Gastrocnemius-plantaris-soleus Research Articles

Exercise, but Not Metformin Prevents Muscle Function Loss due to Doxorubicin in Mice Using an In‐Situ Model

Doxorubicin (DOX) is a clinically relevant chemotherapeutic known to cause severe side effects in heart and skeletal muscle. The deleterious effects DOX elicits on skeletal muscle are concerning due to the high rates of severe fatigue experienced by chemotherapy patients. Identifying treatments that prevent this skeletal muscle dysfunction is critically important. Exercise (EX) and metformin (MET) have both been suggested as treatments to reduce the toxic effects of DOX. However, their effects on muscle function in the gastrocnemius‐plantaris‐soleus (GPS) complex are unknown. Additionally, the mechanism by which DOX decreases muscle function are still under investigation. DOX can cause mitochondrial disruption, but the direct effects of mitochondrial function on muscle function have not been studied in this model. The purpose of this study is three fold: (1) to determine if DOX decreased muscle function of the GPS complex using an in‐situ muscle fatigue protocol, (2) determine if EX or MET treatment can prevent muscle function loss due to DOX, and (3) determine if muscle function loss is due to a decrease in mitochondrial respiratory function. C57BL/6 mice were randomly assigned to 4 groups: CON, DOX, DOX+EX, or DOX+MET. DOX was administered as a single intraperitoneal injection at 15 mg/kg body weight 3 days before sacrifice. Moderate EX treatment was done by treadmill running for 60 minutes for 5 days at 70% max prior to DOX injection. MET treatment was administered via oral gavage at 500 mg/kg body weight for 5 days beginning 2 days before DOX injection. GPS complex muscle function was measured in‐situ using tetanic contractions elicited by direct stimulation in a 6 minute fatigue protocol. Real time mitochondrial respiratory function was measured in gastrocnemius muscle. We report that DOX treatment caused ~20% body weight loss at 3 days, an effect that was partially prevented in DOX+EX (p<0.05) or DOX+MET (p=0.06). A small decrease in total force production was observed with DOX, which trended towards significance (0.08≥p≤0.04). DOX+EX treatment restored force production back to CON levels, but DOX+MET did not. The half relaxation rate was impaired by an average of 18% with DOX treatment (p<0.05). DOX+EX prevented this impairment of the half relaxation rate (p<0.05). DOX+MET did not restore impaired half relaxation rate caused by DOX. Mitochondrial respiration was not different between the groups, suggesting that the changes in muscle function occur independent of mitochondrial function. In conclusion, DOX treatment decreased muscle function in the GPS complex by impairing half relaxation rates. EX but not MET treatment prevented impaired half relaxation rate. These changes occurred independent of mitochondrial respiration.Support or Funding InformationMentoring Environment Grant, Brigham Young UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Partial Pancreatectomized Diabetic Rats Present with Altered Skeletal Muscle Contractility and Phenotype

People with long‐term type 1 diabetes (T1D) often have skeletal muscle complications (myopathy). Studies on T1D and skeletal muscle are limited and often use animal models that induce T1D with streptozotocin, which causes myopathy independent of hyperglycemia. The objective of this study was to characterize diabetic myopathy in a partially pancreatectomized (Px) model of T1D. Male Sprague Dawley rats were randomly assigned to either Px or sham surgery groups. Following 8 weeks of diabetes, Px had significantly lower body and gastrocnemius‐plantaris‐soleus (GPS) mass compared to shams (349.1±21 vs 484.4±10 g; 1.8±0.1 vs 3.3±0.1 g/g mass, respectively). In situ muscle stimulation of the GPS revealed a lower maximal force (Fmax) in Px vs shams (12.1±1.3N vs 22.6±1.8N, p<0.01), but was similar when corrected for muscle mass. During a 2 minute fatigue protocol, stimulating the GPS at ~50% Fmax (3 sec stimulation, 3 sec rest), Px had a lower rate of decline in force than shams (52.8±3.4% vs 69.7±7.2%, p<0.05). Histochemical analysis of the GPS demonstrates that Px have reduced myofiber area irrespective of fiber type and an increase in type IIa fiber number. These data suggest that sustained hyperglycemia/hypoinsulinemia induced by Px is characterized by changes in the skeletal muscle phenotype that result in compromised function. This research was funded by NSERC.

Chronic Treatment with the β2-Adrenoceptor Agonist Prodrug BRL-47672 Impairs Rat Skeletal Muscle Function by Inducing a Comprehensive Shift to a Faster Muscle Phenotype

Discovering approaches to maintain or improve muscle function (fatigue resistance) in patients with cachexia, postoperative weakness, and sarcopenia is of clinical importance. beta(2)-Agonist treatment increases muscle mass, yet it alters fiber proportions such that the net consequences on muscle function remain unclear. In the present study, we focus on the contractile and metabolic consequences of chronic treatment with the beta(2)-agonist prodrug BRL-47672 (BRL). Gastrocnemius-plantaris-soleus (GPS) muscles were harvested at rest and studied for fatigue characteristics during 4 and 20 s of isometric stimulation (30 Hz; 10 V; 200 ms) using the perfused hind limb model. BRL treatment increased GPS mass by 21% (P < 0.05), whereas greater fatigue occurred during 20 s of contraction (45% less work; P < 0.05). Phenotypically, BRL resulted in 17% more type IIb myosin heavy chain protein expression (P < 0.001) and greater adenine nucleotide catabolism during 20 s of contraction (P < 0.05). Chronic BRL treatment impaired maximal lipid oxidation capacity by 30% (P < 0.05) and reduced glutamate dehydrogenase activity by 15% (P < 0.05). We conclude that beta(2)-agonist induced muscle hypertrophy may be clinically limited as impaired energy metabolism and function occur, presumably as a consequence of the shift in muscle phenotype.

Glycogen Phosphorylase Inhibition in Type 2 Diabetes Therapy

Inhibition of hepatic glycogen phosphorylase is a promising treatment strategy for attenuating hyperglycemia in type 2 diabetes. Crystallographic studies indicate, however, that selectivity between glycogen phosphorylase in skeletal muscle and liver is unlikely to be achieved. Furthermore, glycogen phosphorylase activity is critical for normal skeletal muscle function, and thus fatigue may represent a major development hurdle for this therapeutic strategy. We have carried out the first systematic evaluation of this important issue. The rat gastrocnemius-plantaris-soleus (GPS) muscle was isolated and perfused with a red cell suspension, containing 3 micromol/l glycogen phosphorylase inhibitor (GPi) or vehicle (control). After 60 min, the GPS muscle was snap-frozen (rest, n = 11 per group) or underwent 20 s of maximal contraction (n = 8, control; n = 9, GPi) or 10 min of submaximal contraction (n = 10 per group). GPi pretreatment reduced the activation of the glycogen phosphorylase a form by 16% at rest, 25% after 20 s, and 44% after 10 min of contraction compared with the corresponding control. AMP-mediated glycogen phosphorylase activation was impaired only at 10 min (by 21%). GPi transiently reduced muscle lactate production during contraction, but other than this, muscle energy metabolism and function remained unaffected at both contraction intensities. These data indicate that glycogen phosphorylase inhibition aimed at attenuating hyperglycaemia is unlikely to negatively impact muscle metabolic and functional capacity.

An acute decrease in TCA cycle intermediates does not affect aerobic energy delivery in contracting rat skeletal muscle

We tested the hypothesis that an acute decrease in muscle TCA cycle intermediates during contraction would compromise aerobic energy delivery. Male Wistar rats were anaesthetized and the gastrocnemius-plantaris-soleus (GPS) muscle complex from one leg was isolated and perfused with a red cell medium containing either saline (Con) or cycloserine (Cyclo; 0.05 mg g-1), an inhibitor of alanine aminotransferase (AAT). After 1 h of perfusion, the GPS muscle was either snap frozen (Con-Rest, n=11; Cyclo-Rest, n=9) or stimulated to contract for 10 min (1 Hz, 0.3 ms, 2 V) with blood flow fixed at 30 ml min-1 (100 g)-1 and then snap frozen (Con-Stim, n=10; Cyclo-Stim, n=10). Maximal AAT activity was>80% lower (P<0.001) in both Cyclo-treated groups (Rest: 0.61+/-0.02; Stim: 0.63+/-0.01 mmol (kg wet wt)-1 min-1; mean+/-s.e.m.) compared to Con (Rest: 3.56+/-0.16; Stim: 3.92+/-0.29). The sum of five measured TCAI (SigmaTCAI) was reduced by 23% in Cyclo-Rest versus Con-Rest but this was not different (P=0.08). However, after 10 min of contraction, the SigmaTCAI was 25% lower (P=0.006) in Cyclo-Stim compared to Con-Stim (1.88+/-0.15 versus 2.48+/-0.11 mmol (kg dry wt)-1). Despite the acute decrease in TCAI after Cyclo treatment, the contraction-induced changes in markers of non-oxidative energy provision (phosphocreatine, ATP and lactate) and the decline in tension after 10 min of stimulation were similar compared to Con. These data do not support the hypothesis that the total muscle concentration of TCAI is causally linked to the rate of mitochondrial respiration during contraction.

Skeletal muscle contractile performance and ADP accumulation in adenylate kinase-deficient mice

The production of AMP by adenylate kinase (AK) and subsequent deamination by AMP deaminase limits ADP accumulation during conditions of high-energy demand in skeletal muscle. The goal of this study was to investigate the consequences of AK deficiency (-/-) on adenine nucleotide management and whole muscle function at high-energy demands. To do this, we examined isometric tetanic contractile performance of the gastrocnemius-plantaris-soleus (GPS) muscle group in situ in AK1(-/-) mice and wild-type (WT) controls over a range of contraction frequencies (30-120 tetani/min). We found that AK1(-/-) muscle exhibited a diminished inosine 5'-monophosphate formation rate (14% of WT) and an inordinate accumulation of ADP ( approximately 1.5 mM) at the highest energy demands, compared with WT controls. AK-deficient muscle exhibited similar initial contractile performance (521 +/- 9 and 521 +/- 10 g tension in WT and AK1(-/-) muscle, respectively), followed by a significant slowing of relaxation kinetics at the highest energy demands relative to WT controls. This is consistent with a depressed capacity to sequester calcium in the presence of high ADP. However, the overall pattern of fatigue in AK1(-/-) mice was similar to WT control muscle. Our findings directly demonstrate the importance of AMP formation and subsequent deamination in limiting ADP accumulation. Whole muscle contractile performance was, however, remarkably tolerant of ADP accumulation markedly in excess of what normally occurs in skeletal muscle.

Evaluation of disuse atrophy of rat skeletal muscle based on muscle energy metabolism assessed by 31P-MRS.

The purpose of this study was to evaluate disuse atrophy of skeletal muscle using a hind-limb suspension model, with special reference to energy metabolism. Twenty-four Sprague-Dawley rats were divided into four groups: control group (C), hind-limb suspended for 3 days (HS-3), for 7 days (HS-7) and for 14 days (HS-14). The gastrocnemius-plantaris-soleus (GPS) muscles in each group were subjected to the following measurements. After a 2-min rest, contraction of the GPS muscles was induced by electrical stimulation of the sciatic nerve at 0.25 Hz for 10 min, then the frequency was increased to 0.5 and 1.0 Hz every 10 min. During the stimulation, twitch forces were recorded by a strain gauge, and 31P-MRS was performed simultaneously. Maximum tension was measured at the muscle contraction induced at 0.25 Hz; the wet weight of the whole and each muscle in the GPS muscles was also measured. From the 31P-MR spectra during muscle contraction, the oxidative capacity was calculated and compared among the groups. The weights of the whole GPS muscles in C, HS-3, HS-7 and HS-14, were 2.66 +/- 0.09, 2.39 +/- 0.21, 2.34 +/- 0.21 and 2.18 +/- 0.14 (g) respectively. Thus, the muscle mass significantly decreased with time (p < 0.05). Among the GPS muscles, the decrease in weight of the soleus muscle was especially remarkable; in the HS-14 group its weight decreased to 60% of that in the C group. We evaluated maximum tension and oxidative capacity as the muscle function. The maximum tensions in C, HS-3, HS-7 and HS-14 were 519 +/- 43, 446 +/- 66, 450 +/- 23 and 465 +/- 29 (g), respectively. This was significantly greater in the C group than in any other groups, however there were no significant differences among the three HS groups. The oxidative capacity during muscle contraction in the C group was higher than in any HS group and it did not further decrease even if the suspension of the limbs was prolonged beyond 3 days. The present study showed that in disuse atrophy, muscle mass and muscle function did not change simultaneously. Thus, it is necessary to develop countermeasures to prevent muscle atrophy and muscle function deterioration independently.

Effects of ovariectomy on intramuscular energy metabolism in young rats: how does sports-related-amenorrhea affect muscles of young female athletes?

The purpose of this study was to evaluate the effects of ovariectomy on intramuscular energy metabolism in young rats. Twenty-four Sprague-Dawley rats (7 weeks old) were used. Twelve of them underwent ovariectomy (OVX), and the others were sham-operated on. Seven OVX rats were examined 1-week after surgery (OVX-1 group), and the other five, 4 weeks after surgery (OVX-4 group). The gastrocnemius-plantaris-soleus (GPS) muscles group was subjected to the following measurements, and the data were compared with those of the sham group (Sham-1: n = 7, or Sham-4 group: n = 5). From the 31P-MR spectra of the GPS muscles group at rest and during electric stimulation, the muscular oxidative capacity was measured. Maximum tension and wet weight of the whole GPS muscles group were also measured. Body weight in the OVX-4 group was significantly (p < 0.01) larger than that in the Sham-4 group. The weights of the whole GPS muscles group in the Sham-1, Sham-4, OVX-1 and OVX-4 groups were 1.17, 1.51, 1.25 and 1.71 (g), respectively. The muscle weight in the OVX group tended to be greater than that in the Sham group (p < 0.10). The maximum tension and oxidative capacity did not differ significantly among the groups. These data indicated that in young rats, ovariectomy induced an increase in body and muscle weight, but did not affect the maximum tension nor oxidative capacity.

Clenbuterol Induces Hypertrophy of the Latissimus Dorsi Muscle and Heart in the Rat With Molecular and Phenotypic Changes

Skeletal muscle assistance of the circulation for patients in end-stage heart failure requires electrical training of the latissimus dorsi flap to produce fatigue resistance. This process of electrical transformation and the development of postmobilization atrophy results in a profound loss in peak power generated. The beta 2-adrenoceptor agonist clenbuterol was used to investigate its potential to selectively induce skeletal muscle hypertrophy, particularly the latissimus dorsi muscle (LDM), independent of adverse effects on cardiac muscle. Forty-one male Sprague-Dawley rats were divided into four groups and used in this study. Clenbuterol 2 micrograms.g body wt-1.d-1 was administered subcutaneously for a period of either 5 weeks (group A) or 2 weeks (group A1). Groups B and B1 (controls) were injected with 0.5 mL normal saline once daily. At the end of the experimental period, all rats were weighed and terminally anesthetized for removal of the left LDM, left gastrocnemius-plantaris-soleus (GPS) muscles, and heart. The results showed that the increase in body weight did not differ significantly between the clenbuterol-treated and control groups (P > .5). The ratio of LDM to tibial length (hypertrophic index) for groups A and A1 was significantly greater than controls (P < .01), which represented a 20% to 29% increase. The hypertrophy was more pronounced for hindlimb skeletal muscle (21% to 35% for GPS), and the effects of this relatively high dose of clenbuterol on the heart were less marked (18% to 20% hypertrophy). RNA analyses indicate that ventricles of clenbuterol-treated rats express elevated levels of mRNA to atrial natriuretic factor without a concomitant increase in skeletal alpha-actin and beta-myosin heavy chain, consistent with a "physiological" form of cardiac hypertrophy. Clenbuterol induces significant hypertrophy of the LDM associated with specific changes in cardiac gene expression.

Training increases collateral-dependent muscle blood flow in aged rats

The potential for physical training to enhance collateral-dependent blood flow (BF) to the hindlimbs of aged male rats (Fischer 344) was evaluated following bilateral femoral artery ligation at 20.5 mo of age. Rats were either limited to cage activity (sedentary, n = 11) or trained by a mild-intensity treadmill program (trained, n = 14), which involved walking twice a day at 15 m/min (15% grade) to fatigue, 5 days/wk for 8-11 wk. Exercise tolerance of the trained rats increased from approximately 5 to approximately 25 min/bout by week 7, whereas exercise tolerance of the sedentary group changed little (to approximately 8 min/bout) during the training period. At approximately 23 mo age, animals were surgically prepared for hindquarter perfusion (aortic pressure = 100 +/- 1.3 mmHg) and force measurement of the left gastrocnemius-plantaris-soleus (GPS) muscle group during isometric contractions at 4, 8, 15, 30, and 45 tetani/min via sciatic nerve stimulation (approximately 6 V, 0.1-ms square waves at 100 Hz for 100 ms). Although initial force development was similar between groups (12.9 N/g), trained rats maintained tension better at 8, 15, 30, and 45 tetani/min (P < 0.01). BF to the entire hindlimb of the trained group, determined with 85Sr 15-microns microspheres, was 43% greater (P < 0.05) than in the sedentary group. Thus collateral-dependent BF was improved by physical training. The greatest increase in BF was to the distal limb muscles (approximately 78%), the tissues most at risk during intermittent claudication.(ABSTRACT TRUNCATED AT 250 WORDS)

Interaction of glucocorticoids and activity patterns affect muscle function.

The purpose of this study was to determine the effects of glucocorticoids on muscle mass and contractile properties of muscles of similar fiber composition but differing in activity patterns. Rats were divided into two groups and administered prednisolone (5 mg/kg per day) (P; N = 9) or served as controls (C; N = 10) for 10 days. Contractile properties were then determined in the left gastrocnemius-plantaris-soleus (GPS) muscle complex and a strip of costal diaphragm (D). An index of fatigue was also determined in both muscle preparations. Whole-body, GPS, and D weights decreased (P < 0.05) in the P animals (22%, 17%, and 15%, respectively) when compared to C. Specific tension (PO) increased (P < 0.05) in the GPS complex (21%) and decreased in the D (26%). Maximal shortening velocity (Vmax) was not different (P > 0.05) between groups in either the GPS or the D. While the index of fatigue was not different (P > 0.05) between groups in the D, there was a 30% increase (P < 0.05) in the rate of fatigue in the GPS. These data indicate that, although glucocorticoids cause decreased muscle mass in both D and GPS, a change in muscle architecture may prevent a decrease in force-generating ability in some limb muscles. However, glucocorticoids do not increase D fatigability as seen in the GPS.

Effects of the creatine analogue β-guanidinopropionic acid on skeletal muscles of mice deficient in muscle creatine kinase

To evaluate the effects of phosphocreatine (PCr) and creatine (Cr) depletion on skeletal muscles of mice deficient in muscle creatine kinase (M-CK), we have fed mutant mice a diet containing the creatine analogue β-guanidinopropionic acid (βGPA). After 8–10 weeks of feeding, accumulation of the creatine analogue in M-CK-deficient muscles was comparable to that observed in muscles of wild-type mice. Strikingly, and unlike wild types, mutants did not accumulate phosphorylated βGPA, indicating that MM-CK is the only muscle CK isoform which can phosphorylate βGPA. In M-CK-deficient muscles there was respective depletion of PCr, Cr and ATP levels to 31, 41 and 83% of normal. The average cross-sectional area of type 2B fibres in gastrocnemius muscles was very much reduced and was similar to type 1 and type 2A fibres which maintained their normal size. The maximal isometric twitch force developed by gastrocnemius-plantaris-soleus (GPS) muscle complexes of βGPA-treated mutants was reduced by about 30%, but these muscles showed an increased fatigue resistance during 1 and 5 Hz contraction. Mitochondrial enzyme activities in the upper hind limb musculature of null mutants were 20–35% increased by the βGPA diet. Altogether, these results provide evidence that certain functions of the creatine kinase/phosphocreatine (CK/PCr) system are not eliminated solely by the loss of M-CK.

Fatigability and blood flow in the rat gastrocnemius-plantaris-soleus after hindlimb suspension.

The purpose of this study was to test the hypothesis that hindlimb suspension increases the fatigability of the soleus during intense contractile activity and determine whether the increased fatigue is associated with a reduced muscle blood flow. Cage-control (C) and 15-day hindlimb-suspended (HS) rats were anesthetized, and either the gastrocnemius-plantaris-soleus (G-P-S) muscle group or the soleus was stimulated (100 Hz, 100-ms trains at 120/min) for 10 min in situ. In the G-P-S preparation, blood flow was measured with radiolabeled microspheres before and at 2 and 10 min of contractile activity. The G-P-S fatigued markedly at this stimulation frequency, and the differences between C and HS animals were not significant until the 9th min of contractile activity. In contrast, the stimulation resulted in faster rates and significantly larger amounts of fatigue in the soleus from HS than from C animals. The atrophied soleus showed significant differences by 1 min of stimulation (C = 70 +/- 1% vs. HS = 57 +/- 2% of peak train force) and remained different at 10 min (C = 64 +/- 4% vs. HS = 45 +/- 2% peak train force). Relative blood flow to the soleus was similar between groups before and during contractile activity (rest: C = 20 +/- 3 vs. HS = 12 +/- 3; 2 min: C = 128 +/- 6 vs. HS = 118 +/- 4; 10 min: C = 123 +/- 11 vs. HS = 105 +/- 11 ml.min-1.100 g-1). In conclusion, these results established that 15 days of HS increased the fatigability of the soleus, but the effect was not caused by a reduced muscle blood flow.