of eIF4E binding protein 1 (4E-BP1) and ribosomal S6 kinase 1 (S6K1), markers of mammalian target of rapamycin complex 1 (mTORC1) signalling, increased in shams (by two- and ninefold, respectively) and in active-Px (1.5- and fourfold, respectively) rats, but not in sedentary-Px rats. Moderate physical activity in young Px rats normalises impairments in skeletal muscle growth and protein synthesis. These findings illustrate the critical compensatory role that modest physical activity and targeted nutrition can have on skeletal muscle growth during periods of hypoinsulinaemia in adolescent diabetes.
... First of all, the effect of postexercise hyperinsulinaemia has been shown to decrease mixed muscle protein degradation whereas synthesis is unaffected (Biolo et al. 1999) and, presumably, the effect is primarily on lysosomal degradation and not myofibrillar breakdown, which follows on the ubiquitin-proteasome pathway. Second, a recent study on postabsorptive exercise in diabetic/nondiabetic rats observed that insulin only played a permissive role at low concentrations in stimulating protein synthesis (Fedele et al. 2000). Thus, it was concluded that the effect of insulin on protein synthesis was only apparent in the low range of plasma insulin, whereas a further increase in insulin did not enhance net protein synthesis additionally (Fedele et al. 2000). ...
... Second, a recent study on postabsorptive exercise in diabetic/nondiabetic rats observed that insulin only played a permissive role at low concentrations in stimulating protein synthesis (Fedele et al. 2000). Thus, it was concluded that the effect of insulin on protein synthesis was only apparent in the low range of plasma insulin, whereas a further increase in insulin did not enhance net protein synthesis additionally (Fedele et al. 2000). ...
Article
1. Age-associated loss of skeletal muscle mass and strength can partly be counteracted by resistance training, causing a net synthesis of muscular proteins. Protein synthesis is influenced synergistically by postexercise amino acid supplementation, but the importance of the timing of protein intake remains unresolved. 2. The study investigated the importance of immediate (P0) or delayed (P2) intake of an oral protein supplement upon muscle hypertrophy and strength over a period of resistance training in elderly males. 3. Thirteen men (age, 74 +/- 1 years; body mass index (BMI), 25 +/- 1 kg m(-2) (means +/- S.E.M.)) completed a 12 week resistance training programme (3 times per week) receiving oral protein in liquid form (10 g protein, 7 g carbohydrate, 3 g fat) immediately after (P0) or 2 h after (P2) each training session. Muscle hypertrophy was evaluated by magnetic resonance imaging (MRI) and from muscle biopsies and muscle strength was determined using dynamic and isokinetic strength measurements. Body composition was determined from dual-energy X-ray absorptiometry (DEXA) and food records were obtained over 4 days. The plasma insulin response to protein supplementation was also determined. 4. In response to training, the cross-sectional area of m. quadriceps femoris (54.6 +/- 0.5 to 58.3 +/- 0.5 cm(2)) and mean fibre area (4047 +/- 320 to 5019 +/- 615 microm(2)) increased in the P0 group, whereas no significant increase was observed in P2. For P0 both dynamic and isokinetic strength increased, by 46 and 15 %, respectively (P < 0.05), whereas P2 only improved in dynamic strength, by 36 % (P < 0.05). No differences in glucose or insulin response were observed between protein intake at 0 and 2 h postexercise. 5. We conclude that early intake of an oral protein supplement after resistance training is important for the development of hypertrophy in skeletal muscle of elderly men in response to resistance training.
... However, results obtained when using moderately diabetic rats imply that the insulin concentration necessary for the effect may be minimal. To address the question of what concentration is required for the permissive effect of insulin on protein synthesis, Fedele et al. (31) compared muscle protein synthesis to plasma insulin concentration in animals exhibiting a wide range of plasma concentrations of the hormone. The results of that study, illustrated diagrammatically in Fig. 4, show that rates of protein synthesis are related to arterial insulin only in the very low range (Ͻ80 pmol/l for a 5-h fasted rat). ...
... Infusion of essential but not nonessential amino acids after resistance exercise results in a change from a net negative protein balance to a significantly less negative protein Fig. 4. Relationship between plasma insulin concentrations and rates of protein synthesis in rat gastrocnemius. The graph represents an idealized curve derived from data obtained by Fedele et al. (31). In that study, control, moderately diabetic, and severely diabetic rats were subjected to acute resistance exercise, and 16 h later plasma insulin concentration and muscle protein synthesis were measured. ...
Article
Protein synthesis in skeletal muscle is modulated in response to a variety of stimuli. Two stimuli receiving a great deal of recent attention are increased amino acid availability and exercise. Both of these effectors stimulate protein synthesis in part through activation of translation initiation. However, the full response of translation initiation and protein synthesis to either effector is not observed in the absence of a minimal concentration of insulin. The combination of insulin and either increased amino acid availability or endurance exercise stimulates translation initiation and protein synthesis in part through activation of the ribosomal protein S6 protein kinase S6K1 as well as through enhanced association of eukaryotic initiation factor eIF4G with eIF4E, an event that promotes binding of mRNA to the ribosome. In contrast, insulin in combination with resistance exercise stimulates translation initiation and protein synthesis through enhanced activity of a guanine nucleotide exchange protein referred to as eIF2B. In both cases, the amount of insulin required for the effects is low, and a concentration of the hormone that approximates that observed in fasting animals is sufficient for maximal stimulation. This review summarizes the results of a number of recent studies that have helped to establish our present understanding of the interactions of insulin, amino acids, and exercise in the regulation of protein synthesis in skeletal muscle.
... Although insulin concentrations remain relatively unchanged in the recovery period following resistance exercise (Kraemer et al. 1998;Biolo et al. 1999;Hernandez et al. 2000), a permissive amount of insulin is required to mediate increases in skeletal-muscle protein synthesis. This concept is illustrated by the results of studies utilizing diabetic rats in which animals engaged in resistance exercise (Farrell et al. 1999;Fedele et al. 2000;Kostyak et al. 2001). In such studies rats are operantly conditioned to touch an illuminated bar high on a Plexiglas exercise cage while wearing a weighted vest strapped over the scapulae (Fluckey et al. 1995). ...
... The movement performed during the exercise requires the animals to complete concentric and eccentric muscle contractions of the hindlimb musculature (Fluckey et al. 1995). While moderatelydiabetic animals (fed arterial insulin, approximately 180 pM) exhibit an increase in protein synthesis rate as well as augmented eIF2B activity post-exercise, severely-diabetic (fed arterial insulin, approximately 72 pM) animals exhibit depressed protein synthesis rates and eIF2B activity at rest, which fails to change with subsequent exercise (Farrell et al. 1999;Fedele et al. 2000;Kostyak et al. 2001). In contrast with the results of studies using rodents, elevated insulin concentrations induced through local infusions in human subjects have been shown to stimulate skeletal muscle protein synthesis at rest, but hyperinsulinaemia fails to further augment synthesis after resistance exercise (Biolo et al. 1999). ...
Article
Although insulin, amino acids and exercise individually activate multiple signal transduction pathways in skeletal muscle, one pathway, the phosphatidylinositol 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) signalling pathway, is a target of all three. Activation of the PI3K-mTOR signal transduction pathway results in both acute (i.e. occurring in minutes to hours) and long-term (i.e. occurring in hours to days) up-regulation of protein synthesis through modulation of multiple steps involved in mediating the initiation of mRNA translation and ribosome biogenesis respectively. In addition, changes in gene expression through altered patterns of mRNA translation promote cell growth, which in turn promotes muscle hypertrophy. The focus of the present discussion is to review current knowledge concerning the mechanism(s) through which insulin, amino acids and resistance exercise act to activate the PI3K-mTOR signal transduction pathway and thereby enhance the rate of protein synthesis in muscle.
... It is more difficult to assign a role to insulin in the change from net negative protein balance to positive protein balance. After exercise, insulin seems to be necessary for protein synthesis to occur (11,12,14), yet increased insulin does not stimulate muscle protein synthesis (7). However, elevated insulin after resistance exercise does diminish the increase of muscle protein breakdown in response to exercise (7). ...
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The present study was designed to determine whether consumption of an oral essential amino acid-carbohydrate supplement (EAC) before exercise results in a greater anabolic response than supplementation after resistance exercise. Six healthy human subjects participated in two trials in random order, PRE (EAC consumed immediately before exercise), and POST (EAC consumed immediately after exercise). A primed, continuous infusion of L-[ring-(2)H(5)]phenylalanine, femoral arteriovenous catheterization, and muscle biopsies from the vastus lateralis were used to determine phenylalanine concentrations, enrichments, and net uptake across the leg. Blood and muscle phenylalanine concentrations were increased by approximately 130% after drink consumption in both trials. Amino acid delivery to the leg was increased during exercise and remained elevated for the 2 h after exercise in both trials. Delivery of amino acids (amino acid concentration times blood flow) was significantly greater in PRE than in POST during the exercise bout and in the 1st h after exercise (P < 0.05). Total net phenylalanine uptake across the leg was greater (P = 0.0002) during PRE (209 +/- 42 mg) than during POST (81 +/- 19). Phenylalanine disappearance rate, an indicator of muscle protein synthesis from blood amino acids, increased after EAC consumption in both trials. These results indicate that the response of net muscle protein synthesis to consumption of an EAC solution immediately before resistance exercise is greater than that when the solution is consumed after exercise, primarily because of an increase in muscle protein synthesis as a result of increased delivery of amino acids to the leg.
... It has been our experience that animals do not typically complete 100% of the repetitions at the onset of training after hindlimb suspension because of addition of resistance from the operant conditioning paradigm and/or fatigue during the experimental protocol. Data related to maximum force per set and the force profile curve through all repetitions are shown in Fig. 2. Force produced has been demonstrated to elicit a training effect in adult rats (12) and is similar to forces applied during earlier studies (9,10,14,15). ...
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Hindlimb suspension (HS) results in rapid losses of muscle mass, which may in part be explained by attenuated rates of protein synthesis. Mammalian target of rapamycin (mTOR) regulates protein synthesis and has been implicated as a potential mediator of the muscle mass decrement with HS. This study examined the effect of resistance exercise, a muscle hypertrophy stimulant, on rates of protein synthesis after 4 days of HS in mature male Sprague-Dawley rats. Flywheel resistance exercise (2 sets x 25 repetitions) was conducted on days 2 and 4 of HS (HSRE). Sixteen hours after the last exercise bout, soleus muscles were assessed for in vitro rates of protein synthesis, with and without insulin (signaling agonist) and/or rapamycin (mTOR inhibitor). Results demonstrated that soleus mass was reduced (P < 0.05) with HS, but this loss of mass was not observed (P > 0.05) with HSRE. Muscle protein synthesis was diminished (P < 0.05) with HS, with or without insulin. HSRE also had reduced rates of synthesis without insulin; however, insulin administration yielded higher (P < 0.05) rates in HSRE compared with HS or control. Rapamycin diminished protein synthesis in all groups (P < 0.05), but insulin rescued synthesis rates in HS and HSRE to levels similar to insulin alone for each group, suggesting that alternate signaling pathways develop to increase protein synthesis with HS. These results demonstrate that the capacity for an augmented anabolic response to resistance exercise is maintained after 4 days of HS and is independent of a rapamycin-sensitive pathway.
... This was an expected finding because the rats were fed identically before exercise and this type of exercise does not alter circulating insulin concentrations (9), whereas it does increase insulin secretion from isolated islets of Langerhans (12). These data agree with those of our previous work showing that elevations in protein synthesis after similar exercise are independent of insulin concentrations as long as a low but critical amount of insulin is available in vivo (10). Such a concept is compatible with many studies (11,22) demonstrating that severe diabetes or the absence of insulin in an in situ hindlimb perfusion model (20) markedly reduces basal rates of protein synthesis. ...
Article
Translational control of protein synthesis depends on numerous eukaryotic initiation factors (eIFs) and we have previously shown (Am. J. Physiol. Endocrinol. Metab. 276: E721-E727, 1999) that increases in one factor, eIF2B, are associated with increases in rates of protein synthesis after resistance exercise in rats. In the present study we investigated whether the eIF4E family of initiation factors is also involved with an anabolic response to exercise. Male Sprague-Dawley rats either remained sedentary (n = 6) or performed acute resistance exercise (n = 6), and rates of protein synthesis were assessed in vivo 16 h after the last session of resistance exercise. eIF4E complexed to eIF4G (eIF4E x eIF4G), eIF4E binding protein 1 (4E-BP1) complexed to eIF4E, and phosphorylation state of eIF4E and 4E-BP1 (gamma-form) were assessed in gastrocnemius. Rates of protein synthesis were higher in exercised rats compared with sedentary rats [205 +/- 8 (SE) vs. 164 +/- 5.5 nmol phenylalanine incorporated x g muscle(-1) x h(-1), respectively; P < 0.05]. Arterial plasma insulin concentrations were not different between the two groups. A trend (P = 0.09) for an increase in eIF4E x eIF4G with exercise was noted; however, no statistically significant differences were observed in any of the components of the eIF4E family in response to resistance exercise. These new data, along with our previous report on eIF2B, suggest that the regulation of peptide chain initiation after exercise is more dependent on eIF2B than on the eIF4E system.
... Therefore, the increase in whole body protein synthesis and leucine balance observed in PIϩ compared with PIϪ patients resulted presumably from insulin-independent mechanisms. Accordingly, the dose-response curves for protein synthesis as a function of plasma insulin showed that the maximum insulin effect was obtained at very low insulin levels (12), and an additional increase in plasma insulin did not further increase protein synthesis. In other words, insulin appears to be a rather permissive modulator for protein synthesis (34,36). ...
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The present study was carried out to assess the effects of protease inhibitor (PI) therapy on basal whole body protein metabolism and its response to acute amino acid-glucose infusion in 14 human immunodeficiency virus (HIV)-infected patients. Patients treated with PIs (PI+, 7 patients) or without PIs (PI-, 7 patients) were studied after an overnight fast during a 180-min basal period followed by a 140-min period of amino acid-glucose infusion. Protein metabolism was investigated by a primed constant infusion of l-[1-(13)C]leucine. Dual-energy X-ray absorptiometry for determination of fat-free mass (FFM) and body fat mass measured body composition. In the postabsorptive state, whole body leucine balance was 2.5 times (P < 0.05) less negative in the PI+ than in the PI- group. In HIV-infected patients treated with PIs, the oxidative leucine disposal during an acute amino acid-glucose infusion was lower (0.58 +/- 0.09 vs. 0.81 +/- 0.07 micromol x kg FFM(-1) x min(-1) using plasma [(13)C]leucine enrichment, P = 0.06; or 0.70 +/- 0.10 vs. 0.99 +/- 0.08 micromol x kg FFM(-1) x min(-1) using plasma [(13)C]ketoisocaproic acid enrichment, P = 0.04 in PI+ and PI- groups, respectively) than in patients treated without PIs. Consequently, whole body nonoxidative leucine disposal (an index of protein synthesis) and leucine balance (0.50 +/- 0.10 vs. 0.18 +/- 0.06 micromol x kg FFM x (-1) x min(-1) in PI+ and PI- groups respectively, P < 0.05) were significantly improved during amino acid-glucose infusion in patients treated with PIs. However, whereas the response of whole body protein anabolism to an amino acid-glucose infusion was increased in HIV-infected patients treated with PIs, any improvement in lean body mass was detected.
... However, insulin should not be regarded as a primary regulator of muscle protein synthesis, as insulin exerts only a modest effect on muscle protein synthesis in the absence of elevated amino acid concentrations (29). In rodent models, it has been reported that an increase in circulating plasma insulin concentrations does not further enhance mRNA translation initiation during postexercise recovery (38,52,65). In a recent attempt to assess whether carbohydrate coingestion is required to maximize postexercise muscle protein synthesis, we observed no surplus effect of carbohydrate coingestion on postexercise muscle protein synthesis under conditions in which ample protein is ingested (67). ...
Article
Aging is accompanied by a progressive loss of skeletal muscle mass and strength, leading to the loss of functional capacity and an increased risk of developing chronic metabolic disease. The age-related loss of skeletal muscle mass is attributed to a disruption in the regulation of skeletal muscle protein turnover, resulting in an imbalance between muscle protein synthesis and degradation. As basal (fasting) muscle protein synthesis rates do not seem to differ substantially between the young and elderly, many research groups have started to focus on the muscle protein synthetic response to the main anabolic stimuli, i.e., food intake and physical activity. Recent studies suggest that the muscle protein synthetic response to food intake is blunted in the elderly. The latter is now believed to represent a key factor responsible for the age-related decline in skeletal muscle mass. Physical activity and/or exercise stimulate postexercise muscle protein accretion in both the young and elderly. However, the latter largely depends on the timed administration of amino acids and/or protein before, during, and/or after exercise. Prolonged resistance type exercise training represents an effective therapeutic strategy to augment skeletal muscle mass and improve functional performance in the elderly. The latter shows that the ability of the muscle protein synthetic machinery to respond to anabolic stimuli is preserved up to very old age. Research is warranted to elucidate the interaction between nutrition, exercise, and the skeletal muscle adaptive response. The latter is needed to define more effective strategies that will maximize the therapeutic benefits of lifestyle intervention in the elderly.
... Diabetic muscles become atrophic (34) and are more vulnerable to exercise-induced myofiber damage than nondiabetic muscles (8). The ability to increase muscle mass and the rate of protein synthesis depend on the severity of the disease (10,11). Gene expressions of MLP and MARPs are known to increase and that of myostatin to decrease after physical activity (2,36). ...
Article
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In striated muscle, a sarcomeric noncontractile protein, titin, is proposed to form the backbone of the stress- and strain-sensing structures. We investigated the effects of diabetes, physical training, and their combination on the gene expression of proteins of putative titin stretch-sensing complexes in skeletal and cardiac muscle. Mice were divided into control (C), training (T), streptozotocin-induced diabetic (D), and diabetic training (DT) groups. Training groups performed for 1, 3, or 5 wk of endurance training on a motor-driven treadmill. Muscle samples from T and DT groups together with respective controls were collected 24 h after the last training session. Gene expression of calf muscles (soleus, gastrocnemius, and plantaris) and cardiac muscle were analyzed using microarray and quantitative PCR. Diabetes induced changes in mRNA expression of the proteins of titin stretch-sensing complexes in Z-disc (MLP, myostatin), I-band (CARP, Ankrd2), and M-line (titin kinase signaling). Training alleviated diabetes-induced changes in most affected mRNA levels in skeletal muscle but only one change in cardiac muscle. In conclusion, we showed diabetes-induced changes in mRNA levels of several fiber-type-biased proteins (MLP, myostatin, Ankrd2) in skeletal muscle. These results are consistent with previous observations of diabetes-induced atrophy leading to slower fiber type composition. The ability of exercise to alleviate diabetes-induced changes may indicate slower transition of fiber type.
... Mitochondrial degeneration has been associated with altered protein anabolism (51,52), often seen in T2DM (53,54). Specifically, the obese Zucker rat, a common model of genetic obesity and T2DM, has an inherently greater basal protein anabolism (32,55). ...
Article
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microRNAs (miRs) are linked to various human diseases including Type 2 Diabetes Mellitus (T2DM) and emerging evidence suggests miRs may serve as potential therapeutic targets. Lower miR-16 content is consistent across different models of T2DM; however, the role of miR-16 in muscle metabolic health is still elusive. Therefore, the purpose of this study was to investigate how deletion of miR-16 in mice affects skeletal muscle metabolic health and contractile function in both sexes. This study was conducted using both in vitro (1) and in vivo (2) experiments. (1) We utilized C2C12 myoblasts to test if inhibition or overexpression of miR-16 affected insulin-mediated glucose handling. (2) We generated muscle-specific miR-16 knockout (KO) mice fed a high-fat diet (HFD) to assess how miR-16 content impacts metabolic and contractile properties including glucose tolerance, insulin sensitivity, muscle contractile function, protein anabolism, and mitochondrial network health. (1) Although inhibition of miR-16 induced impaired insulin signaling (p=0.002) and glucose uptake (p=0.014), overexpression of miR-16 did not attenuate lipid overload-induced insulin resistance using the diacylglycerol analog 1‐oleoyl‐2‐acetyl‐sn‐glycerol. (2) miR-16 deletion induced both impaired muscle contractility (p=0.031-0.033), and mitochondrial network health (p=0.008-0.018) in both sexes. However, while males specifically exhibited impaired insulin sensitivity following miR-16 deletion (p=0.030), female KO mice showed pronounced glucose intolerance (p=0.046), corresponding with lower muscle weights (p=0.015), and protein hyperanabolism (p=0.023). Our findings suggest distinct sex differences in muscle adaptation in response to miR-16 deletion and miR-16 may serve as a key regulator for metabolic dysregulation in T2DM.
... . Some of these studies, however, used electrical stimulation or food reward to conduct resistance exercise [Tamaki et al., 1992; Farrell et al., 2000; Klitgaard et al, 1988]. So, it is not clear if the observed hypertrophy is due to exercise training itself or other changes caused by these manipulations. ...
... *Significantly different from t ϭ 0 (P Ͻ 0.05). However, while muscle protein synthesis does not occur in the absence of insulin (15), the impact of increasing insulin concentrations on muscle protein turnover is believed to be more related to a decrease in muscle protein breakdown rather than to stimulation of muscle protein synthesis (18,23). We (2) recently showed that ingestion of protein with carbohydrate during exercise stimulates muscle protein synthesis during resistance type exercise activities. ...
Article
This study investigates the impact of protein coingestion with carbohydrate on muscle protein synthesis during endurance type exercise. Twelve healthy male cyclists were studied during 2 h of fasted rest followed by 2 h of continuous cycling at 55% W(max). During exercise, subjects received either 1.0 g·kg(-1)·h(-1) carbohydrate (CHO) or 0.8 g·kg(-1)·h(-1) carbohydrate with 0.2 g·kg(-1)·h(-1) protein hydrolysate (CHO+PRO). Continuous intravenous infusions with l-[ring-(13)C(6)]phenylalanine and l-[ring-(2)H(2)]tyrosine were applied, and blood and muscle biopsies were collected to assess whole body protein turnover and muscle protein synthesis rates at rest and during exercise conditions. Protein coingestion stimulated whole body protein synthesis and oxidation rates during exercise by 22 ± 3 and 70 ± 17%, respectively (P < 0.01). Whole body protein breakdown rates did not differ between experiments. As a consequence, whole body net protein balance was slightly negative in CHO and positive in the CHO+PRO treatment (-4.9 ± 0.3 vs. 8.0 ± 0.3 μmol Phe·kg(-1)·h(-1), respectively, P < 0.01). Mixed muscle protein fractional synthetic rates (FSR) were higher during exercise compared with resting conditions (0.058 ± 0.006 vs. 0.035 ± 0.006%/h in CHO and 0.070 ± 0.011 vs. 0.038 ± 0.005%/h in the CHO+PRO treatment, respectively, P < 0.05). FSR during exercise did not differ between experiments (P = 0.46). We conclude that muscle protein synthesis is stimulated during continuous endurance type exercise activities when carbohydrate with or without protein is ingested. Protein coingestion does not further increase muscle protein synthesis rates during continuous endurance type exercise.
... In untreated diabetic rats, rates of protein synthesis are reduced in the gastrocnemius muscle via reduced mTOR signaling and lowered activity of eukaryotic initiation factor 2B (eIF2B) (56). Interestingly, chronic resistance exercise does not increase rates of protein synthesis in situations of severe diabetes in rats (92), perhaps because some permissive amount of insulin is needed to activate the pathways of protein synthesis. Recently, aerobic-type exercise has been shown to help normalize rates of muscle protein synthesis and growth in partially pancreatectomized diabetic rats, independent of insulin signaling (277). ...
Article
Physical exercise is firmly incorporated in the management of type 1 diabetes (T1DM), due to multiple recognized beneficial health effects (cardiovascular disease prevention being preeminent). When glycemic values are not excessively low or high at the time of exercise, few absolute contraindications exist; practical guidelines regarding amount, type, and duration of age-appropriate exercise are regularly updated by entities such as the American Diabetes Association and the International Society for Pediatric and Adolescent Diabetes. Practical implementation of exercise regimens, however, may at times be problematic. In the poorly controlled patient, specific structural changes may occur within skeletal muscle fiber, which is considered by some to be a disease-specific myopathy. Further, even in well-controlled patients, several homeostatic mechanisms regulating carbohydrate metabolism often become impaired, causing hypo- or hyperglycemia during and/or after exercise. Some altered responses may be related to inappropriate exogenous insulin administration, but are often also partly caused by the "metabolic memory" of prior glycemic events. In this context, prior hyperglycemia correlates with increased inflammatory and oxidative stress responses, possibly modulating key exercise-associated cardio-protective pathways. Similarly, prior hypoglycemia correlates with impaired glucose counterregulation, resulting in greater likelihood of further hypoglycemia to develop. Additional exercise responses that may be altered in T1DM include growth factor release, which may be especially important in children and adolescents. These multiple alterations in the exercise response should not discourage physical activity in patients with T1DM, but rather should stimulate the quest for the identification of the exercise formats that maximize beneficial health effects. © 2013 American Physiological Society. Compr Physiol 3:1309-1336, 2013.
... Recent evidence also implicates accumulation of phosphatidic acid promoting activation of mTOR in muscle hypertrophy [67]. However, it is critical to note that insulin has its full effect on stimulating muscle protein synthesis at very low "permissive" levels (below ~100 pM) [68], and therefore its function here is often inappropriately overlooked. Justification in many conditions is that researchers are looking for "physiologic ranges" to work with insulin concentrations in experiments, however in the diabetic state ranges of insulin concentration nearing zero may well be observed. ...
Article
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Background: Prevalence of Type 2 Diabetes Mellitus (T2DM) has reached pandemic levels in the Western societies. T2DM begins with the development of peripheral insulin resistance which prior research suggests may commonly originate within the skeletal muscle. A number of mechanisms have been proposed for the development of muscle insulin resistance including those of classical glucose handling, and also other cellular derangements observed in this disease which include mitochondrial degeneration, alterations in muscle protein turnover and early evidences for dysregulation of the microRNAs. The purpose of the current review is to examine the current findings on these latter aspects of mitochondrial maintenance, protein turnover and microRNA dysregulation along with the potential implications for these derangements in the development of insulin resistance and hence T2DM. We summarize multiple evidences for the degeneration of mitochondria and known elements of the processes regulating mitochondrial quality. Subsequently, we examine current findings of the alterations in muscle protein synthesis and autophagic protein degradation in T2DM and potential feedback of these systems onto canonical insulin signaling. Finally, evidences have emerged for the dysregulation of microRNAs in muscle insulin resistance. Of note early data point to several microRNAs altered by the insulin resistant state which exhibit relations to classic insulin signaling and the other processes discussed here. Conclusion: Considering that T2DM may be initiated with muscle insulin resistance, improved understanding of the dysregulation of these metabolic parameters of skeletal muscle in the pathogenesis of T2DM may be key to developing efficacious therapeutic modalities to prevent and treat this condition.
... These findings are consistent with in vitro and in vivo studies reporting increased protein breakdown in skeletal muscle from diabetic rats (19,78,129,130,(156)(157)(158). However, unlike in humans with type 1 diabetes, rates of synthesis of muscle protein in rats with alloxan-and streptozotocin-induced diabetes (20,(158)(159)(160)(161)(162) and in eviscerated dogs (163) are consistently decreased compared with rates in control animals. In addition, a recent study demonstrated a significant decrease in protein synthesis in rat muscle during diazoxideinduced postprandial insulin deficiency (56). ...
... The existence of a distinct DCM in human patients is reasonably well established, and has been extensively reviewed in recent papers (Poornima et al. 2006, Boudina et al. 2010. Because insulin signaling is generally required for muscle protein synthesis after exercise (Farrell et al. 1998, Fedele et al. 2000 and is absolutely necessary for physiological hypertrophy in cardiomyocytes (Kim et al. 2008, Ikeda et al. 2009), it is possible that diabetes-associated hypertrophy is completely distinct from other types of pathological hypertrophy. For example, it has been proposed that DCM results from cardiomyocyte atrophy and apoptosis and cardiac fibrosis, rather than cardiomyocyte hypertrophy (Poornima et al. 2006). ...
Article
Aerobic exercise training (AET) attenuates or reverses pathological cardiac remodeling after insults such as chronic hypertension and myocardial infarction. The phenotype of the pathologically hypertrophied heart depends on the insult; therefore, it is likely that distinct types of pathological hypertrophy require different exercise regimens. However, the mechanisms by which AET improves the structure and function of the pathologically hypertrophied heart are not well understood, and exercise research uses highly inconsistent exercise regimens in diverse patient populations. There is a clear need for systematic research to identify precise exercise prescriptions for different conditions of pathological hypertrophy. Therefore, this review synthesizes existing evidence for the distinct mechanisms by which AET benefits the heart in different pathological hypertrophy conditions, suggests strategic exercise prescriptions for these conditions, and highlights areas for future research. Copyright © 2015. Published by Elsevier Inc.
... Although an increase in plasma insulin concentration may not be required for the exercise-induced increase in muscle protein synthesis, evidence from both rodent and human studies suggests that there is a threshold for the hormone whereby plasma concentrations below a certain value result in protein synthesis being refractory to the stimulatory effect of resistance exercise on muscle protein synthesis. For example, muscle protein synthesis is increased after exercise in rats with moderate type 1 diabetes (fed arterial insulin of w180 pmol/L) but not in severely diabetic rats (fed arterial insulin of w72 pmol/L) (42,43). Interestingly, the insulin concentration that is permissive for exercise-induced protein synthesis is at, or below, that observed in a control, fasted animal, suggesting that, in most cases, in-sulin is not limiting for exercise-induced stimulation of muscle protein synthesis. ...
Article
This review focuses on anabolic signaling pathways through which insulin, amino acids, and resistance exercise act to regulate the protein kinase complex referred to as mechanistic target of rapamycin complex (mTORC) 1. Initially, individual pathways through which the 3 anabolic signals act to modulate mTORC1 signaling will be discussed, followed by a summation of evidence showing an additive effect of the regulators. The emphasis will be on mTORC1 signaling in skeletal muscle and its contribution to modulation of rates of protein synthesis. In addition, results from studies using cells in culture will be used to provide a more complete picture of the molecular details of the individual pathways.
... The possibility exists that these body composition changes could be attenuated in elderly individuals consuming an adequate protein diet, and has recently been tested in a randomized trial in which skeletal muscle mass did not change but physical performance improved in 65 frail older adults receiving 15 g of protein at breakfast and lunch (43). Nonetheless it is known that higher fat mass is associated with increased insulin resistance which is a precursor to frailty (44) and is associated with muscle wasting and impaired protein metabolism (45). Older, non-diabetic individuals with evidence of insulin resistance have poor quadriceps muscle performance (46). ...
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Objectives: Objectives Sarcopenia, the involuntary loss of skeletal muscle with age, affects up to onequarter of older adults. Evidence indicates a positive association between dietary protein intake and lean muscle mass and strength among older persons, but information on dietary protein’s effect on physical performance in older adults has received less attention. Design Cross-sectional observational analysis of the relationship of dietary protein on body composition and physical performance. Setting Clinical research center. Participants 387 healthy women aged 60–90 years (mean 72.7 ± 7.0 y). Measurements Measures included body composition (fat-free mass, appendicular skeletal mass and fat mass) via dual x-ray absorptiometry (DXA), physical performance (Physical Performance Test [PPT] and Short Physical Performance Battery [SPPB]), handgrip strength, Physical Activity Scale in the Elderly (PASE), quality of life measure (SF-8), falls, fractures, nutrient and macromolecule intake (four-day food record). Independent samples t-tests determined mean differences between the above or below RDA protein groups. Statistical Analysis Analysis of covariance was used to control for body mass index (BMI) between groups when assessing physical performance, physical activity and health-related quality of life. Results The subjects consumed an average of 72.2 g protein/day representing 1.1 g protein/kg body weight/day. Subjects were categorized as below the recommended daily allowance (RDA) for protein (defined as less than 0.8 g protein/kg) or at or above the RDA (equal to or higher than 0.8 g protein/kg). Ninety-seven subjects (25%) were in the low protein group, and 290 (75%) were in the higher protein group. Women in the higher protein group had lower body mass, including fat and lean mass, and fat-to-lean ratio than those in the lower-protein group (p <0.001). Composite scores of upper and lower extremity strength were impaired in the group with low protein intake; SPPB score was 9.9±1.9 compared to 10.6±1.6 in those with higher protein intake and PPT was 19.8± 2.9 compared to 20.9±2.1 in the low and higher protein groups, respectively. The results were attenuated by correction for BMI, but remained significant. The physical component of the SF-8 was also lower in the low protein group but did not remain significant when controlling for BMI. No significant differences were found in hand grip strength or reported physical activity. Conclusion Healthy, older postmenopausal women consumed, on average, 1.1 g/kg/d protein, although 25% consumed less than the RDA. Those in the low protein group had higher body fat and fat-to-lean ratio than those who consumed the higher protein diet. Upper and lower extremity function was impaired in those who consumed a low protein diet compared to those with a higher protein intake. Protein intake should be considered when evaluating the multi-factorial loss of physical function in older women.
... In diabetic muscle, there is a reduced protein synthesis rate. In mild or moderate diabetes, exercise training can induce a higher rate of protein synthesis (348,349) whereas this could not be observed in severe diabetes (350). A previous animal study showed that exercise caused an increase in muscle mass along with augmented protein synthesis (351). ...
Article
Diabetes mellitus (DM) is an epidemic medical challenge that threatens the health and life quality of people worldwide. DM impairs metabolic, neural and vascular function and thus has profound impacts on different systems and organs in the body. Though continuous endeavour has been made to study its etiology and mechanisms, no cure for DM has yet been found. DM development may be multi-factorial. The skeletal muscle is one of the most important systems, involved in the development of DM, and affected by insulin. DM induces diverse functional, metabolic, and structural changes in the skeletal muscle. DM reduces the functional capacity of skeletal muscle leading to muscle weakness, causes metabolic disturbance characterized by reduced cellular glucose uptake and fatty acid oxidation, and structural changes with muscle atrophy, augmented lipid deposition, decreased mitochondria as well as muscle fiber transformation. DM-induced changes in the skeletal muscle seem to be dependent on types and severity of DM as well as on muscle fibers. The central mechanism underlying these changes is impaired insulin action in the skeletal muscle.
... Thus this altered response of gastrocnemius protein synthesis in adults fed the AA/protein-free meal could be related to this lower plasma insulin. However, based on the dose−response curves for muscle protein synthesis as a function of plasma insulin (Garlick & Grant, 1988; Millward et al. 1988; Davis et al. 1998; Yoshizawa et al. 1998; Fedele et al. 2000; Balage et al. 2001; O'Connor et al. 2003), the insulin levels in adult rats fed the AA/protein-free meal seemed to be high enough to elicit a maximum response of protein synthesis and that any additional increase in plasma insulin would not further increase protein synthesis. Instead, it may be inferred that a portion of postprandial muscle protein synthesis was not driven by insulin in adult rats but by amino acids as previously observed in young rats (Balage et al. 2001 ). ...
Article
The potential roles of insulin and dietary amino acids in the regulation of skeletal muscle protein synthesis were examined in adult and old rats. Animals were fed over 1 h with either a 25% or a 0% amino acid/protein meal. In each nutritional condition, postprandial insulin secretion was either maintained or blocked with diazoxide injections. Protein synthesis in gastrocnemius and soleus muscles was assessed in vivo using the flooding dose method. Insulin suppression decreased protein synthesis in both muscles irrespective of the nutritional condition and age of the rats. Moreover, reduced insulinaemia was associated with 4E-BP1 dephosphorylation, enhanced assembly of the 4E-BP1-eIF4E inactive complex and hypophosphorylation of eIF4E, p70S6k and protein kinase B, key intermediates in the regulation of translation initiation and protein synthesis. Old rats did not differ from adult rats. The lack of amino acids in the meal of insulin-suppressed rats did not result in any additional decrease in protein synthesis. In the presence of insulin secretion, dietary amino acid suppression significantly decreased gastrocnemius protein synthesis in adult but not in old rats. Amino acid suppression was associated with reduced phosphorylation of 4E-BP1 and p70S6k in adults. Along with protein synthesis, only the inhibition of p70S6k phosphorylation was abolished in old rats. We concluded that insulin is required for the regulation of muscle protein synthesis irrespective of age and that the effect of dietary amino acids is blunted in old rats.
... As explained above, insulin should not be regarded as a primary regulator of muscle protein synthesis as insulin exerts only a modest effect on muscle protein synthesis in the absence of elevated amino acid concentrations (Cuthbertson et al. 2005). In rodent models, it has been reported that an increase in circulating plasma insulin concentrations does not further enhance mRNA translation initiation during post-exercise recovery (Fedele et al. 2000;Gautsch et al. 1998;Kimball et al. 2002). In a recent attempt to assess whether carbohydrate co-ingestion is required to maximize post-exercise muscle protein synthesis, we observed no surplus effect of carbohydrate co-ingestion on post-exercise muscle protein synthesis under conditions where ample protein is ingested (Koopman et al. 2007a). ...
Chapter
Aging is accompanied by a progressive loss of skeletal muscle mass and strength, leading to the loss of functional capacity and an increased risk of developing chronic metabolic diseases such as diabetes. The age-related loss of skeletal muscle mass must be due to a chronic disruption in the balance between muscle protein synthesis and degradation. In addition, it has been suggested that a decline in the number of satellite cells (SC) and/or their ability to become activated can contribute to the development of sarcopenia. In healthy active older individuals, there does not seem to be a disturbance in muscle protein metabolism in the fasted (basal) state. Consequently, it has been proposed that older muscle has a deficit in the ability to regulate the protein synthetic response to anabolic stimuli, such as food intake and physical activity. Indeed, recent data suggest that the dose-response relationship between myofibrillar protein synthesis and the availability of essential amino acids and/or resistance exercise intensity is shifted down and to the right in elderly humans. This so-called anabolic resistance is now believed to represent a key factor responsible for the age-related decline in skeletal muscle mass. Although physical activity and/or exercise stimulate muscle protein synthesis in both the young and elderly, the hypertrophic response largely depends on the timed administration of amino acids and/or protein prior to, during, and/or after exercise. However, prolonged resistance type exercise training has been shown to be effective as a therapeutic strategy to augment skeletal muscle mass, increase muscle SC content, and improve functional performance in the elderly. The latter shows that the ability to increase muscle mass is preserved up to very old age. More research is warranted to elucidate the interaction between nutrition, exercise and the skeletal muscle adaptive response. The latter is needed to define more effective strategies that will maximize the therapeutic benefits of lifestyle intervention in the elderly. KeywordsSarcopenia-Nutrition-Exercise training-Muscle hypertrophy
Article
Rosa JS, Galassetti PR. Altered molecular adaptation to exercise in children with type 1 diabetes: beyond hypoglycemia.Pediatric Diabetes 2009: 10: 213–226.
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The contribution of mammalian target of rapamycin (mTOR) signaling to the resistance exercise-induced stimulation of skeletal muscle protein synthesis was assessed by administering rapamycin to Sprague-Dawley rats 2 h prior to a bout of resistance exercise. Animals were sacrificed 16 h postexercise, and gastrocnemius protein synthesis, mTOR signaling, and biomarkers of translation initiation were assessed. Exercise stimulated the rate of protein synthesis; however, this effect was prevented by pretreatment with rapamycin. The stimulation of protein synthesis was mediated by an increase in translation initiation, since exercise caused an increase in polysome aggregation that was abrogated by rapamycin administration. Taken together, the data suggest that the effect of rapamycin was not mediated by reduced phosphorylation of eukaryotic initiation factor 4E (eIF4E) binding protein 1 (BP1), because exercise did not cause a significant change in 4E-BP1(Thr-70) phosphorylation, 4E-BP1-eIF4E association, or eIF4F complex assembly concomitant with increased protein synthetic rates. Alternatively, there was a rapamycin-sensitive decrease in relative eIF2Bepsilon(Ser-535) phosphorylation that was explained by a significant increase in the expression of eIF2Bepsilon protein. The proportion of eIF2Bepsilon mRNA in polysomes was increased following exercise, an effect that was prevented by rapamycin treatment, suggesting that the increase in eIF2Bepsilon protein expression was mediated by an mTOR-dependent increase in translation of the mRNA encoding the protein. The increase in eIF2Bepsilon mRNA translation and protein abundance occurred independent of similar changes in other eIF2B subunits. These data suggest a novel link between mTOR signaling and eIF2Bepsilon mRNA translation that could contribute to the stimulation of protein synthesis following acute resistance exercise.
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To examine the role of both endurance exercise and nutrient supplementation on the activation of mRNA translation signaling pathways postexercise, rats were subjected to a 3-h swimming protocol. Immediately following exercise, the rats were provided with a solution containing either 23.7% wt/vol carbohydrates (CHO), 7.9% wt/vol protein (Pro), 31.6% wt/vol (23.7% wt/vol CHO + 7.9% wt/vol Pro) carbohydrates and Pro (CP), or a placebo (EX). The rats were then killed at 0, 30, and 90 min postexercise, and phosphorylation states of mammalian target of rapamycin (mTOR), ribosomal S6 kinase (p70(S6K)), ribosomal protein S6 (rpS6), and 4E-binding protein 1 (4E-BP1), were analyzed by immunoblot analysis in the red and white quadriceps muscle. Results demonstrated that rat groups provided with any of the three nutritional supplements (CHO, Pro, CP) transiently increased the phosphorylation states of mTOR, 4E-BP1, rpS6, and p70(S6K) compared with EX rats. Although CHO, Pro, and CP supplements phosphorylated mTOR and p70(S6K) after exercise, only CP elevated the phosphorylation of rpS6 above all other supplements 30 min postexercise and 4E-BP1 30 and 90 min postexercise. Furthermore, the phosphorylation states of 4E-BP1 (r(2) = 0.7942) and rpS6 (r(2) = 0.760) were highly correlated to insulin concentrations in each group. These results suggest that CP supplementation may be most effective in activating the mTOR-dependent signaling pathway in the postprandial state postexercise, and that there is a strong relationship between the insulin concentration and the activation of enzymes critical for mRNA translation.
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Resistance exercise is a powerful stimulus to augment muscle protein anabolism, as it can improve the balance between muscle protein synthesis and breakdown. However, the intake of food during post-exercise recovery is necessary for hypertrophy to occur. Therefore, athletes need to ingest protein following exercise to attain a positive protein balance and maximise their skeletal muscle adaptive response. The interaction between exercise and nutrition is not only important for athletes, but is also of important clinical relevance in the elderly. Exercise interventions combined with specific nutritional modulation provide an effective strategy to counteract or reduce the loss of skeletal muscle mass with aging.
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This study examined the effect of a specific acute postprandial leucine deficiency on skeletal muscle protein synthesis in growing and adult rats. Because the anabolic action of dietary leucine supplementation is controversial, except during aging, we hypothesized that the maximum leucine effect might be already achieved for a normal postprandial rise of leucine. Preventing this rise during the 1- to 3-h period after feeding may reveal the leucine regulation. On the day of the experiment, rats were fasted (postabsorptive, PA group) or fed for 1 h a control meal (postprandial, control, PP group) or a leucine-poor meal (postprandial, PP-Leu group). Muscle protein synthesis was assessed in vivo, over the 1- to 3-h period after meal distribution, using the flooding dose method (L-1-(13)C phenylalanine). As expected, the postprandial increase in plasma free leucine was specifically abolished after feeding the leucine-poor meal, whereas all the other plasma free amino acids were roughly at normal postprandial levels. Plasma insulin increased after feeding in young rats but was constant in adult rats. Plasma insulin was similar whatever dietary leucine levels. Rates of muscle protein synthesis were stimulated by feeding in gastrocnemius and soleus muscles from young rats but only in gastrocnemius muscles from adult rats. The PP-Leu group did not differ from the control PP group regarding muscle protein synthesis. The rise in plasma free leucine is not required for the stimulation of muscle protein synthesis during the 1- to 3-h period after feeding young and adult rats, as previously observed in old rats.
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Metabolic risk factors associated with insulin resistance syndrome may attenuate augmentations in skeletal muscle protein anabolism following contractile activity. The purpose of this study was to investigate whether or not the anabolic response, as defined by an increase in cumulative fractional protein synthesis rates (24-h FSR) following resistance exercise (RE), is blunted in skeletal muscle of a well-established rodent model of insulin resistance syndrome. Four-month-old lean (Fa/?) and obese (fa/fa) Zucker rats engaged in four lower body RE sessions over 8 days, with the last bout occurring 16 h prior to muscle harvest. A priming dose of deuterium oxide ((2)H(2)O) and (2)H(2)O-enriched drinking water were administered 24 h prior to euthanization for assessment of cumulative FSR. Fractional synthesis rates of mixed (-5%), mitochondrial (-1%), and cytosolic (+15%), but not myofibrillar, proteins (-16%, P = 0.012) were normal or elevated in gastrocnemius muscle of unexercised obese rats. No statistical differences were found in the anabolic response of cytosolic and myofibrillar subfractions between phenotypes, but obese rats were not able to augment 24-h FSR of mitochondria to the same extent as lean rats following RE (+14% vs. +28%, respectively). We conclude that the mature obese Zucker rat exhibits a mild, myofibrillar-specific suppression in basal FSR and a blunted mitochondrial response to contractile activity in mixed gastrocnemius muscle. These findings underscore the importance of assessing synthesis rates of specific myocellular subfractions to fully elucidate perturbations in basal protein turnover rates and differential adaptations to exercise stimuli in metabolic disease.
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Eukaryotic initiation factor (eIF)2B is a guanine nucleotide exchange factor (GEF) for a second initiation factor, eIF2. During the initiation of messenger RNA (mRNA) translation, eIF2 binds Guanosine-5'-triphosphate (GTP) and initiator methionyl- transfer RNA (tRNAi) (met-tRNAi) and the resulting ternary complex subsequently binds to the 40S ribosomal subunit. Alterations in ternary complex formation due to changes in eIF2B GEF activity not only produce alterations in global rates of protein synthesis but also cause specific changes in the translation of mRNAs encoding certain proteins, such as the transcription factors general control nonrepressed 2 (GCN2) and activating transcription factor 4 (ATF4). Studies have implicated changes in eIF2B GEF activity in alterations in protein synthesis in a number of pathophysiological and physiological conditions, such as diabetes, cancer, sepsis, vanishing white matter disease, and resistance exercise. The chapter discusses the state of knowledge concerning the function of eIF2B and its role in regulating mRNA translation.
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Diabetes induces changes in the structure and function of the extracellular matrix (ECM) in many tissues. We investigated the effects of diabetes, physical training, and their combination on the gene expression of ECM proteins in skeletal muscle. Mice were divided to control (C), training (T), streptozotocin-induced diabetic (D), and diabetic training (DT) groups. Training groups (T, DT) performed 1, 3, or 5 wk of endurance training on a treadmill. Gene expression of calf muscles was analyzed using microarray and quantitative PCR. Training group samples were collected 24 h after the last training session. Diabetes affected the gene expression of several collagens (types I, III, IV, V, VI, and XV), some noncollagenous glycoproteins, and proteoglycans (e.g., elastin, thrombospondin-1, laminin-2, decorin). Reduced gene expression of collagens in diabetic skeletal muscle was partially attenuated as a result of physical training. In diabetes, mRNA expression of the basement membrane (BM) collagens decreased and that of noncollagenous glycoproteins increased. This may change the structure of the BM in a less collagenous direction and affect its properties.
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An infusion of amino acids stimulates heat production in skeletal muscle and then attenuates the anesthesia-induced hypothermia. However, in a clinical setting, some patients have atrophic skeletal muscle caused by various factors. The present study was therefore conducted to investigate the effect of amino acids on the anesthesia-induced hypothermia in the state of muscle atrophy. As the muscle atrophy model, Sprague-Dawley rats were subjected to hindlimb immobilization for 2 wk. Normal rats and atrophy model rats were randomly assigned to one of the two treatment groups: saline or amino acids (n=8 for each group). Test solutions were administered intravenously to the rats under sevoflurane anesthesia for 180 min, and the rectal temperature was measured. Plasma samples were collected for measurement of insulin, blood glucose, and free amino acids. The rectal temperature was significantly higher in the normal-amino acid group than in the muscle atrophy-amino acid group from 75 to 180 min. The plasma insulin level was significantly higher in the rats given amino acids than in the rats given saline in both normal and model groups. In the rats given amino acids, plasma total free amino acid concentration was higher in the model group than in the normal group. These results indicate that skeletal muscle plays an important role in changes in body temperature during anesthesia and the effect of amino acids on anesthesia-induced hypothermia decreases in the muscle atrophy state. In addition, intravenous amino acids administration during anesthesia induces an increase in the plasma insulin level.
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The purpose of the study was to investigate simultaneous effects of energy balance, caloric intake, and the hormonal anabolic-catabolic balance in bodybuilders prior to competition. Fourteen male bodybuilders took part in an 11-week energy-restricted period to reduce body fat. The subjects were divided into the energy-restricted group (ERG) (n = 7), who were preparing for the competition, or the control group (CG) (n = 7) who continued to train regularly and did not change their dietary or training pattern. Participants were tested at 11 weeks (T1), 5 weeks (T2), and 3 days (T3) before competition for diet, body composition, and fasting hormonal assessment. Body mass and body fat percentage of ERG were significantly (p < 0.05) decreased during the study period. In ERG, insulinlike growth factor-1 (IGF-1) and insulin decreased significantly during the 11-week weight-reduction period (p < 0.05). Testosterone was decreased only from week 11 to week 5 (from 20.3 +/- 6.0 to 18.0 +/- 6.8 nmol/L). Changes in IGF-I concentration were significantly related to changes in insulin (r = 0.741), fat mass (r = 0.705), lean body mass (r = 0.696), and body mass (r = 0.652). Changes in insulin concentrations were significantly related to changes in fat mass (r = 0.630) and lean body mass (r = 0.725). These data indicate that severe energy restriction to extremely low body energy reserves decreases significantly the concentrations of 3 anabolic pathways despite high protein intake. Monitoring of insulin and IGF-1 concentration is suggested to prevent losses in muscle mass in energy-restricted conditions. Other nutritional strategies might be needed to prevent possible catabolic effect during preparation of bodybuilders to competition.
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The aim of the present study was to evaluate the influence of moderate physical exercise on muscle, liver, and plasma insulin-like growth factor–1 (IGF-1) levels and to analyze certain protein and carbohydrate metabolism patterns related to this physiological state. Adult male rats were subjected to physical exercise consisting of swimming for a period of 6 weeks. The animals were divided into two groups, a trained (T) and a sedentary (S) group, with both groups receiving a 12% casein diet. The following parameters were analyzed: body weight, food intake, muscle and liver weight, plasma IGF-1, glucose, insulin, urea and creatinine, muscle protein, glycogen, and muscle and liver IGF-1. Group T showed lower body weight, food intake, and glucose and insulin levels than group S. The other parameters did not differ between groups. These results suggest that moderate physical exercise does not alter IGF-1 levels and therefore cannot be considered a regulating factor of IGF-1 under these experimental conditions.
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For the competitive athlete and the serious recreational athlete, nutritional supplementation can have a positive effect on training and on performance. There are many fad supplements on the market, and many that have come and gone. However, two nutrients have withstood the test of time and many tests in research laboratories around the world, and they continue to have positive training- and performance-enhancing effects. Carbohydrates are commonly supplemented to improve energy availability and to replace valuable muscle and liver glycogen stores. Protein supplementation usually is associated with building muscle tissue.
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Infusion of physiological levels of insulin and/or amino acids reproduces the feeding-induced stimulation of muscle protein synthesis in neonates. To determine whether insulin and amino acids independently stimulate skeletal muscle protein synthesis in neonates, insulin secretion was blocked with somatostatin in fasted 7-day-old pigs (n = 8-12/group) while glucose and glucagon were maintained at fasting levels and insulin was infused to simulate either less than fasting, fasting, intermediate, or fed insulin levels. At each dose of insulin, amino acids were clamped at either the fasting or fed level; at the highest insulin dose, amino acids were also reduced to less than fasting levels. Skeletal muscle protein synthesis was measured using a flooding dose of l-[4-(3)H]phenylalanine. Hyperinsulinemia increased protein synthesis in skeletal muscle during hypoaminoacidemia and euaminoacidemia. Hyperaminoacidemia increased muscle protein synthesis during hypoinsulinemia and euinsulinemia. There was a dose-response effect of both insulin and amino acids on muscle protein synthesis. At each insulin dose, hyperaminoacidemia increased muscle protein synthesis. The effects of insulin and amino acids on muscle protein synthesis were largely additive until maximal rates of protein synthesis were achieved. Amino acids enhanced basal protein synthesis rates but did not enhance the sensitivity or responsiveness of muscle protein synthesis to insulin. The results suggest that insulin and amino acids independently stimulate protein synthesis in skeletal muscle of the neonate.
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The purpose of this study was to evaluate the effects of hyperglycemia on skeletal muscle recovery following disuse-induced muscle atrophy in rats. Wistar rats were grouped as streptozotocin-induced diabetic rats and non-diabetic rats. Both ankle joints of each rat were immobilized to induce atrophy of the gastrocnemius muscles. After two weeks of immobilization and an additional two weeks of recovery, tail blood and gastrocnemius muscles were isolated. Serial cross sections of muscles were stained for myosin ATPase (pH 4.5) and alkaline phosphatase activity. Serum insulin and muscle insulin-like growth factor-1 (IGF-1) levels were also measured. Serum insulin levels were significantly reduced in the diabetic rats compared to the non-diabetic controls. The diameters of type I, IIa, and IIb myofibers and capillary-to-myofiber ratio in the isolated muscle tissue were decreased after immobilization in both treatments. During the recovery period, these parameters were restored in the non-diabetic rats, but not in the diabetic rats. In addition, muscle IGF-1 levels after recovery increased significantly in the non-diabetic rats, but not in the diabetic rats. We conclude that decreased levels of insulin and IGF-1 and impairment of angiogenesis associated with diabetes might be partly responsible for the inhibition of regrowth in diabetic muscle.
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For over 10 years, we have known that the activation of the mammalian target of rapamycin complex 1 (mTORC1) has correlated with the increase in skeletal muscle size and strength that occurs following resistance exercise. Initial cell culture and rodent models of muscle growth demonstrated that the activation of mTORC1 is common to hypertrophy induced by growth factors and increased loading. The further observation that high loads increased the local production of growth factors led to the paradigm that resistance exercise stimulates the autocrine production of factors that act on membrane receptors to activate mTORC1, and this results in skeletal muscle hypertrophy. Over the last few years, there has been a paradigm shift. From both human and rodent studies, it has become clear that the phenotypic and molecular responses to resistance exercise occur in a growth factor-independent manner. Although the mechanism of load-induced mTORC1 activation remains to be determined, it is clear that it does not require classical growth factor signaling.
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Sarcopenia, the age-related loss of muscle mass and strength, is a fundamental cause of frailty, functional decline and disability. In the year 2000, $18.5 billion in health care costs were directly attributable to sarcopenia. This economic burden will increase dramatically as the elderly population grows over the next decade. The primary causes of sarcopenia include a sedentary lifestyle and malnutrition. While resistance training appears to be a promising intervention, older individuals exhibit a blunted hypertrophic response to exercise stimuli. It has been posited that this decrement in regenerative capacity may be due to the loss of postprandial anabolism as well as an increase in reactive oxygen species. As such, a combination of resistance training and nutritional interventions may be a promising candidate in combating sarcopenia. Nevertheless, the mechanisms by which the manipulation of dietary variables may improve the sarcopenic condition are not well understood. To address this gap in extant knowledge, this review will examine the effects of protein, amino acid and/or antioxidant intake on sarcopenia both at rest and following resistance training exercise.
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The aim of the present study was to evaluate the influence of moderate physical exercise on muscle, liver, and plasma insulin-like growth factor–1 (IGF-1) levels and to analyze certain protein and carbohydrate metabolism patterns related to this physiological state. Adult male rats were subjected to physical exercise consisting of swimming for a period of 6 weeks. The animals were divided into two groups, a trained (T) and a sedentary (S) group, with both groups receiving a 12% casein diet. The following parameters were analyzed: body weight, food intake, muscle and liver weight, plasma IGF-1, glucose, insulin, urea and creatinine, muscle protein, glycogen, and muscle and liver IGF-1. Group T showed lower body weight, food intake, and glucose and insulin levels than group S. The other parameters did not differ between groups. These results suggest that moderate physical exercise does not alter IGF-1 levels and therefore cannot be considered a regulating factor of IGF-1 under these experimental conditions.
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IntroductionInsulin action in skeletal muscleExercise mimics insulin action in skeletal muscleInsulin and exercise stimulate MAPK signalingClinical implicationsSummaryReferences
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The respective roles of insulin and amino acids in regulation of skeletal muscle protein synthesis and degradation after feeding were examined in rats fasted for 17 h and refed over 1 h with either a 25 or a 0% amino acid/protein meal. In each nutritional condition, postprandial insulin secretion was either maintained (control groups: C(25) and C(0)) or blocked with diazoxide injections (diazoxide groups: DZ(25) and DZ(0)). Muscle protein metabolism was examined in vitro in epitrochlearis muscles. Only feeding the 25% amino acid/protein meal in the presence of increased plasma insulin concentration (C(25) group) stimulated protein synthesis and inhibited proteolysis in skeletal muscle compared with the postabsorptive state. The stimulation of protein synthesis was associated with increased phosphorylation of eukaryotic initiation factor (eIF)4E binding protein-1 (4E-BP1), reduced binding of eIF4E to 4E-BP1, and increased assembly of the active eIF4E. eIF4G complex. The p70 S6 kinase (p70(S6k)) was also hyperphosphorylated in response to the 25% amino acid/protein meal. Acute postprandial insulin deficiency induced by diazoxide injections totally abolished these effects. Feeding the 0% amino acid/protein meal with or without postprandial insulin deficiency did not stimulate muscle protein synthesis, reduce proteolysis, or regulate initiation factors and p70(S6k) compared with fasted rats. Taken together, our results suggest that both insulin and amino acids are required to stimulate protein synthesis, inhibit protein degradation, and regulate the interactions between eIF4E and 4E-BP1 or eIF4G in response to feeding.
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We have examined the effects of infusing recombinant human growth hormone (hGH), insulin-like growth factor-I (IGF-I), the truncated IGF-I analogue, des(1-3)IGF-I, and insulin over a 7-day period in streptozotocin-induced diabetic rats. IGF-I at a dose of 1.05 or 1.08 mg/kg per day in two experiments increased body weight and nitrogen retention above those of vehicle-infused controls to about 30% of the improvement achieved with 25 or 30 units of insulin/kg per day, but only in the second experiment were the differences statistically significant (P less than 0.05). A 2.5-fold higher IGF-I dose, or des(1-3)IGF-I at 1.08 mg/kg per day, gave effects that were approx. 70% of those obtained with insulin. hGH at 1.38 mg/kg per day was not effective. The IGF peptides, unlike insulin, did not ameliorate the diabetic glucosuria. The improvements in nitrogen balance could be accounted for in part by increases in muscle protein synthesis. Muscle protein breakdown, as assessed by 3-methylhistidine excretion, was inhibited by insulin, but not by the IGF peptides. Carcass fat increased substantially following insulin administration. This did not occur with the IGF peptides, suggesting that IGF predominantly stimulates the growth of lean tissue. IGF-I concentrations and IGF-I-binding proteins in plasma were increased by IGF-I, especially at the higher dose, whereas hGH produced only a transient increase in IGF-I. Des(1-3)IGF-I induced binding proteins, but had only a slight effect on measured IGF-I concentrations. We conclude that IGF peptides stimulate muscle protein synthesis and improve nitrogen balance in diabetes without obviously influencing the abnormal carbohydrate metabolism. Moreover, des(1-3)IGF-I is at least as potent as the full-length IGF-I.
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I. PHYSIOLOGICAL SIGNIFICANCE A. Significance of Adaptations to Environment The ability of an animal to adapt to repeated bouts of physical exercise over a period of weeks such that exercise capacity is improved is termed physical training. Some of the inherent mechanisms that are crucial for adaptation to changes in the environment likely carry over to some of the adaptations occurring because of physical training. Prosser (316, 317) has written that cellular, organ, and systemic alterations that favor survival of an animal to an environmental change are said to be adaptive. Physical exercise, like environmental change, disrupts the milieu interieur. Fisher (103) has emphasized that biochemical and physiological adaptations to a changed environment or physiological stimulus fall into two categories based on their duration . Cellular, organ, or systemic alterations that occur on the same time scale as a single exercise bout are said to be acute exercise responses. On the other hand, changes in cells, organs, or systems that persist for appreciable periods after or as a consequence of physical training are said to be exercise adaptations. A function of exercise adaptation seems to be to minimize disruption of homeostasis during an exercise bout. It is this better maintenance of the milieu interieur by the exercise adaptations that favors the functional effectiveness of the animal beyond the resting state. Less disruption in homeostasis permits the animal or human to undergo physical work for longer durations at the same absolute power before fatigue. This review considers molecular and cellular responses to exercise that may signal molecular and cellular adaptations during physical training. B. Significance of Adaptations to Exercise This review is organized to use some of the known causes of fatigue during physical exercise as links between molecular and cellular changes that occur as a result of physical training and the chronic adaptations that are characteristic of physical training. One can speculate that adaptations that improved an animal's work capacity enhanced its survival. The genetic ability to alter exercise performance through physical training has not been lost along the evolutionary scale. Consequently, not all known molecular and cellular changes to exercise are considered here ; this is because their function may not yet be recognized to be associated with adaptations that ameliorate fatigue. In addition, this review does not repeat in great detail material that is available in other reviews. There are 11,689 documents for the Medline MESH word "exertion," Medline's term for exercise, between 1984-1989, inclusive. To the reader unfamiliar with the causes of fatigue during physical exertion, enough description has been given (Table 1) to permit understanding of the physiological significance of the molecular and cellular events. A more detailed review on fatigue is available (108) . In addition, the reader is referred to earlier reviews that have comprehensively documented biochemical responses to a single exercise bout and biochemical adaptations of muscle to physical training.
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This study examined whether insulin stimulation of protein synthesis and inhibition of protein degradation is enhanced after exercise. The isolated perfused rat hindquarter preparation was used to evaluate net protein breakdown, myofibrillar protein degradation, and protein synthesis. Thirty minutes after treadmill exercise of high and moderate intensity, rates of tyrosine release were increased by 58 and 25%, respectively. Insulin at 75 microU/ml had no effect on these increases after intense exercise; however, 20,000 microU/ml of insulin totally inhibited this increase. Cycloheximide increased the tyrosine release in both control and exercised rat muscle. It also abolished the difference between them, suggesting that the increase in tyrosine release after exercise is caused by an inhibition of protein synthesis. Phenylalanine incorporation into protein was marginally depressed (22%, P = NS) in the white gastrocnemius muscle after intense exercise. Insulin at 200 microU/ml stimulated protein synthesis in these rats, but no more than it did in a nonexercised control group. Failure to observe a greater effect of insulin on protein metabolism was also noted when rat muscle was studied 150 min after intense exercise and after contractions induced by electrical stimulation of the sciatic nerve. These findings suggest that after exercise or electrically induced contractions the enhanced ability of insulin to stimulate hexose and amino acid transport is not paralleled by an increase in its ability to stimulate protein synthesis or inhibit protein degradation.
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The response of muscle and liver protein metabolism to either a single or three successive daily injections of an endotoxin (Escherichia coli lipopolysaccharide, serotype 0127 B8; 1 mg/ml, 0.3 mg/100 g body wt.) was studied in vivo in the fed rat, and at 24 and 30 h after endotoxin treatment during fasting. In the fed rats there was a catabolic response in muscle, owing to a 60-100% increase in muscle protein degradation rate, and a 52% fall in the synthesis rate. Although there was a 20% decrease in food intake, the decrease in protein synthesis was to some extent independent of this, since rats treated with endotoxin and fasted also showed a lower rate of muscle protein synthesis, which was in excess of the decrease caused by fasting alone. The mechanism of this decreased protein synthesis involved decreased translational activity, since in both fed and fasted rats there was a decreased rate of synthesis per unit of RNA. This occurred despite the fact that insulin concentrations were either maintained or increased, in the fasted rats, to those observed in fed rats. In the liver total protein mass was increased in the fed rats by 16% at 24 h, and the fractional synthesis rate at that time was increased by 35%. In rats fasted after endotoxin treatment the liver protein mass was not decreased as it was in the control fasted rats, and the fractional synthesis rate was increased by 22%. In both cases the increased synthesis rate reflected an elevated hepatic RNA concentration. The extent of this increase in hepatic protein synthesis was sufficient at one point to compensate for the fall in estimated muscle protein synthesis, so that the sum total in the two tissues was maintained.
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Constant infusion of [14C]tyrosine was used to measure the fractional rates of protein synthesis in vivo in skeletal muscle, heart and liver of normal, streptozotocin diabetic and insulin treated rats. Diabetes resulted in a 70% reduction in the rate of protein synthesis in skeletal muscle, and a 44% reduction in that in heart. Insulin treatment increased the protein synthesis rate in both tissues in diabetic, but not in normal animals. Changes in the RNA to protein ratio in skeletal muscle and heart in response to diabetes and insulin treatment were qualitatively similar to those in the rate of protein synthesis, but were less pronounced. No effects of diabetes, insulin treatment, or fasting on the rate of synthesis of hepatic tissue proteins in vivo could be observed. Diabetes resulted in a slight fall in the RNA to protein ratio in liver. The results are discussed in the light of data from various laboratories on the effect of diabetes on protein synthesis in subcellular systems.
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The avian skeletal alpha-actin gene was used as a template for construction of a myogenic expression vector that was utilized to direct expression of a human IGF-I cDNA in cultured muscle cells and in striated muscle of transgenic mice. The proximal promoter region, together with the first intron and 1.8 kilobases of 3'-noncoding flanking sequence of the avian skeletal alpha-actin gene directed high level expression of human insulin-like growth factor I (IGF-I) in stably transfected C2C12 myoblasts and transgenic mice. Expression of the actin/IGF-I hybrid gene in C2C12 muscle cells increased levels of myogenic basic helix-loop-helix factor and contractile protein mRNAs and enhanced myotube formation. Expression of the actin/IGF-I hybrid gene in mice elevated IGF-I concentrations in skeletal muscle 47-fold resulting in myofiber hypertrophy. IGF-I concentrations in serum and body weight were not increased by transgene expression, suggesting that the effects of transgene expression were localized. These results indicate that sustained overexpression of IGF-I in skeletal muscle elicits myofiber hypertrophy and provides the basis for manipulation of muscle physiology utilizing skeletal alpha-actin-based vectors.
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The role of insulin in the regulation of muscle protein synthesis in adult humans has been investigated with intravenous infusion of insulin at levels comparable with those observed after normal feeding. Glucose was also infused to maintain euglycemia. Muscle protein synthesis was measured in six healthy subjects before and during insulin and glucose infusion from the incorporation of L-[2H5]phenylalanine into the protein of vastus lateralis sampled by percutaneous biopsy. L-[2H5]phenylalanine was given as a single injection of a flooding amount (45 mg/kg). The relatively low levels of enrichment of phenylalanine in protein (0.005 atom%) were measured by modified gas chromatography-mass spectrometry and verified by comparison with incorporation of L-[2,6-3H]phenylalanine. Similarity of enrichment in tissue-free and plasma pools (flooding) and linear incorporation over the period of measurement were also verified. The fractional rate of muscle protein synthesis in the group of postabsorptive subjects was 1.65 +/- 0.11% (SE)/day. The rate was unaltered by insulin and glucose infusion, 1.66 +/- 0.16%/day.
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This paper reviews the evidence for and against the adoption of methods for the measurement of human tissue protein synthesis based upon the incorporation of stable isotopically labeled amino acids administered either as a continuous infusion or as a flooding dose. The practical advantages of the flooding dose method are the relative ease of application of the tracer and the ability to make a repeat measurement within approximately 2 h. For the method depending upon continuous infusion of labeled amino acid, the advantages include the use of labeled amino acids at true tracer doses (i.e., with no disturbance of metabolism) and the ability to make simultaneous measurements of whole body turnover and limb or organ turnover (given appropriate sampling techniques). The crucial question concerning the accuracy of the two methods (e.g., the 2-fold difference in the rate of skeletal muscle protein synthesis) remains unresolved, but in our opinion more evidence exists in favor of the values obtained from the continuous infusion method. Furthermore, as techniques for measurement of stable isotopically labelled amino acids improve, the length of time necessary for tracer infusion will fall, and the practical advantages of the flooding dose protocol will lessen in comparison.
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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.
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We examined the association of the mRNA cap binding protein eIF4E with the translational inhibitor 4E-BP1 in the acute modulation of skeletal muscle protein synthesis during recovery from exercise. Fasting male rats were run on a treadmill for 2 h at 26 m/min and were realimented immediately after exercise with either saline, a carbohydrate-only meal, or a nutritionally complete meal (54.5% carbohydrate, 14% protein, and 31.5% fat). Exercised animals and nonexercised controls were studied 1 h postexercise. Muscle protein synthesis decreased 26% after exercise and was associated with a fourfold increase in the amount of eIF4E present in the inactive eIF4E.4E-BP1 complex and a concomitant 71% decrease in the association of eIF4E with eIF4G. Refeeding the complete meal, but not the carbohydrate meal, increased muscle protein synthesis equal to controls, despite similar plasma concentrations of insulin. Additionally, eIF4E.4E-BP1 association was inversely related and eIF4E.eIF4G association was positively correlated to muscle protein synthesis. This study demonstrates that recovery of muscle protein synthesis after exercise is related to the availability of eIF4E for 48S ribosomal complex formation, and postexercise meal composition influences recovery via modulation of translation initiation.
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The purpose of the present study was to investigate the regulation of plasma and tissue levels of insulin-like growth factor-I (IGF-I) and IGF-binding protein-1, -2, and -3 (IGFBP-1, -2, and -3) in rats injected with Escherichia coli lipopolysaccharide (LPS), a component of the outer cell wall of gram-negative bacteria. When injected iv into conscious overnight fasted rats, plasma IGF-I levels were initially decreased within 1 h, maximally depressed at 4 h, and still only 35-45% of control values at 24 h. GH levels were reduced as early as 30 min after LPS, averaged 80-90% of control values between 1-4 h, but had returned to basal levels by 24 h. The magnitude and duration of these changes were similar regardless of whether 100 or 10 μgrams/100 g BW (LD20 and LD0, respectively) LPS were injected. Plasma levels of IGFBP-1 and a 28K mol wt BP (BP-28K) were elevated 2- to 3-fold 4 h after LPS treatment, whereas IGFBP-3 and -2 levels were unchanged. The elevation in plasma IGFBP-1 and IGFBP-28K was observed as early as 1 h and was sustained for up to 24 h after LPS treatment. IGF-I levels were decreased 30-50% in liver, pituitary, and skeletal muscle, unchanged in brain, and elevated 5-fold in kidney in response to LPS. Of the tissues sampled, IGFBP-3 and -2 were selectively elevated in liver after LPS treatment. IGFBP-1 was increased in liver, muscle, and kidney in response to LPS. The level of the 28, 000 mol wt BP was increased in liver (83%) and not changed in muscle or brain. These data indicate that LPS produces both rapid and sustained alterations in circulating levels of GH, IGF-I, and IGFBPs. Furthermore, there were marked tissue-specific changes in levels of IGF-I and IGFBPs. LPS-induced changes in plasma and tissue IGFBP-3 were not regulated by changes in GH, and changes in insulin could not explain the alterations in IGFBP-1 and -2. These results suggest that after the injection of LPS, changes in IGF-I and IGFBP levels are regulated by a mechanism(s) different from those previously described.
Article
To ascertain if IGF-1 is a regulator of local muscle growth, total RNA was extracted from rabbit muscle induced to undergo rapid hypertrophy using active stretch and from control muscles. This was analysed by Northern hybridization with a 280 base pair probe containing sequences derived from exons 3 and 4 of the insulin-like growth factor 1 gene. Two types of insulin-like growth factor 1 mRNA were shown to be strong expressed in the stretched muscles. In situ hybridization using the same probe (280 base pair) showed that IGF-1 is strongly expressed in muscle that is induced to grow rapidly and is expressed in the muscle fibres themselves. Using RT-PCR a single insulin-like growth factor 1 isoform cDNA (IGF-1Ea) could be cloned from the normal resting muscles. However, an additional isoform of insulin-like growth factor 1 (insulin-like growth factor 1Eb) was found to be expressed in stretched muscle undergoing hypertrophy. The E domain sequence of the additional isoform differs from the liver insulin-like growth factor 1Ea by the presence a 52 base pair insert. This changes the reading frame of the derived carboxyl-terminal resulting in a different precursor insulin-like growth factor 1 isoform. This insulin-like growth factor 1 mRNA probably encodes the precursor insulin-like growth factor 1 isoform that is responsible for local muscle growth regulation in response to mechanical stimulation. To confirm that alternative splicing of the insulin-like growth factor 1 gene occurs in muscle in response to physical activity, oligonucleotide primers were made which specifically amplify the cDNAs of two isoforms (insulin-like growth factors 1Ea and Eb) in the human as well as the rabbit. Following altered physical activity for 2 h to 6 days, appreciable levels of insulin-like growth factor 1Eb (in human the Ec) isoform were detected in skeletal muscle by using RT-PCR. In contrast very little if any of this splice variant could be detected in control muscle not subjected to stretch or extra physical activity.
Article
Treatment of normal rats with cortisone acetate for 5 days reduced the rate of synthesis of skeletal muscle protein 55% as measured in the perfused rat hemicorpus. Loss of tissue RNA accounted for 15% of this decrease; the remaining reduction resulted from a block in peptide-chain initiation that developed in vivo. In contrast, protein synthesis in perfused heart muscle was not affected by steroid treatment, nor was tissue RNA content or peptide-chain initiation altered. eIF-2-like activity in postribosomal supernatants from psoas was reduced 35% by cortisone treatment. This difference was not abolished by overnight dialysis and was not due to accelerated rates of deacylation of Met-tRNA(f)(Met), or to changes in nucleotide levels during incubation. No difference in activity was observed in supernatants from hearts of normal and cortisone-treated animals. Changes in RNA content in skeletal muscle, but not in heart, of hormone-treated rats suggested a relationship between initiation factor activity and RNA content. In other experiments, ribosomal subunits accumulated in psoas muscles of rats treated 4 h with dexamethasone, whereas tissue RNA content and Met-tRNA(f)(Met) binding activity remained unchanged. These results suggested that adrenocortical steroids may reduce skeletal muscle protein synthesis in part by regulating the activity of factors involved in peptide initiation and that factor activity may be linked to tissue RNA content.
Article
In another study (J. Appl. Physiol. 69: 1709-1717, 1990) we reported that gastrocnemius (GAST) muscle enlargement failed to occur after 10 wk of 192 contractions performed every 3rd or 4th day. This result was surprising because increased protein synthesis rates were determined after an initial acute exercise bout with the same paradigms. In the same set of animals, tibialis anterior (TA) muscles were enlarged 16-30% compared with sedentary control muscles after the same chronic training regimen. This indicated that the regulation of protein expression may be different between the GAST and TA muscles. The present experiment attempted to define and explain these differences by comparing changes in various indexes of protein metabolism in TA with the same parameters determined in the accompanying study for the GAST. As in the GAST, results showed that TA protein synthesis rates are increased by acute exercise and principally regulated by translational and possibly posttranslational mechanisms. The differential response in muscle mass between the GAST and TA muscles after training may be due, in part, to greater relative resistances imposed on the TA than on the GAST that result in a more-prolonged effect on protein synthesis rates, with lower numbers of stimulated contractions required to stimulate increases in protein synthesis. Data also revealed that although as little as 1 min of total contractile duration (24 repetitions) increased TA protein synthesis rate by 30%, 8 min of total contractile duration (192 repetitions) further increased TA protein synthesis rates to only 45% above control.
Article
Previous results by use of a model of resistance exercise consisting of nonvoluntary electrical contraction of rat skeletal muscle have shown that significant gastrocnemius muscle enlargement was produced after 16 wk of chronic concentric resistance training with progressively increased weights but not after the same training program without weights (J. Appl. Physiol. 65: 950-954, 1988). In the present study we examined whether this differential effect on muscle mass between high- and low-resistance exercise is mediated through differential actions on muscle protein synthesis rates. In addition, we determined whether accumulation of specific mRNA quantities had a primary role in the protein synthesis response to this type of exercise. The data revealed that as little as 8 min of total contractile duration increased gastrocnemius protein synthesis rates by nearly 50%. Contrary to our hypothesis, post-exercise protein synthesis rates do not appear to be differentially regulated by the resistance imposed on the muscle during exercise but rather by the number of repetitions performed during the acute bout. This observation, the failure of high-frequency chronic training to produce gastrocnemius enlargement, and the relatively minor effects on mRNA levels collectively suggest that translational and posttranslational mechanisms, including protein degradation, may be the principal processes by which gastrocnemius protein expression is regulated in this model of stimulated concentric exercise.
Article
The effects of exercise training on glucose-stimulated insulin secretion (GSIS) were studied in male Sprague-Dawley rats made mildly to severely diabetic by partial pancreatectomy. Exercise trained (10 wk treadmill; T) and untrained (Unt) rats were grouped according to posttraining fed-state hyperglycemia as follows: T less than 200 and Unt less than 200 (glucose concn less than 200 mg/dl), T 200-300 and Unt 200-300 (glucose concn 200-300 mg/dl), and T greater than 300 and Unt greater than 300 (glucose concn greater than 300 mg/dl). After exercise training, hyperglycemic glucose clamps were performed in awake rats by elevation of arterial blood glucose concentration 126 mg/dl above fasting basal levels for 90 min. Exercise training significantly increased muscle citrate synthase activity. Prevailing hyperglycemia was reduced during the 10-wk exercise training period in all T rats with fed-state glucose concentrations less than 300, and only 53% of Unt rats in these groups had reduced glycemia. GSIS was significantly higher in T less than 200 [2.4 +/- 0.7 (SD) ng/ml at 90 min] than in Unt less than 200 (1.5 +/- 0.3). A similar response was found for T 200-300 (1.1 +/- 0.3 ng/dl) vs. Unt 200-300 (0.7 +/- 0.1) but not T greater than 300 (0.36 +/- 0.2) vs Unt greater than 300 (0.44 +/- 0.05). Sham-operated control rats had insulin concentrations of 6.6 +/- 1.6 ng/ml at the 90th min of the clamp. Acute exercise reduced fed-state glycemia in rats with mild-to-moderate (less than 300 mg/dl) diabetes.(ABSTRACT TRUNCATED AT 250 WORDS)
Article
Despite its anabolic effects on protein balance, acute administration of insulin has been reported to have no effect on skeletal muscle or whole body protein synthesis in man. However, insulin also reduces plasma and intramuscular amino acid availability, which may limit protein synthesis. We have therefore measured the acute effects of insulin on skeletal muscle (anterior tibialis) protein synthesis and whole body leucine turnover in eight insulin-withdrawn Type 1 (insulin-dependent) diabetic patients. They were studied initially when insulin deficient, but during infusion of mixed amino acids at a rate sufficient to raise plasma amino acids by 30% i.e. to 4 mmol/l in total; measurements were continued when insulin was infused together with an increased rate of amino acids to maintain insulinopoenic plasma amino acid concentrations. Using 13C-alpha-ketoisocaproate in plasma as an index of the intracellular precursor labelling, incorporation of [1-13C]leucine into skeletal muscle protein was 0.068 +/- 0.007%/h during insulin withdrawal and was unaltered during insulin infusion. The value is higher than observed in muscle of healthy man, possibly because of a stimulatory effect of endogenous intramuscular amino acids. Also, calculated on the basis of alpha-ketoisocaproate labelling, non-oxidised whole body leucine disappearance (i.e. whole body protein synthesis) was 110 +/- 4 mumol.kg-1.h-1 during insulin withdrawal; this also was unchanged during insulin infusion. Despite stable or increased plasma concentrations of most amino acids, the intramuscular concentrations of a number of amino acids decreased during insulin infusion.(ABSTRACT TRUNCATED AT 250 WORDS)
Article
A nitrocellulose gel transfer technique has been adapted to study the insulin-like growth factor (IGF) binding proteins of human serum. Normal and hypopituitary sera were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by electroblotting to nitrocellulose or nylon membrane. Nonidet-P40 (3%) and Tween 20 (0.1%) were required for quenching and to allow detection of the IGF binding proteins by autoradiography after overlay with either 125I-labeled IGF I or IGF II. Several forms of IGF binding protein have been identified with molecular weights of 41,500, 38,500, 34,000, 30,000, and 24,000. Titration and competitive binding studies with IGF were performed on the transferred IGF binding proteins, indicating that binding proteins isolated by this technique can be characterized.
Article
We evaluated the influence of insulin on fractional mixed skeletal muscle protein synthesis (FMPS) in eight type I (insulin-dependent) diabetic patients in the postabsorptive state. FMPS was calculated from the increment in [13C]leucine in mixed skeletal muscle protein obtained by serial percutaneous needle biopsy during a continuous 8-h intravenous infusion of L-[13C]leucine. We used the plasma [13C]-alpha-ketoisocaproate (representing intracellular leucine labeling) as the precursor pool of protein synthesis for our calculations. FMPS during the insulin treatment (0.0472 +/- 0.0046%/h; plasma glucose 4.6 +/- 1.0 mM) was not different from FMPS during insulin deprivation (0.0499 +/- 0.0046%/h; plasma glucose 16.4 +/- 0.5 mM). Using plasma [13C]-alpha-ketoisocaproate at isotopic plateau for calculation of leucine flux and as the precursor for leucine oxidation, we further confirmed the findings of our group and others that insulin treatment decreases leucine flux, leucine oxidation, and the nonoxidative portion of leucine flux. Our data on direct measurement of FMPS provide further evidence that the anabolic effect of insulin in the postabsorptive type I diabetic patient is mediated via reduction of proteolysis rather than by increasing protein synthesis.
Article
Although insulin stimulates protein synthesis and inhibits protein breakdown in skeletal muscle in vitro, the actual contribution of these actions to its anabolic effects in man remains unknown. Using the forearm perfusion method together with systemic infusion of L-[ring-2,6-3H]phenylalanine and L-[1-14C]leucine, we measured steady state amino acid exchange kinetics across muscle in seven normal males before and in response to a 2-h intraarterial infusion of insulin. Postabsorptively, the muscle disposal (Rd) of phenylalanine (43 +/- 5 nmol/min per 100 ml forearm) and leucine (113 +/- 13) was exceeded by the concomitant muscle production (Ra) of these amino acids (57 +/- 5 and 126 +/- 9 nmol/min per dl, respectively), resulting in their net release from the forearm (-14 +/- 4 and -13 +/- 5 nmol/min per dl, respectively). In response to forearm hyperinsulinemia (124 +/- 11 microU/ml), the net balance of phenylalanine and leucine became positive (9 +/- 3 and 61 +/- 8 nmol/min per dl, respectively (P less than 0.005 vs. basal). Despite the marked increase in net balance, the tissue Rd for both phenylalanine (42 +/- 2) and leucine (124 +/- 9) was unchanged from baseline, while Ra was markedly suppressed (to 33 +/- 5 and 63 +/- 9 nmol/min per dl, respectively, P less than 0.01). Since phenylalanine is not metabolized in muscle (i.e., its only fates are incorporation into or release from protein) these results strongly suggest that in normal man, physiologic elevations in insulin promote net muscle protein anabolism primarily by inhibiting protein breakdown, rather than by stimulating protein synthesis.
Article
A method is described for the estimation of serum free fatty acids based on the ultramicro method of Novák. The difficulty of obtaining an accurate standard graph has been overcome and the interfering effects of phospholipids have been reduced. The method is simple to perform and gives accurate, precise results in 50 mul samples; it is thus suitable for work on newborn babies and small laboratory animals.
Article
The effects of acute (2-day) and long-term (7-day) diabetes on rates of protein synthesis, peptide-chain initiation, and levels of RNA were examined in rat skeletal muscles that are known to have differing proportions of the three fiber types: fast-twitch white, fast-twitch red, and slow-twitch red. Short-term diabetes resulted in a 15% reduction in the level of RNA in all the muscles studied and an impairment in peptide-chain initiation in muscles with mixed fast-twitch fibers. In contrast, the soleus, a skeletal muscle with high proportions of slow-twitch red fibers, showed little impairment in initiation. When the muscles were perfused as a part of the hemicorpus preparation, addition of insulin to the medium caused a rapid reversal of the block in initiation in mixed fast-twitch muscles but had no effect in the soleus. The possible role of fatty acids in accounting for these differences is discussed. Long-term diabetes caused no further reduction in RNA, but resulted in the development of an additional impairment to protein synthesis that also affected the soleus and that was not corrected by perfusion with insulin. The defect resulting from long-term diabetes may involve elongation or termination reactions.
Article
1. Insulin was infused into young male rats in the postabsorptive state. Rates of protein synthesis in skeletal muscle were determined during the final 10 min of infusion from the incorporation of label into protein after intravenous injection of a massive dose of [3H]phenylalanine. Rates of synthesis were not altered during the first 10 min of insulin infusion, but were increased significantly between 10 and 60 min. 2. Rats were infused with different amounts of insulin for 30 min. When concentrations were increased from 10 to 40 microunits/ml of plasma there was no change in muscle protein synthesis, but concentrations higher than 70 microunits/ml caused a significant stimulation. Concentrations below 10 microunits/ml, obtained by infusion of anti-insulin serum, did not depress synthesis below that found in the postabsorptive rat. 3. Infusion of glucose for 30 or 60 min led to an increase in plasma insulin to 40 microunits/ml, but this also failed to stimulate muscle protein synthesis. 4. Rates of synthesis in postabsorptive rats, even when stimulated maximally by insulin, were not so high as those in fed rats or in postabsorptive rats refed for 60 min. However, in fed and refed rats insulin concentrations were below that required to stimulate synthesis in postabsorptive animals. Despite this, infusion of large amounts of insulin into fed rats did not increase synthesis further. 5. The sensitivity of plasma glucose to insulin infusion was different from that of protein synthesis. A decrease in glucose concentration preceded the increase in synthesis and occurred at lower insulin concentrations. 6. It is concluded that changes in circulating insulin may have been partly responsible for the increase in muscle protein synthesis brought about by feeding, but that other factors must also play a part.
Article
The purpose of this study was to estimate the absolute and relative masses of the three types of skeletal muscle fibers in the total hindlimb of the male Sprague-Dawley rat (Rattus norvegicus). For six rats, total body mass was recorded and the following weights taken from dissection of one hindlimb: 32 individual major muscles or muscle parts, remaining skeletal musculature (small hip muscles and intrinsic foot muscles), bone, inguinal fat pad, and skin. The fibers from the 32 muscles or muscle parts (which constituted 98% of the hindlimb skeletal muscle mass) were classified from histochemistry as fast-twitch oxidative glycolytic (FOG), fast-twitch glycolytic (FG), or slow-twitch oxidative (SO), and their populations were determined. Fiber cross-sectional areas from the same muscles were measured with a digitizer. Mass of each of the fiber types within muscles and in the total hindlimb was then calculated from fiber-type population, fiber-type area, and muscle-mass data. Skeletal muscle made up 71% of the total hindlimb mass. Of this, 76% was occupied by FG fibers, 19% by FOG fibers, and 5% by SO fibers. Thus, the FG fiber type is clearly the predominant fiber type in the rat hindlimb in terms of muscle mass. Fiber-type mass data are compared with physiological (blood flow) and biochemical (succinate dehydrogenase activities) data for the muscles taken from previous studies, and it is demonstrated that these functional properties are closely related to the proportions of muscle mass composed of the various fiber types.
Article
A rapid procedure for measuring the specific radioactivity of phenylalanine in tissues was developed. This facilitates the accurate determination of rates of protein synthesis in a wide range of tissues by injection of 150 mumol of L-[4-(3)H]phenylalanine/100 g body wt. The large dose of amino acid results in a rapid rise in specific radioactivity of free phenylalanine in tissues to values close to that in plasma, followed by a slow but linear fall. This enables the rate of protein synthesis to be calculated from measurements of the specific radioactivity of free and protein-bound phenylalanine in tissues during a 10 min period after injection of radioisotope.