Assessment of biochemical viability of isolated incubated muscle preparations

Newsholme, Eric A.; Leighton, Brendan; Challiss, R. A. John; Lozeman, Fred J.

doi:10.1042/bj2380621

Cited by 23 publications

(14 citation statements)

References 4 publications

(7 reference statements)

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“…Oxygenation of the incubating medium generally occurs either by continuous gassing of 95% O 2 and 5% CO 2 through the medium or by saturating the oxygen content of the buffer. Both methods have been shown to maintain adequate oxygen levels during an in vitro experiment (Newsholme et al, 1986;Van Breda et al, 1990;Bonen et al, 1994). For in vitro experiments, the surgical preparation of both the superior oblique muscle and lateral gastrocnemius muscles were similar to the in situ preparation until blood flow was interrupted.…”

Section: Surgical Preparation Of Animalsmentioning

confidence: 99%

Measurement of contractile force of skeletal and extraocular muscles: Effects of blood supply, muscle size and in situ or in vitro preparation

Croes

Bartheld

2007

Journal of Neuroscience Methods

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Contractile forces can be measured in situ and in vitro. To maintain metabolic viability with sufficient diffusion of oxygen, established guidelines for in vitro skeletal muscle preparations recommend use of relatively thin muscles (≤1.25 mm thick). Nevertheless, forces of thin extraocular muscles vary substantially between studies. Here, we examined parameters that affect force measurements of in situ and in vitro preparations, including blood supply, nerve stimulation, direct muscle stimulation, muscle size, oxygenated or non-oxygenated buffer solutions, and the time after interruption of vascular circulation. We found that the absolute forces of extraocular muscle are substantially lower when examined in vitro. In vitro preparation of 0.58 mm thick extraocular muscle from 3-week old birds underestimated contractile function, but not of thinner (0.33 mm) muscle from 2-day old birds. Our study shows that the effective criteria for functional viability, tested in vitro, differ between extraocular and other skeletal muscle. We conclude that contractile force of extraocular muscles will be underestimated by between 10 and 80%, when measurements are made after cessation of blood supply (at 5 -40 min). The mechanisms responsible for the declining values for force measurements are discussed, and we make specific recommendations for obtaining valid measurements of contractile force.

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Section: Surgical Preparation Of Animalsmentioning

confidence: 99%

Measurement of contractile force of skeletal and extraocular muscles: Effects of blood supply, muscle size and in situ or in vitro preparation

Croes

Bartheld

2007

Journal of Neuroscience Methods

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“…Previous experiments have established that, at this size, this muscle preparation is in a biochemically viable state for periods of 60-90 min at 37 ЊC (22).…”

Section: Resultsmentioning

confidence: 92%

Furosemide decreases the sensitivity of glucose transport to insulin in skeletal muscle in vitro

et al. 1998

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The effects of the diuretic furosemide on the sensitivity of glucose disposal to insulin were investigated in rat soleus muscle in vitro. At basal levels of insulin, the rates of 3-O-methylglucose transport, 2-deoxyglucose phosphorylation and lactate formation were not affected significantly by furosemide (0.5 mmol/l). However, furosemide significantly decreased these rates at physiological and maximal levels of insulin. The contents of 2-deoxyglucose and glucose 6-phosphate in the presence of furosemide were not significantly different from those in control muscles at all levels of insulin studied. It is concluded that furosemide decreases the sensitivity of glucose utilization to insulin in skeletal muscle by directly inhibiting the glucose transport process.

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“…This method of muscle preparation also allows for the determination of the absolute rate of glutamine uptake, in contrast with the net rate of transport determined from arteriovenous difference studies 24 . In addition, this technique provides a viable in vitro muscle preparation with which it is possible to investigate rates of glutamine transport at rest and during exercise 25,42 …”

Section: Discussionmentioning

confidence: 99%

“…Glutamine transport was determined in whole soleus muscle from one leg incubated according to a method described by Parry‐Billings et al 24 Briefly, the muscle was dissected longitudinally and the distal and proximal tendons were ligated and attached to stainless steel holders such that the muscle was maintained at in situ resting tension. Several lines of evidence have demonstrated that this setup is a viable in vitro muscle preparation with which to study the rate of glutamine release: after 60 min of incubation, mitochondrial morphology is normal, no significant hypoxia is observed, the glycogenolytic rate is low and ATP, ADP and AMP are maintained at near in vivo levels 25 . After 30 min incubation at 37°C in incubation medium containing PBS, 1.5% BSA, 5 mmol/L glutamine and 10 µUI/mL insulin, the rate of glutamine uptake from incubated soleus muscle was determined by enzymatic analysis of the change in glutamine concentration in the incubation medium over the incubation period.…”

Section: Methodsmentioning

confidence: 97%

Effect of Exercise on Glutamine Synthesis and Transport in Skeletal Muscle From Rats

Santos

Caperuto

Mello

et al. 2009

Clin Exp Pharma Physio

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1. Reductions in plasma glutamine are observed after prolonged exercise. Three hypotheses can explain such a decrease: (i) high demand by the liver and kidney; (ii) impaired release from muscles; and (iii) decreased synthesis in skeletal muscle. The present study investigated the effects of exercise on glutamine synthesis and transport in rat skeletal muscle. 2. Rats were divided into three groups: (i) sedentary (SED; n = 12); (ii) rats killed 1 h after the last exercise bout (EX-1; n = 15); and (iii) rats killed 24 h after the last exercise bout (EX-24; n = 15). Rats in the trained groups swam 1 h/day, 5 days/week for 6 weeks with a load equivalent to 5.5% of their bodyweight. 3. Plasma glutamine and insulin were lower and corticosterone was higher in EX-1 compared with SED rats (P < 0.05 and P < 0.01, respectively). Twenty-four hours after exercise (EX-24), plasma glutamine was restored to levels seen in SED rats, whereas insulin levels were higher (P < 0.001) and corticosterone levels were lower (P < 0.01) than in EX-1. In the soleus, ammonia levels were lower in EX-1 than in SED rats (P < 0.001). After 24 h, glutamine, glutamate and ammonia levels were lower in EX-24 than in SED and EX-1 rats (P < 0.001). Soleus glutamine synthetase (GS) activity was increased in EX-1 and was decreased in EX-24 compared with SED rats (both P < 0.001). 4. The decrease in plasma glutamine concentration in EX-1 is not mediated by GS or glutamine transport in skeletal muscle. However, 24 h after exercise, lower GS may contribute to the decrease in glutamine concentration in muscle.

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Assessment of biochemical viability of isolated incubated muscle preparations

Cited by 23 publications

References 4 publications

Measurement of contractile force of skeletal and extraocular muscles: Effects of blood supply, muscle size and in situ or in vitro preparation

Measurement of contractile force of skeletal and extraocular muscles: Effects of blood supply, muscle size and in situ or in vitro preparation

Furosemide decreases the sensitivity of glucose transport to insulin in skeletal muscle in vitro

Effect of Exercise on Glutamine Synthesis and Transport in Skeletal Muscle From Rats

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