Regeneration of pigeon fast and slow muscle fiber types after partial excision and mincing.

The pigeon's metapatagialis muscles, containing fast fibers in two slips and slow fibers in another slip, were excised for a third of their length, minced, and replaced into their previous sites. After regeneration, the pattern of fiber types and their ATPase and oxidative enzymes were examined histochemically. Ultrastructural examination was carried out on the fast fibers. After 4-17 wk the muscles had regenerated into patterns histochemically similar to the controls only within the slip containing fast fibers. The slow slip was much less regenerated, and had a histochemically embryonic composition. Fiber types were characterized and their cross-sectional areas measured, and the degree of atrophy was greatest in the large fast fibers and the slow fibers. Ultrastructural studies revealed a number of alterations of the mitochondria, including dense and light areas in the matrix and an altered pattern of the cristae into parallel tubular or vesicular aggregations. Other changes included dilated sarcoplasmic reticulum, myofibril disorganization, and a compaction of filaments. The slow fibers were thought to be slower in their regeneration rates because of the pattern of multiple innervation's producing a more complex regenerative pattern.

Protein-protein interactions of proteolytic fragments of actin.

Proteolytic fragments of actin, prepared by removal of up to sixty-eight residues from the N-terminal end of the molecule, can form filamentous structures after denaturation in urea solution. The filaments have a diameter similar to F-actin filaments and interact with myosin and tropomyosin. A fragment comprising residues 1 to 207 of the actin sequence did not form filaments or interact with myosin after the urea treatment.

Histochemical, biochemical, and ultrastructural analyses of single human muscle fibers, with special reference to the C-fiber population.

A muscle biopsy from the vastus lateralis muscle of a strength-trained woman was found to contain an unusual fiber type composition and was analyzed by histochemical, biochemical, and ultrastructural techniques. Special attention was given to the C-fibers, which comprised over 15% of the total fiber number in the biopsy. The mATPase activity of the C-fibers remained stable to varying degrees over the pH range normally used for routine mATPase histochemistry. Although a continuum existed, the C-fibers were histochemically subdivided into three main fiber types: IC, IIC, and IIAC. The IC fibers were histochemically more similar to the Type I, the IIAC were more similar to the Type IIA, and the IIC were darkly stained throughout the pH range. Biochemical analysis revealed that all C-fibers coexpressed myosin heavy chains (MHC) I and IIa in variable ratios. The histochemical staining intensity correlated with the myosin heavy chain composition such that the Type IC fibers contained a greater ratio of MHCI/MHCIIa, the IIAC contained a greater ratio of MHCIIa/MHCI, and the Type IIC contained equal amounts of these two heavy chains. Ultrastructural data of the C-fiber population revealed an oxidative capacity between fiber Types I and IIA and suggested a range of mitochondrial volume percent from highest to lowest such that I greater than IC greater than IIC greater than IIA-C greater than IIA. Under physiological conditions, it appears that the IC fibers represent Type I fibers that additionally express some fast characteristics, whereas the Type IIAC are Type IIA fibers that additionally express some slow characteristics. Fibers expressing a 50:50 mixture of MHCI and MHCIIa (IIC fibers) were rarely found. It is not known whether C-fibers represent a distinct population between the fast- and slow-twitch fibers that is specifically adapted to a particular usage or whether they are transforming fibers in the process of going from fast to slow or slow to fast.

Human skeletal muscle fiber type adaptability to various workloads.

Muscle biopsy specimens were removed from the vastus lateralis muscles of three groups of human subjects: controls, weight lifters, and distance runners. The runners proved to be a unique group with respect to the variables measured (low body weight and percentage body fat, and high VO2 max). Additionally, a histochemical analysis of the biopsy specimens revealed that the runners had a significantly higher percentage of fiber types I and IIC than either the controls or the weight lifters. Using a cryostatic retrieval method, each of the fibers identified histochemically was then analyzed morphometrically using electron microscopy. The results of volume-percent mitochondria demonstrated a strong relationship between the ATPase activity and oxidative potential of the fiber types for all three groups such that the oxidative activity would be ranked I greater than IIA greater than IIB. Irrespective of fiber type, there were significant differences between the groups with regard to muscle-fiber mitochondrial (runners greater than lifters greater than controls) and lipid content (runners greater than controls greater than lifters). The lifters had a significantly greater content of mitochondria than the controls, which may suggest that inactivity rather than the lifting exercise contributes to a low volume-percent mitochondria and a high percentage of type IIB fibers.

Fiber type composition of the vastus lateralis muscle of young men and women.

This study presents data collected over the past 10 years on the muscle fiber type composition of the vastus lateralis muscle of young men and women. Biopsies were taken from the vastus lateralis muscle of 55 women (21.2+/-2.2 yr) and 95 men (21.5+/-2.4 yr) who had volunteered to participate in various research projects. Six fiber types (I, IC, IIC, IIA, IIAB, and IIB) were classified using mATPase histochemistry, and cross-sectional area was measured for the major fiber types (I, IIA, and IIB). Myosin heavy chain (MHC) content was determined electrophoretically on all of the samples from the men and on 26 samples from the women. With the exception of fiber Type IC, no significant differences were found between men and women for muscle fiber type distribution. The vastus lateralis muscle of both the men and women contained approximately 41% I, 1% IC, 1% IIC, 31% IIA, 6% IIAB, and 20% IIB. However, the cross-sectional area of all three major fiber types was larger for the men compared to the women. In addition, the Type IIA fibers were the largest for the men, whereas the Type I fibers tended to be the largest for the women. Therefore, gender differences were found with regard to the area occupied by each specific fiber type: IIA>I>IIB for the men and I>IIA>IIB for the women. These data establish normative values for the mATPase-based fiber type distribution and sizes in untrained young men and women.

Effects of high-intensity resistance training on untrained older men. I. Strength, cardiovascular, and metabolic responses.

Most resistance training studies of older subjects have emphasized low-intensity, short-term training programs that have concentrated on strength measurements. The purpose of this study was, in addition to the determination of strength, to assess intramuscular and transport factors that may be associated with strength increments. Eighteen untrained men ages 60-75 years volunteered for the study; 9 were randomly placed in the resistance-training group (RT), and the other half served as untrained (UT) or control subjects. RT subjects performed a 16-week high-intensity (85-90% 1 repetition maximum (RT]) resistance training program (2 x/wk) consisting of 3 sets each to failure (6-8 repetitions based on 1 RM of 3 exercises): leg press (LP), half squat (HS), and leg extension (LE) with 1-2 minutes rest between sets. Pre- and post- training strength was measured for the 3 training exercises using a 1 RM protocol. Body fat was calculated using a 3-site skinfold method. Biopsies from the vastus lateralis m. were obtained for fiber type composition, cross-sectional area, and capillarization measurements. Exercise metabolism, electrocardiography, and arterial blood pressure were observed continuously during a progressive treadmill test, and resting echocardiographic data were recorded for all subjects. Pre- and post-training venous blood samples were analyzed for serum lipids. Resistance training caused significant changes in the following comparisons: % fat decreased in the RT group by almost 3%, strength improved for all exercises: LE = + 50.4%, LP = + 72.3%, HS = + 83.5%; type IIB fibers decreased and IIA fibers increased; cross-sectional areas of all fiber types (I, IIA, IIB) increased significantly, and capillary to fiber ratio increased but not significantly. No differences were noted for ECG and echocardiographic data. The RT group significantly improved treadmill performance and VO2max. Pre- and post-training serum lipids improved but not significantly. No significant changes occurred in any pre- to post-tests for the UT group. The results show that skeletal muscle in older, untrained men will respond with significant strength gains accompanied by considerable increases in fiber size and capillary density. Maximal working capacity, VO2max, and serum lipid profiles also benefited from high-intensity resistance training, but no changes were observed for HR max, or maximal responses of arterial blood pressure. Older men may not only tolerate very high intensity work loads but will exhibit intramuscular, cardiovascular, and metabolic changes similar to younger subjects.

Effects of high-intensity resistance training on untrained older men. II. Muscle fiber characteristics and nucleo-cytoplasmic relationships.

During growth and repair of skeletal muscle fibers, satellite cells become activated, undergo mitosis, and a daughter nucleus becomes incorporated into the muscle fiber to increase myonuclear numbers. An increase in myonuclei appears to be required for this postnatal growth. This study examined whether muscle fibers of elderly men can hypertrophy with strength training and, if so, whether they have the capacity to incorporate nuclei into the fibers. The sarcoplasmic area associated with each myonucleus was calculated in nine elderly men before and after 16 weeks of strength training, and compared to nine elderly control men. Muscle fiber type changes and myosin heavy chain composition were also compared. All major fiber types (I, IIA, IIB) became significantly larger after training, and a transition of type IIB fibers to IIA occurred with training. The area occupied by each fiber type correlated with myosin heavy chain percentage, and both of these changed similarly with strength training. The cytoplasm-to-myonucleus ratio increased, but not significantly (p = .07), with muscle fiber hypertrophy. Number of myonuclei per fiber and myonuclei per unit length of muscle fiber increased, but not significantly. Cross-sectional areas of the muscle fibers in untrained elderly men were much smaller than in untrained young men (when compared with our earlier studies). Training increased the sizes of the elderly muscle fibers to that of the untrained young men. This hypertrophy of muscle fibers by 30% with training resulted in no change in the cytoplasm-to-myonucleus ratio. This suggests that the myonuclear population continues to adapt to growth stimuli in the elderly muscles.

Strength and skeletal muscle adaptations in heavy-resistance-trained women after detraining and retraining.

Six women who had participated in a previous 20-wk strength training study for the lower limb detrained for 30-32 wk and subsequently retrained for 6 wk. Seven untrained women also participated in the 6-wk "retraining" phase. In addition, four women from each group volunteered to continue training an additional 7 wk. The initial 20-wk training program caused an increase in maximal dynamic strength, hypertrophy of all three major fiber types, and a decrease in the percentage of type IIb fibers. Detraining had relatively little effect on fiber cross-sectional area but resulted in an increased percentage of type IIb fibers with a concomitant decrease in IIa fibers. Maximal dynamic strength decreased but not to pretraining levels. Retraining for 6 wk resulted in significant increases in the cross-sectional areas of both fast fiber types (IIa and IIab + IIb) compared with detraining values and a decrease in the percentage of type IIb fibers. The 7-wk extension accentuated these trends such that cross-sectional areas continued to increase (nonsignificant) and no IIb fibers could be found. Similar results were found for the nonpreviously trained women. These data suggest that rapid muscular adaptations occur as a result of strength training in previously trained as well as non-previously trained women. Some adaptations (fiber area and maximal dynamic strength) may be retained for long periods during detraining and may contribute to a rapid return to "competitive" form.

Effects of sprint cycle training on human skeletal muscle.

Eleven men sprint trained two to three times per week for 6 wk to investigate possible exercise-induced slow-to-fast fiber type conversions. Six individuals served as controls. Both groups were tested at the beginning and end of the study to determine anaerobic performance and maximal oxygen consumption. In addition, pre- and postbiopsies were extracted from the vastus lateralis muscle and were analyzed for fiber type composition, cross-sectional area, and myosin heavy chain (MHC) content. No significant changes were found in anaerobic or aerobic performance variables for either group. Although a trend was found for a decrease in the percentage of type IIb fibers, high-intensity sprint cycle training caused no significant changes in the fiber type distribution or cross-sectional area. However, the training protocol did result in a significant decrease in MHC IIb with a concomitant increase in MHC IIa for the training men. These data appear to support previous investigations that have suggested exercise-induced adaptations within the fast fiber population (IIb-->IIa) after various types of training (endurance and strength).

Skeletal muscle adaptations during early phase of heavy-resistance training in men and women.

An 8-wk progressive resistance training program for the lower extremity was performed twice a week to investigate the time course for skeletal muscle adaptations in men and women. Maximal dynamic strength was tested biweekly. Muscle biopsies were extracted at the beginning and every 2 wk of the study from resistance-trained and from nontrained (control) subjects. The muscle samples were analyzed for fiber type composition, cross-sectional area, and myosin heavy chain content. In addition, fasting blood samples were measured for resting serum levels of testosterone, cortisol, and growth hormone. With the exception of the leg press for women (after 2 wk of training) and leg extension for men (after 6 wk of training), absolute and relative maximal dynamic strength was significantly increased after 4 wk of training for all three exercises (squat, leg press, and leg extension) in both sexes. Resistance training also caused a significant decrease in the percentage of type IIb fibers after 2 wk in women and 4 wk in men, an increase in the resting levels of serum testosterone after 4 wk in men, and a decrease in cortisol after 6 wk in men. No significant changes occurred over time for any of the other measured parameters for either sex. These data suggest that skeletal muscle adaptations that may contribute to strength gains of the lower extremity are similar for men and women during the early phase of resistance training and, with the exception of changes in the fast fiber type composition, that they occur gradually.

The effects of detraining on an elite power lifter. A case study.

Muscle biopsies were taken from the vastus lateralis muscle of an elite power lifter during training and following a 7-month detraining period. The effects of detraining were investigated by combining ultrastructure, histochemistry, and pertinent metabolic data. Muscle fibers were classified ultrastructurally as fast-twitch and slow-twitch. Fast-twitch fibers were histochemically subdivided into a fast-twitch glycolytic (FG) and a fast-twitch oxidative glycolytic (FOG) in order to compare oxidative capacities between biopsy 1 and biopsy 2. The high intensity level of strength training prior to biopsy 1 caused an apparent hypertrophy of all fiber types. Detraining and weight loss resulted in a reversal of the training effect toward "control" values and adjustments in the oxygen delivery system. Atrophy occurred in all of the fiber types and altered the fiber composition of the muscle. A shifting of fibers classified as FG to the more oxidative FOG fibers caused a significant increase in the percent distribution of oxidative fibers (slow-twitch + FOG). The hypothesis is presented that the mitochondrial content (both number and form) of the detrained fiber remains constant (for a strength trained muscle), but as the volume of the fiber decreases the mitochondrial volume percent increases.

Patterns of avian muscle fiber type regeneration. Evidence for a myotrophic influence.

Serratus superficialis metapatagialis (SSM) and posterior biventer cervicis (BVC) muscles were free-grafted into the site of the slow tonic anterior latissimus dorsi (ALD) of adult pigeons. Grafts were prepared for histochemical examination at 1, 3, 7, 12 and 15 days; at monthly intervals from 1 to 4 months; and at various intervals up until 23 months after surgery. Few original muscle fibers survived the trauma of grafting and contributed to the regenerated muscle. The arrangement of fast and slow fibers and the percentage of slow fibers were recorded for each graft at least one month of age. Young grafts of the SSM (one to four months of age) regenerated a segregated pattern of fast and slow fibers that resembled control SSM muscles, whereas early grafts of the BVC regenerated a mosaic arrangement characteristic of normal BVC muscles. The percentages of slow fibers in both SSM and BVC grafts up to 4 months after surgery did not significantly differ from control muscles. Both SSM and BVC grafts older than 4 months of age had a significantly higher percentage of slow fibers than control muscles. These data indicate that the donor muscle influences the pattern of fiber types that is originally manifested and this pattern is maintained relatively intact for 4 months. Eventually the nerve at the host site modifies this pattern, and the fibers of the graft assume the histochemical characteristics of the muscle removed from the host site. In addition, the time required for histochemical stabilization of muscle fibers of avian heterotopic autografts is longer than that for whole muscle grafts in rats or avian minced muscle preparations, and is comparable to tenotomized avian muscle.

Muscle fiber necrosis associated with human marathon runners.

This study describes the events occurring in exercise-induced muscular necrosis. Biopsies of the gastrocnemius muscles of volunteer human marathon runners were extracted prior to and at intervals for 7 days following a marathon, and investigated ultrastructurally. Most of the preparations, including the pre-marathon samples, showed evidence of muscle fiber necrosis and inflammation. These preparations had many mitochondria, erythrocytes, leukocytes and other phagocytic cells within the extracellular and extravascular spaces. Less frequently observed were Z-line streaming and degeneration, contracture knots, disrupted sarcolemma, presence of erythrocytes within the muscle fibers, and empty basal lamina tubes in which the contents of the fibers and the sarcolemma had broken down to leave only the basal lamina outlining the former fiber. These abnormal conditions were most prevalent at 1 and 3 days after the marathon. These ultrastructural changes are compared and correlated with the reports of clinical manifestations of rhabdomyolysis and myoglobinuria. Because the abnormalities persist for the 7 day duration of these observations, and because many of these were observed in the pre-marathon biopsies, we conclude that both the intensive training for, and the marathon itself, induce inflammation and fiber necrosis which are manifested in the clinical symptoms for rhabdomyolysis and myoglobinuria. The inflammatory reaction that accompanies these activities may be a major factor in post-exercise soreness. The combined influences of training and necrosis are discussed in relation to muscle fiber type compositions of endurance athletes.

Lipid depletion and repletion in skeletal muscle following a marathon.

Intramuscular lipid content was investigated in muscle biopsies from 10 well-trained endurance athletes before, immediately after, and 1, 3, 5, and 7 days after a marathon. Diets were controlled throughout the entire period of the study. Triglyceride content was ultrastructurally determined by the use of stereological methods. The volume percent lipid significantly decreased after the marathon and was lowest at 3 days post-marathon, rising slightly but still 35% lower than the pre-marathon value by 7 days post-marathon. Glycogen granules were abundant and tightly packed in the pre-marathon biopsies, scarce immediately post-marathon, and abundant, but less tightly packed, 7 days post-marathon. Post-marathon fluctuations in the volume percentages of mitochondria indicated possible fluid shifts within the muscle fibers: dehydration immediately post-marathon followed by rehydration with possible edema. Assuming the content of mitochondria remained constant throughout the recovery period, the ratio of the volume percentage of lipid to the volume percentage of mitochondria indicated that lipid content may have reached pre-marathon levels after 7 days post-marathon.

A morphometric analysis of human muscle fibers with relation to fiber types and adaptations to exercise.

Muscle biopsies were obtained from the vastus lateralis of 14 male subjects: 3 long distance runners, 2 world class power lifters and 9 active, although not highly trained, individuals used as controls. The fibers were investigated by electron microscopy and the mitochondrial volume percent, lipid volume percent and Z-line width were analyzed morphometrically. With the combined data a direct correlation was found between mitochondrial volume percent and lipid volume percent, lipid volume percent and Z-line width and mitochondrial volume percent and Z-line width. The muscle fibers were classified as slow-twitch oxidative (SO), fast-twitch-oxidative-glycolytic (FOG) and fast-twitch-glycolytic (FG) based on relationships found in the data and well established properties of muscle fiber types. Although no distinct patterns emerged, a good approximation of fiber type characteristics was obtained, and values for volume percent of central mitochondria, volume percent lipid and Z-line width are reported. The fibers classified as SO were characterized by having wide Z-lines, a high mitochondrial volume percent and high lipid volume percent. The fast-twitch fibers (fibers with narrow Z-lines) were separated into 2 groups, those with high mitochondrial volume percent (FOG) and those with low mitochondrial volume percent (FG). No distinction could be made between the fast-twitch subgroups with regard to Z-line width. The fibers from distance runners differed from those from controls by exhibiting a greater capacity for aerobic activity as evidenced by the increased volume percent of mitochondria and lipid in both slow- and fast-twitch fibers. The high strength, anaerobic activity of the world class power lifters was reflected by the low mitochondrial volume percent of many fast-twitch fibers (FG) and the decreased lipid stores in all fibers.

Skeletal muscle-smooth muscle interaction: an unusual myoelastic system.

The serratus superficialis metapatagialis (SSM) of pigeons is a skeletal muscle with unusual properties. It lies between the ribs and the trailing edge of the wing, where it is attached to the skin by a system of smooth muscles having elastic tendons. Wing movements during flight induce marked changes in this muscle's length. The SSM inserts onto the deep fascia, and at its termination the skeletal muscle contains large numbers of microtubules. Many myofibrils attach to leptomeric organelles, which then attach to the terminal end of the skeletal muscle fiber. The deep fascia next connects to the dermis of the skin by bundles of smooth muscles that have elastic tendons at both ends. This system allows large movements of the muscle while preventing its fibers from overstretching. The movements and presumed forces acting at this muscle make the presence of sensory receptors such as muscle spindles unlikely. Spindles are absent in this muscle.

Analysis of fiber types in the pigeon's metapatagialis muscle. II. Effects of denervation.

The pigeon's metapatagialis muscle consists of three slips, two twitch and one tonic, and these slips are distinguishable at the gross anatomical level. Comparative studies of denervation are facilitated because the two fiber types are under the same mechanical forces, can be denervated as one muscle, and can be distinguished after denervation. Both fiber types atrophied after denervation, with the twitch fibers having a more variable response. Pathological alterations observed by light microscopy suggested that the twitch fibers were more affected by denervation than the tonus fibers. Ultrastructurally, both fiber types showed the same changes, with the twitch fibers again being more consistently altered. Proliferation of the transverse tubular system and sarcoplasmic reticulum were more marked in the tonus than twitch fibers, and the sarcoplasmic reticulum proliferated prior to the transverse tubules. Filament and fibril degeneration, peripheral and central degeneration, lysosomes and their derivatives, and satellite cell proliferation were common to both fiber types. Contracture knots were common to the denervated fibers, and were suggested to be characteristic of degenerating fibers. Degenerating motor end plates were observed, and most neurons in the fibers were naked, lacking myelin sheaths. The results are discussed in relation to the function of the neuron in maintaining the muscle, and the possibility of denervation inducing a transformation of tonic to twitch fibers.

Structural and metabolic characteristics of human skeletal muscle following 30 days of simulated microgravity.

Percutaneous needle biopsy samples were obtained from the vastus lateralis and soleus muscles before and after 30 d of 6 degree head-down bedrest to determine the influence of this model of simulated microgravity on human skeletal muscle. Fiber atrophy was evident in both muscles with both fast-twitch and slow-twitch fiber cross-sectional areas decreasing. Predominant atrophy of slow-twitch fibers was not evident. The soleus had a greater proportion of slow-twitch fibers than the vastus lateralis before bedrest. Neither muscle showed a change in fiber type percentage with bedrest. Phosphofructokinase and lactate dehydrogenase activities in the soleus and vastus lateralis muscles were similar before and after bedrest. The activities of beta-hydroxyacyl-CoA dehydrogenase and citrate synthase, however, were reduced during bedrest with these responses being somewhat greater in the soleus. While the ultrastructure of most of the fibers of the soleus and vastus lateralis appeared normal after bedrest, evidence of remodeling was present in both muscles. The proliferation of core/targetoid lesions, honeycomb networks, regenerating satellite cells, necrotic foci and myofibrillar disorganization after bedrest indicates that force development is an important factor in determining the organization of the fine structure of muscle. The results indicate that short-duration exposure to simulated microgravity decreases fiber size and the capacity for aerobic energy supply of human skeletal muscle. Moreover, disorganization of the contractile machinery occurs. Thus, it appears that bedrest alters the "normal" load-time constraints imposed on skeletal muscle sufficiently to change its inherent structural and metabolic characteristics.

Effects of bovine growth hormone analogs on mouse skeletal muscle structure.

Skeletal muscles of transgenic mice expressing altered bovine growth hormones (bGH) have been compared with those of nontransgenic mice to determine whether muscle fiber type-specific responses or histopathologies are associated with the altered gene. The slow soleus and predominantly fast gastrocnemius muscles were prepared for myofibrillar ATPase activity (to determine muscle fiber type) and histological examination from mice that were either giant (M4 line), larger than normal (M11 line), dwarf (G119K line), or nontransgenic (NTC). No histopathology was observed in any of the muscles. Although body weights were significantly different between all four lines of mice, only the giant M4 mice had significantly larger muscle fibers than the other lines of mice, while neither the G119K nor M11 lines were significantly different from the NTC for either muscle. No fiber type-specific differences were noted. These results suggest that the different muscles are the product of differences in numbers of muscle fibers expressed in the G119K and M11 lines of mice; the increase in body mass matched the fiber size growth only in the giant M4 line. Therefore, the altered bGH genes may be acting on fetal liver and myoblast/myotube GH receptors to change the GH and IGF-I regulated pattern of muscle development, and eventually, to determine the adult muscle fiber numbers.