Functional and structural changes after disuse of human muscle – first study to quantify disuse muscle atrophy at fibre level in humans

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Clinical Science and Molecular Medicine (1977) 52, 337-342. Functional and structural changes after disuse of human muscle – Authors: ANTHONY J SARGEANT,* C. T. M. DAVIES,* R. H. T. EDWARDS, C. MAUNDER AND A. YOUNG *Medical Research Council Environmental Physiology Unit, London School of Hygiene and Tropical Medicine, University of London, and Jerry Lewis Muscle Research Centre, Royal Postgraduate Medical School, Hammersmith Hospital, London

Summary

1. Seven patients who had suffered unilateral leg fracture were studied after removal of immobilizing plaster casts.

2. Leg volume measured anthropometrically was reduced by 12% in the injured leg (5.68 f 1.05 litres) compared with the uninjured (6.43 f 0.87 litres). Associated with this loss was a similar reduction in the net maximum oxygen uptake achieved in one-leg cycling, from 1.89 k 0.21 l/min in the uninjured leg to 1.57+0.18 l/min in the injured.

3. Measured by a percutaneous needle biopsy technique, a reduction of 42% was found in the cross-sectional area of the muscle fibres sampled from the vastus lateralis of the injured compared with the uninjured leg.

4. Staining for myosin adenosine triphosphatase activity showed that both type I and I1 fibres were affected, being reduced respectively from 3410 to 1840 pm2 and from 3810 to 2390 pm2 cross-sectional area.

5. Possible reasons and implications are discussed for the discrepancy between the magnitude of the difference observed in the gross measurement of leg function (maximum oxygen uptake) and structure (leg volume) as compared with the cellular level (cross-sectional fibre area).

 

Correspondence: Dr A. J. Sargeant, MRC Environmental Physiology Unit, London School of Hygiene and Tropical Medicine, University of London, Keppel Street (Gower Street), London WClE 7HT.

Introduction

Atrophy of the affected limb and loss of muscle power follows bone fracture and subsequent immobilization. Years of experience have enabled the rehabilitation professions to develop empirical programmes to reverse these changes. However, the efficacy of such programmes may be further improved if we can increase our understanding of the atrophic response to disuse in human muscle. Recent studies showed that 15 weeks immobilization in a long-leg plaster cast after fracture reduced the fat-free volume of the affected leg by 12%, which was accompanied by a similar fall in the maximum oxygen uptake ( ~oz,,,,=.) achieved with oneleg pedalling (Davies & Sargeant, 1975a,b). However, it was not known how far these changes in gross structure and function were reflected at a cellular level within the affected muscles. Since the work of pedalling is performed mainly by the leg extensors (A. J. Sargeant & C. T. M. Davies, unpublished work) needle biopsy was used (Edwards, Maunder, Lewis & Pearse, 1973) to study fibre atrophy in the quadriceps femoris muscle and to compare this with measurements of the gross leg volume and maximal oxygen uptake of patients recovering from unilateral leg fracture.

http://www.clinsci.org/content/ppclinsci/52/4/337.full.pdf

Structural and functional determinants of human muscle power – Review by Anthony J Sargeant

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Experimental PhysiologyExperimental Physiology

Abstract

Measurements of human power need to be interpreted in relation to the movement frequency, since that will determine the velocity of contraction of the active muscle and hence the power available according to the power–velocity relationship. Techniques are described which enable movement frequency to be kept constant during human exercise under different conditions. Combined with microdissection and analysis of muscle fibre fragments from needle biopsies obtained pre- and postexercise we have been able ‘to take the muscle apart’, having measured the power output, including the effect of fatigue, under conditions of constant movement frequency. We have shown that fatigue may be the consequence of a metabolic challenge to a relatively small population of fast fatigue-sensitive fibres, as indicated by [ATP] depletion to ∼30% of resting values in those fibres expressing myosin heavy chain isoform IIX after just 10 s of maximal dynamic exercise. Since these same fibres will have a high maximal velocity of contraction, they also make a disproportionate contribution to power output in relation to their number, especially at faster movement rates. The microdissection technique can also be used to measure phosphocreatine concentration ([PCr]), which is an exquisitely sensitive indicator of muscle fibre activity; thus, in just seven brief maximal contractions [PCr] is depleted to levels < 50% of rest in all muscle fibre types. The technique has been applied to study exercise at different intensities, and to compare recruitment in lengthening, shortening and isometric contractions, thus yielding new information on patterns of recruitment, energy turnover and efficiency.

Within a single muscle there can be large differences in fatiguability and other physiological properties

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This research carried out in Amsterdam under the direction of Professor Anthony J Sargeant demonstrated how within the same anatomical muscle there can be quiet different physiological properties in different areas of the same muscle. This work was part of the PhD research of Jo de Ruiter supervised by Professor Tony Sargeant and Arnold de Haan.

Repeated force production and metabolites in two medial gastrocnemius muscle compartments of the rats

De Ruiter CJArnold de HaanAnthony J Sargeant.

Journal of Applied Physiology
J Appl Physiol. 1995 Dec;79(6):1855-61
  • The most proximal and distal motor nerve branches in the rat medial gastrocnemius innervate discrete muscle compartments dominated by fast-twitch oxidative and fast-twitch glycolytic fibers, respectively. The functional consequences of the difference in oxidative capacity between these compartments were investigated. Wistar rats were anesthetized with pentobarbital sodium (90 mg/kg ip). Changes in force of both compartments during 21 isometric contractions (train duration 200 ms, stimulation frequency 120 Hz, 3 s between contractions) were studied in situ with and without blood flow. Without blood flow, force and phosphocreatine declined to a greater extent in the proximal than the distal compartment compared with the run with intact flow. After the protocol without blood flow, when flow was restored, the time constants for force recovery (which were closely associated to the recovery of phosphocreatine) were 37 +/- 7 (SD) (proximal compartment) and 148 +/- 20 s (distal compartment). It was concluded that the proximal compartment had a four times higher oxidative capacity and, therefore, a superior ability for repeated force production.

Human Muscle Power

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The ability of generating muscle power power is important whether you are an Olympic athlete, a ballet dancer, or an elderly person wanting to climb the stairs to go to bed. In this comprehensive review of his research Anthony Sargeant points out the importance of different types of muscle fibres that make up the human skeletal muscles that produce power in legs and arms. Tony also points out that in research seeking to measure human muscle power it is essential to measure or control the speed at which the power is generated (this is because power is the product of work and velocity).

Structural and functional determinants of human muscle power
by Anthony J Sargeant

Experimental Physiology
Exp Physiol. 2007 Mar;92(2):323-31

Measurements of human power need to be interpreted in relation to the movement frequency, since that will determine the velocity of contraction of the active muscle and hence the power available according to the power-velocity relationship. Techniques are described which enable movement frequency to be kept constant during human exercise under different conditions. Combined with microdissection and analysis of muscle fibre fragments from needle biopsies obtained pre- and postexercise we have been able ‘to take the muscle apart’, having measured the power output, including the effect of fatigue, under conditions of constant movement frequency. We have shown that fatigue may be the consequence of a metabolic challenge to a relatively small population of fast fatigue-sensitive fibres, as indicated by [ATP] depletion to approximately 30% of resting values in those fibres expressing myosin heavy chain isoform IIX after just 10 s of maximal dynamic exercise. Since these same fibres will have a high maximal velocity of contraction, they also make a disproportionate contribution to power output in relation to their number, especially at faster movement rates. The microdissection technique can also be used to measure phosphocreatine concentration ([PCr]), which is an exquisitely sensitive indicator of muscle fibre activity; thus, in just seven brief maximal contractions [PCr] is depleted to levels < 50% of rest in all muscle fibre types. The technique has been applied to study exercise at different intensities, and to compare recruitment in lengthening, shortening and isometric contractions, thus yielding new information on patterns of recruitment, energy turnover and efficiency.

Anthony J Sargeant

Muscle Fibre can co-express different isoforms of Myosin Heavy Chain

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Anthony Sargeant directed this work on skeletal muscle as Head of the Amsterdam research group. The meticulous work was carried out by Jose Sant’Ana Pereira who was one of Tony Sargeant’s PhD students.

The mATPase histochemical profile of rat type IIX fibres: correlation with myosin heavy chain immunolabelling

Jose A A Sant’Ana PereiraArnold de HaanWessels AAnton F MoormanAnthony J Sargeant.

Journal of Histochemistry
Histochem J. 1995 Sep;27(9):715-22
In the present study we report a novel histochemical method which, by sequential pre-incubations in alkaline and acidic media, selectively differentiates muscle fibres expressing myosin heavy chain IIX, on the basis of a specific profile for myofibrillar actomyosin ATPase (mATPase) activity. The enzyme reactions were tested for specificity by means of anti-myosin heavy chain monoclonal antibodies, which were characterized on Western blots of muscle homogenates. Enzyme histochemical reactions with the traditional pH buffers were compared to those of the new method and, in conjunction with the immunoreactions, used to confirm the relationship between MyHC expression and the distinct profiles for mATPase. Immunohistochemical reactions demonstrated that the new method only differentiates those fibres expressing myosin heavy chain IIX. The method revealed a continuum in which the intermediate staining intensities corresponded to hybrid fibres expressing myosin heavy chain IIX in combination with either the IIA or IIB forms. Quantitative histochemistry and immunohistochemistry (by image analysis), used to examine the relationship between staining intensities for mATPase and amounts of myosin heavy chain IIX expression, revealed that the new method discriminates well between hybrid fibres expressing variable amounts of the IIX isoform (r2 = 0.93)

Human muscle fatigue

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Anita Beelen presented this research as part of her PhD thesis supervised by Professor Anthony Sargeant. Uniquely the study used electrical stimulation superimposed upon on maximal voluntary activation in dynamic exercise.

Fatigue and recovery of voluntary and electrically elicited dynamic force in humans

Anita BeelenAnthony J SargeantDavid A JonesC. J. de Ruiter

Journal of Physiology
J Physiol. 1995 Apr 1;484 ( Pt 1):227-235
1. Percutaneous electrical stimulation of the human quadriceps muscle has been used to assess the loss of central activation immediately after a bout of fatiguing exercise and during the recovery period.
2. Fatigue was induced in eight healthy males by a maximal effort lasting 25 s performed on an isokinetic cycle ergometer at a constant pedal frequency of 60 revolutions per minute. The cranks of the ergometer were driven by an electric motor. Before and after the sprint, subjects allowed their legs to be passively taken round by the motor. During the passive movement the knee extensors were stimulated (4 pulses; 100 Hz). Peak voluntary force (PVF) during the sprint and peak stimulated forces (PSF) before and in recovery were recorded via strain gauges in the pedals. Recovery of voluntary force was assessed in a series of separate experiments in which subjects performed a second maximal effort after recovery periods of different durations.
3. Peak stimulated forces were reduced to 69f8 + 9 3 % immediately after the maximal effort, (P< 0 05), but had returned to pre-exercise values after 3 min. The maximum rate of force development (MRFD) was also reduced following fatigue to 68f8 + 11 0% (P < 0’05) of control and was fully recovered after 2 min. PVF was reduced to 72-0 + 9 4% (P< 0 05) of the control value following the maximal effort. After 3 min voluntary force had fully recovered.
4. The effect of changing the duration of the fatiguing exercise (10, 25 and 45 s maximal effort) resulted in an increased degree of voluntary force loss as the duration of the maximal effort increased. This was associated with an increased reduction in PSF measured immediately after the exercise.
5. The close association between the changes in stimulated force and voluntary force suggests that the fatigue in this type of dynamic exercise may be due to changes in the muscle itself and not to failure of central drive.

Different regions within the same muscle can have very different properties

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This research was part of the PhD thesis of CJ (Jo) de Ruiter. It shows how within a single muscle there may be marked regional differences in physiological characteristics implying task dependent differences in recruitment patterns of motor units. The work was carried out under the direction of Anthony J Sargeant and Arnold de Haan.

Physiological characteristics of two extreme muscle compartments in gastrocnemius medialis of the anaesthetized rat

De Ruiter CJArnold de HaanAnthony J Sargeant
Acta Physiologica Scandavica
Acta Physiol Scand. 1995 Apr;153(4):313-24
Rat medial gastrocnemius (GM) muscle is a compartmentalized muscle. The functional properties and fibre type composition of the most proximal and most distal compartment were studied in in situ preparations. The proximal compartment contained predominantly fast twitch oxidative fibres. The distal compartment was mainly composed of fast twitch glycolytic fibres. With the use of two small electrodes placed around the primary nerve branches, both compartments could be separately stimulated within the same muscle. The length-force relationship was less broad and maximal twitch and tetanic forces were obtained at lower muscle lengths for the proximal compartment. The differences (mm) were 0.9 +/- 0.2 and 1.2 +/- 0.2 for maximal twitch and tetanic force (120 Hz) production, respectively (P < 0.001). The shortening velocity for maximal power production was lower (P < 0.001) for the proximal compartment (proximal: 57.1 +/- 2.7 mm s-1, distal: 73.1 +/- 3.0 mm s-1). During a standard fatigue test the fatiguability was significantly lower for the proximal compared with the distal fibres. Our findings suggest that the proximal compartment is likely to be activated in vivo during activities requiring relatively low power outputs for longer time periods. In contrast the distal compartment is probably recruited only during high power demanding short lasting activities. The presented model makes it possible to study fatigue related changes in power production of the ‘red’ and ‘white’ areas of the GM separately in a way that is probably meaningful with respect to in vivo function.