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Effects of zero gravity on myofibril content and isomyosin distribution in rodent skeletal muscle.

The purpose of this experiment was to investigate the effects of 12.5 days of zero gravity (0 g) exposure (Cosmos 1887 Biosputnik) on the enzymatic properties, protein content, and isomyosin distribution of the myofibril fraction of the slow-twitch vastus intermedius (VI) and the fast-twitch vastus lateralis (VL) muscles of adult male rats. Measurements were obtained on three experimental groups (n = 5 each group) designated as flight group (FG), vivarium control (VC), and synchronous control (SC). Body weight of the FG was significantly lower than that of the two control groups (P less than 0.05). Compared with the two control groups, VI weight was lower by 23% (P less than 0.10), whereas no such pattern was apparent for the VL muscle. Myofibril yields (mg protein/g muscle) in the VI were 35% lower in the FG than in controls (P less than 0.05), whereas no such pattern was apparent for the VL muscle. When myofibril yields were expressed on a muscle basis (mg/g x muscle weight), the loss of myofibril protein was more exaggerated and suggests that myofibril protein degradation is an early event in the muscle atrophy response to 0 g. Analysis of myosin isoforms indicated that slow myosin (Sm) was the primary isoform lost in the calculated degradation of total myosin. No evidence of loss of the fast isomyosins was apparent for either muscle following spaceflight. Myofibril ATPase activity of the VI was increased in the FG compared with controls, which is consistent with the observation of preferential Sm degradation. These data suggest that muscles containing a high percentage of slow-twitch fibers undergo greater degrees of myofibril protein degradation than muscles containing predominantly fast-twitch fibers in response to a relatively short period of 0 g exposure, and the primary target appears to be the Sm molecule.

Adenosine Triphosphatases↗

Substrate oxidation capacity in rodent skeletal muscle: effects of exposure to zero gravity.

A study was conducted, as part of the integrated National Aeronautics and Space Administration Space Life Sciences 1 mission flown in June of 1991, to ascertain the effects of 9 days of exposure to zero gravity on the capacity of rodent skeletal muscle fiber types to oxidize either [14C]pyruvate or [14C]palmitate under state 3 metabolic conditions, i.e., nonlimiting amounts of substrate and cofactors. In addition, activity levels of marker enzymes of the tricarboxylic acid cycle, malate shuttle, and beta-oxidation were measured. Results showed that significant differences in muscle weight occurred in both the predominantly slow vastus intermedius and predominantly fast vastus lateralis of flight vs. control groups (P < 0.05). Total protein content of the muscle samples was similar between groups. Both pyruvate oxidation capacity and the marker oxidative enzymes were not altered in the flight relative to control animals. However, the capacity to oxidize long-chain fatty acids was significantly reduced by 37% in both the high- and low-oxidative regions of the vastus muscle (P < 0.05). Although these findings of a selective reduction in fatty acid oxidation capacity in response to spaceflight are surprising, they are consistent with previous findings showing 1) an increased capacity to take up glucose and upregulate glucose transporter proteins and 2) a marked accumulation of triglycerides in the skeletal muscles of rats subjected to states of unloading. Thus, skeletal muscle of animals exposed to non-weight-bearing environments undergo subcellular transformations that may preferentially bias energy utilization to carbohydrates.

3-Hydroxyacyl CoA Dehydrogenases↗

Effect of spaceflight on human protein metabolism.

Nitrogen balance and the whole body protein synthesis rate were measured before, during, and after a 9.5-day spaceflight mission on the space shuttle Columbia. Protein synthesis was measured by the single-pulse [15N]glycine method. Determinations were made 56, 26, and 18 days preflight, on flight days 2 and 8, and on days 0, 6, 14, and 45 postflight. We conclude that nitrogen balance was decreased during spaceflight. The decrease in nitrogen balance was greatest on the 1st day when food intake was reduced and again toward the end of the mission. An approximately 30% increase in protein synthesis above the preflight baseline was found for flight day 8 for all 6 subjects (P < 0.05), indicating that the astronauts showed a stress response to spaceflight.

Body Mass Index↗

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↗

Comparative morphometry of fibers and capillaries in soleus following weightlessness (SL-3) and suspension.

This work is a continuation of efforts to assess responses of rat skeletal muscle to weightlessness and earthside laboratory experiments with unloading of hind limbs. The soleus is a slow twitch, load bearing (antigravity) muscle. Both exposure to weightlessness (W) and to the hypokinesia/hypodynamia of whole body suspension (WBS) results in soleus atrophy. Cross sectional areas of both slow and fast twitch fibers decrease during 7 days of W, and 7 or 14 days of WBS. Density and area changes tended to reverse to control levels during 7 days of recovery (R) following WBS. Capillary density was increased with 7 days of W, and 7 or 14 days of WBS. During 7 days of R the capillary density returned toward control levels. In summary, the reduction in fiber cross sectional areas and increase in fiber and capillary densities support the hypothesis that in both forms of disuse, i.e., W and WBS, there is a loss in soleus muscle cell mass and not in fiber numbers.

Animals↗

Human mononuclear cell function after 4 degrees C storage during 1-G and microgravity conditions of spaceflight.

Future space missions of long duration may require that autologous leukocytes be stored in flight for infusion to restore normal immune competence in crewmembers. Peripheral blood mononuclear cells (PBMNCs), as leukocyte concentrates in autologous plasma and 2% dextrose, were stored in the microgravity conditions provided by the U.S. Space Shuttle Columbia (Mission 61-C). Activity of PBMNC after space flight was compared with that from a series of preflight ground control experiments, which demonstrated in culture a progressive daily loss in mitogen-stimulated protein synthesis at 24 h and thymidine uptake at 72 h after storage for 7 d at 4 degrees C. Post-storage viabilities were at least 90% as determined by trypan dye exclusion. A progressive reduction in the percentage of PBMNC expressing cell-surface phenotype markers, which was similar for monocytes, B cells, and T-cell subsets, also occurred after storage. The ability of PBMNC, stored for 8 d in Columbia's middeck, to become activated and proliferate in vitro was similar to that of cells that remained in identical flight lockers on the ground as 1-G controls, thus indicating that PBMNCs were not adversely affected by storage under microgravity conditions.

Benzoates↗

Histomorphometric and electron microscopic analyses of tibial epiphyseal plates from Cosmos 1887 rats.

Previous studies have shown that the changes seen in the bones of growing rats exposed to microgravity are due in part to changes that occur in the growth plate during spaceflight. In this study, growth plates of rats flown aboard Cosmos 1887 (12.5-day flight plus 53.5-h recovery at 1 g) were analyzed using light and electron microscopy and computerized planimetry. The proliferative zone of flight animals was found to be significantly (P less than or equal to 0.01) larger than that of controls, while the reserve and hypertrophic/calcification zones were significantly reduced. Flight animals also had more cells per column in the proliferative zone than did controls and less in the hypertrophic/calcification region. The total number of cells, however, was significantly greater in flight animals. No difference was found in perimeter or in shape factor, but area was significantly less in flight animals. Electron microscopy showed that collagen fibrils in flight animals were wider than in controls. Since the time required for a cell to cycle through the growth plate is 2-3 days at 1 g, the results reported here represent both the effects of exposure to microgravity and the initial stages of recovery from that exposure.

Animals↗

Altered carbohydrate, lipid, and xenobiotic metabolism by liver from rats flown on Cosmos 1887.

To determine the possible biochemical effects of prolonged weightlessness on liver function, samples of liver from rats that had flown aboard Cosmos 1887 were analyzed for protein, glycogen, and lipids as well as the activities of a number of key enzymes involved in metabolism of these compounds and xenobiotics. Among the parameters measured, the major differences were elevations in the glycogen content and hydroxymethylglutaryl-CoA (HMG-CoA) reductase activities for the rats flown on Cosmos 1887 and decreases in the amount of microsomal cytochrome P-450 and the activities of aniline hydroxylase and ethylmorphine N-demethylase, cytochrome P-450-dependent enzymes. These results support the earlier finding of differences in these parameters and suggest that altered hepatic function could be important during spaceflight and/or the postflight recovery period.

Aniline Hydroxylase↗

Ecdysteroids affect in vivo protein metabolism of the flight muscle of the tobacco hornworm (Manduca sexta).

Ecdysteroid growth promotion of the dorsolongitudinal flight muscle of Manduca sexta was studied by measuring in vivo protein metabolism using both "flooding-dose" and "non-carrier" techniques. These procedures differ in that the former method includes injection of non-labelled phenylalanine (30 micromoles/insect) together with the [3H]amino acid. Injected radioactivity plateaued in the haemolymph within 7 min. With the flooding-dose method, haemolymph and intramuscular specific radioactivities were similar between 15 min and 2 h. Incorporation of [3H]phenylalanine into muscle protein was linear with either method between 30 and 120 min. Fractional rates (%/12 h) of synthesis with the flooding-dose technique were best measured after 1 h because of the initial delay in radioactivity equilibration. Estimation of body phenylalanine turnover with the non-carrier method showed 24-53%/h which was negligible with the flooding-dose method. Since the two methods yielded similar rates of protein synthesis, the large injection of non-labelled amino acid did not alter the rate of synthesis. Because the flooding-dose technique requires only a single time point measurement, it is the preferred method. The decline and eventual cessation of flight-muscle growth was mostly a consequence of declining protein synthesis though degradation increased between 76-86 h before eclosion and was relatively rapid. This decline in muscle growth could be prevented by treating pupae with 20-hydroxyecdysone (10 micrograms/insect). Protein accretion was promoted by a decline of up to 80% in protein breakdown, which was offset in part by a concurrent though much smaller decrease in protein synthesis. Therefore, ecdysteroids may increase flight-muscle growth by inhibiting proteolysis.

Animals↗

The prevention of adverse physiological change in Space Station crewmembers.

Various physiological countermeasures, consisting primarily of isotonic and isometric exercises but also including prescribed nutrient intake, have been used in all manned spaceflights exceeding about one month in duration. So consistent has been this practice that the effects of weightlessness on the human, unconfounded by the use of countermeasures, are difficult to discern. Equally elusive, in the absence of control studies conducted in weightlessness, is an accurate assessment of the efficacy of the countermeasures themselves. Changes in body composition occurring during and following flights from Gemini through Shuttle, when compared with changes during and following bedrest, demonstrate certain mitigating effects that may be attributable to countermeasures and which provide some rationale for the choice of countermeasures in the Space Station.

Adaptation, Physiological↗

Pulmonary function in microgravity: Spacelab 4 and beyond.

This paper refers principally to the composition gradient of gases within the lung in various conditions of gravity, as revealed by exhaled breath. A rapid gas analyzer-based system has been developed for tests in Spacelab 4. The test sequence and expected results are presented.

Breath Tests↗

Properties of electrophoretic fractions of human embryonic kidney cells separated on Space Shuttle flight STS-8.

Suspensions of cultured primary human embryonic kidney cells were subjected to continuous flow electrophoresis on Space Shuttle flight STS-8. The objectives of the experiments were to obtain electrophoretically separated fractions of the original cell populations and to test these fractions for the amount and kind of urokinase (a kidney plasminogen activator that is used medically for digesting blood clots), the morphologies of cells in the individual fractions, and their cellular electrophoretic mobilities after separation and subsequent proliferation. Individual fractions were successfully cultured after return from orbit, and they were found to differ substantially from one another and from the starting sample with respect to all of these properties.

Cell Line↗

Canadian medical experiments on Shuttle flight 41-G.

During the 41-G mission, two payload specialist astronauts took part in six Canadian medical experiments designed to measure how the human nervous system adapts to weightlessness, and how this might contribute to space motion sickness. Similar tests conducted pre-flight provided base-line data, and post-flight experiments examined re-adaptation to the ground. No changes were detected in the vestibulo-ocular reflex during this 8-day mission. Pronounced proprioceptive illusions were experienced, especially immediately post-flight. Tactile acuity was normal in the fingers and toes, but the ability to judge limb position was degraded. Estimates of the locations of familiar targets were grossly distorted in the absence of vision. There were no differences in taste thresholds or olfaction. Despite pre-flight tests showing unusual susceptibility to motion sickness, the Canadian payload specialist turned out to be less susceptible than normal on-orbit. Re-adaptation to the normal gravity environment occurred within the first day after landing.

Adaptation, Physiological↗

Vestibular ataxia following shuttle flights: effects of microgravity on otolith-mediated sensorimotor control of posture.

Orbital spaceflight exposes astronauts to an environment in which gravity is reduced to negligible magnitudes of 10(-3) to 10(-6) G. Upon insertion into earth orbit, the abrupt loss of the constant linear acceleration provided by gravity removes the otolith stimulus for vestibular sensation of vertical orientation constantly present on Earth. Since the central nervous system (CNS) assesses spatial orientation by simultaneously interpreting sensory inputs from the vestibular, visual, and proprioceptive systems, loss of the otolith-mediated vertical reference input results in an incorrect estimation of spatial orientation, which, in turn, causes a degradation in movement control. Over time, however, the CNS adapts to the loss of gravitational signals. Upon return to Earth, the vertical reference provided by gravitational stimulation of the otolith organ reappears. As a result, a period of CNS readaptation must occur upon return to terrestrial environment. Among the physiological changes observed during the postflight CNS readaptation period is a disruption of postural equilibrium control. Using a dynamic posturography system (modified NeuroCom EquiTest), 16 astronauts were tested at 60, 30, and 10 days preflight and retested at 1 to 5 hours, and 8 days postflight. All astronauts tested demonstrated decreased postural stability immediately upon return to Earth. The most dramatic increases in postural sway occurred during those sensory conditions in which both the visual and proprioceptive feedback information used for postural control were altered by the dynamic posturography system, requiring reliance primarily upon vestibular function for control of upright stance. Less marked but statistically significant increases in sway were observed under those conditions in which visual and foot support surface inputs alone were altered.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Spaceflight on STS-48 and earth-based unweighting produce similar effects on skeletal muscle of young rats.

Our knowledge of the effects of unweighting on skeletal muscle of juvenile rapidly growing rats has been obtained entirely by using hindlimb-suspension models. No spaceflight data on juvenile animals are available to validate these models of simulated weightlessness. Therefore, eight 26-day-old female Sprague-Dawley albino rats were exposed to 5.4 days of weightlessness aboard the space shuttle Discovery (mission STS-48, September 1991). An asynchronous ground control experiment mimicked the flight cage condition, ambient shuttle temperatures, and mission duration for a second group of rats. A third group of animals underwent hindlimb suspension for 5.4 days at ambient temperatures. Although all groups consumed food at a similar rate, flight animals gained a greater percentage of body mass per day (P < 0.05). Mass and protein data showed weight-bearing hindlimb muscles were most affected, with atrophy of the soleus and reduced growth of the plantaris and gastrocnemius in both the flight and suspended animals. In contrast, the non-weight-bearing extensor digitorum longus and tibialis anterior muscles grew normally. Earlier suspension studies showed that the soleus develops an increased sensitivity to insulin during unweighting atrophy, particularly for the uptake of 2-[1,2-3H]deoxyglucose. Therefore, this characteristic was studied in isolated muscles within 2 h after cessation of spaceflight or suspension. Insulin increased uptake 2.5- and 2.7-fold in soleus of flight and suspended animals, respectively, whereas it increased only 1.6-fold in control animals. In contrast, the effect of insulin was similar among the three groups for the extensor digitorum longus, which provides a control for potential systemic differences in the animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗