Release of Intracellular Enzymes from An Isolated Mammalian Skeletal Muscle Preparation

1983 ◽  
Vol 65 (2) ◽  
pp. 193-201 ◽  
Author(s):  
D. A. Jones ◽  
M. J. Jackson ◽  
R. H. T. Edwards
Keyword(s):  
Skeletal Muscle ◽  
Contractile Activity ◽  
Atp Depletion ◽  
Mammalian Skeletal Muscle ◽  
Enzyme Release ◽  
Muscle Preparation ◽  
Muscle Enzymes ◽  
Hypoxic Conditions ◽  
Detergent Treatment

1. An isolated skeletal muscle preparation is described which has been used to study the efflux of enzymes in response to contractile activity, metabolic poisons and detergent treatment. 2. In both fast and slow muscles contractile activity caused a release of lactate dehydrogenase and creatine kinase that reached a peak 1–2 h after the end of stimulation. There was very little release during the 30 min stimulation period whether the muscles were under aerobic or hypoxic conditions. 3. Incubation of the muscles with cyanide and iodoacetate caused a similar delayed release of enzyme. 4. Disruption of cell membranes with detergent treatment caused an entirely different and very rapid pattern of enzyme release. 5. Enzyme release from the fatigued isolated muscle preparations appears to be initiated as a consequence of phosphorylcreatine or ATP depletion. The relevance of this to release of muscle enzymes after activity in vivo is discussed.

A malonyl-CoA fuel-sensing mechanism in muscle: effects of insulin, glucose, and denervation

1995 ◽  
Vol 269 (2) ◽  
pp. E283-E289 ◽  
Author(s):  
A. K. Saha ◽  
T. G. Kurowski ◽  
N. B. Ruderman
Keyword(s):  
Skeletal Muscle ◽  
Plasma Insulin ◽  
Contractile Activity ◽  
High Plasma ◽  
Muscle Contractions ◽  
Glucose Levels ◽  
Malonyl Coa ◽  
Ad Libitum ◽  
Sciatic Nerve Stimulation

Increases in the concentration of malonyl-CoA in skeletal muscle have been observed in the KKAy mouse, an obese rodent with high plasma insulin and glucose levels [Saha et al. Am. J. Physiol. 267 (Endocrinol. Metab. 30): E95-E101, 1994]. To assess whether insulin and glucose directly regulate malonyl-CoA in muscle, soleus muscles from young rats were incubated with insulin and glucose at various concentrations, and their content of malonyl-CoA was determined. In addition, the effect on malonyl-CoA of denervation and electrically induced muscle contractions was assessed. The concentration of malonyl-CoA in the soleus, taken directly from a rat fed ad libitum, was 2.0 +/- 0.2 nmol/g. In muscles incubated for 20 min in a medium devoid of added insulin and glucose, the concentration was decreased to 0.8 +/- 0.2 nmol/g. When the medium contained 0.5, 7.5, or 30 mM glucose, malonyl-CoA levels were 1.3 +/- 0.1, 1.8 +/- 0.1, or 2.4 +/- 0.2 nmol/g, respectively, in the absence of insulin and 1.7 +/- 0.1, 4.6 +/- 0.3, or 5.5 +/- 0.6 nmol/g in its presence (10 mU/ml). Compared with its level in a control muscle, the concentration of malonyl-CoA was increased threefold in the soleus 6-8 h after denervation and remained twofold higher for > or = 48 h. In contrast, muscle contractions induced by sciatic nerve stimulation, in vivo, acutely decreased the concentration of malonyl-CoA by 30-35%. The results indicate that insulin and glucose, and probably contractile activity, regulate the concentration of malonyl-CoA in muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Skeletal muscle phenotype signaling with ex vivo endurance-type dynamic contractions in rat muscle

2021 ◽  
Author(s):  
Jesper Emil Jakobsgaard ◽  
Jacob Andresen ◽  
Frank V. de Paoli ◽  
Kristian Vissing
Keyword(s):  
Skeletal Muscle ◽  
Translation Initiation ◽  
Ex Vivo ◽  
Contractile Activity ◽  
Specific Protein ◽  
Signaling Proteins ◽  
Dependent Manner ◽  
Metabolic Signaling ◽  
Dynamic Endurance

Skeletal muscle phenotype may influence the response sensitivity of myocellular regulatory mechanisms to contractile activity. To examine this, we employed an ex vivo endurance-type dynamic contraction model to evaluate skeletal muscle phenotype-specific protein signaling responses in rat skeletal muscle. Preparations of slow-twitch soleus and fast-twitch extensor digitorum longus skeletal muscle from 4-wk old female Wistar rats were exposed to an identical ex vivo dynamic endurance-type contraction paradigm consisting of 40 minutes of stretch-shortening contractions under simultaneous low-frequency electrostimulation delivered in an intermittent pattern. Phosphorylation of proteins involved in metabolic signaling and signaling for translation initiation was evaluated at 0, 1, and 4 hours after stimulation by immunoblotting. For both muscle phenotypes, signaling related to metabolic events was upregulated immediately after stimulation, with concomitant absence of signaling for translation-initiation. Signaling for translation-initiation was then activated in both muscle phenotypes at 1-4 hours after stimulation, coinciding with attenuated metabolic signaling. The recognizable pattern of signaling responses support how our ex vivo dynamic muscle contraction model can be utilized to infer a stretch-shortening contraction pattern resembling stretch-shortening contraction of in vivo endurance exercise. Moreover, using this model, we observed that some specific signaling proteins adhering to metabolic events or to translation initation exhibited phosphorylation changes in a phenotype-dependent manner, whereas other signaling proteins exhibited phenotype-independent changes. These findings may aid the interpretation of myocellular signaling outcomes adhering to mixed muscle samples collected during human experimental trials.

In vivo supraspinatus muscle contractility and architecture in rabbit

2020 ◽  
Vol 129 (6) ◽  
pp. 1405-1412
Author(s):  
Sydnee A. Hyman ◽  
Mackenzie B. Norman ◽  
Shanelle N. Dorn ◽  
Shannon N. Bremner ◽  
Mary C. Esparza ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Isometric Force ◽  
Muscle Physiology ◽  
Mammalian Skeletal Muscle ◽  
Improved Method ◽  
Muscle Contractility ◽  
Supraspinatus Muscle ◽  
Contractile Stress ◽  
Maximum Isometric Force

We introduce an improved method to assess rabbit supraspinatus muscle physiology. Maximum isometric force measured for the rabbit supraspinatus was dramatically greater than previous reports in the literature. Consequently, the isometric contractile stress reported is almost 10 times greater than previous reports of rabbit supraspinatus, but similar to available literature of other mammalian skeletal muscle. We show that previous reports of peak supraspinatus isometric force were subphysiological by ∼90%

THE APPARENT EXTRACELLULAR SPACE OF MAMMALIAN SKELETAL MUSCLE: A COMPARISON OF THE INULIN SPACE IN NORMAL AND DYSTROPHIC MOUSE TISSUES

1960 ◽  
Vol 38 (8) ◽  
pp. 829-835 ◽  
Author(s):  
L. H. Burr ◽  
H. McLennan
Keyword(s):  
Skeletal Muscle ◽  
Normal Tissue ◽  
Muscle Size ◽  
Mammalian Skeletal Muscle ◽  
Normal Tissues ◽  
Mouse Tissues ◽  
Inulin Space ◽  
Excised Tissues ◽  
Dystrophic Mice

The apparent extracellular volumes of the muscles from young normal and dystrophic mice have been estimated, using inulin dilution techniques. The inulin spaces were measured in the muscle both following injection of inulin in vivo and after soaking of excised tissues in a solution containing inulin. Comparisons were made between muscles of different size from the same animal as well as from different animals whose age, and consequently muscle size, varied. In all cases it has been found that the inulin space decreases with increasing muscle size. Similar results have been obtained by others with toad sartorii. The inulin space in muscles from dystrophic mice is larger than that of comparable normal tissues, and the dependence on muscle size, although similar to normal, is more pronounced. The results suggest that the dystrophic cells are permeable to inulin, and the question that some small permeability may be present also in normal tissue is considered.

Effect of initial length on relations between oxygen uptake and load in dog muscle

1976 ◽  
Vol 230 (4) ◽  
pp. 1008-1012 ◽  
Author(s):  
WN Stainsby ◽  
JK Barclay
Keyword(s):  
Skeletal Muscle ◽  
Oxygen Uptake ◽  
The Other ◽  
Mammalian Skeletal Muscle ◽  
Initial Length ◽  
Muscle Preparation ◽  
Optimal Length ◽  
Muscle Energetics ◽  
Isotonic Contractions ◽  
Fenn Effect

Oxygen uptake for brief tetanic contractions was calculated from measurements of blood flow and blood arteriovenous oxygen content differences. Each muscle preparation was pretested under isometric conditions to establish optimal length, Lo. After this one group of preparations performed afterload isotonic contractions at several loads with initial length, Li, less than Lo. The other groups of preparations performed similar contractions with Li greater than Lo. When Li was less than Lo, oxygen uptake for the highest load was always greater than oxygen uptake at the lowest load whereas intermediate loads were usually higher than both extremes. However, when Li was greater than Lo, oxygen uptake at the highest load was always less than oxygen uptake at the lowest load; again the intermediate loads were usually higher than both extremes. The data confirm and extend similar effects of initial length on heat production for contractions by amphibian muscles (7). It seems likely that the differences in initial lengths may account for the fact that the Fenn effect has not previously been observed in studies of mammalian skeletal muscle energetics.

scholarly journals Muscle-Derived Extracellular Signal-Regulated Kinases 1 and 2 Are Required for the Maintenance of Adult Myofibers and Their Neuromuscular Junctions

2015 ◽  
Vol 35 (7) ◽  
pp. 1238-1253 ◽  
Author(s):  
Bonnie Seaberg ◽  
Gabrielle Henslee ◽  
Shuo Wang ◽  
Ximena Paez-Colasante ◽  
Gary E. Landreth ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Tibialis Anterior ◽  
Neuromuscular Junctions ◽  
Germ Line ◽  
Fiber Type ◽  
Postnatal Growth ◽  
Mammalian Skeletal Muscle ◽  
Neuromuscular Synapses ◽  
Extracellular Signal

The Ras–extracellular signal-regulated kinase 1 and 2 (ERK1/2) pathway appears to be important for the development, maintenance, aging, and pathology of mammalian skeletal muscle. Yet no gene targeting ofErk1/2in muscle fibersin vivohas been reported to date. We combined a germ lineErk1mutation with Cre-loxPErk2inactivation in skeletal muscle to produce, for the first time, mice lacking ERK1/2 selectively in skeletal myofibers. Animals lacking muscle ERK1/2 displayed stunted postnatal growth, muscle weakness, and a shorter life span. Their muscles examined in this study, sternomastoid and tibialis anterior, displayed fragmented neuromuscular synapses and a mixture of modest fiber atrophy and loss but failed to show major changes in fiber type composition or absence of cell surface dystrophin. Whereas the lack of only ERK1 had no effects on the phenotypes studied, the lack of myofiber ERK2 explained synaptic fragmentation in the sternomastoid but not the tibialis anterior and a decrease in the expression of the acetylcholine receptor (AChR) epsilon subunit gene mRNA in both muscles. A reduction in AChR protein was documented in line with the above mRNA results. Evidence of partial denervation was found in the sternomastoid but not the tibialis anterior. Thus, myofiber ERK1/2 are differentially required for the maintenance of myofibers and neuromuscular synapses in adult mice.

scholarly journals Purification of desmin from adult mammalian skeletal muscle

1981 ◽  
Vol 195 (2) ◽  
pp. 345-356 ◽  
Author(s):  
J M O'Shea ◽  
R M Robson ◽  
M K Hartzer ◽  
T W Huiatt ◽  
W E Rathbun ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Average Diameter ◽  
Intermediate Filament Protein ◽  
Mammalian Skeletal Muscle ◽  
Cytoskeletal Function ◽  
10 Nm Filaments ◽  
And Function ◽  
Filament Protein

A method has been developed for preparation of purified desmin from mature mammalian (porcine) skeletal muscle. A crude desmin-containing fraction was prepared by modification of procedures used for isolation of smooth-muscle intermediate-filament protein [Small & Sobieszek (1977) J. Cell Sci. 23, 243-268]. The desmin was extracted in 1 M-acetic acid/20 mM-NaCl at 4 degrees C for 15h from the residue remaining after actomyosin extraction from washed myofibrils. Successive chromatography on hydroxyapatite and DEAE-Sepharose CL-6B in 6M-urea yielded desmin that was routinely more than 97% 55 000-dalton protein and that had no detectable actin contamination. Removal of urea by dialysis against 10mM-Tris/acetate (pH 8.5)/1 mM dithioerythritol and subsequent clarification at 134 000 g (rav. 5.9 cm) for 1 h results in a clear desmin solution. Dialysis of purified desmin against 100 mM-NaCl/1 mM-MgCl2/10 mM-imidazole/HCl, pH 7.0, at 2 degrees C resulted in the formation of synthetic desmin filaments have an average diameter of 9-11.5 nm. The present studies demonstrate that the relatively small amount of desmin in mature skeletal muscle can be isolated in sufficient quantity and purity to permit detailed studies of its properties and function. Although 10nm filaments have not been unequivocally demonstrated in mature muscle in vivo, that the purified skeletal-muscle desmin will form 10 nm filaments in vitro lends support to their possible existence and cytoskeletal function in mature skeletal-muscle cells.

scholarly journals Long-chain n-3 DHA reduces the extent of skeletal muscle fatigue in the rat in vivo hindlimb model

2013 ◽  
Vol 111 (6) ◽  
pp. 996-1003 ◽  
Author(s):  
Gregory E. Peoples ◽  
Peter L. McLennan
Keyword(s):  
Skeletal Muscle ◽  
Fish Oil ◽  
Muscle Fatigue ◽  
Muscle Membrane ◽  
Mammalian Skeletal Muscle ◽  
Skeletal Muscle Fatigue ◽  
Arterial Blood ◽  
Oxygen Efficiency ◽  
Long Chain

Dietary fish oil modifies skeletal muscle membrane fatty acid composition and oxygen efficiency similar to changes in the myocardium. Oxygen efficiency is a key determinant of sustained force in mammalian skeletal muscle. Therefore, in the present study, we tested the effects of a fish-oil diet on skeletal muscle fatigue under the stress of contraction using the rat in vivo autologous perfused hindlimb model. For 8 weeks, male Wistar rats were fed a diet rich in saturated fat (SF), a diet rich in n-6 PUFA or a diet rich in long-chain (LC) n-3 PUFA DHA derived from fish oil. In anaesthetised, mechanically ventilated rats, with their hindlimbs perfused with arterial blood at a constant flow, the gastrocnemius–plantaris–soleus muscle bundle was stimulated via sciatic nerve (2 Hz, 6–12 V, 0·05 ms) to contract repetitively for 30 min. Rats fed the n-3 PUFA diet developed higher maximum twitch tension than those fed the SF and n-6 PUFA diets (P< 0·05) and sustained twitch tension through more repetitions before the tension declined to 50 % of the maximum twitch tension (P< 0·05). The n-3 PUFA group used less oxygen for tension developed and produced higher venous lactate concentrations with no difference in glycogen utilisation compared with the SF and n-6 PUFA groups. These results further support that incorporation of DHA into skeletal muscle membranes increases the efficiency of oxygen use over a range of contractile force and this is expressed as a higher sustained force and prolonged time to fatigue.

Release of sarcoplasmic enzymes from frog skeletal muscle

1981 ◽  
Vol 241 (3) ◽  
pp. C98-C105 ◽  
Author(s):  
G. Suarez-Kurtz ◽  
A. B. Eastwood
Keyword(s):  
Skeletal Muscle ◽  
Creatine Kinase ◽  
Lactate Dehydrogenase ◽  
Enzyme Release ◽  
Frog Skeletal Muscle ◽  
Free Medium ◽  
Release Rates ◽  
Brij 58 ◽  
Order Of Magnitude ◽  
Detergent Treatment

Isolated, intact frog muscles bathed in control saline release creatine kinase (CK) and lactate dehydrogenase (LDH) at constant rates for several hours. The basal rates of release from “toe” muscles (CK 0.087%/min; LDH 0.105%/min) were one order of magnitude greater than those from semitendinosus muscles. This is attributed to differences in muscle mass and geometry, and to the smaller diameter of toe muscle fibers. Enzyme release rates were not affected by Na-free or Cl-free solutions, whereas LDH release rate doubled during exposure to Ca-free (EGTA-containing) saline or in the presence of isosmotic solutions containing 120 mM KCl or potassium propionate. Following mechanical injury or detergent treatment (Brij 58), the enzyme release rates into Ca-free medium reached peak values 4 and 16 times (toe muscle), and 16 and 20–30 times (semitendinosus), respectively, the control rates. The greater effect of detergent treatment is ascribed to a larger area of sarcolemmal damage plus possible changes in the state of the enzymes in the sarcoplasm.

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