RESUMEN
Based on studies of fast skeletal muscles, hibernating black and brown bears resist skeletal muscle atrophy during months of reduced physical activity and not feeding. The present study examined atrophy sparing in the slow soleus muscle, known to be highly prone to disuse atrophy in humans and other mammals. We demonstrated histochemically that the black bear soleus is rich in slow fibers, averaging 84.0 ± 6.6%. The percentages of slow fibers in fall (87.3 ± 4.9%) and during hibernation (87.1 ± 5.6%) did not differ ( P = 0.3152) from summer. The average fiber cross-sectional area to body mass ratio (48.6 ± 11.7 µm2/kg) in winter hibernating bears was not significantly different from that of summer (54.1 ± 11.8 µm2/kg, P = 0.4186) and fall (47.0 ± 9.7 µm2/kg, P = 0.9410) animals. The percentage of single hybrid fibers containing both slow and fast myosin heavy chains, detected biochemically, increased from 2.6 ± 3.8% in summer to 24.4 ± 24.4% ( P = 0.0244) during hibernation. The shortening velocities of individual hybrid fibers remained unchanged from that of pure slow and fast fibers, indicating low content of the minority myosins. Slow and fast fibers in winter bears exhibited elevated specific tension (kN/m2; 22%, P = 0.0161 and 11%, P = 0.0404, respectively) and maintained normalized power. The relative stability of fiber type percentage and size, fiber size-to-body mass ratio, myosin heavy chain isoform content, shortening velocity, power output, and elevated specific tension during hibernation validates the ability of the black bear to preserve the biochemical and performance characteristics of the soleus muscle during prolonged hibernation.
Asunto(s)
Hibernación , Contracción Muscular , Fuerza Muscular , Músculo Esquelético/fisiología , Atrofia Muscular/prevención & control , Ursidae/fisiología , Animales , Complejo IV de Transporte de Electrones/metabolismo , Metabolismo Energético , Femenino , Glucógeno/metabolismo , Masculino , Mitocondrias Musculares/metabolismo , Fibras Musculares de Contracción Rápida/fisiología , Fibras Musculares de Contracción Lenta/fisiología , Músculo Esquelético/citología , Músculo Esquelético/metabolismo , Atrofia Muscular/metabolismo , Atrofia Muscular/fisiopatología , Cadenas Pesadas de Miosina/metabolismo , Fenotipo , Factores de Tiempo , Ursidae/metabolismoRESUMEN
Our primary goal was to determine the effects of 6-mo flight on the International Space Station (ISS) on selected anaerobic and aerobic enzymes, and the content of glycogen and lipids in slow and fast fibers of the soleus and gastrocnemius. Following local anesthesia, biopsies were obtained from nine ISS crew members â¼45 days preflight and on landing day (R+0) postflight. We subdivided the crew into those who ran 200 min/wk or more (high treadmill, HT) in-flight from those who ran <100 min/wk (low treadmill, LT). In the LT group, there was a loss of lipid in soleus type I fibers, and muscle glycogen significantly increased in soleus fiber types postflight. Soleus cytochrome oxidase (CO) activity was significantly depressed postflight in the type I fiber. This was attributed to the LT group where CO activity was reduced 59%. Otherwise, there was no change in the crew mean for type I or IIa fiber glycolytic or mitochondrial enzyme activities pre- vs. postflight in either muscle. However, two of the three HT subjects (Subjects E and H) showed significant increases in both ß-hydroxyacyl-CoA dehydrogenase and citrate synthase in the soleus type I fibers, and Subject E, exhibiting the largest increase in soleus oxidative enzymes, was the only subject to show a significant decrease in glycolytic enzyme activity. It is apparent that crew members performing adequate treadmill running can maintain calf muscle enzymes, which suggests that increased fatigue with weightlessness cannot be directly caused by a decline in muscle enzyme capacity.
Asunto(s)
Fibras Musculares de Contracción Rápida/enzimología , Fibras Musculares de Contracción Rápida/metabolismo , Fibras Musculares de Contracción Lenta/enzimología , Fibras Musculares de Contracción Lenta/metabolismo , 3-Hidroxiacil-CoA Deshidrogenasas/metabolismo , Citrato (si)-Sintasa/metabolismo , Ejercicio Físico/fisiología , Prueba de Esfuerzo/métodos , Glucógeno/metabolismo , Humanos , Lípidos , Fatiga Muscular/fisiología , Fibras Musculares de Contracción Rápida/fisiología , Fibras Musculares de Contracción Lenta/fisiología , Vuelo Espacial , IngravidezRESUMEN
The primary goal of this study was to determine the effects of prolonged space flight (180 days) on the structure and function of slow and fast fibres in human skeletal muscle. Biopsies were obtained from the gastrocnemius and soleus muscles of nine International Space Station crew members 45 days pre- and on landing day (R+0) post-flight. The main findings were that prolonged weightlessness produced substantial loss of fibre mass, force and power with the hierarchy of the effects being soleus type I > soleus type II > gastrocnemius type I > gastrocnemius type II. Structurally, the quantitatively most important adaptation was fibre atrophy, which averaged 20% in the soleus type I fibres (98 to 79 µm diameter). Atrophy was the main contributor to the loss of peak force (P(0)), which for the soleus type I fibre declined 35% from 0.86 to 0.56 mN. The percentage decrease in fibre diameter was correlated with the initial pre-flight fibre size (r = 0.87), inversely with the amount of treadmill running (r = 0.68), and was associated with an increase in thin filament density (r = 0.92). The latter correlated with reduced maximal velocity (V(0)) (r = 0.51), and is likely to have contributed to the 21 and 18% decline in V(0) in the soleus and gastrocnemius type I fibres. Peak power was depressed in all fibre types with the greatest loss (55%) in the soleus. An obvious conclusion is that the exercise countermeasures employed were incapable of providing the high intensity needed to adequately protect fibre and muscle mass, and that the crew's ability to perform strenuous exercise might be seriously compromised. Our results highlight the need to study new exercise programmes on the ISS that employ high resistance and contractions over a wide range of motion to mimic the range occurring in Earth's 1 g environment.
Asunto(s)
Adaptación Fisiológica/fisiología , Fibras Musculares Esqueléticas/fisiología , Vuelo Espacial , Adulto , Atrofia , Fenómenos Biomecánicos , Ejercicio Físico , Humanos , Persona de Mediana Edad , Fibras Musculares Esqueléticas/ultraestructura , Factores de TiempoRESUMEN
Single skinned fibers from soleus and adductor longus (AL) muscles of weight-bearing control rats and rats after 14-day hindlimb suspension unloading (HSU) were studied physiologically and ultrastructurally to investigate how slow fibers increase shortening velocity (V0) without fast myosin. We hypothesized that unloading and shortening of soleus during HSU reduces densities of thin filaments, generating wider myofilament separations that increase V0 and decrease specific tension (kN/m2). During HSU, plantarflexion shortened soleus working length 23%. AL length was unchanged. Both muscles atrophied as shown by reductions in fiber cross-sectional area. For AL, the 60% atrophy accounted fully for the 58% decrease in absolute tension (mN). In the soleus, the 67% decline in absolute tension resulted from 58% atrophy plus a 17% reduction in specific tension. Soleus fibers exhibited a 25% reduction in thin filaments, whereas there was no change in AL thin filament density. Loss of thin filaments is consistent with reduced cross bridge formation, explaining the fall in specific tension. V0 increased 27% in soleus but was unchanged in AL. The V0 of control and HSU fibers was inversely correlated (R = -0.83) with thin filament density and directly correlated (R = 0.78) with thick-to-thin filament spacing distance in a nonlinear fashion. These data indicate that reduction in thin filament density contributes to an increased V0 in slow fibers. Osmotically compacting myofilaments with 5% dextran returned density, spacing, and specific tension and slowed V0 to near-control levels and provided evidence for myofilament spacing modulating tension and V0.
Asunto(s)
Citoesqueleto de Actina/ultraestructura , Fibras Musculares Esqueléticas/fisiología , Músculo Esquelético/fisiología , Atrofia Muscular/etiología , Animales , Suspensión Trasera/efectos adversos , Masculino , Microscopía Electrónica de Transmisión , Fibras Musculares Esqueléticas/ultraestructura , Músculo Esquelético/ultraestructura , Atrofia Muscular/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Hindlimb suspension unloading (HSU) is a ground-based model simulating the effects of microgravity unloading on the musculoskeletal system. In this model, gravity causes the hind foot of the rat to drop, opening the front of the ankle to 90-105 degrees plantar flexion at rest. As HSU proceeds, the normal weight-bearing angle of 30 degrees dorsiflexion is achieved progressively less, and the contraction range of soleus is abbreviated. Our laboratory reported that 12 days of HSU caused central corelike lesions (CCLs) of myofibril breakdown (Riley DA, Slocum GR, Bain JL, Sedlak FR, Sowa TE, and Mellender JW. J Appl Physiol. 69: 58-66, 1990). The present study investigated whether daily stretch of the calf muscles prevents CCL formation. The soleus muscles of HSU Sprague-Dawley male rats (approximately 287 g) were lengthened by unilateral ankle splinting at 30 degrees. Compared with the nonsplinted side, splinting for 10 or 20 min per day in awake rats significantly decreased CCLs in soleus by 88 and 91%, respectively (P < 0.01). Compared with control muscle wet weight, 20-min splinting reduced atrophy by 33%, whereas 10-min splinting ameliorated atrophy by 17% (P < 0.01). Bilateral soleus electromyograph recording revealed higher levels of contractile activity on the splinted side during splinting. To isolate the effects of stretch from isometric contractile activity, contractions were eliminated by whole animal anesthesia with isoflurane during 10-min daily splinting. The percentage of fibers with CCLs was reduced by 57%, and the average lesion size was 29% smaller in the stretched muscle (P < 0.05). Soleus muscle wet weight and fiber area were unaltered by stretch alone. Loaded contractions during splinting are necessary to prevent muscle fiber atrophy. Passive muscle stretch acts to maintain myofibril structural integrity.
Asunto(s)
Suspensión Trasera/efectos adversos , Suspensión Trasera/métodos , Músculo Esquelético/patología , Músculo Esquelético/fisiopatología , Atrofia Muscular/etiología , Atrofia Muscular/prevención & control , Estimulación Física/métodos , Animales , Tirantes , Masculino , Contracción Muscular , Atrofia Muscular/patología , Atrofia Muscular/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
We used Ca2+-activated skinned muscle fibers to test the hypothesis that unilateral lower leg suspension (ULLS) alters cross-bridge mechanisms of muscle contraction. Soleus and gastrocnemius biopsies were obtained from eight subjects before ULLS, immediately after 12 days of ULLS (post-0 h), and after 6 h of reambulation (post-6 h). Post-0 h soleus fibers expressing type I myosin heavy chain (MHC) showed significant reductions in diameter, absolute and specific peak Ca2+-activated force, unloaded shortening velocity, and absolute and normalized peak power. Fibers obtained from the gastrocnemius were less affected by ULLS, particularly fibers expressing fast MHC isoforms. Post-6 h soleus fibers produced less absolute and specific peak force than did post-0 h fibers, suggesting that reambulation after ULLS induced cell damage. Like bed rest and spaceflight, ULLS primarily affects soleus over gastrocnemius fibers. However, in contrast to these other models, slow soleus fibers obtained after ULLS showed a decrease in unloaded shortening velocity and a greater reduction in specific force.
Asunto(s)
Reposo en Cama , Pierna/fisiología , Fibras Musculares Esqueléticas/fisiología , Vuelo Espacial , Simulación de Ingravidez , Adulto , Calcio/fisiología , Femenino , Lateralidad Funcional/fisiología , Humanos , Masculino , Contracción Muscular/fisiología , Fibras Musculares de Contracción Lenta/fisiología , Músculo Esquelético/citología , Músculo Esquelético/fisiología , Músculo Esquelético/ultraestructuraRESUMEN
The purpose of this investigation was to study the effects of a 17-day spaceflight on the contractile properties of individual fast- and slow-twitch fibers isolated from biopsies of the fast-twitch gastrocnemius muscle of four male astronauts. Single chemically skinned fibers were studied during maximal Ca2+-activated contractions with fiber myosin heavy chain (MHC) isoform expression subsequently determined by SDS gel electrophoresis. Spaceflight had no significant effect on the mean diameter or specific force of single fibers expressing type I, IIa, or IIa/IIx MHC, although a small reduction in average absolute force (P(o)) was observed for the type I fibers (0.68 +/- 0.02 vs. 0.64 +/- 0.02 mN, P < 0.05). Subject-by-flight interactions indicated significant intersubject variation in response to the flight, as postflight fiber diameter and P(o) where significantly reduced for the type I and IIa fibers obtained from one astronaut and for the type IIa fibers from another astronaut. Average unloaded shortening velocity [V(o), in fiber lengths (FL)/s] was greater after the flight for both type I (0.60 +/- 0.03 vs. 0.76 +/- 0.02 FL/s) and IIa fibers (2.33 +/- 0.25 vs. 3.10 +/- 0.16 FL/s). Postflight peak power of the type I and IIa fibers was significantly reduced only for the astronaut experiencing the greatest fiber atrophy and loss of P(o). These results demonstrate that 1) slow and fast gastrocnemius fibers show little atrophy and loss of P(o) but increased V(o) after a typical 17-day spaceflight, 2) there is, however, considerable intersubject variation in these responses, possibly due to intersubject differences in in-flight physical activity, and 3) in these four astronauts, fiber atrophy and reductions in P(o) were less for slow and fast fibers obtained from the phasic fast-twitch gastrocnemius muscle compared with slow and fast fibers obtained from the slow antigravity soleus [J. J. Widrick, S. K. Knuth, K. M. Norenberg, J. G. Romatowski, J. L. W. Bain, D. A. Riley, M. Karhanek, S. W. Trappe, T. A. Trappe, D. L. Costill, and R. H. Fitts. J Physiol (Lond) 516: 915-930, 1999].
Asunto(s)
Fibras Musculares de Contracción Rápida/fisiología , Fibras Musculares de Contracción Lenta/fisiología , Músculo Esquelético/fisiología , Vuelo Espacial , Atrofia , Calcio/farmacología , Electroforesis en Gel de Poliacrilamida , Humanos , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Fibras Musculares de Contracción Rápida/ultraestructura , Fibras Musculares de Contracción Lenta/ultraestructura , Músculo Esquelético/citología , Músculo Esquelético/ultraestructura , Miofibrillas/fisiologíaRESUMEN
Slow oxidative (SO) fibers of the adductor longus (AL) were predominantly damaged during voluntary reloading of hindlimb unloaded (HU) rats and appeared explainable by preferential SO fiber recruitment. The present study assessed damage after eliminating the variable of voluntary recruitment by tetanically activating all fibers in situ through the motor nerve while applying eccentric (lengthening) or isometric contractions. Muscles were aldehyde fixed and resin embedded, and semithin sections were cut. Sarcomere lesions were quantified in toluidine blue-stained sections. Fibers were typed in serial sections immunostained with antifast myosin and antitotal myosin (which highlights slow fibers). Both isometric and eccentric paradigms caused fatigue. Lesions occurred only in eccentrically contracted control and HU muscles. Fatigue did not cause lesions. HU increased damage because lesioned- fiber percentages within fiber types and lesion sizes were greater than control. Fast oxidative glycolytic (FOG) fibers were predominantly damaged. In no case did damaged SO fibers predominate. Thus, when FOG, SO, and hybrid fibers are actively lengthened in chronically unloaded muscle, FOG fibers are intrinsically more susceptible to damage than SO fibers. Damaged hybrid-fiber proportions ranged between these extremes.
Asunto(s)
Contracción Muscular/fisiología , Fibras Musculares Esqueléticas/fisiología , Músculo Esquelético/fisiología , Animales , Miembro Posterior , Contracción Isométrica/fisiología , Masculino , Neuronas Motoras/fisiología , Fatiga Muscular , Fibras Musculares de Contracción Lenta/fisiología , Músculo Esquelético/inervación , Músculo Esquelético/patología , Ratas , Ratas Sprague-Dawley , Sarcómeros/fisiología , Sarcómeros/ultraestructura , Factores de TiempoRESUMEN
STUDY OBJECTIVE: We describe 9 patients who underwent ultrasound-guided transvenous cardiac pacing in which ultrasonographic imaging was used to assist and confirm the placement of electrode catheters within the right ventricle. METHODS: We prospectively enrolled consecutive patients with complete heart block who received emergency ultrasound-assisted transvenous cardiac pacing (TVCP). Emergency physicians performed both ultrasound scanning and placement of the TVCP electrodes at a busy urban teaching medical center. RESULTS: Real-time ultrasound-guided TVCP was successful in 8 (88.9%) of the 9 patients studied. The pacing catheter was not adequately visualized in 1 patient who ultimately required placement by a cardiologist. Echocardiography was useful in identifying pacing catheter misplacement and subsequent successful repositioning in 3 patients. CONCLUSION: Emergency physicians should be aware that ultrasound technology could be useful in assisting TVCP in the emergency department setting. Further investigation is required to adequately evaluate this modality as a new indication for ED echocardiography.
Asunto(s)
Estimulación Cardíaca Artificial/métodos , Ecocardiografía , Marcapaso Artificial , Ultrasonografía Intervencional , Anciano , Anciano de 80 o más Años , Presión Sanguínea/fisiología , Cateterismo Venoso Central , Electrodos Implantados , Servicio de Urgencia en Hospital , Femenino , Bloqueo Cardíaco/terapia , Frecuencia Cardíaca/fisiología , Ventrículos Cardíacos/diagnóstico por imagen , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Tasa de Supervivencia , Factores de Tiempo , Resultado del TratamientoRESUMEN
We have developed a method of fixing, embedding, sectioning, and staining that allows high-resolution detection of myofibrillar structure and myosin immunocytochemical muscle fiber typing in serial semithin sections of LR White plastic embedded muscle at the light microscopic level. Traditional approaches, such as cryostat sections, permit fiber typing, but small myofibrillar lesions (1-3 sarcomeres) are difficult to detect because of section thickness. Semithin sections of hydrophobic resins do not stain well either histochemically or immunocytochemically. Electron microscopy can resolve lesions and discriminate fiber types based on morphology, but the sampling area is small. Our goal was to develop a rapid method for defining both fiber type and high-resolution primary myofibrillar lesion damage. Mild fixation (1-4% paraformaldehyde, 0. 05-0.1% glutaraldehyde) and embedment in a hydrophilic resin (LR White) were used. Myofibrillar structure was extremely well preserved at the light microscopic (LM) level, and lesions could be readily resolved in Toluidine blue stained 500-nm sections. Fiber type was defined by LM immunomyosin staining of serial plastic semithin sections, which demonstrated reciprocal staining patterns for "fast (Sigma M4276) and "total" (skeletal muscle) myosins (Sigma M7523).
Asunto(s)
Fibras Musculares Esqueléticas/clasificación , Adhesión en Plástico/métodos , Resinas Acrílicas , Animales , Inmunohistoquímica , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/ultraestructura , Miosinas/análisis , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Fijación del Tejido , Cloruro de TolonioRESUMEN
The traditional approach to the study of selective attention in animal discrimination learning has been to ask if animals are capable of the central selective processing of stimuli, such that certain aspects of the discriminative stimuli are partially or wholly ignored while their relationships to each other, or other relevant stimuli, are processed. A notable characteristic of this research has been that procedures involve the acquisition of discriminations, and the issue of concern is whether learning is selectively determined by the stimulus dimension defined by the discriminative stimuli. Although there is support for this kind of selective attention, in many cases, simpler nonattentional accounts are sufficient to explain the results. An alternative approach involves procedures more similar to those used in human information-processing research. When selective attention is studied in humans, it generally involves the steady state performance of tasks for which there is limited time allowed for stimulus input and a relatively large amount of relevant information to be processed; thus, attention must be selective or divided. When this approach is applied to animals and alternative accounts have been ruled out, stronger evidence for selective or divided attention in animals has been found. Similar processes are thought to be involved when animals search more natural environments for targets. Finally, an attempt is made to distinguish these top-down attentional processes from more automatic preattentional processes that have been studied in humans and other animals.
Asunto(s)
Atención/fisiología , Aprendizaje Discriminativo/fisiología , Animales , Conducta Animal/fisiología , Condicionamiento Psicológico/fisiología , Señales (Psicología) , Generalización de la Respuesta , RatasRESUMEN
Soleus muscle fibers were examined electron microscopically from pre- and postflight biopsies of four astronauts orbited for 17 days during the Life and Microgravity Sciences Spacelab Mission (June 1996). Myofilament density and spacing were normalized to a 2. 4-microm sarcomere length. Thick filament density ( approximately 1, 062 filaments/microm(2)) and spacing ( approximately 32.5 nm) were unchanged by spaceflight. Preflight thin filament density (2, 976/microm(2)) decreased significantly (P < 0.01) to 2,215/microm(2) in the overlap A band region as a result of a 17% filament loss and a 9% increase in short filaments. Normal fibers had 13% short thin filaments. The 26% decrease in thin filaments is consistent with preliminary findings of a 14% increase in the myosin-to-actin ratio. Lower thin filament density was calculated to increase thick-to-thin filament spacing in vivo from 17 to 23 nm. Decreased density is postulated to promote earlier cross-bridge detachment and faster contraction velocity. Atrophic fibers may be more susceptible to sarcomere reloading damage, because force per thin filament is estimated to increase by 23%.
Asunto(s)
Citoesqueleto de Actina/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/metabolismo , Atrofia Muscular/fisiopatología , Vuelo Espacial , Astronautas , Humanos , Masculino , Microscopía Electrónica , Fibras Musculares Esqueléticas/química , Músculo Esquelético/química , Miofibrillas/metabolismo , Miofibrillas/ultraestructura , Sarcómeros/metabolismo , Sarcómeros/ultraestructura , IngravidezRESUMEN
We hypothesized that hindlimb suspension unloading of 8-day-old neonatal rats would disrupt the normal development of muscle fiber types and the motor innervation of the antigravity (weightbearing) soleus muscles but not extensor digitorum longus (EDL) muscles. Five rats were suspended 4.5 h and returned 1.5 h to the dam for nursing on a 24 h cycle for 9 days. To control for isolation from the dam, the remaining five littermates were removed on the same schedule but not suspended. Another litter of 10 rats housed in the same room provided a vivarium control. Fibers were typed by myofibrillar ATPase histochemistry and immunostaining for embryonic, slow, fast IIA and fast IIB isomyosins. The percentage of multiple innervation and the complexity of singly-innervated motor terminal endings were assessed in silver/cholinesterase stained sections. Unique to the soleus, unloading accelerated production of fast IIA myosin, delayed expression of slow myosin and retarded increases in standardized muscle weight and fiber size. Loss of multiple innervation was not delayed. However, fewer than normal motor nerve endings achieved complexity. Suspended rats continued unloaded hindlimb movements. These findings suggest that motor neurons resolve multiple innervation through nerve impulse activity, whereas the postsynaptic element (muscle fiber) controls endplate size, which regulates motor terminal arborization. Unexpectedly, in the EDL of unloaded rats, transition from embryonic to fast myosin expression was retarded. Suspension-related foot drop, which stretches and chronically loads EDL, may have prevented fast fiber differentiation. These results demonstrate that neuromuscular development of both weightbearing and non-weightbearing muscles in rats is dependent upon and modulated by hindlimb loading.
Asunto(s)
Placa Motora/crecimiento & desarrollo , Placa Motora/fisiología , Desarrollo de Músculos , Fibras Musculares Esqueléticas/fisiología , Músculo Esquelético/crecimiento & desarrollo , Músculo Esquelético/inervación , Animales , Animales Recién Nacidos , Diferenciación Celular/fisiología , Miembro Posterior/inervación , Miembro Posterior/fisiología , Neuronas Motoras/fisiología , Fibras Musculares Esqueléticas/química , Fibras Musculares Esqueléticas/citología , Músculo Esquelético/citología , Miosinas/fisiología , Tamaño de los Órganos , Ratas , Ratas Sprague-Dawley , Soporte de Peso/fisiología , IngravidezRESUMEN
1. Soleus biopsies were obtained from four male astronauts 45 days before and within 2 h after a 17 day spaceflight. 2. For all astronauts, single chemically skinned post-flight fibres expressing only type I myosin heavy chain (MHC) developed less average peak Ca2+ activated force (Po) during fixed-end contractions (0.78 +/- 0. 02 vs. 0.99 +/- 0.03 mN) and shortened at a greater mean velocity during unloaded contractions (Vo) (0.83 +/- 0.02 vs. 0.64 +/- 0.02 fibre lengths s-1) than pre-flight type I fibres. 3. The flight-induced decline in absolute Po was attributed to reductions in fibre diameter and/or Po per fibre cross-sectional area. Fibres from the astronaut who experienced the greatest relative loss of peak force also displayed a reduction in Ca2+ sensitivity. 4. The elevated Vo of the post-flight slow type I fibres could not be explained by alterations in myosin heavy or light chain composition. One alternative possibility is that the elevated Vo resulted from an increased myofilament lattice spacing. This hypothesis was supported by electron micrographic analysis demonstrating a reduction in thin filament density post-flight. 5. Post-flight fibres shortened at 30 % higher velocities than pre-flight fibres at external loads associated with peak power output. This increase in shortening velocity either reduced (2 astronauts) or prevented (2 astronauts) a post-flight loss in fibre absolute peak power (microN (fibre length) s-1). 6. The changes in soleus fibre diameter and function following spaceflight were similar to those observed after 17 days of bed rest. Although in-flight exercise countermeasures probably reduced the effects of microgravity, the results support the idea that ground-based bed rest can serve as a model of human spaceflight. 7. In conclusion, 17 days of spaceflight decreased force and increased shortening velocity of single Ca2+-activated muscle cells expressing type I MHC. The increase in shortening velocity greatly reduced the impact that impaired force production had on absolute peak power.
Asunto(s)
Fibras Musculares Esqueléticas/fisiología , Músculo Esquelético/fisiología , Vuelo Espacial , Ingravidez/efectos adversos , Adulto , Señalización del Calcio/fisiología , Electroforesis en Gel de Poliacrilamida , Humanos , Masculino , Microscopía Electrónica , Contracción Muscular/fisiología , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/ultraestructura , Músculo Esquelético/citología , Músculo Esquelético/metabolismo , Cadenas Pesadas de Miosina/metabolismoRESUMEN
Sarcomere disruptions are observed in the adductor longus (AL) muscles following voluntary reloading of spaceflown and hindlimb suspension unloaded (HSU) rat, which resemble lesions in eccentrically challenged muscle. We devised and tested an eccentric contraction (ECCON) test system for the 14-day HSU rat AL. Six to 7 hours following ECCON, ALs were fixed to allow immunostaining and electron microscopy (EM). Toluidine blue-stained histology semithin sections were screened for lesion density (#/mm2). Serial semithin sections from the ECCON group were characterized for myosin immunointensity of lesions. Five myofibrillar lesion types were identified in histological semithin sections: focal contractions; wide A-bands; opaque areas; missing A-bands; and hyperstretched sarcomeres. Lesion density by type was greater for ECCON than NonECCON ALs (P< or =0.05; focal contractions and opaque regions). Lesion density (#-of-all-five-types/mm2) was significantly different (ECCON: 23.91+/-10.58 vs. NonECCON: 5.48+/-1.28, P< or =0.05; ECCON vs. SHAM: 0.00+/-0.00; P< or = 0.025). PostECCON optimal tension decreased (Poi-drop, 17.84+/-4.22%) and was correlated to lesion density (R2=0.596), but prestretch tension demonstrated the highest correlation with lesion density (R2=0.994). In lesions, the darkly staining A-band lost the normally organized thick filament alignment to differing degrees across the different lesion types. Ranking the five lesion types by a measure of lesion length deformation (hypercontracted to hyperstretched) at the light microscopy level, related to the severity of thick filament registry loss across the lesion types at the electron microscopic level. This ranking suggested that the five lesion types seen in semithin sections at the light level represented a lesion progression sequence and paralleled myosin immunostaining loss as the distorted A-band filaments spread across the hyperlengthening lesion types. Lesion ultrastructure indicated damage involved calcium homeostasis loss (focal contraction lesions) and "thick-filament-centering" failure of titin (wide A-band lesions) in the early stages of lesion development.
Asunto(s)
Músculo Esquelético/patología , Músculo Esquelético/fisiopatología , Miofibrillas/patología , Miofibrillas/fisiología , Simulación de Ingravidez , Animales , Estimulación Eléctrica , Suspensión Trasera/efectos adversos , Suspensión Trasera/fisiología , Inmunohistoquímica , Microscopía Electrónica , Contracción Muscular/fisiología , Atrofia Muscular/patología , Atrofia Muscular/fisiopatología , Miosinas/metabolismo , Necrosis , Ratas , Estrés MecánicoRESUMEN
It is now clear that prevention of muscle debilitation during spaceflight will require a broader approach than simple exercise aimed at strengthening of the muscle fibers. The levels of several hormones and receptors are altered by unloading and must be returned to homeostasis. Pharmacotherapy and gene transfer strategies to raise the relative level of structural proteins may minimize the problems faced by astronauts in readapting to Earth-gravity. Up to now, we have only minimally exploited microgravity for advancing our understanding of muscle biology. A research laboratory in the space station with a centrifuge facility (gravity control) is essential for conducting basic research in this field. Microgravity has proven an excellent tool for noninvasively perturbing the synthesis of muscle proteins in the search for molecular signals and gene regulatory factors influencing differentiation, growth, maintenance and atrophy of muscle. Understanding the relation between blood flow and interstitial edema and between workload and subsequent structural failure are but two important problems that require serious attention. The roles of hormones and growth factors in regulating gene expression and their microgravity-induced altered production are other urgent issues to pursue. These types of studies will yield information that advances basic knowledge of muscle biology and offers insights into countermeasure design. This knowledge is likely to assist rehabilitation of diseased or injured muscles in humans on Earth, especially individuals in the more vulnerable aging population and persons participating in strenuous sports. Will the skeletal muscle system be prepared for the increased exposure to microgravity and the return to gravity loading without injury when space station is operational? The answer depends in large part on continued access to space and funding of ground-based models and flight experiments. The previous two decades of spaceflight research have described the effects of microgravity on multiple systems. The next generation of experiments promises to be even more exciting as we are challenged to define the cellular and molecular mechanisms of microgravity-induced changes.
Asunto(s)
Gravitación , Músculo Esquelético/fisiología , Vuelo Espacial , Animales , Humanos , Factores de TiempoRESUMEN
Histochemical staining for carbonic anhydrase and cholinesterase (CE) activities was used to analyze sensory and motor axon regeneration, respectively, during neuroma formation in transected and tube-encapsulated peripheral nerves. Median-ulnar and sciatic nerves in the rodent model permitted testing whether a 4 cm greater distance of the motor neuron soma from axotomy site or intrinsic differences between motor and sensory neurons influenced regeneration and neuroma formation 10, 30, and 90 days later. Ventral root radiculotomy confirmed that CE-stained axons were 97% alpha motor axons. Distance significantly delayed axon regeneration. When distance was negligible, sensory axons grew out sooner than motor axons, but motor axons regenerated to a greater quantity. These results indicate regeneration differences between axon subtypes and suggest more extensive branching of motor axons within the neuroma. Thus, both distance from injury site to soma and inherent motor and sensory differences should be considered in peripheral nerve repair strategies.
Asunto(s)
Axones/fisiología , Neuronas Motoras/fisiología , Fibras Nerviosas Mielínicas/fisiología , Regeneración Nerviosa/fisiología , Neuroma/fisiopatología , Neuronas Aferentes/fisiología , Neoplasias del Sistema Nervioso Periférico/fisiopatología , Animales , Anhidrasas Carbónicas/metabolismo , Colinesterasas/metabolismo , Masculino , Neuroma/patología , Nervios Periféricos/enzimología , Neoplasias del Sistema Nervioso Periférico/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Sarcomere lesions were previously observed with reloading of rat adductor longus muscles after spaceflight and hindlimb unloading (HU). Spaceflown rats displayed more lesioned fibers in the "slow-fiber" region, suggesting a damage-susceptible fiber type. Unloading induces fast myosin expression in some slow fibers, generating hybrid fibers. We examined whether lesion damage differed among slow-, hybrid-, and fast-fiber types in HU-reloaded adductor longus muscles. Temporal HU for 5, 8, 11, 14, and 17 days revealed that hybrid fiber percent, detected by antimyosin immunostaining, peaked at 29 +/- 12% by 14 days. A 14-day HU followed by 12-14 h of voluntary reloading was performed to induce lesions. chi2 analysis showed that slow fibers were preferentially damaged, accounting for 92 +/- 5% of lesioned fibers; hybrid and fast fibers accounted for 7 +/- 4 and <0.5%, respectively. Atrophy did not explain differential lesion damage across fiber types, as slow and hybrid fibers atrophied to a similar extent. Because active myofiber contractions are requisite for lesion formation, selective recruitment of slow fibers most likely explains their damage susceptibility.
Asunto(s)
Fibras Musculares de Contracción Lenta/fisiología , Músculo Esquelético/fisiología , Sarcómeros/fisiología , Simulación de Ingravidez , Animales , Atrofia/patología , Miembro Posterior/fisiología , Inmunohistoquímica , Masculino , Contracción Muscular/fisiología , Fibras Musculares de Contracción Rápida/fisiología , Fibras Musculares de Contracción Lenta/ultraestructura , Músculo Esquelético/citología , Músculo Esquelético/ultraestructura , Ratas , Ratas Sprague-Dawley , Sarcómeros/ultraestructuraRESUMEN
Previously we reported that, after 17-day bed rest unloading of 8 humans, soleus slow fibers atrophied and exhibited increased velocity of shortening without fast myosin expression. The present ultrastructural study examined fibers from the same muscle biopsies to determine whether decreased myofilament packing density accounted for the observed speeding. Quantitation was by computer-assisted morphometry of electron micrographs. Filament densities were normalized for sarcomere length, because density depends directly on length. Thick filament density was unchanged by bed rest. Thin filaments/microm2 decreased 16-23%. Glycogen filled the I band sites vacated by filaments. The percentage decrease in thin filaments (Y) correlated significantly (P < 0.05) with the percentage increase in velocity (X), (Y = 0.1X + 20%, R2 = 0.62). An interpretation is that fewer filaments increases thick to thin filament spacing and causes earlier cross-bridge detachment and faster cycling. Increased velocity helps maintain power (force x velocity) as atrophy lowers force. Atrophic muscles may be prone to sarcomere reloading damage because force/microm2 was near normal, and force per thin filament increased an estimated 30%.
Asunto(s)
Citoesqueleto de Actina/ultraestructura , Reposo en Cama , Músculo Esquelético/ultraestructura , Adulto , Atrofia , Biopsia , Humanos , Masculino , Persona de Mediana Edad , Fibras Musculares Esqueléticas/ultraestructura , Músculo Esquelético/patología , Factores de TiempoRESUMEN
Human carboxypeptidase N is a 280-kDa tetrameric enzyme consisting of two 83-kDa regulatory subunits and two catalytic 50-kDa subunits. The 83-kDa subunit is a member of the leucine-rich repeat family of proteins and has been localized to chromosome 8p22-p23. The 50-kDa subunit is a member of the regulatory B-type carboxypeptidase family, which includes carboxypeptidases M, E/H, AEBP1, and a newly described member, carboxypeptidase D, which has three tandem active site domains. The human genes for carboxypeptidase D (HGMW-approved symbol CPD) and the 50-kDa subunit of carboxypeptidase N (HGMW-approved symbol CPN1) were localized to chromosomes 17 and 10, respectively, using the polymerase chain reaction with gene-specific primers and DNAs derived from somatic cell hybrids. The carboxypeptidase D gene was further localized to the centromeric region 17p11.1-q11.1/11.2 by use of a regional mapping panel derived from somatic cell hybrids containing different portions of chromosome 17.