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At least 955 records · Page 53

Time course of myosin heavy chain transitions in neonatal rats: importance of innervation and thyroid state

During the postnatal period, rat limb muscles adapt to weight bearing via the replacement of embryonic (Emb) and neonatal (Neo) myosin heavy chains (MHCs) by the adult isoforms. Our aim was to characterize this transition in terms of the six MHC isoforms expressed in skeletal muscle and to determine the importance of innervation and thyroid hormone status on the attainment of the adult MHC phenotype. Neonatal rats were made hypothyroid via propylthiouracil (PTU) injection. In normal and PTU subgroups, leg muscles were unilaterally denervated at 15 days of age. The MHC profiles of plantaris (PLN) and soleus (Sol) muscles were determined at 7, 14, 23, and 30 days postpartum. At day 7, the Sol MHC profile was 55% type I, 30% Emb, and 10% Neo; in the PLN, the pattern was 60% Neo and 25% Emb. By day 30 the Sol and PLN had essentially attained an adult MHC profile in the controls. PTU augmented slow MHC expression in the Sol, whereas in the PLN it markedly repressed IIb MHC by retaining neonatal MHC expression. Denervation blunted the upregulation of IIb in the PLN and of Type I in the Sol and shifted the pattern to greater expression of IIa and IIx MHCs in both muscles. In contrast to previous observations, these findings collectively suggest that both an intact thyroid and innervation state are obligatory for the attainment of the adult MHC phenotype, particularly in fast-twitch muscles.

Non-NASA Center↗

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.

Non-NASA Center↗

Interstitial space and collagen alterations of the developing rat diaphragm

The effect of growth on the relative interstitial space [%total cross-sectional area (CSA)] and collagen content of the rat diaphragm muscle was examined at postnatal ages of 0, 7, 14, and 21 days as well as in adult males. The proportion of interstitial space relative to total muscle CSA was determined by computerized image analysis of lectin-stained cross sections of diaphragm muscle. To assess collagen content and extent of collagen maturation (i.e., cross-linking), high-pressure liquid chromatography analysis was used to measure hydroxyproline concentration and the nonreducible collagen cross-link hydroxylysylpyridinoline (HP), respectively. At birth, interstitial space accounted for approximately 47% of total diaphragm muscle CSA. During postnatal growth, the relative contribution of interstitial space decreased such that by adulthood the interstitial space accounted for approximately 18% of total muscle CSA. The change in relative interstitial space occurred without a concomitant change in hydroxyproline concentration. However, the concentration of HP markedly increased with age such that the adult diaphragm contained approximately 17 times more HP than at birth. These results indicate that during development the relative CSA occupied by interstitial space decreases as muscle fiber size increases. However, the reduction in relative interstitial space is not associated with a change in collagen concentration. Thus collagen density in the interstitial space may increase with age. It is possible that the observed changes in relative interstitial space and collagen influence the passive length-force properties of the diaphragm.

NASA Program Space Physiology and Countermeasures↗

Responses of motor and sensory neurons of rodents to spaceflight

Spinal motoneurons innervating skeletal muscles comprised predominantly of high oxidative fibers, i.e. slow oxidative and fast oxidative glycolytic, have higher oxidative enzyme activities than motoneurons innervating skeletal muscles comprised primarily of low oxidative fibers, i.e. fast glycolytic. These findings suggest that there is a close relationship between the oxidative phosphorylation capacity of a motoneuron and of the muscle fibers that it innervates. Since some skeletal muscles become faster and less oxidative after 4-14 days of spaceflight, it might be expected that oxidative enzyme activities in some motoneurons also may decrease after spaceflight. In addition, there is significant muscular atrophy after even short spaceflights and, therefore, it may be expected that some motoneurons associated with these muscles also would atrophy. In the present paper, we examine the issue of whether spaceflight induces changes in the oxidative enzyme activity and/or size of spinal motoneurons.

NASA Discipline Musculoskeletal↗

Nitric-oxide synthase is a mechanical signal transducer that modulates talin and vinculin expression

Mechanical stimuli can cause changes in muscle mass and structure which indicate that mechanisms exist for transducing mechanical stimuli into signals that influence gene expression. Myotendinous junctions show adaptations to modified muscle loading which suggest that these are transcriptionally distinct domains in muscle fibers that may experience local regulation of expression of structural proteins that are concentrated at these sites. Vinculin and talin are cytoskeletal proteins that are highly enriched at myotendinous junctions that we hypothesize to be subject to local transcriptional regulation. Our findings show that mechanical stimulation of muscle cells in vivo and in vitro causes an increase in the expression of vinculin and talin that is mediated by nitric oxide. Furthermore, nitric oxide-stimulated increases in vinculin and talin expression occur through a protein kinase G-dependent pathway and therefore differ from other mechanisms through which nitric oxide has been shown previously to modulate transcription. Analysis of vinculin mRNA distribution in mechanically stimulated muscle fibers shows that the mRNA is highly concentrated at myotendinous junctions, which supports the hypothesis that myotendinous junctions are distinct domains in which the expression of cytoskeletal proteins is modulated by mechanical stimuli through a nitric oxide and protein kinase G-dependent pathway.

NASA Discipline Musculoskeletal↗

Structural OFF/ON transitions of myosin in relaxed porcine myocardium predict calcium-activated force

Contraction in striated muscle is initiated by calcium binding to troponin complexes, but it is now understood that dynamic transition of myosin between resting, ordered OFF states on thick filaments and active, disordered ON states that can bind to thin filaments is critical in regulating muscle contractility. These structural OFF to ON transitions of myosin are widely assumed to correspond to transitions from the biochemically defined, energy-sparing, super-relaxed (SRX) state to the higher ATPase disordered-relaxed (DRX) state. Here, in this study, we examined the effect of 2’-deoxy-ATP (dATP), a naturally occurring energy substrate for myosin, on the structural OFF to ON transitions of myosin motors in porcine cardiac muscle thick filaments. Small-angle X-ray diffraction revealed that titrating dATP in relaxation solutions progressively moves the myosin heads from ordered OFF states on the thick filament backbone to disordered ON states closer to thin filaments. Importantly, we found that the structural OFF to ON transitions are not equivalent to the biochemically defined SRX to DRX transitions and that the dATP-induced structural OFF to ON transitions of myosin motors in relaxed muscle are strongly correlated with submaximal force augmentation by dATP. These results indicate that structural OFF to ON transitions of myosin in relaxed muscle can predict the level of force attained in calcium-activated cardiac muscle. Computational modeling and stiffness measurements suggest a final step in the OFF to ON transition may involve a subset of DRX myosins that form weakly bound cross-bridges prior to becoming active force-producing cross-bridges.

59 BASIC BIOLOGICAL SCIENCES↗

A study of the effects of prolonged simulated microgravity on the musculature of the lower extremities in man - An introduction

The experimental approach and the protocol are described for a study designed to investigate the effect of prolonged microgravity (simulated by a continuous exposure of subjects to 30-d-long 6-deg headdown tilt) on the musculature of the lower extremities in humans. The objectives of this study are as follows: (1) to determine changes in the functional characteristics of knee joint flexor and extensor muscle group; (2) to examine changes in the histochemical, biochemical, electron microscopic, and computed tomographic characteristics of skeletal muscle; (3) to determine if functional characteristics of skeletal muscle would be normalized four weeks after the bedrest; and (4) to compare these results to those of spaceflight. Percutaneous muscle biopsy, computed tomography, anthropometry, and in vivo muscle strength measurements are to be used to assess mechanical, structural, and metabolic characteristics of skeletal muscle.

Buchanan, Paul↗

Techniques for studying the effects of microgravity on model particle/cell systems

In an effort to learn more about the effects of a simulated low gravity environment on skeletal muscle, skeletal muscle cell cultures were grown within the lumen of XM-80 hollow fibers (i.d. = 0.5 mm) in a Clinostat rotating at 100 rpm. Cells were isolated from the thigh muscle tissue of 12 day embryos and were cultured for up to 14 days in the hollow fiber environment. Cells proliferated to confluency within several days, and fusion into multinucleated myotubes was then apparent. Fibers were stretched by a built-in spring mechanism to hold the fiber tightly at the center of rotation, and sections of the fiber were removed at 3, 7 and 14 days for electron microscopic analysis. When the Clinostat is rotated in the horizontal position, the gravity vector approaches zero and the cells are in an environment that simulates microgravity. Control experiments consist of one fiber rotated in the vertical position in the clinostat and another fiber that is held in a horizontal configuration in a comparable sized tube that is not rotated at all. Examination of skeletal muscle cells by electron microscopy revealed that myoblast fusion and myofibril accumulation were extensive. Two general conclusions were apparent. First, muscle cells undergo the normal progression of proliferation, fusion, and myofibril assembly in the presence of simulated microgravity for the first week in culture. After 14 days, however, many muscle fibers undergo degeneration such that myofibrillar structures are not extensive or well organized. Second, although no major abnormalities in myofibril assembly were detected in Clinostat-rotated cultures in comparison to controls that were not rotated in a Clinostat, the myofibrils in non-rotated controls tend to be more highly organized than those in either horizontally or vertically rotated Clinostat samples.

Young, Ronald B.↗

Alterations in glucose and protein metabolism in animals subjected to simulated microgravity

Reduction of physical activity due to disease or environmental restraints, such as total bed rest or exposure to spaceflight, leads to atrophy of skeletal muscle and is frequently accompanied by alterations in food intake and the concentration of metabolic regulatory hormones such as insulin. Hindlimb suspension of laboratory rats, as a model for microgravity, also shows marked atrophy of gravity-dependent muscles along with a reduced gain in body weight. Suspended rats exhibit enhanced sensitivity to insulin-induced glucose uptake when compared with normal control rats and resistance to insulin action when compared with control rats matched similarly for reduced body weight gain. These changes are accompanied by decreased insulin binding and tyrosine kinase activity in soleus but not plantaris muscle, unchanged glucose uptake by perfused hindlimb and decreased sensitivity but not responsiveness to insulin-induced suppression of net proteolysis in hindlimb skeletal muscle. These findings suggest that loss of insulin sensitivity during muscle atrophy is associated with decreased insulin binding and tyrosine kinase activity in atrophied soleus muscle along with decreased sensitivity to the effects of insulin on suppressing net protein breakdown but not on enhancing glucose uptake by perfused hindlimb.

Mondon, C. E.↗

Neuromuscular aspects in development of exercise countermeasures

The paper reviews data obtained at the Human Life Sciences Research Program at the Kennedy Space Center on the structural and functional characteristics of the human muscle and on the effects of simulated microgravity (head-down bed rest) and exercise on the muscle. It is shown that prolonged exposure to microgravity induced ultrastructural abnormalities and the atrophy of muscle and impaired muscle functioning (especially upon return to terrestrial gravity) and that ground-based models can be effectively used to study these changes. Results indicate that the incorporation of resistive exercise with a strong emphasis on the use of eccentric muscle action, in addition to concentric action, may provide a very effective countermeasure against muscle atrophy and dysfunction caused by long-duration exposures to microgravity.

Convertino, Victor A.↗

Intramuscular pressure and electromyography as indexes of force during isokinetic exercise

A direct method for measuring force production of specific muscles during dynamic exercise is presently unavailable. Previous studies indicate that both intramuscular pressure (IMP) and electromyography (EMG) correlate linearly with muscle contraction force during isometric exercise. The objective of this study was to compare IMP and EMG as linear assessors of muscle contraction force during dynamic exercise. IMP and surface EMG activity were recorded during concentric and eccentric isokinetic plantarflexion and dorsiflexion of the ankle joint from the tibialis anterior (TA) and soleus (SOL) muscles of nine male volunteers. Ankle torque was measured using a dynamometer, and IMP was measured via catheterization. IMP exhibited better linear correlation than EMG with ankle joint torque during concentric contractions of the SOL and the TA, as well as during eccentric contractions. IMP provides a better index of muscle contraction force than EMG during concentric and eccentric exercise through the entire range of torque. IMP reflects intrinsic mechanical properties of individual muscles, such as length-tension relationships, which EMG is unable to assess.

Aratow, M.↗

Specification list and function structure for a full-body dynamometer to be used aboard Space Station Freedom

NASA has a need for a machine which can be used as an exercise device and as an instrument to measure an astronaut's muscle performance. The purpose of the exercise device is to work various muscle groups of the astronaut to prevent muscle atrophy, the loss of muscle strength and mass from prolonged exposure to a microgravity environment. The measurement part of the machine will be used to collect data on the strength and power of the astronaut's muscle groups to be used in studies examining the effects of prolonged space inhabitation. The principle device used in this machine to both exercise and measure muscle performance is the dynamometer. The dynamometer converts electrical energy to mechanical energy and mechanical energy to electrical energy or signals. The task of the designer will be to incorporate a dynamometer into a device which can meet all of the needs discussed above. This memorandum has two sections which clarify the design task of producing a full-body dynamometer. The first section is a specification list. The specification list provides the requirements that the designer must meet in his/her design. The second part is a function structure. The function structure shows graphically the flow of material, energy, and information through the machine. These two items will be used by the designer in the design process for the full-body dynamometer.

Mcmahan, Robert↗

Intramuscular pressure: A better tool than EMG to optimize exercise for long-duration space flight

A serious problem experienced by astronauts during long-duration space flight is muscle atrophy. In order to develop countermeasures for this problem, a simple method for monitoring in vivo function of specific muscles is needed. Previous studies document that both intramuscular pressure (IMP) and electromyography (EMG) provide quantitative indices of muscle contraction force during isometric exercise. However, at present there are no data available concerning the usefulness of IMP versus EMG during dynamic exercise. Methods: IMP (Myopress catheter) and surface EMG activity were measured continuously and simultaneously in the tibalis anterior (TA) and soleus (SOL) muscles of 9 normal male volunteers (28-54 years). These parameters were recorded during both concentric and eccentric exercises which consisted of plantarflexon and dorsiflexon of the ankle joint. A Lido Active Isokinetic Dynamometer concurrently recorded ankle joint torque and position. Results: Intramuscular pressure correlated linearly with contraction force for both SOL (r exp 2 = 0.037) and TA (R exp 2 = 0.716 and r exp 2 = 0.802, respectively). During eccentric exercises, SOL and TA IMP also correlated linearly with contraction force (r(exp 2) = 0.883 and r(exp 2) = 0.904 respectively), but SOL and TA EMG correlated poorly with force (r(exp 2) = 0.489 and r(exp 2) = 0.702 respectively). Conclusion: IMP measurement provides a better index of muscle contraction force than EMG during concentric and eccentric exercise. IMP reflects intrinsic mechanical properties of individual muscles, such as length tension relationships. Although invasive, IMP provides a more powerful tool and EMG for developing exercise hardware and protocols for astronauts exposed to long-duration space flight.

Hargens, A. R.↗

Intramuscular Pressure Measurement During Locomotion in Humans

To assess the usefulness of intramuscular pressure (IMP) measurement for studying muscle function during gait, IMP was recorded in the soleus and tibialis anterior muscles of ten volunteers during, treadmill walking, and running using transducer-tipped catheters. Soleus IMP exhibited single peaks during late-stance phase of walking (181 +/- 69 mmHg, mean +/- S.E.) and running (269 +/- 95 mmHg). Tibialis anterior IMP showed a biphasic response, with the largest peak (90 +/- 15 mmHg during walking and 151 +/- 25 mmHg during running) occurring shortly after heel strike. IMP magnitude increased with gait speed in both muscles. Linear regression of soleus IMP against ankle joint torque obtained by a dynamometer in two subjects produced linear relationships (r = 0.97). Application of these relationships to IMP data yielded estimated peak soleus moment contributions of 0.95-165 Nm/Kg during walking, and 1.43-2.70 Nm/Kg during running. IMP results from local muscle tissue deformations caused by muscle force development and thus, provides a direct, practical index of muscle function during locomotion in humans.

Ballard, Ricard E.↗

The Hematopoietic Stem Cell Therapy for Exploration of Space

Astronauts experience severe/invasive disorders caused by space environments. These include hematological/cardiac abnormalities, bone and muscle losses, immunodeficiency, neurological disorders and cancer. While the cause of these symptoms are not yet fully delineated, one possible explanation could be the inhibition of hematopoietic stem cell (HSC) growth and hematopoiesis in space. HSCs differentiate into all types of blood cells, and growing evidence indicates that the HSCs also have the ability to transdifferentiate to various tissues, including muscle, skin, liver, neuronal cells and possibly bone. Therefore, a hypothesis was advanced in this laboratory that the hematopoietic stem cell-based therapy, herein called the hematopoietic stem cell therapy (HSCT), could mitigate some of the disorders described above. Due to the magnitude of this project our laboratory has subdivided it into 3 sections: a) HSCT for space anemia; b) HSCT for muscle and bone losses; and c) HSCT for immunodeficiency. Toward developing the HSCT protocol for space anemia, the HSC transplantation procedure was established using a mouse model of beta thalassemia. In addition, the NASA Rotating Wall Vessel (RWV) culture system was used to grow HSCs in space condition. To investigate the HSCT for muscle loss and bone loss, donor HSCs were genetically marked either by transfecting the beta-galactosidase-containing plasmid, pCMV.SPORT-beta-gal or by preparing from b-galactosidase transgenic mice. The transdifferentiation of HSCs to muscle is traced by the reporter gene expression in the hindlimb suspended mice with some positive outcome, as studied by the X-gal staining procedure. The possible structural contribution of HSCs against muscle loss is being investigated histochemically.

Roach, Allana Nicole↗

Role of microtubules in the contractile dysfunction of hypertrophied myocardium

OBJECTIVES: We sought to determine whether the ameliorative effects of microtubule depolymerization on cellular contractile dysfunction in pressure overload cardiac hypertrophy apply at the tissue level. BACKGROUND: A selective and persistent increase in microtubule density causes decreased contractile function of cardiocytes from cats with hypertrophy produced by chronic right ventricular (RV) pressure overloading. Microtubule depolymerization by colchicine normalizes contractility in these isolated cardiocytes. However, whether these changes in cellular function might contribute to changes in function at the more highly integrated and complex cardiac tissue level was unknown. METHODS: Accordingly, RV papillary muscles were isolated from 25 cats with RV pressure overload hypertrophy induced by pulmonary artery banding (PAB) for 4 weeks and 25 control cats. Contractile state was measured using physiologically sequenced contractions before and 90 min after treatment with 10(-5) mol/liter colchicine. RESULTS: The PAB significantly increased RV systolic pressure and the RV weight/body weight ratio in PAB; it significantly decreased developed tension from 59+/-3 mN/mm2 in control to 25+/-4 mN/mm2 in PAB, shortening extent from 0.21+/-0.01 muscle lengths (ML) in control to 0.12+/-0.01 ML in PAB, and shortening rate from 1.12+/-0.07 ML/s in control to 0.55+/-0.03 ML/s in PAB. Indirect immunofluorescence confocal microscopy showed that PAB muscles had a selective increase in microtubule density and that colchicine caused complete microtubule depolymerization in both control and PAB papillary muscles. Microtubule depolymerization normalized myocardial contractility in papillary muscles of PAB cats but did not alter contractility in control muscles. CONCLUSIONS: Excess microtubule density, therefore, is equally important to both cellular and to myocardial contractile dysfunction caused by chronic, severe pressure-overload cardiac hypertrophy.

NASA Discipline Cardiopulmonary↗

Glucose transporters and maximal transport are increased in endurance-trained rat soleus

Voluntary wheel running induces an increase in the concentration of the regulatable glucose transporter (GLUT4) in rat plantaris muscle but not in soleus muscle (K. J. Rodnick, J. O. Holloszy, C. E. Mondon, and D. E. James. Diabetes 39: 1425-1429, 1990). Wheel running also causes hypertrophy of the soleus in rats. This study was undertaken to ascertain whether endurance training that induces enzymatic adaptations but no hypertrophy results in an increase in the concentration of GLUT4 protein in rat soleus (slow-twitch red) muscle and, if it does, to determine whether there is a concomitant increase in maximal glucose transport activity. Female rats were trained by treadmill running at 25 m/min up a 15% grade, 90 min/day, 6 days/wk for 3 wk. This training program induced increases of 52% in citrate synthase activity, 66% in hexokinase activity, and 47% in immunoreactive GLUT4 protein concentration in soleus muscles without causing hypertrophy. Glucose transport activity stimulated maximally with insulin plus contractile activity was increased to roughly the same extent (44%) as GLUT4 protein content in soleus muscle by the treadmill exercise training. In a second set of experiments, we examined whether a swim-training program increases glucose transport activity in the soleus in the presence of a maximally effective concentration of insulin. The swimming program induced a 44% increase in immunoreactive GLUT4 protein concentration. Glucose transport activity maximally stimulated with insulin was 62% greater in soleus muscle of the swimmers than in untrained controls. Training did not alter the basal rate of 2-deoxyglucose uptake.(ABSTRACT TRUNCATED AT 250 WORDS).

Non-NASA Center↗

Neuromuscular Adaptations to Reduced Use

This viewgraph presentation reviews the studies done to reduce neuromuscular strength loss during unilateral lower limb suspension (ULLS). Since there are animals that undergo fairly long periods of muscular disuse without any or minimal muscular atrophy, there is an answer to that might be applicable to human in situations that require no muscular use to diminish the effects of muscular atrophy. Three sets of ULLS studies were reviewed indicated that muscle strength decreased more than the muscle mass. The study reviewed exercise countermeasures to combat the atrophy, including: ischemia maintained during Compound muscle action potential (CMAP), ischemia and low load exercise, Japanese kaatsu, and the potential for rehabilitation or situations where heavy loading is undesirable. Two forms of countermeasures to unloading have been successful, (1) high-load resistance training has maintained muscle mass and strength, and low load resistance training with blood flow restriction (LL(sub BFR)). The LL(sub BFR) has been shown to increase muscle mass and strength. There has been significant interest in Tourniquet training. An increase in Growth Hormone(GH) has been noted for LL(sub BFR) exercise. An experimental study with 16 subjects 8 of whom performed ULLS, and 8 of whom performed ULLS and LL(sub BFR) exercise three times per week during the ULLS. Charts show the results of the two groups, showing that performing LL(sub BFR) exercise during 30 days of ULLS can maintain muscle size and strength and even improve muscular endurance.

Ploutz-Snyder, Lori↗