Effects of Growth hormone on rat skeletal muscle after hindlimb suspension

To examine the effects of growth hormone (GH) on the preferential atrophy of the soleus muscle (SOL) occurring after hindlimb suspension (HS), two groups of male rats received daily injections of 2 IU · kg ^–1 body mass of recombinant human growth hormone (rhGH). Rats were either suspended by the tail for 21 days (HS-GH, n = 5) or nonsuspended (CGH, n=5). The effects of rhGH treatment on SOL and extensor digitorum longus muscles (EDL) were compared in two groups of animals receiving daily injections of saline, either suspended by the tail (HS-SA, n = 5) or nonsuspended (C-SA, n = 5). The results showed that the SOL hypertrophy in response to rhGH administration was mostly observed in C rats (+33%, P<0.01). This increase in muscle mass was correlated with a concomitant increase in the size of type I fibres (+21%, P<0.05). Although SOL mass decreased during HS in rhGH treated animals (–44%, P<0.001), the mean normalized mass of this muscle did not significantly differ between C-SA and HS-GH groups. A statistically significant increase in the absolute mass of EDL occurred with rhGH treatment in CGH (+12%, P<0.05). The HS-induced decrease in the percentage distribution of type I fibres in SOL was unaffected by the rhGH treatment. In addition, a decrease in the citrate synthase activity in the whole SOL was observed in the two groups of tail-suspended rats (–31%, P<0.05; –21%, P<0.05 in SA and GH animals, respectively). The activity of 3-hydroxyacyl coenzyme A dehydrogenase was enhanced by the rhGH treatment (P<0.05) with similar magnitude in both C (+25%) and HS rats (+24%). Therefore, GH prevented only slightly the atrophy of SOL, occurring after 21 days of HS. The effects of rhGH treatment appeared most effective in C rats, suggesting that HS impaired the growth-promoting effects of this hormone on skeletal muscle.     

Repositioning of fast and slow skeletal muscle

Summary The origins of the rat fast plantaris and slow soleus muscles were surgically reversed and their fibre types and contractile properties examinedin vitro up to 12 weeks post surgery. Muscles in which the origins had been severed and then immediately sutured back in place served as one control group. Unoperated animals served as a second control group. As compared to these groups, no significant differences in the histochemical or mechanical properties of the repositioned muscles were detected. Under the conditions of the experiments, no evidence was obtained to indicate that the intrinsic properties of a muscle could be altered by changing its site of origin and thus its functional environment.     

Collagen of slow twitch and fast twitch muscle fibres in different types of rat skeletal muscle

Summary The appearance of collagen around individual fast twitch (FT) and slow twitch (ST) muscle fibres was investigated in skeletal muscles with different contractile properties using endurance trained and untrained rats as experimental animals. The collagenous connective tissue was analyzed by measuring hydroxyproline biochemically and by staining collagenous material histochemically in M. soleus (MS), M. rectus femoris (MRF), and M. gastrocnemius (MG). The concentration of hydroxyproline in the ST fibres dissected from MS (2.72±0.35 g·mg^–1 d.w.) was significantly higher than that of the FT fibres dissected from MRF (1.52±0.33 g·mg^–1 d.w.). Similarly, the concentration of hydroxyproline was higher in ST (2.54±0.51 g·mg^–1 d.w.) than in FT fibres (1.60±0.43 g·mg^–1 d.w.), when the fibres were dissected from the same muscle, MG. Histochemical staining of collagenous material agreed with the biochemical evidence that MS and the slow twitch area of MG are more collagenous than MRF and the fast twitch area of MG both at the level of perimysium and endomysium. The variables were not affected by endurance training. When discussing the role of collagen in the function of skeletal muscle it is suggested that the different functional demands of different skeletal muscles are also reflected in the structure of intramuscular connective tissue, even at the level of endomysial collagen. It is supposed that the known differences in the elastic properties of fast tetanic muscle compared to slow tonic muscle as, e.g., the higher compliance of fast muscle could at least partly be explained in terms of the amount, type, and structure of intramuscular collagen.This study was supported by grants from the Finnish Research Council for Physical Education and Sport (Ministry of Education) and the Academy of Finnland     

Changes in contractile activation characteristics of rat fast and slow skeletal muscle fibres during regeneration

Damaged skeletal muscle fibres are replaced with new contractile units via muscle regeneration. Regenerating muscle fibres synthesize functionally distinct isoforms of contractile and regulatory proteins but little is known of their functional properties during the regeneration process. An advantage of utilizing single muscle fibre preparations is that assessment of their function is based on the overall characteristics of the contractile apparatus and regulatory system and as such, these preparations are sensitive in revealing not only coarse, but also subtle functional differences between muscle fibres. We examined the Ca²⁺- and Sr²⁺-activated contractile characteristics of permeabilized fibres from rat fast-twitch (extensor digitorum longus) and slow-twitch (soleus) muscles at 7, 14 and 21 days following myotoxic injury, to test the hypothesis that fibres from regenerating fast and slow muscles have different functional characteristics to fibres from uninjured muscles. Regenerating muscle fibres had ∼10% of the maximal force producing capacity ( P _o) of control (uninjured) fibres, and an altered sensitivity to Ca²⁺ and Sr²⁺ at 7 days post-injury. Increased force production and a shift in Ca²⁺ sensitivity consistent with fibre maturation were observed during regeneration such that P _o was restored to 36–45% of that in control fibres by 21 days, and sensitivity to Ca²⁺ and Sr²⁺ was similar to that of control (uninjured) fibres. The findings support the hypothesis that regenerating muscle fibres have different contractile activation characteristics compared with mature fibres, and that they adopt properties of mature fast- or slow-twitch muscle fibres in a progressive manner as the regeneration process is completed.     

Changes in CNTF receptor α expression in rat skeletal muscle during the recovery period after hindlimb suspension

This study investigated whether modifications in the contractile function of rat muscle after 3 weeks of hindlimb suspension followed by different recovery periods were associated with changes in ciliary neurotrophic factor (CNTF) receptor α. CNTF receptor α was stronger in control extensor digitorum longus (EDL) than soleus. CNTF receptor α was increased in unweighted as compared to control soleus muscle, while no significant changes occurred in EDL muscle. Furthermore, CNTF level was not significantly modified in adult sciatic nerve.     

Changes in stiffness induced by hindlimb suspension in rat soleus muscle

Soleus muscle atrophy was induced by hindlimb suspension of rats for 3 weeks with the intention of inducing a relative increase in the percentage of fast twitch fibres and assessing modifications in muscle stiffness. A method of dual controlled releases was used to obtain tension/extension curves and force/velocity relationships characterizing the mechanical behaviour of the soleus. Fibre typing was achieved by myofibrillar adenosine 5-triphosphatase staining. Results showed that hindlimb suspension decreased the percentage of slowtwitch fibres (–31%) to the profit of fast-twitch fibres (+370%) and intermediate fibres (+255%). This led to an increase in maximal shortening velocity. Tension/extension curves indicated a decrease in soleus stiffness after 3 weeks of unloading. Changes in elastic properties are interpreted in terms of modifications occurring in the active part and the passive part of the so-called series elastic component. These changes also suggest that the parameters derived from a twitch are inappropriate to account for modifications in speed-related properties of muscle.     

Comparative effects of hindlimb suspension and exercise on skeletal muscle myosin isozymes in rats

Summary The purpose of this study was to ascertain the time course of changes, whilst suspending the hindlimb and physical exercise training, of myosin light chain (LC) isoform expression in rat soleus and vastus lateralis muscles. Two groups of six rats were suspended by their tails for 1 or 2 weeks, two other groups of ten rats each were subjected to exercise training on a treadmill for 9 weeks, one to an endurance training programme (1-h running at 20 m · min^–1 5 days · week^–1), and the other to a sprint programme (30-s bouts of running at 60 m · min^–1 with rest periods of 5 min). At the end of these experimental procedures, soleus and vastus lateralis superficialis muscles were removed for myosin LC isoform determination by two-dimensional gel electrophoresis. Hindlimb suspension for 2 weeks significantly increased the proportion of fast myosin LC and decreased slow myosin LC expression in the soleus muscle. The pattern of myosin LC was unchanged in the vastus lateralis muscle. Sprint training or endurance training for 9 weeks increased the percentage of slow myosin LC in vastus lateralis muscle, whereas soleus muscle myosin LC was not modified. These data indicate that hindlimb suspension influences myosin LC expression in postural muscle, whereas physical training acts essentially on phasic muscle. There were no differences in myosin LC observed under the influence of sprint- or endurance-training programme.     

Dynamic properties of fast and slow skeletal muscles of the rat after nerve cross-union

1. The properties of self-innervated (S-EDL, S-SOL) and cross-innervated (X-EDL, X-SOL) extensor digitorum longus (EDL) and soleus (SOL) muscles have been determined at various times between 25 and 490 days after operations, and these are compared with the properties of normal muscles from unoperated animals of about the same age.2. The muscle fibres of X-SOL were 1.15 times longer than fibres of N-SOL and S-SOL at about 480 days after operations but the diameter of fibres was the same in the three muscles.3. The length of muscle fibres was the same in X-EDL, N-EDL and S-EDL 480 days after operations but the fibres of X-EDL failed to grow in cross-sectional area after innervation by soleus nerve fibres.4. The twitch: tetanus ratio was altered transitorily in both X-EDL and X-SOL but returned to near normal values within about 300 days after operations.5. The time courses of isometric contractions and the force: velocity properties were virtually the same in normal and self-innervated muscles 480 days after operations. The isometric twitch contraction time was about 13 msec for N-EDL, 25 msec for X-EDL, 34 msec for N-SOL and 15 msec for X-SOL, and the intrinsic speed of shortening of sarcomeres was about 45.1 mu/sec for N-EDL, 22.5 mu/sec for X-EDL, 19.8 mu/sec for N-SOL and 33.8 mu/sec for X-SOL; in these respects there was incomplete transformation of EDL to a slow muscle and of SOL to a fast muscle.6. There was a high degree of correlation between the intrinsic speed of shortening of sarcomeres and the isometric twitch contraction time but there was no correlation between isometric twitch contraction time and twitch: tetanus ratio of self-innervated and cross-innervated muscles 200-480 days after operations, and normal muscles from unoperated animals of the same age.7. The relation between intrinsic speed of shortening of sarcomeres and isometric twitch contraction time was approximately hyperbolic for normal, self-innervated and cross-innervated EDL and SOL muscles 480 days after operations.8. The maximum speed of shortening of whole muscle fibres was the same for X-SOL, S-EDL and N-EDL, and the same for X-EDL, S-SOL and N-SOL. The possibility that a neural influence determines the speed of shortening of whole fibres is discussed.     

Effect of disuse on sarcoplasmic reticulum in fast and slow skeletal muscle

The effect of 6 wk of hindlimb immobilization on rat skeletal muscle sarcoplasmic reticulum (SR) was determined in the slow-twitch, type I soleus (SOL), the fast-twitch, type IIA deep region of the vastus lateralis (DVL), and the fast-twitch, type IIB superficial region of the vastus lateralis (SVL). Immobilization produced a significant decline in the Ca2+ uptake rate (Vmax) of SR vesicles from the slow SOL (0.930 +/- 0.116 to 0.365 +/- 0.071 mumol Ca2+ . mg-1 . min-1), while the SR Vmax increased in the fast SVL (2.763 +/- 0.133 to 5.209 +/- 0.687) and was unaltered in the DVL. Vesicles from the fast SVL and DVL also exhibited a higher total Ca2+ uptake capacity following immobilization. An evaluation of the time course of the immobilization-mediated effect revealed an increased Ca2+ uptake capacity in all three samples after 1 wk. In the SOL total Ca2+ uptake returned to control level after 2 wk, while in the fast-twitch muscles the higher capacities were maintained. The Ca2+-stimulated SR ATPase activity was not altered in any of the muscles studies, although the total SR ATPase activity increased twofold in the slow SOL.     

Non-crossbridge calcium-dependent stiffness in slow and fast skeletal fibres from mouse muscle

We showed previously that force development in frog and FDB mouse skeletal muscle fibres is preceded by an increase of fibre stiffness occurring well before crossbridge attachment and force generation. This stiffness increase, referred to as static stiffness, is due to a Ca²⁺-dependent stiffening of a non-crossbridge sarcomere structure which we suggested could be attributed to the titin filaments. To investigate further the role of titin in static stiffness, we measured static stiffness properties at 24 and 35°C in soleus and EDL mouse muscle fibres which are known to express different titin isoforms. We found that static stiffness was present in both soleus and EDL fibres, however, its value was about five times greater in EDL than in soleus fibres. The rate of development of static stiffness on stimulation increased with temperature and was slightly faster in EDL than in soleus in agreement with previously published data on the time course of the intracellular Ca²⁺ transients in these muscles. The present results show that the presence of a non-crossbridge Ca²⁺-dependent stiffening of the muscle fibre is a physiological general characteristic of skeletal muscle. Static stiffness depends on fibre type, being greater and developing faster in fast than in slow fibres. Our observations are consistent with the idea that titin stiffening on contraction improves the sarcomere structure stability. Such an action in fact seems to be more important in EDL fast fibre than in soleus slow fibres.     

Human skeletal muscle: transition between fast and slow fibre types

Human skeletal muscles consist of different fibre types: slow fibres (slow twitch or type I) containing the myosin heavy chain isoform (MHC)-I and fast fibres (fast twitch or type II) containing MHC-IIa (type IIA) or MHC-IId (type IID). The following order of decreasing kinetics is known: type IID?>?type IIA?>>?type I. This order is especially based on the kinetics of stretch activation, which is the most discriminative property among fibre types. In this study we tested if hybrid fibres containing both MHC-IIa and MHC-I (type C fibres) provide a transition in kinetics between fast (type IIA) and slow fibres (type I). Our data of stretch activation kinetics suggest that type C fibres, with different ratios of MHC-IIa and MHC-I, do not provide a continuous transition. Instead, a specialized group of slow fibres, which we called ??transition fibres??, seems to provide a transition. Apart of their kinetics of stretch activation, which is most close to that of type IIA, the transition fibres are characterized by large cross-sectional areas and low maximal tensions. The molecular cause for the mechanical properties of the transition fibres is unknown. It is possible that the transition fibres contain an unknown slow MHC isoform, which cannot be separated by biochemical methods. Alternatively, or in addition, isoforms of myofibrillar proteins, other than MHC, and posttranslational modifications of myofibrillar proteins could play a role regarding the characteristics of the transition fibres.     

Structure, distribution and innervation of muscle spindles in avian fast and slow skeletal muscle

Muscle spindles in 2 synergistic avian skeletal muscles, the anterior (ALD) and posterior (PLD) latissimus dorsi, were studied by light and electron microscopy to determine whether morphological or quantitative differences existed between these sensory receptors. Differences were found in the density, distribution and location of muscle spindles in the 2 muscles. They also differed with respect to the morphology of their capsules and intracapsular components. The slow ALD possessed muscle spindles which were evenly distributed throughout the muscle, whereas in the fast PLD they were mainly concentrated around the single nerve entry point into the muscle. The muscle spindle index (number of spindles per gram wet muscle weight) in the ALD was more than double that of its fast-twitch PLD counterpart (130.5±2.0 vs 55.4±2.0 respectively, n=6). The number of intrafusal fibres per spindle ranged from 1 to 8 in the ALD and 2 to 9 in the PLD, and their diameters varied from 5.0 to 16.0 μm and 4.5 to 18.5 μm, respectively. Large diameter intrafusal fibres were more frequently encountered in spindles of the PLD. Unique to the ALD was the presence of monofibre muscle spindles (12.7% of total spindles observed in ALD) which contained a solitary intrafusal fibre. In muscle spindles of both the ALD and PLD, sensory nerve endings terminated in a spiral fashion on the intrafusal fibres in their equatorial regions. Motor innervation was restricted to either juxtaequatorial or polar regions of the intrafusal fibres. Outer capsule components were extensive in polar and juxtaequatorial regions of ALD spindles, whereas inner capsule cells of PLD spindles were more numerous in juxtaequatorial and equatorial regions. Overall, muscle spindles of the PLD exhibited greater complexity with respect to the number of intrafusal fibres per spindle, range of intrafusal fibre diameters and development of their inner capsules. It is postulated that the differences in muscle spindle density and structure observed in this study reflect the function of the muscles in which they reside.     

Effects of hindlimb suspension and androgen treatment on testosterone receptors in rat skeletal muscles

This study tested the specific and combined effects of testosterone treatment and hindlimb suspension (HS) on the properties of steroid receptors in skeletal muscle. Male rats were either administered weekly high doses of testosterone heptylate (10?mg?·?kg^?1) or olive oil placebo, and were either tail-suspended or acted as controls. After 3 weeks of treatment, three muscles were excised from each animal, soleus (SOL), extensor digitorum longus (EDL), and plantaris. The results showed that the testosterone treatment was unable to minimise the HS-induced atrophy of skeletal muscle. As expected, HS altered the fibre-type composition of SOL muscles (?33% of type I, +188% and +161% of type IIa and intermediate fibres respectively, P?P?相似文献   

Na channel density in extrajunctional sarcolemma of fast and slow twitch mouse skeletal muscle fibres: functional implications and plasticity after fast motoneuron transplantation on to a slow muscle

Summary Na channel densities were measured in fast and slow twitch mouse skeletal muscle fibres using the loose patch voltage clamp technique. It was found that Na channel density was approximately four times greater in fast twitch fibres than in slow. Computer simulations of action potential propagation in these fibres strongly suggest that the higher channel densities in fast twitch fibres are necessary to maintain action potential amplitude and fidelity of transmission across the neuromuscular junction, especially during the periods of rapid stimulation that these fibres are subjected to by their motoneurons. Transplantation of a foreign nerve containing axons which had previously innervated fast twitch fibres on to a slow twitch muscle resulted in an approximate doubling of the Na channel density in fibres innervated by the foreign nerve. These results suggest that motoneurons may exert considerable control over Na channel density in the muscle fibres they innervate.     

Quantitative assessment of degenerative changes in soleus muscle after hindlimb suspension and recovery

The aim of this study was to quantify the degenerative and regenerative changes in rat soleus muscle resulting from 3-week hindlimb suspension at 45° tilt (HS group, n?=?8) and 4-week normal cage recovery (HS-R group, n?=?7). Degenerative changes were quantified by microscope examination of muscle cross sections, and the myosin heavy chain (MHC) composition of soleus muscles was studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis. At the end of 3-week hindlimb suspension, histological signs of muscle degenerative changes were detected in soleus muscles. There was a significant variability in the percentage of fibres referred to as degenerating (%dg) in individual animals in the HS group [%dg?=?8.41 (SEM 0.5)%, range 4.66%–14.08%]. Moreover, %dg varied significantly along the length of the soleus muscle. The percentage of fibres with internal nuclei was less than %dg in HS-soleus muscles [4.12 (SEM 0.3)%, range 1.24%–8.86%]. In 4-week recovery rats, the greater part of the fibres that were not referred to as normal, retained central nuclei [15.8 (SEM 2.2)%, range 6.2%–21.1%]. A significant increase in the slow isoform of MHC was recorded in the HS-R rats, compared to muscles from age-matched rats (P?相似文献   

Effects of eccentric and concentric training on capillarization and myosin heavy chain contents in rat skeletal muscles after hindlimb suspension

We studied the effects of different protocols of post-disuse rehabilitation on angiogenesis and myosin heavy chain (MHC) content in rat hindlimb muscles after caudal suspension. Thirty female Wistar rats were divided into five groups: (1) Control I, (2) Control II, (3) Suspended, (4) Suspended trained on declined treadmill, and (5) Suspended trained on flat treadmill. Fragments of the soleus and tibialis anterior (TA) muscles were frozen and processed by electrophoresis and immunohistochemistry (CD31 antibody). Hindlimb suspension caused reduction of capillary/fiber (C/F) ratios and contents of MHC type I (MHCI) in the soleus in parallel to increased capillary density. Flat treadmill protocols increased the content of the MHCI isoform. The C/F ratio was increased by concentric training after hypokinesis, but was not modified by eccentric training, which caused a greater reduction of capillary density compared to the other protocols. In the TA muscle, hindlimb suspension caused a non-significant increase in capillary density and C/F ratio with limited changes in MHC. The present data demonstrate that the different training protocols adopted and the functional performance of the muscles analyzed caused specific changes in capillarization and in the content of the various MHC types.     

Effect of hyperthyroidism on the contractile and histochemical properties of fast and slow twitch skeletal muscle in the rat

Male Wistar rats were made hyperthyroid by intraperitoneal injections on alternate days for 1–6 weeks, of 200 g/kg triiodothyronine (T₃). The effects of this treatment on the contractile properties of the soleus, a slow twitch and the extensor digitorum longus (EDL), a fast twitch skeletal muscle, were studied in vivo in the anaesthetized animal. Post mortem, serial frozen sections of both muscles were stained histochemically for myosin ATPase, succinic dehydrogenase and phosphorylase. Muscle fibres were classified as either slow twitch (SO), fast twitch oxidative glycolytic (FOG) or fast twitch glycolytic (FG).Elevation of plasma T₃ levels is associated with progressive alterations in the muscle fibre populations of both muscles. In the soleus there is conversion of SO to FOG fibres while in the EDL the main effect is FG to FOG conversion. There are also marked changes, mainly confined to the soleus muscle, in contractile properties; progressive increases in isometric twitch and tetanic tension and in the rates of contraction and relaxation during both twitch and tetanus.The effect of T₃ on slow muscle contractility may be related to its effect on muscle phenotype. However, changes in the former precede detectable fibre population changes. T₃ may influence properties such as the Ca²⁺ binding activity of sarcoplasmic reticulum of existing slow twitch fibres before the later changes associated with the interconversion of fibre types occur.     

Changes in fibre-type composition and myosin heavy-chain IId isoform in rat soleus muscle during recovery period after hindlimb suspension

We examined the changes in myosin heavy-chain (HC) isoforms and fibre-type composition in rat soleus muscle using both myosin adenosine triphosphatase staining and sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS-PAGE) analyses during the recovery period after 4 weeks of hindlimb suspension. Although there was no change in type IIc fibres after the suspension, an increase in this type of fibres was observed during the 1- to 4-week recovery period. The increase in type Ilc fibres was considered to be due to a shift from type Ila to IIc fibres. The SDS-PAGE analysis revealed the presence of the HC IId isoform, which was not observed in the control muscle, after a 4-week hindlimb suspension. The HC IId isoform gradually decreased over 3 weeks of recovery and disappeared in the 4th week of recovery after the suspension. These results suggest that the hypogravity conditions induced by hindlimb suspension stimulated the synthesis of the HC IId isoform, whereas an increase in mechanical load to the muscle accelerated the degradation of the HC IId isoform and the synthesis of type Ilc fibres during the recovery period after hindlimb suspension.     

Work and metabolic patterns of fast and slow twitch skeletal muscle contractingin situ

Summary A study was conducted to compare the work capacity and aerobic and anaerobic metabolic profiles of the fast-twitch tibialis anterior and slow-twitch soleus muscles of the rat stimulated to contractin situ. Results showed that the soleus could sustain greater relative and absolute work levels than the tibialis anterior. Metabolic profiles revealed that the tibialis anterior depended on anaerobic pathways to a greater extent than the soleus, since greater amounts of phosphocreatine were depleted and greater amounts of glycogen were broken down and metabolized to lactate in the tibialis anterior. The ability of both muscles to sustain prolonged work was attributed to the high capacity of red and intermediate muscle for fatty acid oxidation as evidenced by the higher levels of resting and exercise acyl-carnitine and the greater activity levels of carnitine palmityltransferase measured in the soleus.