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Behaviour of adult hamsters subjected to hypergravity.

We studied vestibular function in 20 adult hamsters (3 months old) subjected to either prolonged hypergravity (n = 10) or normal gravity (n = 10) for 2 months. Locomotion and swimming of the hypergravity hamsters under light conditions were normal. Equilibrium maintenance was severely disturbed; only 6 of 10 hypergravity hamsters managed to walk on the small tube after 2 months, whereas all 10 controls were able to walk on the tube. The air-righting reflex was severely disturbed; the hypergravity hamsters made 30% correct air-righting responses, while the control hamsters made 88% correct responses. Finally, 5 of 8 hypergravity hamsters had to be saved from drowning when swimming in total darkness. Histological examination of the utricular otoconial layers afterwards, using energy dispersive X-ray element (EDAX) analysis and scanning electron microscopy, did not reveal any differences in calcium content, shape and size distribution of the otoconia between hypergravity hamsters and controls. We suggest that adult hamsters adapt to hypergravity, leading to problems in normal functioning when tested in 1 G, especially in tasks in which sensory input of the vestibular system is important for spatial orientation. These disturbances were more severe in adult hamsters than in young ones, tested in previous experiments. Therefore, we assume that age is a factor for adaptation to altered gravity conditions.

Adaptation, Physiological↗

Effects of hypergravity on the elongation growth in radish and cucumber hypocotyls.

The elongation growth of the hypocotyls of radish and cucumber seedlings was examined under hypergravity in a newly developed centrifuge (Kasahara et al. 1995). The effects of hypergravity on elongation growth differed between the two species. The rate of elongation of radish hypocotyls was reduced under basipetal hypergravity (H+2O g) but not under acropetal hypergravity (H-13 g), as compared to growth under the control conditions (C+1 g and C-1 g). In cucumber hypocotyls, elongation growth was inhibited not only by basipetal but also by acropetal hypergravity. Under these conditions, the reduction in the elongation growth of both radish and cucumber hypocotyls was accompanied by an increase in their thickness. Although no distinct differences in relative composition of neutral sugars were found, the amounts of cell-wall components (pectic substances, hemicelluloses and cellulose) per unit length of hypocotyls were increased by exposure to hypergravity.

Brassicaceae↗

Increased molecular mass of hemicellulosic polysaccharides is involved in growth inhibition of maize coleoptiles and mesocotyls under hypergravity conditions.

Elongation growth of dark grown maize (Zea mays L cv. Cross Bantam T51) coleoptiles and mesocotyls was suppressed by hypergravity at 30 g and above. Acceleration at 300 g significantly decreased the mechanical extensibility of cell walls of both organs. Hypergravity increased the amounts of hemicellulose and cellulose per unit length in mesocotyl walls, but not in coleoptile walls. The weight average molecular masses of hemicellulosic polysaccharides were also increased by hypergravity in both organs. On the other hand, the activities of beta-glucanases extracted from coleoptile and mesocotyl cell walls were decreased by hypergravity. These results suggest that the decreased activities of beta-glucanases by hypergravity cause an increase in the molecular mass of hemicellulosic polysaccharides of both organs. The upshift of molecular mass of hemicellulosic polysaccharides as well as the thickening of cell walls under hypergravity conditions seems to be involved in making the cell wall mechanically rigid, thereby inhibiting elongation growth of maize coleoptiles and mesocotyls.

Acceleration↗

Hypergravity induces reorientation of cortical microtubules and modifies growth anisotropy in azuki bean epicotyls.

We examined the changes in the orientation of cortical microtubules during the hypergravity-induced modification of growth anisotropy (inhibition of elongation growth and promotion of lateral growth) in azuki bean (Vigna angularis Ohwi et Ohashi) epicotyls. The percentage of cells with transverse microtubules was decreased, while that with longitudinal microtubules was increased, in proportion to the logarithm of the magnitude of gravity. The percentage of cells with longitudinal microtubules showed an increase within 0.5 h of transfer of the 1g-grown seedlings to a 300g-hypergravity condition. Lanthanum and gadolinium, blockers of calcium channels, nullified the modification of growth anisotropy and reorientation of microtubules by hypergravity. Horizontal and acropetal hypergravity modified growth anisotropy and reorientation of microtubules, as did basipetal hypergravity, and these changes were not seen in the presence of lanthanum or gadolinium. These results suggest that hypergravity changes activities of lanthanum- and gadolinium-sensitive calcium channels independently of its direction, which may lead to reorientation of cortical microtubules and modification of growth anisotropy in azuki bean epicotyls.

Fabaceae↗

CNS development under altered gravity: cerebellar glial and neuronal protein expression in rat neonates exposed to hypergravity.

The future of space exploration depends on a solid understanding of the developmental process under microgravity, specifically in relation to the central nervous system (CNS). We have previously employed a hypergravity paradigm to assess the impact of altered gravity on the developing rat cerebellum. The present study addresses the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of selected glial and neuronal cerebellar proteins in rat neonates exposed to hypergravity (1.5 G) from embryonic day (E)11 to postnatal day (P)6 or P9 (the time of maximal cerebellar changes) comparing them against their expression in rat neonates developing under normal gravity. Proteins were analyzed by quantitative Western blots of cerebellar homogenates; RNA analysis was performed in the same samples using quantitative PCR. Densitometric analysis of Western blots suggested a reduction in glial (glial acidic protein, GFAP) and neuronal (neuronal cell adhesion molecule, NCAM-L1, synaptophysin) proteins, but the changes in individual cerebellar proteins in hypergravity-exposed neonates appeared both age- and gender-specific. RNA analysis suggested a reduction in GFAP and synaptophysin mRNAs on P6. These data suggest that exposure to hypergravity may interfere with the expression of selected cerebellar proteins. These changes in protein expression may be involved in mediating the effect of hypergravity on the developing rat cerebellum.

Animals↗

Mechanoreceptors rather than sedimentable amyloplasts perceive the gravity signal in hypergravity-induced inhibition of root growth in azuki bean.

Elongation of primary roots of azuki bean (Vigna angularis Ohwi et Ohashi) was suppressed under hypergravity conditions produced by centrifugation, such that the growth rate decreased in proportion to the logarithm of the magnitude of the gravity. The removal of the root cap did not influence the hypergravity-induced inhibition of root growth, although it completely inhibited the gravitropic root curvature. Lanthanum and gadolinium, blockers of mechanoreceptors, nullified the growth-inhibitory effect of hypergravity. These results suggest that the gravity signal for the hypergravity-induced inhibition of root growth is perceived independently from that of gravitropism, which involves amyloplasts as statoliths. Horizontal and basipetal hypergravity suppressed root growth as did acropetal hypergravity, all of which were nullified by the presence of lanthanum or gadolinium. These findings suggest that mechanoreceptors on the plasma membrane perceive the gravity signal independently of the direction of the stimuli and roots may utilise it to regulate their growth rate.

Centrifugation↗

Effects of hypergravity on growth and cell wall properties of cress hypocotyls.

Elongation growth of etiolated hypocotyls of cress (Lepidium sativum L.) was suppressed when they were exposed to basipetal hypergravity at 35 x g and above. Acceleration at 135 x g caused a decrease in the mechanical extensibility and an increase in the minimum stress-relaxation time of the cell wall. Such changes in the mechanical properties of the cell wall were prominent in the lower regions of hypocotyls. The amounts of cell wall polysaccharides per unit length of hypocotyls increased under the hypergravity condition and, in particular, the increase in the amount of cellulose in the lower regions was conspicuous. Hypergravity did not influence the neutral sugar composition of either the pectin or the hemicellulose fraction. The amount of lignin was also increased by hypergravity treatment, although the level was low. The data suggest that hypergravity modifies the metabolism of cell wall components and thus makes the cell wall thick and rigid, thereby inhibiting elongation growth of cress hypocotyls. These changes may contribute to the plants' ability to sustain their structures against hypergravity.

Acceleration↗

Bidirectional effects of hypergravity on the cell growth and differentiated functions of osteoblast-like ROS17/2.8 cells.

A low level of hypergravity (1.5-2.0 G) stimulated the proliferation of ROS17/2.8 cells, whereas it inhibited the differentiated functions of alkaline phosphatase activity and osteocalcin synthesis. These results were just the opposite of our results obtained when the cells were exposed to a high level of hypergravity (40-80 G): inhibition of cell growth and the stimulation of the differentiated functions. The direction of change in the cAMP contents of the cells was also reversed, with a low level of hypergravity causing a decrease in the cAMP content and a high level of hypergravity an increase in it. Therefore, bidirectional effects of hypergravity on the growth and differentiated functions exist in ROS17/2.8 cells according to the magnitude of the hypergravity.

Alkaline Phosphatase↗

Short-term hypergravity does not affect protein-ubiquitination and proliferation in rat L6 myoblastic cells.

We previously reported that spaceflight (STS-90) and tail-suspension stimulated muscle protein ubiquitination and accumulated the degradation fragments. However, in space experiments the side-effects of hypergravity on samples are inevitable during the launch of a space shuttle into space or the reentry. To examine whether hypergravity also caused protein-ubiquitination in skeletal muscle cells, we exposed rat myoblastic L6 cells to various hypergravity conditions. Immunoblot analysis showed that the centrifugation at 2, 3, 30 or 100 G for 10 min did not increase the amount of ubiquitinated proteins in L6 cells, whereas the centrifugation at 100 G for 1 or 2 hrs significantly induced the protein-ubiquitination. In contrast, heat shock protein 70 (HSP70), another stress-responsive protein, in L6 cells was accumulated only by centrifugation at 100 G for more than 10 min. Short-term (10 min) hypergravity including 3 or 100 G did not affect the proliferation and morphological changes in L6 cells. Our present results suggest that the ubiquitination of muscle proteins is less sensitive to hypergravity than the induction of HSP70, and that the effect of hypergravity on protein-ubiquitination and proliferation of skeletal muscle cells may be negligible, as far as its duration is short-term.

Animals↗

Altered behaviour of hamsters by prolonged hypergravity: adaptation to 2.5 G and re-adaptation to 1 G.

We studied the functional adaptation process in 40 hamsters subjected to either prolonged hypergravity to normal gravity. Subadult golden hamsters (n = 20) exposed to a hypergravity condition of 2.5 G for 6 months were tested to investigate the effect of hyper gravity on the perceptive motor skills and compared with control hamsters (n = 20). The motor coordination of the hypergravity hamsters hardly changed; locomotion was normal and swimming was possible. Equilibrium maintenance was disturbed during the first 3 months as was shown by the higher crossing time (p < 0.001) and higher fall frequency (p < 0.001) for the hypergravity group. Significant differences were also found in orientation during swimming (p = 0.007) and turning behaviour in the rotation task (p < 0.001) and in the no-rotation task (p = 0.029). After 6 months, 10 hamsters of both groups were tested for another 4 months, also the hypergravity hamsters were living at 1 G. Differences in orientation in the two groups did not change during swimming and turning behaviour during the rotation task (p = 0.026). Based on our findings, we conclude that the hamsters functionally adapted to hypergravity, which led to an altered performance of several tasks. The condition continued after 4 months of normal gravity.

Animals↗

No effect of hypergravity on adult rat ventral horn neuron size or SDH activity.

BACKGROUND: Spaceflights of short duration (approximately 2 wk) result in adaptations in the size and/or metabolic properties of a select population of motoneurons located in the lumbosacral region of the rat spinal cord. A decrease in succinate dehydrogenase (SDH, an oxidative marker enzyme) activity of moderately sized (500-800 microm2) motoneurons in the retrodorsolateral region of the spinal cord (L6) has been observed after a 14-d flight. HYPOTHESIS: Our hypothesis was that exposure to short-term hypergravity would result in adaptations in the opposite direction, reflecting a continuum of morphological and biochemical responses in the spinal motoneurons from zero gravity to hypergravity. METHODS: Young, male rats were centrifuged at either 1.5 or 2.0 G for 2 wk. The size and SDH activity of a population of motoneurons in the retrodorsolateral region of the spinal cord (L5) were determined and compared with age-matched rats maintained at 1.0 G. The absolute and relative (to body weight) masses of the soleus, gastrocnemius, adductor longus and tibialis anterior muscles were compared among the three groups. RESULTS: There were no effects of either hypergravity intervention on the motoneuron properties. Rats maintained under hypergravity conditions gained less body mass than rats kept at 1.0 G. For the 1.5 and 2.0 G groups, the muscle absolute mass was smaller and relative mass similar to that observed in the 1.0 G rats, except for the adductor longus. The adductor longus absolute mass was similar to and the relative mass larger in both hypergravity groups than in the 1.0 G group. CONCLUSIONS: Our hypothesis was rejected. The findings suggest that rat motoneurons are more responsive to short-term chronic exposure to spaceflight than to hypergravity conditions.

Adaptation, Physiological↗

Hypergravity effects on osteoclasts activity.

The long-term space flight induces a loss of bone density. However, the mechanism has not been well understood, especially about gravity effect on osteoclast. To elucidate the gravitational effect on osteoclasts, we examined the rabbit primary osteoclasts applied to hypergravity and compared the mRNA expression of two kinds of osteoclast marker enzymes, TRAP (tartrate-resistant acid phosphatase) and cathepsin K at the transcription level. Rabbit osteoclasts isolated according to the modified method of Kakudo et al. were cultured on ivory and exposed to 30 x g for 2 hr or 18 hr by placing the culture tubes in a swinging bucket rotor. Results by RT-PCR suggested that hypergravity enhanced the mRNA expression of both enzymes with different manner; the expression of the TRAP showed a smaller increase, that of the cathepsin K showed a non-monotonous time course with maximum hypergravity effect for 2 hr incubation. Moreover we examined the hypergravity effect on actin ring formation in osteoclasts; however, we got no hypergravity effect on the numbers of activated osteoclasts with actin ring formation. These results might suggest that hypergravity has no effect on the number of osteoclasts with resorption activity, but enhances the resorption activity of activated osteoclasts.

Acid Phosphatase↗

[Study on the effect of hypergravity stress on L-glutamate uptake by rat brain nerve endings].

Using rat brain synaptosomes, we have investigated the effect of hypergravity on the kinetic parameters of Na(+)-dependent, high-affinity L-glutamate transport activity. The time-course of L-[14C]-glutamate uptake and dependence of L-[14C]-glutamate uptake velocity on glutamate concentrations were analyzed. K(m) and Vmax of this process have been determined. The hypergravity stress was created by centrifugation of rats for 1 hour at 10 g. We observed no differences in K(m) values between the control rats (10.7 +/- 2.5 microM) and animals exposed to hypergravity (6.7 +/- 1.5 microM). The similarity of this parameter for the two studied groups of animals showed that affinity of glutamate transporter to substrate was not sensitive to hypergravity stress. In contrast, the maximal velocity of glutamate uptake changed in hypergravity conditions. Vmax reduced from 12.5 +/- +/- 3.2 nmol/min per 1 mg of protein (control group) to 5.6 +/- 0.9 nmol/min per 1 mg of protein (animals, exposed to hypergravity stress). The possible mechanisms of attenuation of the glutamate transporter activity without modifying K(m) of glutamate uptake were discussed.

Animals↗

Short-term hypergravity influences NGF and BDNF expression, and mast cell distribution in the lungs and heart of adult male mice.

In this study we investigate the effects of short-term hypergravity on lung and heart neurotrophins and mast cell distribution. Our results showed that brain derived-neurotrophic factor (BDNF) protein and mRNA expression are increased in the lungs of mice exposed to hypergravity while in the heart hypergravity causes a marked reduction in BDNF mRNA expression, and a decrease in BDNF protein. Compared to controls, nerve growth factor (NGF) protein was expressed more in the heart of rotated mice. These observations demonstrate that altered hypergravity can affect, though differentially, the local expression of NGF and BDNF proteins and their mRNAs in the lung and heart and indicates that short-term exposure to hypergravity causes a marked increase in BDNF, but not in NGF in the lungs of adult mice. Moreover, mast cells, which are NGF-producing cells and implicated in cardiac and respiratory activity, increased in number in proximity to blood vessels in the heart and in lung airway epithelium of rotated mice. This study indicates that hypergravity influences cardiovascular and respiratory tissue and suggests a neurotrophin involvement in the reaction to this environmental exposure.

Animals↗

Effect of a hypergravity environment on cortical bone elasticity in rats.

There is considerable interest in determining whether hypergravity can be used as a countermeasure for microgravity-induced bone loss. This study was conducted on 20 immature male rats in order to investigate possible elastic adaptations of cortical bone in rapidly growing rats exposed to chronic hypergravity. Ten rats were continuously centrifuged for 14 days at twice gravitational acceleration (2G) on a 12.75 foot radius centrifuge and 10 rats concurrently acted as stationary controls. The effect of hypergravity on the elastic characteristics of cortical bone was quantified via ultrasonic wave propagation. Propagation velocities of longitudinal and shear waves were measured through cubic cortical specimens from the posterior femoral diaphyses. Density was measured with an Archimedes' technique. The orthotropic elastic properties were calculated and used to compare the difference between groups. Results showed an average increase in both the Young's moduli (Eii, + 2.2%) and shear moduli (Gij, + 4.3%) with a statistically significant increase only in G12 (+15.7%, P = 0.046). The ratio of transverse to axial strain (Poisson's ratio, nuij) demonstrated statistically significant changes in nu12, nu21, nu13, and nu31 (P < 0.05). These findings suggest that although slight elastic changes were incurred via a hypergravity environment, the treatment level or duration in this study do not dramatically perturb the normal elastic behavior of cortical bone and that dramatic biomechanical differences noted in previous studies were due more to structural changes than material elasticity changes. Hypergravity applied post facto to a microgravity environment would offer further illucidation of this method as treatment for a degenerative spaceflight experience.

Animals↗

Hypergravity speeds up the development of T-lymphocyte motility.

The effect of altered gravity on single cells has been reported in a number of studies. From the investigation of the immune system response to spaceflight conditions, interest has focused on the influence of gravity on single lymphocytes. Microgravity has been shown to decrease lymphocyte activation and to influence motility. On the other hand, the effect of hypergravity on lymphocyte motility has not been explored. We studied the migration of human peripheral blood T lymphocytes cultured in vitro in a hypergravity environment (10g). After hypergravity culture for 1-11 days, T cells were seeded on a fibronectin-coated glass surface, observed by time-lapse bright-field microscopy, and tracked by a computer program. We found that T cells, activated and then cultured in hypergravity, become motile earlier than cells cultured at normal gravity. These results suggest that hypergravity stimulates T cell migration.

Adaptation, Physiological↗

Hypergravity exercise against bed rest induced changes in cardiac autonomic control.

An intermittent exposure to artificial hypergravity with physical exercise by a human centrifuge may provide a countermeasure against various physiological problems after space flight. To test the effects of hypergravity with ergometric exercise on dynamic regulation of heart rate during weightlessness, we quantified autonomic cardiovascular control before and after head-down-tilt bed rest (HDBR) with and without the countermeasure. Twelve male subjects underwent a 14-day period of HDBR. Six of them were exposed to a hypergravity (+1.2 Gz acceleration at heart level) for 30 min with ergometric exercise (60 W, n=4; 40 W, n=2) as a countermeasure on day 1, 2, 3, 5, 7, 9, 11, 12, 13 and 14, during HDBR (CM group). The remaining six were not exposed to a hypergravity exercise during HDBR (control group). Blood pressure and ECG were recorded at a supine position before and after HDBR. The high frequency power of R-R interval (HFRR; 1,008+/-238 to 353+/-56 ms(2) P<0.05) as an index of cardiac parasympathetic activity, and transfer function gain between BP and R-R interval in the high frequency range (GainHF; 21.9+/-5.4 to 14.5+/-4.2 ms/mmHg, P<0.01) as an index of vagally mediated arterial-cardiac baroreflex, decreased significantly after HDBR in the control group. However, these changes were not statistically significant in the CM group (HFRR, 1,150+/-344 to 768+/-385 ms(2); GainHF, 21.5+/-3.3 to 18.6+/-3.4 ms/mmHg). Moreover, baroreflex gain by sequence analysis showed similar results. This observation suggests that the intermittent exposure to hypergravity with ergometric exercise may attenuate the decreases in the parasympathetic activity and the spontaneous arterial-cardiac baroreflex function after weightlessness.

Adult↗

Effects of hypergravity conditions on elongation growth and lignin formation in the inflorescence stem of Arabidopsis thaliana.

The effects of hypergravity on elongation growth and lignin deposition in secondary cell walls of the Arabidopsis thaliana (L.) Heynh. inflorescence stem were examined in plants grown for 3 days after exposure to hypergravity in the direction from shoot to root at 300 g for 24 h. The content of acetylbromide-extractable lignins in a secondary cell wall fraction prepared by enzyme digestion of inflorescence stem segments removing primary cell wall components was significantly increased by the hypergravity stimulus. Xylem vessels, particularly in a region closer to the base of the inflorescence stem, increased in number. Gadolinium chloride at 0.1 mM, a blocker of mechanoreceptors, partially suppressed the effect of hypergravity on lignin deposition in the secondary cell wall fraction. These results suggest that mechanoreceptors are responsible for hypergravity-induced lignin deposition in secondary cell walls in A. thaliana inflorescence stems.

Arabidopsis↗