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The reaction of Xenopus laevis Daudin (South African toad) to linear accelerations.

Preparing the German Spacelab Mission D-2 project "Gravity Perception and Neuronal Plasticity"--STATEX II--ground based experiments have been performed with larvae of the amphibian vertebrate Xenopus laevis Daud. to study the reactions to different levels of acceleration forces and profiles. The larvae have been exposed to accelerations of up to 5 g for different time periods using a modified laboratory centrifuge and the NIZEMI (Niedergeschwindigkeits-Zentrifugen-Mikroskop) which allows direct observation and video documentation. The results will be discussed and compared with those of the D1-Mission, parabolic flights, and simulated weightlessness.

Acceleration↗

Behavioural and biochemical investigations of the influence of altered gravity on the CNS of aquatic vertebrates during ontogeny.

Quantitative data are presented on the influences of hyper-gravity (3 +/- 1g) and of simulated weightlessness (approximately 0g) during early ontogeny of cichlid fish (Oreochromis mossambicus) and clawed toad (Xenopus laevis, Daudin) demonstrating changes in the swimming behaviour and the brain energy and plasma membrane metabolism. After return to 1g conditions, hyper-g reared fish and toads express the well known "loop-swimming" behaviour. By means of a computer based video analyzing system different types of swimming movements and velocities were quantitatively determined. Analyses of the brain energy and plasma-membrane metabolism of hyper-g fish larvae demonstrated an increase in energy availability (glucose 6Pi dehydrogenase, G-6P-DH), a decrease of cellular energy transformation (creatine kinase activity, CK) but no changes in energy consumptive processes (e.g. ATPases) and cytochrome oxidase activity (Cyt.-Ox). In contrast hypo-g fish larvae showed a slight increase in brain CK activity. In addition, unlike 1g controls, hyper-g fish larvae showed pronounced variations in the composition (=polarity) of sialoglycosphingolipids (=gangliosides), typical constituents of the nerve cell membranes, and a slight increase in the activity of sialidase, the enzyme responsible for ganglioside degradation.

Animals↗

Influence of longitudinal whole animal clinorotation on lens, tail, and limb regeneration in urodeles.

Two species of newts (Urodela) and two types of clinostats for fast clinorotation (60 rpm) were used to investigate the influence of simulated weightlessness on regeneration and to compare results obtained with data from spaceflight experiments. Seven or fourteen days of weightlessness in Russian biosatellites caused acceleration of lens and limb regeneration by an increase in cell proliferation, differentiation, and rate of morphogenesis in comparison with ground controls. After a comparable time of clinorotation the results obtained with Triturus vulgaris using a horizontal clinostat were similar to those found in spaceflight. In contrast, in Pleurodeles waltl using both horizontal and radial clinostats the results were contradictory compared to Triturus. We speculate that different levels of gravity or/and species specific thresholds for gravitational sensitivity could be responsible for these contradictory results.

Animals↗

Influence of altered gravity on the cytochemical localization of cytochrome oxidase activity in central and peripheral gravisensory systems in developing cichlid fish.

Cichlid fish larvae were reared from hatching to active free swimming under different gravity conditions: natural environment, increased acceleration in a centrifuge, simulated weightlessness in a clinostat and near weightlessness during space flight. Cytochrome oxidase activity was analyzed semiquantitatively on the ultrastructural level as a marker of regional neuronal activity in a primary, vestibular brainstem nucleus and in gravity receptive epithelia in the inner ear. Our results show, that gravity seems to be positively correlated with cytochrome oxidase activity in the magnocellular nucleus of developing fish brain. In the inner ear the energy metabolism is decreased under microgravity concerning utricle but not saccule. Hypergravity has no effect on cytochrome oxidase activity in sensory inner ear epithelia.

Animals↗

Dynamic change of ERPs related to selective attention to signals from left and right visual field during head-down tilt.

To study further the effect of head-down tilt (HDT) on slow positive potential in the event-related potentials (ERPs), the temporal and spatial features of visual ERPs changes during 2 hour HDT (-10 degrees) were compared with that during HUT (+20 degrees) in 15 normal subjects. The stimuli were consisted of two color LED flashes appeared randomly in left or right visual field (LVF or RVF) with same probability. The subjects were asked to make switch response to target signals (T) differentially: switching to left for T in LVF and to right for T in RVF, ignoring non-target signals(N). Five sets of tests were made during HUT and HDT. ERPs were obtained from 9 locations on scalp. The mean value of the ERPs in the period from 0.32-0.55 s was taken as the amplitude of slow positive potential(P400). The main results were as follows. 1) The mean amplitude of P400 decreased during HDT which was more significant at the 2nd, 3rd and 5th set of tests; 2) spatially, the reduction of mean P400 amplitude during HDT was more significant for signals from RVF and was more significant at posterior and central brain regions than that on frontal locations. As that the positive potential probably reflects the active inhibition activity in the brain during attention process, these data provide further evidence showing that the higher brain function was affected by the simulated weightlessness and that this effect was not only transient but also with interesting spatial characteristics.

Adult↗

Kinetics of amyloplast movement in cress root statocytes under different gravitational loads.

In order to investigate the movement of a statolith complex along the longitudinal axis of root cap statocytes under different mass accelerations, a series of experiments with Lepidium sativum L. in an automatically operating centrifuge during the Bion-11 satellite flight and on a centrifuge-clinostat have been performed. During spaceflight, roots were grown for 24 h under root-tip-directed centrifugal 1-g acceleration, then exposed to microgravity for 6, 12 and 24 min and chemically fixed. During the first 6 min of microgravity, the statoliths moved towards the cell center with a mean velocity of 0.31 +/- 0.04 micrometers/min, which decreased to 0.12 +/- 0.01 micrometers/min within subsequent 12-24 min period. The mean relative position of the statolith complex in respect to the distal cell wall (% of total cell length) increased from 24.0 +/- 0.5% in 1 g-grown roots to 38.8 +/- 0.8% in roots exposed for 24 min to microgravity, but remained smaller than in roots grown continuously in microgravity (48.0 +/- 0.7%). The properties of the statolith movement away from the distal pole of the statocyte were studied in roots grown for 24 h vertically under 1 g and then placed for 6 min on a fast rotating clinostat (50 rpm) or 180 degrees inverted. After 2 min of both treatments, the mean relative position of the statoliths increased by about 10% versus its initial position. Later on, the proximal displacement of amyloplasts slowed down under simulated weightlessness, while it proceeded at a constant velocity under 1 g inversion. In roots grown on the clinostat and then exposed to 1 g in the longitudinal direction, amyloplast sedimentation away from the central region of statocyte was similar at the beginning of distal and proximal 6-min 1-g stimulation. However, at the end of this period statolith displacement was more pronounced in proximal direction as compared to distal. It is proposed that statolith position in the statocyte of a vertical root is controlled by the force of gravity, however, the intracellular forces, first of all those generated by the network of the cytoskeleton, are manifested when an usual orientation of the organ is changed or the statocytes are exposed to microgravity and clinorotation.

Centrifugation↗

Influence of clinorotation and fettering stress on tail regeneration of Triturus vulgaris (Urodela).

Tail-amputated adult Triturus vulgaris, fettered in cuvettes of a fast-rotating clinostat were exposed to simulated weightlessness (60 rpm; equiv. to 10(-3)-10(-4) g), during a 14-day period. To feed and clean the animals rotation was stopped once a day for approx. 10 min. To test the influence of the fettering stress, a second series of animals was kept separately under normal earth conditions without rotation. A further control series was kept in a dark container without any handicap. While tail regeneration of the rotated animals was markedly accelerated, the fettered-only animals showed a considerably less marked acceleration effect. At the end of the 14-day period, all regenerates were reamputated together with an additional 5 mm of the tail stump. Although this second level of amputation was distant from the first, the regenerative growth rate of the rotated series was accelerated 123% in contrast to both the control and the fettered-only series. Our results demonstrate that the growth acceleration is induced by clinorotation. Fettering stress has no comparable influence. The growth promoting effect is not limited to the regenerating area.

Amputation, Surgical↗

Space flight nutrition research: platforms and analogs.

Conducting research during actual or simulated weightlessness is a challenging endeavor, where even the simplest activities may present significant challenges. This article reviews some of the potential obstacles associated with performing research during space flight and offers brief descriptions of current and previous space research platforms and ground-based analogs, including those for human, animal, and cell-based research. This review is intended to highlight the main issues of space flight research analogs and leave the specifics for each physiologic system for the other papers in this section.

Animals↗

Effect of prolonged head-down bed rest on complex cardiovascular dynamics.

We postulated that a change in complex dynamics of the cardiovascular system could be involved in the orthostatic intolerance observed after simulated weightlessness. Supine recordings of 1024 consecutive pulse intervals and systolic blood pressures were obtained on 7 subjects adapted to a 42 day head-down bed rest (day 22 and 42) but also before and 6 days after head-down bed rest (-6 degrees). Coarse graining spectral analysis was used to extract the non-harmonic (fractal) component from each time series. The power spectral densities of this fractal component are inversely proportional to their frequency (1/f beta). We fitted an inverse power law estimate to the fractal component to determine the spectral exponent beta. The complex dynamics of blood pressure and heart rate variability were also analyzed by correlation dimension and non-linear prediction. Bed rest induced orthostatic intolerance in 4 subjects. There was a significant increase in the spectral exponent beta of RR-interval variability during and after head-down bed rest (before: 1.039 +/- 0.090; during: 1.552 +/- 0.080 and 1.547 +/- 0.100; after: 1.428 +/- 0.040). Analysis of the blood pressure dynamics indicated lower correlation dimensions during head-down bed rest and higher coefficients of predictability after head-down bed rest. Complexity alterations of RR-interval and blood pressure variability were not linked with one another during head-down bed rest. These alterations seemed to be correlated with the orthostatic intolerance observed after bed rest. These results suggest a change of the integration level of cardiovascular autonomic regulation.

Adult↗

Hormonal, water balance, and electrolyte changes during sixty-day confinement.

The EXEMSI experiment has made it clear that it is difficult to perform psychological and physiological protocols satisfactorily in the same study. It is, therefore, essential that the objectives of study be defined clearly before the start. While behavioral and psychological studies may be possible and provide valid results for a small group of mixed gender, it is more difficult to conduct valid physiological studies due to large differences between individuals and even in the same individual over time. As stated before, it is unusual in space research on humans and even during space simulation studies to have large and homogeneous groups of subjects. The consequence is that the results remain tentative. For a better understanding of the physiological data collected during the ISEMSI ad EXEMSI experiments, they should be correlated with the results of the psychological studies. One of the conclusions drawn from the ISEMSI experiment was that confinement provides a valuable parallel to other simulations of weightlessness, such as bedrest. The same pattern of changes in parameters like the blood volume regulating hormones renin and aldosterone was observed as in bedrest. After the EXEMSI study we can say that the conditions imposed by confinement, high work load, and stress, potentiate these effects. This implies that in using head-down bedrest as a weightlessness simulation the confinement effects must be identified by setting adequate control conditions for the head-down position, for short-term as well as for long-term simulations. Indeed, we have seen in the two isolation studies that confinement may have its effects at the beginning of the isolation period (EXEMSI) as well as during the entire isolation period (ISEMSI). In planning for EXEMSI we wanted to obtain more insight in some of the phenomena observed during ISEMSI by the introduction of new techniques such as the doubly labeled water method for determination of total body water. However, in some cases the opposite effects of those encountered in ISEMSI were found. This was probably due to the many changes in the experimental scenario, like number of subjects, mixed gender, living space per subject, and workload. Thus, for future isolation studies the operational scenario should be better examined and preferably standardized. Nevertheless, in such studies as well as in long-term sojourns in a space station, the crew size will not be larger than that of the EXEMSI crew. Physiologists will, therefore, have to become familiar with the study of small groups of subjects and to try to overcome the problems of large individual differences and statistical analysis of data from small groups.

Adult↗

Ultrastructure of gravity-perceiving cells in plant roots.

The root cap is the gravity-perceiving organ of plant roots. The central statenchyma consists of polarly organized statocytes which are characterized by sedimentable amyloplasts and by a striking distal endoplasmic reticulum (e.r. complex). During the normal downwards orientated growth of the root, the amyloplasts are sedimented onto the e.r. complex. Some observations indicate that the amyloplasts stress the e.r. complex by their sedimentation. The stress possibly influences the structural and functional state of the e.r. membranes. Therefore, graviperception is probably a function of differential stress on the e.r. complexes. The amyloplasts are partly or totally separated from the e.r. complex under experimental conditions such as deviations of the roots from the perpendicular between 30 and 180 degrees, rotation of the roots on the horizontal klinostat at 2 rev/min and at 55-120 rev/min, and placing the roots into an electric field at 2000-3000 V/cm. These are important facts for an understanding of the nature of graviperception. Spacelab experiments may help to verify the interpretation of results obtained in the experiments in simulated weightlessness.

Brassicaceae↗

Enhanced insulin action on glucose transport and insulin signaling in 7-day unweighted rat soleus muscle.

Hindlimb suspension (HS), a model of simulated weightlessness, enhances insulin action on glucose transport in unweighted rat soleus muscle. In the present study, we tested the hypothesis that these changes in glucose transport in 3- and 7-day HS soleus of juvenile, female Sprague-Dawley rats were due to increased functionality of insulin signaling factors, including insulin receptor (IR), IR substrate-1 (IRS-1), phosphatidylinositol 3-kinase (PI3-kinase), and Akt. Insulin-stimulated (2 mU/ml) glucose transport was significantly (P < 0.05) enhanced in 3- and 7-day HS soleus by 59 and 113%, respectively, compared with weight-bearing controls. Insulin-stimulated tyrosine phosphorylation of IR and Ser(473) phosphorylation of Akt was not altered by unweighting. Despite decreased (34 and 64%) IRS-1 protein in 3- and 7-day HS soleus, absolute insulin-stimulated tyrosine phosphorylation of IRS-1 was not diminished, indicating relative increases in IRS-1 phosphorylation of 62 and 184%, respectively. In the 7-day HS soleus, this was accompanied by increased (47%) insulin-stimulated IRS-1 associated with the p85 subunit of PI3-kinase. Interestingly, the enhanced insulin-stimulated glucose transport in the unweighted soleus was not completely inhibited (89-92%) by wortmannin, a PI3-kinase inhibitor. Finally, protein expression and activation of p38 MAPK, a stress-activated serine/threonine kinase associated with insulin resistance, was decreased by 32 and 18% in 7-day HS soleus. These results indicate that the increased insulin action on glucose transport in the 7-day unweighted soleus is associated with increased insulin signaling through IRS-1 and PI3-kinase and decreased p38 MAPK protein expression. However, PI3-kinase-independent mechanisms must also play a small role in this adaptive response to HS.

Animals↗

Twenty four hours head-down tilt decreases arterial baroreflex function in conscious rat.

In human zero gravity induces a cardiovascular deconditioning. The transfer of blood from the periphery to the thoracic compartment, occuring immediately at the beginning of the exposure to microgravity seems to be involved in this phenomenon. We have previously shown that LBNP (-40 mm Hg) produced an exaggerated heart rate response at the end of 24 hours of bed rest without modification of the cardiopulmonary receptors in human. Some other studies have shown that microgravity induces an impairment of the baroreflex sensitivity in human, but its involvement in the cardiovascular deconditioning is not completely understood. The aim of this study was to determine the modification of the arterial baroreflex using a model of simulated weightlessness in conscious rats during the exposure to microgravity.

Animals↗

Effects of hindlimb suspension training on the central and regional hemodynamic responses during 24 hours antiorthostatic hypokinesia in the awake rat.

To date the hindlimb suspension model utilizing rats has a wide application to simulate weightlessness. In our previous study we have examined the cardiovascular responses in the tail suspension model using the radiolabeled microspheres technique. We have reported increases of cardiac output (CO) and decreases of total peripheral vascular resistance (TPVR) after 24 hours of head-down tilt (HDT). However, there is no comparison of arterial and venous systems parameters with blood flow changes in organs and tissues. Apart from the preliminary HDT training influence on these parameters is unknown. Thus we aimed to evaluate the role of HDT training in central circulation responses, venous tone and regional blood flow.

Animals↗

Studies on atrophic change of soleus muscle and its countermeasures in suspended rat.

In the environment of microgravity, the disused atrophy of skeletal muscle, especially leg's muscle, would occur. The three purposes of this study were: 1. To observe the dynamic changes of disused atrophy of skeletal muscle under simulated weightlessness; 2. To approach the mechanism of disused atrophy of muscle; 3. To approach the countermeasures for reducing the degree of atrophy of muscle.

Animals↗

Effectiveness of a cytokine restraining agent (CRA (TM)) in attenuating disuse deconditioning induced by hindlimb unloading in rats.

Spaceflight alters many immune responses and among the regulatory components of an organisms response system that have been to be affected by spaceflight is the cytokine network. Spaceflight, as well as ground-based model systems of spaceflight, have been shown to affect the production and activation of various cytokines including interleukins (IL) and tumor necrosis factor (TNF). Levels of urinary IL-2 are elevated on the first day of spaceflight and again after returning from space. Most results from ground-based studies in rodents indicate either no alterations in cytokines or decreased levels. Results from this experiment indicate that HP 228, a potent cytokine restraining agent (CRA (TM)) was effective in attentuating many of the disuse deconditioning changes induced by the ground-based hindlimb suspension model that simulates weightlessness in rats. HP 228 is a novel heptapeptide with unnatural amino acids and can effectively restrain lipopolysaccharide (LPS)-induced increased levels of several key cytokines, including plasma TNF alpha, IL-1 beta and IL-6. HP 228 has also been shown to be effective in several rodent models of pain, inflammation and LPS-induced lethality, as well as in reducing inducible nitric oxide synthase.

Animals↗

Alterations in glomerular and tubular dynamics at 1 and 14 days simulated microgravity and after acute return to orthostasis.

Head-down tilt (HDT) is utilized to simulate microgravity and produces a cephalad fluid shift, which results in alterations in fluid and electrolyte balance. These changes in volume homeostasis are due, in part, to alterations in multiple volume control mechanisms in which renal function is a major participant. We have previously demonstrated that glomerular filtration rate increases early in HDT and eventually returns to values not different from non-tilt measurements. This early increase in glomerular filtration rate was also demonstrated during days 2 and 8 of the SLS-1 mission. However, urine flow and electrolyte excretion does not parallel the alterations in glomerular filtration rate and the site of this change in nephron fluid reabsorption pattern has not been previously examined. Through determination of the location of alterations in tubular fluid reabsorption within the nephron, a more detailed hypothesis can be forwarded as to which specific neuro-humoral agents participating in control of renal function in microgravity conditions. The importance of this type of examination is that measurements in circulating neuro-humoral agents and urinary excretion patterns alone are not accurate predictors of how renal functional response may alter to head-down tilt or other models of simulated weightlessness. To examine this issue, renal micropuncture techniques were utilized in Munich-Wistar rats submitted 24 hours and 14 day head-down tilt, measuring all the determinants of glomerular ultrafiltration and obtaining data regarding segmental tubular fluid reabsorption. Following these measurements, the rats were returned to an orthostatic position and after 60 min, the measurements were repeated.

Animals↗

Changes in bone noncollagenous proteins and bone mineral loss in lumbar vertebrae of tail-suspended rats.

The purpose of the study is to clarify whether the noncollagenous proteins play a role in bone mineral loss and whether the quality of osteocalcin (OC) is deteriorated in simulated weightlessness. Noncollagenous proteins, such as OC and albumin, have been shown to be effective in promoting bone mineralization. In this study, the contents of OC, albumin, and mineral in the third lumbar vertebra (L3) were determined in rats suspended by tail for 21 days. The concentration of serum-immunoreactive osteocalcin (irOC) were also measured before and after adsorption to hydroxylapatite. The results showed that the contents of mineral, osteocalcin, and albumin in L3 were significantly decreased in suspended rats. The concentration of serum irOC and serum irOC with a high affinity to hydroxylapatite (irOCbound) were significantly decreased, but the low affinity (irOCfree) remained unaffected. The data suggest that the mineral loss in L3 of suspended rats is related to the decreased contents of osteocalcin, and albumin in bone mass. The incomplete gamma-carboxylation of osteocalcin in suspended rats is also one of the important causes of bone loss.

Albumins↗