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Effects of hypergravity on the morphological properties of the vestibular sensory epithelium. I. Long-term exposure of rats after full maturation of the labyrinths.

The effect of prolonged exposure to hypergravity on the morphology of vestibular epithelia of rats was investigated. At the age of 1 month, i.e., when vestibular end organs are fully maturated, three rats were transferred to a hypergravity environment of 2.5 g inside a large radius centrifuge. After 9 months, vestibular epithelia of these animals and of three control animals were immunohistochemically labeled for actin and tubulin. The apical cross-sectional area of epithelial cells of hypergravity exposed rats appeared to be smaller in all end organs. Area reduction was 1.9% in the saccule (not significant), 5.0% in the utricle (p < 0.005), and 11.6% in the crista (p<<0.001). No indications for a deterioration of vestibular functioning were observed.

Actins↗

Altered behaviour in hamsters conceived and born in hypergravity.

We studied vestibular function in 37 hamsters (1 month old) conceived and born in either hypergravity (n = 21) or normal gravity (n = 16). Four groups were made: (1) HL group: 20 weeks in 2.5 G and 14 weeks in 1 G; (2) HS group: 4 weeks in 2.5 G and 30 weeks in 1 G; (3) CON group: 34 weeks in 1 G; and (4) ROT group: 4 weeks in 1 G, 16 weeks in rotation in 1 G, at the centre of the centrifuge and 14 weeks 1 G. When the hamsters were 4 weeks old, their locomotor activity, swimming ability, and air-righting was assessed. We found that HL and HS hamsters had no disturbances during locomotion in 1 G but their swimming ability was disturbed (swimming underwater, circling, and decreased speed of swimming). The HL hamsters showed less activity during 2.5 G and showed fewer correct air-rightings than the other groups. Differences between groups in swimming ability and the number of correct air-righting responses remained even after 3 months of normal gravity. Based on these findings, we suggest that the persistent behavioural disturbances are caused by the embryonal development of the hamsters in a hypergravity environment. Furthermore, hypergravity and rotation each have a different effect on behaviour.

Age Factors↗

A mild stress, hypergravity exposure, postpones behavioral aging in Drosophila melanogaster.

Flies were submitted to two weeks of hypergravity in a centrifuge (3 or 5 g), starting at the second day of imaginal life, and their behavior (spontaneous locomotor activity, patterns of movement, and climbing activity) was observed from removal of the centrifuge to an older age; the usual effects of age on these behaviors were generally observed. Hypergravity-kept flies had worse behavioral scores on removal of centrifuge than those always kept at 1 g. When they aged, they got either similar or better scores than 1 g flies, which indicates that their behavioral aging may be slower. These results show that a mild stress such as hypergravity, which has been previously shown to increase the longevity of males and resistance to heat shock in both sexes, is an environmental manipulation postponing aging in flies.

Aging↗

Protective effect of prone posture against hypergravity-induced arterial hypoxaemia in humans.

Patients with acute respiratory distress syndrome have increased lung tissue weight and therefore an increased hydrostatic pressure gradient down the lung. Also, they have a better arterial oxygenation in prone (face down) than in supine (face up) posture. We hypothesized that this effect of the direction of gravity also existed in healthy humans, when increased hydrostatic gradients were induced by hypergravity. Ten healthy subjects were studied in a human centrifuge while exposed to 1 or 5 G in anterio-posterior (supine) or posterio-anterior (prone) direction. We measured blood gases using remote-controlled sampling and gas exchange by mass spectrometry. Hypergravity led to marked impairments of arterial oxygenation in both postures and more so in supine posture. At 5 G, the arterial oxygen saturation was 84.6 +/- 1.2 % (mean +/- S.E.M.) in supine and 89.7 +/- 1.4 % in prone posture (P < 0.001 for supine vs. prone). Ventilation and alveolar PO2 were increased at 5 G and did not differ between postures. The alveolar-to-arterial PO2 difference increased at 5 G to 8.0 +/- 0.2 kPa and 6.6 +/- 0.3 kPa in supine and prone postures (P = 0.003). Arterial oxygenation was less impaired in prone during hypergravity due to a better-preserved alveolo-arterial oxygen transport. We speculate that mammals have developed a cardiopulmonary structure that favours function with the gravitational vector in the posterio-anterior direction.

Adult↗

Effects of hypergravity on mammary metabolic function: gravity acts as a continuum.

Mammary metabolic activity in pregnant rats is significantly increased in response to spaceflight. To determine whether changes in mammary metabolism are related to gravity load, we exposed pregnant rats to hypergravity and measured mammary metabolic activity. From days 11-20 of gestation (G), animals were centrifuged (20 rpm; 1.5, 1.75, or 2.0 x gravity) or were maintained at 1 G. On G20, five rats from each group were removed from the centrifuge and euthanized. The remaining dams (n = 5/treatment) were housed at 1 G until parturition. After 2 h of nursing by the pups, the postpartum dams were euthanized (G22). Glucose oxidation to CO2 and incorporation into lipids was measured. Mammary glands from dams euthanized on G20 revealed a strong negative correlation between metabolic rate and increased G load. Approximately 98% of the variation in glucose oxidation and 94% of the variation in glucose incorporation into lipids can be accounted for by differences in G load. Differences in metabolic activity disappeared in the postpartum dams. When we combined previous data from the microgravity with hypergravity environments and plotted the ratio of mammary metabolic rate vs. G load, there was a significant exponential relationship (r2 = 0.99). These data demonstrate a remarkable continuum of response across the microgravity and hypergravity environments and support the concept that gravitational load influences mammary tissue metabolism.

Animals↗

Effect of hypergravity on vestibular compensation in guinea pigs.

The effect of hypergravity on vestibular compensation was studied in guinea pigs. Pharmacological labyrinthectomy was performed by injecting chloroform into the middle ear cavity under ether anesthesia. The guinea pigs were exposed to hypergravity on a centrifuge. The animals were divided into four groups: a group stimulated with 2G after labyrinthectomy of the right ear, a group stimulated with 2G after labyrinthectomy of the left ear to evaluate the influence of the centrifugal rotation, a group stimulated with acceleration and deceleration alone, and a control group which was maintained under similar conditions, but without centrifugation. Head deviation and nystagmus were recorded and analysed to assess the process of compensation at 1, 3, 5, 7 and 9 h after labyrinthectomy. The 2G-stimulated group showed faster compensation in head deviation than the control group. In this study, the hypergravity stimulation seemed to facilitate the compensation in head deviation.

Adaptation, Physiological↗

[Altered behaviour and expression of Fos in rats born in hypergravity and their re-adaptation to the normal gravity].

Changes in behaviour relevant to the vestibular system were studied in Long-Evans rats which were fertilized, born and housed in 2 acceleration of gravity for 4 months and thereafter exposed to 1 acceleration of gravity, and expression of Fos protein in the brain stem was examined. Data from the hypergravity rats were compared respectively with those from the rotation group and the labyrinthectomized group. Static and locomotion modes of the hypergravity rats were changed, tension of extensor was enhanced and the abilities in locomotion equalization and orientation in swimming and air-righting response were reduced. The adaptation process varied with different behaviours. The time for recovery of the ability of orientating in swimming was the longest, taking more than 1 month. The Fos protein expression provides a useful tool for mapping brain functional activities after sensory stimulation, showing a low basal level in normal and labyrinthectomized groups. The hypergravity rats, on the other hand, exhibited more Fos-positive cells in the superior colliculus, inferior colliculus, periaqueductal gray, raphe dorsal nucleus and solitary nucleus. In contrast, the inferior olivary nuclei, locus coeruleus and vestibular nuclei were not strongly labeled. These spatial patterns of Fos expression suggest that a decrease in gravity-inertial force may activate a neural pathway different from the vestibulo-olivar pathways activated by an increase in gravity-inertial force.

Adaptation, Physiological↗

Activity of ornithine decarboxylase in muscle of Japanese quail in hypergravity conditions.

Ornithine decarboxylase (ODC) is the rate limiting enzyme in the synthesis of polyamines which are generally required for animal cell proliferation. Because of many different stimuli changing the activity of ODC, the effect of hypergravity on muscle ODC activity in Japanese quail was investigated. Adult Japanese quail cockerels were exposed to 2G in the centrifuge permanently for 96 hours. The activity of ODC was determined by radioisotopic method in supernatant of homogenised breast and fibular muscles (m. pectoralis minor, m.superficialis fibularis). The activity of ODC was higher in breast muscle than in the fibular muscle in both, control and experimental groups. Exposition of quail to hypergravity evokes approximately 2 fold increase of ODC activity in both breast and fibular muscles. The results suggest that 96 hours of permanent hypergravitation (2G) caused activation of polyamine synthesis and their involvement in biochemical processes of adaptation to hypergravity.

Adaptation, Physiological↗

Microgravity and hypergravity effect on survival and reproduction of microinvertebrates.

Preliminary to carry out long-term experiments on the International Space Station (ISS) using living orgainisms, the capacity of the experimental organisms to cope with perturbations of gravity should be tested. Actually, animals have evolved under gravity, because on earth gravity force cannot be eluded, and several features that influence life-history traits may be affected by the presence of gravity. Among the other features, feeding efficiency may be affected by gravity if the animals feed by filtering suspended particles, creating currents that carry the particulate food to their mouth opening. In presence of gravity the food particles tend to sink to the bottom and filter-feeders must be able to suspend and collect the particles with some apparatus such as ciliary wreaths. It can be predicted that hypergravity, increasing the particle sedimentation rate, will reduce the animal filtering efficiency, while microgravity will increase filtering rate. Differently, some bacteriophagous animals do not possess structures to collect their food, but commonly live and move into sediment and feed on the bacteria upon encounter. Hypergravity will apply higher pressure on their bodies, and could force them to adhere to some surface and to reduce their displacement, and microgravity could impede adhesion to the surface and make food item encounters improbable. Thus, gravity perturbations may affect animal life-history traits, such as survival or fecundity, by influencing their feeding efficiency. In this study we exposed a filter-feeding organism (Macrotrachela quadricornifera, Rotifera Bdelloidea) and a bacteriophagous one (Panagrolaimus rigidus, Nematoda) to both microgravity and hypergravity to test their reproduction capacity under such stressful conditions, and their suitability as models for experiments on the ISS.

Animals↗

Resting energy expenditure of rats acclimated to hypergravity.

BACKGROUND: The use of centrifugation at 1 G has been advocated as a control condition during spaceflight and as a countermeasure to compensate for the adverse effects of spaceflight. Rodents are the primary animal model for the study of the effects of spaceflight and will be used in the evaluation of centrifugation as a countermeasure and means of control at 1 G during flight. HYPOTHESIS: The present study was designed to assess whether resting energy expenditure (EER) of male rats was increased in relation to the magnitude of the level of gravity to which the animals were exposed. The influence of body mass and age on resting energy expenditure (EER) of male rats (n = 42, age 40-400 d) was determined following 2 wk of acclimation to 1, 2.3, or 4.1 G. Hypergravity environments were created by centrifugation. Measurements were made at the gravity level to which the animal was acclimated and during the lights-on period. RESULTS: In rats matched for body mass (approximately 400 g), mean O2 consumption and CO2 production were higher (18% and 27%, respectively) in the 2.3- and 4.1 -G groups than controls. Mean respiratory exchange ratio (RER) increased from 0.80 to 0.87. EER was increased from 47 +/- 0.1 kcal x d(-1) at 1 G, to 57 +/- 1.5 and 58 +/- 2.2 kcal x d(-1) at 2.3 and 4.1 G, respectively. There was no difference in EER between the hypergravity groups. When age differences were considered, EER (kcal x kg(-1) x d(-1)) with increased gravity was 40% higher than at 1 G. The increase in EER was not proportional over gravity levels. CONCLUSION: Acclimation of rats to hypergravity increases their EER, dependent on body mass and age, and may alter substrate metabolism. The increase in EER was not related to the level of gravity increase.

Adaptation, Physiological↗

Hypergravity and opioid-mediated pain suppression in rats.

It is known that pain suppression in animals is induced by certain environmental stimulus. However, little is known about the effects of gravitational alteration on the nociceptive responses in rats. A recent study indicated that Fos protein expression was strongly induced in the vestibular-related brainstem regions of rats that were exposed to 2 G hypergravity (Gustave Dit Duflo et al., 2000). A number of studies indicate that Fos expression is induced in the brain by various kinds of stress. We showed that either long-term exposure or short-term exposure to 2 G hypergravity elevated the nociceptive threshold in the rat skin surfaces, in concomitant with Fos induction in the hypothalamus including the arcuate nucleus and paraventricular nucleus (Kumei et al., 2000). We have examined the possible involvement of beta-endorphin, an endogenous opioid, in the hypergravity-induced analgesic effects on rats and its counteraction by naloxone, an opioid receptor antagonist.

Animals↗

The behavioral response of zebrafish to hypergravity conditions.

Previous reports of the behavior of aquatic organisms in the microgravity environment of space (~10(-6) g) or during the brief weightless period of parabolic flight indicate that most species display a dramatic "looping" or "circling" response (De Jong et al. 1996, Anken, Ibsch and Rahmann 1998). However, the behavior of aquatic species under hypergravity conditions is less clear. Our objectives in the present study were to examine the behavioral response of adult zebrafish (Danio rerio) to hypergravity conditions (2-g), quantify changes in adult swimbladder volume, and to determine if the larvae of zebrafish are capable of accessing the air-water interface for initial swimbladder inflation under hypergravity conditions.

Air Sacs↗

Stimulation of DNA polymerase alpha by hypergravity generated by centrifugal acceleration.

Gravity alteration is known to influence cell proliferation. Here we tested the effects of hypergravity on the action of DNA polymerase alpha, one of the DNA replication enzymes in eukaryotes. Hypergravity was produced by horizontal centrifugal acceleration with a hand-made rotator. The reaction rate of DNA polymerase alpha in centrifuge tubes increased along with the acceleration up to 4g, when a plateau was reached. In contrast, no stimulation was observed with primase, DNA polymerase epsilon, and the E. coli DNA polymerase I Klenow fragment. Kinetic analysis of DNA polymerase alpha reactions revealed that, under high gravity conditions, the K(m) value for template DNA decreased while the V(max) stayed constant. In contrast, the centrifugal acceleration did not affect the K(m) values for deoxyribonucleoside triphosphates. These results suggest that the hypergravity enhances the activity of DNA polymerase alpha by increasing the affinity of the enzyme for template DNA. Such enhancement was more prominent with a low concentration of DNA polymerase alpha under low ionic conditions.

Animals↗

Hypergravity signal transduction in HeLa cells with concomitant phosphorylation of proteins immunoprecipitated with anti-microtubule-associated protein antibodies.

We have shown that hypergravity (35g) stimulates production of inositol 1,4,5-trisphosphate (IP3) and decreases adenosine 3',5'-cyclic monophosphate (cAMP) levels in HeLa cells. IP3 production rapidly increased 1.5- and 2.1-fold greater (P less than 0.05) than the control after 2- and 5-min exposures to 35g, respectively. The intracellular cAMP levels, determined in the presence of isobutylmethylxanthine, were decreased by 11% (P less than 0.05) and 16% (P less than 0.01) relative to the control after 10- and 20-min exposures to 35g, respectively. The phosphorylation of proteins which were immunoprecipitated by antibodies recognizing microtubule-associated proteins (ipMAPs) was also apparent after exposure of these cells to hypergravity. In the detergent-insoluble fraction, phosphorylation of a 115-kDa protein was significantly enhanced compared to the control after a 5-min exposure to 35g. In the detergent-soluble fraction, phosphorylation of a 200-kDa protein was observed served after a 20-min exposure to 35g. Our study suggests that IP3 and cAMP may act as second messengers in hypergravity signal transduction. Phosphorylation of ipMAPs in both the detergent-soluble and -insoluble fractions suggests that cytoskeletal structures may be influenced by gravity.

1-Methyl-3-isobutylxanthine↗

Effects of hypergravity environment on the parathyroid gland of the propranolol-treated golden hamster.

The fine structure of the parathyroid glands of propranolol-treated hamsters subjected to 5 x gravity environment was studied. In the parathyroid glands of the propranolol-treated hamsters exposed to hypergravity environment, the volume density occupied by the Golgi complexes and cisternae of the granular endoplasmic reticulum was increased as compared to that of propranolol-treated hamsters and was decreased as compared to that of hamsters exposed to a hypergravity environment but was almost similar to that of control hamsters. In addition, many chief cells contained rich free ribosomes, abundant mitochondria and some secretory granules located in the peripheral cytoplasm. These findings suggest that the parathyroid gland which may be suppressed by treatment of propranolol and stimulated in response to a hypergravity environment indicates the secretory activity of the control parathyroid gland.

Animals↗

Electron-microscopic study of the parathyroid gland of epinephrine-treated golden hamsters subjected to hypergravity environment.

The ultrastructure of the parathyroid glands of golden hamsters subjected to 5-gravity environment after administration of epinephrine was studied. In the epinephrine-treated animals exposed to a hypergravity environment, the Golgi complexes associated with numerous prosecretory granules were significantly increased compared with those of the control, centrifuged and epinephrine-treated animals, as well as the cisternae of the granular endoplasmic reticulum compared with those of the control and centrifuged animals. In addition, many secretory granules were situated close to the the plasma membrane of the chief cells in the epinephrine-treated animals exposed to a hypergravity environment. Those observations suggest that the secretory activity of the parathyroid gland may be markedly stimulated in the epinephrine-treated animals exposed to a hypergravity environment.

Animals↗

Stereological studies of the parathyroid gland of phosphate-treated golden hamsters subjected to a hypergravity environment.

The ultrastructure of the parathyroid glands of phosphate-treated golden hamsters exposed to a 5-G environment was studied. In the phosphate-treated animals exposed to a hypergravity environment, the Golgi complexes associated with numerous prosecretory granules, and the enlarged intercellular spaces containing floccular or finely particulate material showed a significant increase compared to those of the control, centrifuged, and phosphate-treated groups, and the cisternae of the granular endoplasmic reticulum showed a significant increase compared to those of the control and phosphate-treated groups. In addition, numerous secretory granules were situated close to the plasma membrane of chief cells in the phosphate-treated animals exposed to a hypergravity environment. These findings suggest that the synthesis, and to a greater extent the release of secretory granules may be markedly stimulated, in the parathyroid glands of phosphate-treated animals exposed to a hypergravity environment.

Animals↗

Hypergravity and aging in Drosophila melanogaster. 4. Climbing activity.

Drosophila melanogaster flies climb up the sides of their vial after having been submitted to a mechanical stimulation; that ability is impaired at older ages. The climbing activity (CLI) of flies kept at various gravity levels (1, 3 and 5 g) has been measured throughout life, in cross-sectional studies. Hypergravity had no effect on CLI at young age, but older flies kept in hypergravity displayed lower scores than flies kept at 1 g. Results are discussed in relation with the hypothesis of increased aging rate in hypergravity.

Aging↗