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Biomedical subjects

R R Roy

Publications and source records attributed to R R Roy.

At least 19 recordsLinked to original sources

Three-dimensional structure of cat tibialis anterior motor units.

The motor unit is the basic unit for force production in a muscle. However, the position and shape of the territory of a motor unit within the muscle have not been defined precisely. The territories of five motor units in the cat tibialis anterior muscle were reconstructed three-dimensionally (3-D) from tracings of the glycogen-depleted fibers belonging to each unit. The motor unit territories did not span the entire length of the muscle and their cross-sectional areas tapered along the proximodistal axis producing a conical shape. In addition, the position of the territory of each unit shifted in an anterior-posterior plane along the longitudinal axis of the muscle, presumably as a consequence of the pinnation of the fibers. The area of the motor unit territory at any given level along the proximodistal axis was highly correlated with the number of fibers within the territory at that level. Connective tissue boundaries (outlining fascicles) appeared to have a strong influence on the shape of the territory, territories showed abrupt changes at connective tissue boundaries as groups of motor unit fibers within a fascicle often terminated together while motor unit fibers in neighboring fascicles did not terminate. It is likely that the mechanical impact of the recruitment of a motor unit is affected by the location and shape of motor units within the same muscle area.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Response of mouse plantaris muscle to functional overload: comparison with rat and cat.

Functional overload (FO) of a muscle by removing its synergists results in a compensatory hypertrophy of the muscle. However, the extent of the response appears to be dependent, at least in part, on the activity and/or loading levels of the muscle following surgery. Thus, differences in the inherent physical activity levels across species may be an important factor to consider. In the present study, the effects of 8 weeks of FO on the isometric mechanical properties of the plantaris of mice (highly active) were determined and the findings compared with the results from previous studies performed on the plantaris of rats (highly active) and cats (less active). FO resulted in approximately a doubling of the mass, the physiological cross-sectional area and the maximum tetanic tension per unit cross-sectional area, was similar in the plantaris of control and FO mice. Isometric twitch speed properties were unaffected, but the tension enhancement in response to an increase in the rate of stimulation showed the pattern of a "faster" muscle following FO. The fatigue resistance of the plantaris in FO mice was significantly higher than in control mice. Although the degree of hypertrophy that occurred in the mouse plantaris was similar to that observed after FO in rats and in cats that are exercised intermittently at high intensities, there were differences in the mechanical properties that may be related to the adaptability of species and/or the behavioral responses to the overload.

Animals

Prominence of myosin heavy chain hybrid fibers in soleus muscle of spinal cord-transected rats.

The effect of a midthoracic spinal cord transection (ST) on myosin heavy chain (MHC) isoform expression in the rat soleus muscle was studied. Electrophoretic analyses demonstrated that 15 days after ST there were significant proportional increases in type IIx, decreases in type IIa, and no change in type I MHC composition. Thirty days after ST, some type IIb MHC was expressed, there were further proportional increases in type IIx, an increase in type IIa (compared with 15-day ST), and a decrease in type I MHC. At both time periods after ST, many fibers expressed multiple MHCs, as demonstrated by immunohistochemistry where a battery of monoclonal antibodies specific to MHC isoforms was used. Fibers were observed containing types I and II together or multiple type II MHC isoforms. These data suggest that the expression of the normal complement of MHC isoforms in the adult rat soleus muscle is dependent, in part, on normal neuromuscular activation.

Animals

Human fiber size and enzymatic properties after 5 and 11 days of spaceflight.

Biopsies from the vastus lateralis muscle were obtained from three astronauts before and after two 5-day flights and from five astronauts before and after one 11-day flight (space shuttle flights: STS-32, -33, and -34). Muscle fibers from two separate samples from each biopsy were classified as type I and II or as type I, IIA, and IIB by using qualitative myofibrillar adenosinetriphosphatase (ATPase) staining. Cross-sectional area (CSA), number of capillaries per fiber, and the activities of succinate dehydrogenase (SDH), alpha-glycerophosphate dehydrogenase (GPD), and myofibrillar ATPase were determined from one sample of fibers of each myofibrillar ATPase type. Postflight biopsies had 6-8% fewer type I fibers than preflight. Mean fiber CSAs were 16-36% smaller after the 11-day flight with the relative effect being type IIB > IIA > I. Mean fiber CSAs were 11 and 24% smaller in type I and II fibers after 5 days of flight. Myofibrillar ATPase activities increased in type II but not in type I fibers after flight, whereas SDH activity was unaffected in either fast or slow fibers. GPD activity in type I fibers was approximately 80% higher (P > 0.05) postflight compared with preflight. Myofibrillar ATPase/SDH ratios in type II fibers were higher after than before flight, suggesting that some fast fibers were more susceptible to fatigue after flight. The GPD/SDH ratios were elevated in some type I fibers after spaceflight. The number of capillaries per fiber was 24% lower after than before flight, whereas the number of capillaries per unit CSA of muscle tissue was unchanged. These data suggest that adaptations in the size, metabolic properties, and vascularity of muscle fibers can occur rapidly in the space environment. These adaptations were qualitatively similar to those observed in animals after actual or simulated spaceflight conditions for short periods.

Adenosine Triphosphatases

Myosin heavy chain isoforms of human muscle after short-term spaceflight.

The influence of microgravity on the myosin phenotype of skeletal muscle fibers in the vastus lateralis of eight crew members was studied before and after 5-day (n = 3) and 11-day (n = 5) spaceflights (space shuttle flights: STS-32, -33 and -34). Single-fiber electrophoresis analyses showed that the proportion of fibers expressing only slow (type I) myosin heavy chain (MHC) in the vastus lateralis was significantly lower after than before 11 days of spaceflight. Although the family of type II MHC isoforms was elevated post- compared with preflight, the distribution among the isoforms of type II MHC was not statistically different. Based on monoclonal and polyclonal antibodies specific for three adult MHC isoforms and single-fiber electrophoresis, approximately 3% of the fibers analyzed coexpressed all three adult MHC isoforms. The results from immunohistochemical staining with two different sets of antibodies indicate a reduction in the percentage of fibers expressing type I MHC as a result of spaceflight. The mean difference, however, was significant only when the fibers were categorized simply as type I or II. These changes appeared to be highly individualized among the astronauts. These results suggest that a rapid change in MHC isoform expression can occur in some muscle fibers after a relatively brief exposure to spaceflight.

Adult

Plasticity of myonuclear number in hypertrophied and atrophied mammalian skeletal muscle fibers.

Although a mammalian skeletal muscle fiber may contain thousands of myonuclei, the importance of this number or the potential to modulate it in adult muscle has not been clearly demonstrated. Using immunohistochemistry and confocal microscopy, we examined the plasticity of myonuclear number and fiber size in isolated fast and slow fiber segments from adult cat hindlimb muscles in response to chronic alterations in neuromuscular activity and loading. Compared with slow fibers in the soleus of control cats, myonuclear number in presumably transformed fast fibers was 32% lower and fiber size was decreased 73% after elimination of neuromuscular activation for 6 mo by spinal isolation. Slow fibers in the soleus of spinal-isolated cats had smaller cross-sectional areas, whereas myonuclear number was not significantly different than that in the control cats. Myonuclear number in fast plantaris fibers was more than threefold higher and fiber size was 2.8-fold higher after 3 mo of functional overload compared with the plantaris of control cats. Compared with control slow plantaris fibers, myonuclear number and fiber size also increased in overloaded slow plantaris fibers. These results demonstrate that changes in myonuclear number are associated with changes in myosin type and suggest that modulations in the amount of available DNA may be a factor in regulating cytoplasmic volume of muscle fibers in response to chronic changes in neuromuscular activity.

Animals

Myofibrillar ATPase activity of feline muscle fibers expressing slow and fast myosin heavy chains.

The interrelationships among myofibrillar ATPase activity (Quant-mATPase), qualitative myofibrillar ATPase staining after acid (Acid-mATPase) and alkaline (Alk-mATPase) preincubations, and myosin heavy chain (MHC) composition were determined in frozen sections of soleus (Sol) and medial gastrocnemius (MG) muscle fibers from adult control cats and cats 6 months after complete spinal cord transection (Sp). Fibers were categorized as either fast, slow, or fast and slow (Fast-Slow) based on monoclonal antibody labeling. Slow fibers had low Quant-mATPase activity and stained lightly with Alk-mATPase and darkly with Acid-mATPase. Fast fibers had high Quant-mATPase activity (approximately twice that of slow fibers) and stained darkly with Alk-mATPase and lightly with Acid-mATPase. Fast-Slow fibers had intermediate Quant-mATPase activity and stained intermediately for Acid-mATPase and darkly for Alk-mATPase. There was a positive linear relationship between Alk-mATPase and Quant-mATPase for all fibers of Sol and MG from control and Sp cats. There was a negative linear relationship between Acid-mATPase and Quant-mATPase for all fibers of Sol and MG. However, within the fast fiber population of the MG there was a positive relationship between these two measures of mATPase. In summary, quantitative and qualitative measures of mATPase are highly correlated with the types of MHC expressed by single fibers from control and Sp cat muscles.

Animals

Neuromuscular adaptation to actual and simulated weightlessness.

The chronic "unloading" of the neuromuscular system during spaceflight has detrimental functional and morphological effects. Changes in the metabolic and mechanical properties of the musculature can be attributed largely to the loss of muscle protein and the alteration in the relative proportion of the proteins in skeletal muscle, particularly in the muscles that have an antigravity function under normal loading conditions. These adaptations could result in decrements in the performance of routine or specialized motor tasks, both of which may be critical for survival in an altered gravitational field, i.e., during spaceflight and during return to 1 G. For example, the loss in extensor muscle mass requires a higher percentage of recruitment of the motor pools for any specific motor task. Thus, a faster rate of fatigue will occur in the activated muscles. These consequences emphasize the importance of developing techniques for minimizing muscle loss during spaceflight, at least in preparation for the return to 1 G after spaceflight. New insights into the complexity and the interactive elements that contribute to the neuromuscular adaptations to space have been gained from studies of the role of exercise and/or growth factors as countermeasures of atrophy. The present chapter illustrates the inevitable interactive effects of neural and muscular systems in adapting to space. It also describes the considerable progress that has been made toward the goal of minimizing the functional impact of the stimuli that induce the neuromuscular adaptations to space.

Adaptation, Physiological

Adaptability of the oxidative capacity of motoneurons.

Previous studies have demonstrated that a chronic change in neuronal activation can produce a change in soma oxidative capacity, suggesting that: (i) these 2 variables are directly related in neurons and (ii) ion pumping is an important energy requiring activity of a neuron. Most of these studies, however, have focused on reduced activation levels of sensory systems. In the present study the effect of a chronic increase or decrease in motoneuronal activity on motoneuron oxidative capacity and soma size was studied. In addition, the effect of chronic axotomy was studied as an indicator of whether cytoplasmic volume may also be related to the oxidative capacity of motoneurons. A quantitative histochemical assay for succinate dehydrogenase activity was used as a measure of motoneuron oxidative capacity in experimental models in which chronic electromyography has been used to verify neuronal activity levels. Spinal transection reduced, and spinal isolation virtually eliminated lumbar motoneuron electrical activity. Functional overload of the plantaris by removal of its major synergists was used to chronically increase neural activity of the plantaris motor pool. No change in oxidative capacity or soma size resulted from either a chronic increase or decrease in neuronal activity level. These data indicate that the chronic modulation of ionic transport and neurotransmitter turnover associated with action potentials do not induce compensatory metabolic responses in the metabolic capacity of the soma of lumbar motoneurons. Soma oxidative capacity was reduced in the axotomized motoneurons, suggesting that a combination of axoplasmic transport, intracellular biosynthesis and perhaps neurotransmitter turnover represent the major energy demands on a motoneuron. While soma oxidative capacity may be closely related to neural activity in some neural systems, e.g. visual and auditory, lumbar motoneurons appear to be much less sensitive to modulations in chronic activity levels.

Adaptation, Physiological

Physiological cross-sectional area of human leg muscles based on magnetic resonance imaging.

Magnetic resonance imaging techniques were used to determine the physiological cross-sectional areas (PCSAs) of the major muscles or muscle groups of the lower leg. For 12 healthy subjects, the boundaries of each muscle or muscle group were digitized from images taken at 1-cm intervals along the length of the leg. Muscle volumes were calculated from the summation of each anatomical CSA (ACSA) and the distance between each section. Muscle length was determined as the distance between the most proximal and distal images in which the muscle was visible. The PCSA of each muscle was calculated as muscle volume times the cosine of the angle of fiber pinnation divided by fiber length, where published fiber length:muscle length ratios were used to estimate fiber lengths. The mean volumes of the major plantarflexors were 489, 245, and 140 cm3 for the soleus and medial (MG) and lateral (LG) heads of the gastrocnemius. The mean PCSA of the soleus was 230 cm2, about three and eight times larger than the MG (68 cm2) and LG (28 cm2), respectively. These PCSA values were eight (soleus), four (MG), and three (LG) times larger than their respective maximum ACSA. The major dorsiflexor, the tibialis anterior (TA), had a muscle volume of 143 cm2, a PCSA of 19 cm2, and an ACSA of 9 cm2. With the exception of the soleus, the mean fiber length of all subjects was closely related to muscle volume across muscles. The soleus fibers were unusually short relative to the muscle volume, thus potentiating its force potential.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Enzyme and size profiles in chronically inactive cat soleus muscle fibers.

Spinal isolation (SI), i.e., the isolation of the lumbar spinal cord via a rostral and a caudal cord transection and bilateral dorsal rhizotomy, was used to determine the effects of chronic (6 months) inactivity on the size and metabolic properties of fibers in the cat soleus. Fibers were classified as dark or light, based on their staining reactions to myosin ATPase, alkaline preincubation, and immunohistochemically as expressing fast and/or slow myosin heavy chains (MHC). Succinate dehydrogenase (SDH) and alpha-glycerophosphate dehydrogenase (GPD) activities were assessed histochemically. Following SI, both the light and the dark ATPase fibers in the SI cats were significantly smaller than the light ATPase fibers in the controls. Normally 100% of the fibers were light ATPase and reacted exclusively with the slow antibody. After SI, approximately 45% of the fibers were dark ATPase fibers, many reacting with both fast and slow MHC antibodies. The total amount and concentration of GPD were higher in the light and dark ATPase fibers in SI compared with light ATPase fibers in controls. In contrast, although the total amount of SDH per fiber was decreased, reflecting the decrease in fiber size, the mean SDH concentration per fiber was unchanged following SI. These data indicate that there is a close coordination in the regulation of GPD activity and the type of myosin. SDH activity, on the other hand, appears to be resistant to decreased levels of activity and unloading, i.e., there seems to be a minimum level of oxidative potential in the soleus that is independent of activity level. Fiber sizes, however, are very sensitive to less-than-normal amounts of neuromuscular activity and/or loading.

Adenosine Triphosphatases

Variation and limitations in fiber enzymatic and size responses in hypertrophied muscle.

The present study was designed to determine whether the degree and kind of adaptation of a muscle fiber to a functional overload (FO) are determined by properties that are intrinsic to that fiber. The study also addresses the question of the capability of fibers to maintain a normal level of coordination of proteins per fiber as fiber volume changes dramatically. The plantaris muscle of six adult female cats was overloaded for 12 wk by bilateral synergist removal. Plantaris muscle fiber mean size doubled after FO, although some very small fibers that stained dark for adenosinetriphosphatase (ATPase) were observed in some of the FO muscles. There appeared to be no change in total succinate dehydrogenase activity per fiber. A reduction in succinate dehydrogenase activity per unit volume was observed in a substantial number of fibers, reflecting a disproportionate increase in fiber volume relative to mitochondrial volume. In contrast, total alpha-glycerophosphate dehydrogenase activity and actomyosin ATPase activity increased as fiber size increased, whereas there was no change in alpha-glycerophosphate dehydrogenase and ATPase activities per unit volume. Control and FO muscle fibers generally expressed either a fast or slow myosin heavy chain type, but in some cases FO muscle fibers expressed both fast and slow myosin heavy chains. The persistence of variability in fiber sizes and enzyme activities in fibers of overloaded muscles suggests a wide range in the adaptive potential of individual fibers to FO. These data indicate that a severalfold increase in cell size may occur without significant qualitative changes in the coordination of protein regulation associated with metabolic pathways and ATP utilization.

Adenosine Triphosphatases

Ventral horn cell responses to spaceflight and hindlimb suspension.

Spaceflight or hindlimb suspension results in a loss of mass and alterations of the metabolic and contractile protein profiles of skeletal muscles toward that resembling faster muscles. Given the influence of motoneurons on muscle properties, ventral horn cells of the lumbosacral enlargement of the spinal cord were studied to determine whether similar adaptations were present. Three groups of rats (5 per group) were studied: control, 14-day spaceflight, and 14-day hindlimb suspension. Spinal cords were quick-frozen, and the succinate dehydrogenase (SDH) activity and cross-sectional area of the soma of ventral horn cells were measured using a computer-enhanced image-processing system. Briefly, the optical density for SDH activity was determined after 8 min of incubation in a medium that gave a steady-state enzymatic reaction. Soma sizes were determined in cells with a visible nucleus. Although there were no significant differences among the three groups in mean cross-sectional area and SDH activity, the population distributions of both variables shifted significantly. In the flight rats, there was a shift toward smaller cells. Compared with control, the population distribution of SDH activities in the flight rats shifted toward higher activities, whereas the distribution shifted toward lower activities in the suspended rats. When considering the interactive effects within individual cells, there was a higher percentage of small cells with high SDH activities in the flight than in the control or suspended rats. These contrasting effects of spaceflight and hindlimb suspension suggest that the changes observed in ventral horn cells were due to factors other than simply the absence of weight support.

Animals

Rat soleus muscle fiber responses to 14 days of spaceflight and hindlimb suspension.

Morphological and enzymatic responses in fibers expressing fast, slow, or both types of myosin heavy chain (MHC) were studied in rats after 14 days of spaceflight (COSMOS 2044) or hindlimb suspension. Although the percentage of slow-twitch fibers was unchanged, a higher percentage of fibers that expressed both slow and fast MHC was observed in flight and suspended rats than in synchronous ground-based controls. The soleus was 25 and 34% smaller than control after 14 days of flight and suspension, with the reduction in fiber cross-sectional area (CSA) being greater in slow- than in fast-twitch fibers in both experimental groups. The activities of succinate dehydrogenase (SDH) and alpha-glycerophosphate dehydrogenase (GPD) were not significantly affected by flight or suspension. The total SDH activity (i.e., SDH activity x CSA) decreased significantly in the slow-twitch fibers of the flight and the fast-twitch fibers of the suspended rats, in large part due to fiber atrophy. A shift in MHC expression in 14 and 9% of the fibers in flight and suspended rats occurred without a change in myosin adenosinetriphosphatase activity. The SDH and GPD activities of the fibers that expressed both slow and fast MHC were slightly higher than the slow-twitch fibers and slightly lower than the fast-twitch fibers. These data indicate that events were initiated within 14 days of spaceflight or suspension that began to reconfigure the protein profiles of 9-14% of the slow-twitch fibers from typical slow-twitch toward those of fast-twitch fibers, while all fibers were dramatically losing total protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases

Adaptation of fibers in fast-twitch muscles of rats to spaceflight and hindlimb suspension.

The adaptation of single fibers in medial gastrocnemius (MG), a fast-twitch extensor, and tibialis anterior (TA), a fast-twitch flexor, was studied after 14 days of spaceflight (COSMOS 2044) or hindlimb suspension. Cross-sectional area (CSA) and succinate dehydrogenase (SDH), alpha-glycerophosphate dehydrogenase (GPD), and myofibrillar adenosinetriphosphatase (ATPase) activities were determined in fibers identified in frozen serial cross sections. Fibers were categorized as light, dark, or intermediate on the basis of myosin ATPase staining and alkaline preincubation and immunohistochemically as reacting with slow, fast, or both slow and fast myosin heavy chain monoclonal antibodies. Because there was a close relationship between these two means of categorizing fibers, all fibers were categorized on the basis of the immunohistochemical reaction. The percentage of slow- and fast-twitch fibers of the MG and TA were unchanged in either group. Mean fiber size of all fibers, irrespective of type, was unaffected in either muscle after flight or suspension. The fibers that expressed both fast and slow myosin heavy chains were smaller than control in the MG of both experimental groups. Compared with control, the SDH and total SDH activities in the MG were significantly less in suspended rats, with the fast-twitch fibers showing the largest difference. The ATPase activity in the MG was higher in flight than in control or suspended rats. There were no significant effects of flight on fibers of the TA. In contrast, the TA in suspended rats had higher GPD activities than either control or flight rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological

Spaceflight and growth effects on muscle fibers in the rhesus monkey.

Spaceflight causes considerable atrophy in hindlimb muscles of the rat. The purpose of this study was to investigate the effect of a 14-day spaceflight (COSMOS 2044) on selected morphological and metabolic properties of single muscle fibers in a nonhuman primate, Macaca mulatta. Biopsies were taken from the soleus (Sol), medial gastrocnemius (MG), and tibialis anterior (TA) muscles of two rhesus monkeys 107 days before flight and 24 h after return from flight. Muscle biopsies were taken from two independent sites in each muscle by use of a small (3-mm OD) Bergstrom biopsy needle. The biopsies weighed 8-14 mg and contained 100-200 fibers, of which an average of 40 fibers were acceptable for metabolic and size analyses. The 14-day spaceflight had little effect on fiber size in the Sol and MG muscles, whereas there appeared to be a slight decrease in size in the TA. In each of the flight animals, the mean fiber size in the postflight biopsies increased relative to preflight values. An increase in fiber size over the same period of time was also observed in four control monkeys that were the same age and approximately the same weight as the flight monkeys. The relative increase in size was related to the body weight of the monkey at the time of the pre- and postflight biopsies. The mean fiber succinate dehydrogenase activity appeared to decrease in the MG, whereas there was no apparent effect of spaceflight on the Sol and TA muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Fibre size and type adaptations to spinal isolation and cyclical passive stretch in cat hindlimb.

Impulse activity is known to have a strong influence in determining the characteristics that distinguish skeletal muscle fibres into types. The control of muscle proteins by the neural systems that innervate the muscles, however, is not complete (Edgerton et al. 1985, 1990). The purpose of the present study, therefore, was to determine the effects of inactivity for 6 months on the size and fibre type composition of selected cat hindlimb muscles. Inactivity was produced by isolating the lumbar region of the spinal cord, i.e. transecting the cord at T12-T13 and again at L7-.S1 and then performing a bilateral dorsal rhizotomy between the transection sites (SI). In each SI cat, one hindlimb was passively manipulated for 30 min per day through a range of motion at the ankle mimicking a step cycle. SI resulted in an atrophic response in most muscles, with predominantly slow extensors showing the largest effect. In general, the predominant fibre type, which also had the largest mean size, in each muscle atrophied the most. The mean fibre size of all fibre types were similar after SI, suggesting that there may be a minimal size for inactive intact fibres. In comparison with control animals, all muscles in the SI cats had a higher proportion of fast fibres. Further, the relative contribution of the slow fibres to the total cross-sectional area of the muscle was decreased following SI. Some slow fibres in each muscle, however, were resistant to change. These data demonstrate the extent to which size and myosin type of mammalian muscle fibres are independent of activation characteristics.

Adaptation, Physiological