Orientation of Paramecium under the conditions of weightlessness

A cell culture of Paramecium with a precise negative gravitaxis was exposed to 4 x l0(-6) g during a parabolic flight of a sounding rocket for 6 min. Computer image analysis revealed that without gravity stimulus the individual swimming paths remained straight. In addition, three reactions could be distinguished. For about 30 s, paramecia maintained the swimming direction they had before onset of low gravity. During the next 20 s, an approximate reversal of the swimming direction occurred. This period was followed by the expected random swimming pattern. Similar behavior was observed under the condition of simulated weightlessness on a fast-rotating clinostat. Control experiments on the ground under hyper-gravity on a low-speed centrifuge microscope and on a vibration test facility proved that the observed effects were caused exclusively by the reduction of gravity.     

Variable acceleration influences cyclic AMP levels in Paramecium biaurelia

Paramecium is used as a model system to analyse the gravity signal transduction pathway, that leads to gravitaxis and gravikinesis. In order to prove whether gravistimulation is coupled with second messenger production (cyclic AMP: hyperpolarization, cyclic GMP: depolarization) Paramecium was fixated under variable accelerations (1 x g, 9 x g and 10(-4) x g) on a centrifuge and during a sounding rocket flight (TEXUS 39). The analysis of cAMP and cGMP levels revealed an acceleration-dependent change in cAMP, while cGMP-levels showed gravity-independent variations. Hypergravity did not only induce an amplification of gravitaxis and gravikinesis, but also an increase in cAMP compared to the 1 x g-data. We conclude that the increased pressure of the cytoplasm on the lower membrane of upward swimming cells enhance the number of open K+(-)channels, thus causing hyperpolarization and change in cAMP concentration. Consequently, transition to microgravity declines gravitaxis and gravikinesis, and decreases cAMP concentration due to the loss of pressure on the cell membrane.     

Oriented movement of statoliths studied in a reduced gravitational field during parabolic flights of rockets

During five rocket flights (TEXUS 18, 19, 21, 23 and 25), experiments were performed to investigate the behaviour of statoliths in rhizoids of the green alga Chara globularia Thuill. and in statocytes of cress (Lepidium sativum L.) roots, when the gravitational field changed to approx. 10^–4 · g (i.e. microgravity) during the parabolic flight (lasting for 301–390 s) of the rockets. The position of statoliths was only slightly influenced by the conditions during launch, e.g. vibration, acceleration and rotation of the rocket. Within approx. 6 min of microgravity conditions the shape of the statolith complex in the rhizoids changed from a transversely oriented lens into a longitudinally oriented spindle. The center of the statolith complex moved approx. 14 m and 3.6 m in rhizoids and root statocytes, respectively, in the opposite direction to the originally acting gravity vector. The kinetics of statolith displacement in rhizoids demonstrate that the velocity was nearly constant under microgravity whereas it decreased remarkably after inversion of rhizoids on Earth. It can be concluded that on Earth the position of statoliths in both rhizoids and root statocytes depends on the balance of two forces, i.e. the gravitational force and the counteracting force mediated by microfilaments.Abbreviations ER endoplasmic reticulum - g 9.806 m · s^–2 - MF microfilament - TEXUS Technologische Experimente unter Schwerelosigkeit (technological experiments under reduced gravity) Dedicated to Professor Wolfgang Haupt on the occasion of his 70th birthday     

Recognizing and optimizing flight opportunities with hardware and life sciences limitations

The availability of orbital space flight opportunities to conduct life sciences research has been limited. It is possible to use parabolic flight and sounding rocket programs to conduct some kinds of experiments during short episodes (seconds to minutes) of reduced gravity, but there are constraints and limitations to these programs. Orbital flight opportunities are major undertakings, and the potential science achievable is often a function of the flight hardware available. A variety of generic types of flight hardware have been developed and tested, and show great promise for use during NSTS flights. One such payload configuration is described which has already flown.     

Physiological parameters of gravitaxis in the flagellate Euglena gracilis obtained during a parabolic flight campaign

The unicellular freshwater flagellate Euglena gracilis and its close relative Astasia longa show a pronounced negative gravitaxis. Previous experiments revealed that gravitaxis is most likely mediated by an active physiological mechanism in which changes of the internal calcium concentration and the membrane potential play an important role. In a recent parabolic flight experiment on board an aircraft (ESA 29th parabolic flight campaign), changes of graviorientation, membrane potential and the cytosolic calcium concentration upon changes of the acceleration (between 1 x g(n), 1.8 x g(n), microgravity) were monitored by image analysis and photometric methods using Oxonol VI (membrane potential) and Calcium Crimson (cytosolic calcium concentration). The parabolic flight maneuvers performed by the aircraft resulted in transient phases of 1.8 x g(n) (about 20 s), microgravity (about 22 s) followed by 1.8 x g(n) (about 20 s). A transient increase in the intracellular calcium concentration was detected from lower to higher accelerations (1 x g(n) to 1.8 x g(n) or microgravity to 1.8 x g(n)). Oxonol VI-labeled cells showed a signal, which indicates a depolarization during the transition from 1 x g(n) to 1.8 x g(n), a weak repolarization in microgravity followed by a rapid repolarization in the subsequent 1 x g(n) phase. The results show good coincidence with observations of recent terrestrial and space experiments.     

Enzyme catalysis in microgravity: steady-state kinetic analysis of the isocitrate lyase reaction

Two decades of research in microgravity have shown that certain biochemical processes can be altered by weightlessness. Approximately 10 years ago, our team, supported by the European Space Agency (ESA) and the Agenzia Spaziale Italiana, started the Effect of Microgravity on Enzyme Catalysis project to test the possibility that the microgravity effect observed at cellular level could be mediated by enzyme reactions. An experiment to study the cleavage reaction catalyzed by isocitrate lyase was flown on the sounding rocket MASER 7, and we found that the kinetic parameters were not altered by microgravity. During the 28th ESA parabolic flight campaign, we had the opportunity to replicate the MASER 7 experiment and to perform a complete steady-state analysis of the isocitrate lyase reaction. This study showed that both in microgravity and in standard g controls the enzyme reaction obeyed the same kinetic mechanism and none of the kinetic parameters, nor the equilibrium constant of the overall reaction were altered. Our results contrast with those of a similar experiment, which was performed during the same parabolic flight campaign, and showed that microgravity increased the affinity of lipoxygenase-1 for linoleic acid. The hypotheses suggested to explain this change effect of the latter were here tested by computer simulation, and appeared to be inconsistent with the experimental outcome.     

Gravity-dependent changes in bioconvection of Tetrahymena and Chlamydomonas during parabolic flight: increases in wave number induced by pre- and post-parabola hypergravity

Bioconvection emerges in a dense suspension of swimming protists as a consequence of their negative-gravitactic upward migration and later settling as a blob of density greater than that of water. Thus, gravity is an important parameter governing bioconvective pattern formation. However, inconsistencies are found in previous studies dealing with the response of bioconvection patterns to increased gravity acceleration (hypergravity); the wave number of the patterns has been reported to decrease during the hypergravity phases of parabolic aircraft flight, while it increases in centrifugal hypergravity. In this paper, we reassess the responses of bioconvection to altered gravity during parabolic flight on the basis of vertical and horizontal observations of the patterns formed by Tetrahymena thermophila and Chlamydomonas reinhardtii. Spatiotemporal analyses of the horizontal patterns revealed an increase in the pattern wave number in both pre- and post-parabola hypergravity. Vertical pattern analysis was generally in line with the horizontal pattern analysis, and further revealed that hypergravity-induced changes preceded at the top layer of the suspensions while microgravity-induced changes appeared to occur from the bottom part of the settling blobs. The responses to altered gravity were rather different between the two sample species: T. thermophila tended to drastically modify its bioconvection patterns in response to changes in gravity level, while the patterns of C. reinhardtii responded to a much lesser extent. This difference can be attributed to the distinct physical and physiological properties of the individual organisms, suggesting a significant contribution of the gyrotactic property to the swimming behavior of some protists.     

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.     

Graviresponses of Paramecium biaurelia during parabolic flights

The thresholds of graviorientation and gravikinesis in Paramecium biaurelia were investigated during the 5th DLR (German Aerospace Center) parabolic-flight campaign at Bordeaux in June 2003. Parabolic flights are a useful tool for the investigation of swimming behaviour in protists at different accelerations. At normal gravity (1 g) and hypergravity (1 g to 1.8 g), precision of orientation and locomotion rates depend linearly on the applied acceleration as seen in earlier centrifuge experiments. After transition from hypergravity to decreased gravity (minimal residual acceleration of <10(-2) g), graviorientation as well as gravikinesis show a full relaxation with different kinetics. The use of twelve independent cell samples per flight guarantees high data numbers and secures the statistical significance of the obtained data. The relatively slow change of acceleration between periods of microgravity and hypergravity (0.4 g/s) enabled us to determine the thresholds of graviorientation at 0.6 g and of gravikinesis at 0.4 g. The gravity-unrelated propulsion rate of the sample was found to be 874 microm/s, exceeding the locomotion rate of horizontally swimming cells (855 microm/s). The measured thresholds of graviresponses were compared with data obtained from earlier centrifuge experiments on the sounding rocket Maxus-2. Measured thresholds of gravireactions indicate that small energies, close to the thermal noise level, are sufficient for the gravitransduction process. Data from earlier hypergravity experiments demonstrate that mechanosensitive ion channels are functioning over a relative wide range of acceleration. From this, we may speculate that gravireceptor channels derive from mechanoreceptor channels.     

Effects of long-term space flights on organization of horizontal gaze fixation reaction

Results of Russian-Austrian space experiment "Monimir" which was a part of international space program "Austromir" are presented in this paper. Characteristics of horizontal gaze fixation reaction (hGFR) to visual targets were analyzed. Seven crewmembers of "Mir" space station expeditions took part in the experiment. Experiments were carried out 4 times before space flight, 5 times in flight and 3-4 times after landing. There were revealed significant alterations in characteristics of gaze fixation reaction during flight and after its accomplishing, namely: an increase of the time of gaze fixation to the target, changes of eye and head movements' velocity and increase of the gain of vestibular-ocular reflex, that pointed out to the disturbances of the control mechanisms of vestibular-ocular reflex in weightlessness caused by changes of vestibular input's activity. There was discovered also the difference in the strategies of adaptation to microgravity conditions among the cosmonauts of flight and non-flight occupation: in the first group exposure to weightlessness was accompanied by gaze hypermetry and inhibition of head movements; in the second one--on the contrary--by increase of head movement velocity and decrease of saccades' velocity.     

How to activate a plant gravireceptor. Early mechanisms of gravity sensing studied in characean rhizoids during parabolic flights

Early processes underlying plant gravity sensing were investigated in rhizoids of Chara globularis under microgravity conditions provided by parabolic flights of the A300-Zero-G aircraft and of sounding rockets. By applying centrifugal forces during the microgravity phases of sounding rocket flights, lateral accelerations of 0.14 g, but not of 0.05 g, resulted in a displacement of statoliths. Settling of statoliths onto the subapical plasma membrane initiated the gravitropic response. Since actin controls the positioning of statoliths and restricts sedimentation of statoliths in these cells, it can be calculated that lateral actomyosin forces in a range of 2 x 10(-14) n act on statoliths to keep them in place. These forces represent the threshold value that has to be exceeded by any lateral acceleration stimulus for statolith sedimentation and gravisensing to occur. When rhizoids were gravistimulated during parabolic plane flights, the curvature angles of the flight samples, whose sedimented statoliths became weightless for 22 s during the 31 microgravity phases, were not different from those of in-flight 1g controls. However, in ground control experiments, curvature responses were drastically reduced when the contact of statoliths with the plasma membrane was intermittently interrupted by inverting gravistimulated cells for less than 10 s. Increasing the weight of sedimented statoliths by lateral centrifugation did not enhance the gravitropic response. These results provide evidence that graviperception in characean rhizoids requires contact of statoliths with membrane-bound receptor molecules rather than pressure or tension exerted by the weight of statoliths.     

Head out of water immersion as simulation study: an heart rate variability study

During head out of water immersion (HOI), the hydrostatic pressure on the tissues of the lower limbs causes an increase in thoracic blood volume and a high vascular perfusion. This blood shift results in changes in autonomic balance. The aim of this study was to evaluate the activity of the autonomic nervous system, as obtained from the analysis of heart rate variability (HRV), during HOI at 2 different temperatures and compare these results with data obtained during parabolic flight. In HOI, two different positions (sitting and standing) were compared. Results showed a shift to vagal activity as well during HOI as in microgravity during parabolic flight.     

Graviperception in the flagellate Euglena gracilis during a shuttle space flight

During a recent space flight, gravitaxis of the unicellular photosynthetic flagellate, Euglena gracilis, was studied on board of the American shuttle Columbia. Accelerations were varied between 0 and 1.5 x g using a slow rotating centrifuge microscope (NIZEMI). The cells showed a sigmoidal response curve for the dependence of the precision of gravitaxis on acceleration which is indicative of the involvement of an active, physiological gravireceptor with a threshold at g-values < or = 0.16 x g and a saturation at g-values > or = 1 x g. No adaptation to microgravity was found during the prolonged space mission. After return the cells showed a normal gravitactic behavior at 1 x g. Since the cells are heavier than water, their swimming velocity is affected by sedimentation. The velocity distribution at different accelerations closely follows Stokes' law for sedimentation indicating that, in contrast to the ciliate Paramecium, E. gracilis, does not show any gravikinesis.     

Age and serotonin effects on locomotion in marine trematode cercariae

The effect of serotonin solutions on the locomotor activity of cercariae with different swimming patterns, Cryptocotyle lingua and Himasthla elongate, was investigated during their lifespan using motion track analysis and a frame-by-frame count of behavioral elements. Serotonin caused a noticeable alteration of several locomotion parameters in both species studied, while the activity per se decreased significantly with age. In intermittently swimming C. lingua cercariae, serotonin induced an increase in the frequency of active swimming phases (spurts) without significant changes in the duration and velocity of the spurts themselves. In continuously swimming H. elongata cercariae, serotonin increased the number of stops during the first hour after the emission from the host mollusk under a constant average swimming velocity. Immunocytochemical visualization of serotonin revealed a decline in its level with age (time after emission) and different dynamics of this process in neurons located in the tail and body of C. lingua cercariae. The data are interpreted in the light of the modulatory role of serotonin known for other animals.     

Effect of propelling surface size on the mechanics and energetics of front crawl swimming

In swimming the propulsive force is generated by giving a velocity change to masses of water. In this process energy is transferred from the swimmer to the water, which cannot be used to propel the swimmer. Theoretical considerations indicated that an increase of the propelling surface size should lead to a reduced loss of energy to the water. Thus, in this study, the effect of artificially enlarging the propelling surface of the hand was examined. The effect was examined in terms of the propelling efficiency during front crawl swimming using the arms alone. The legs were floated with a small buoy as previously described (Toussaint et al., J. appl. Physiol. 65, 2506-2512, 1988a). In ten competitive swimmers (six male, four female) the rate of energy expenditure (power input, Pi), power output (Po), work per stroke cycle (As), distance per stroke cycle (d), work per unit distance (Ad), and propelling efficiency (ep) were determined at various swimming speeds once with and once swimming without paddles. At the same average velocity the effect of swimming with paddles was to reduce Pi, Po, and Ad by 6, 7.6, and 7.5% respectively, but to increase ep and As by 7.8 and 7%. The increase in distance per stroke cycle and the decrease in stroke cycle frequency matched the predicted values based on the theoretical considerations in which the actual increase in propelling surface size was taken into account.     

Cardiovascular adaptations to parabolic flight in rats: a radio-telemetry feasibility study

This experiment was a feasibility study which consisted in investigating arterial blood pressure and heart rate to transient and repeated exposure to microgravity in eight unrestrained rats previously implanted with radio-telemetry transmitter. The aim was to perform such recordings throughout all the phases of a parabola during parabolic flights. This study revealed that it was possible to collect the radio-signal without any interference with electronic or magnetic environment. We observed in microgravity a significant reduction in heart rate (6%) and a significant increase in arterial blood pressure (7%). In conclusion, such a study seems to be feasible during longer exposure to microgravity (space flight) in order to study the cardiovascular adaptation in rat.     

The kinematics of Heteroptera in free flight

The kinematics of six species of Heteroptera in free flight are analysed and compared.

• (1) 

  Using nested analysis of variance techniques, statistically significant variation was detected between species for several of the flight parameters measured: mean angular velocity; pronation/supination ratio; upstroke/downstroke ratio; and wing beat frequency. In each case this is discussed in terms of variation in flight behaviour.
• (2) 

  Beneficial aerodynamic forces are generated during the upstroke and the downstroke, in both fast forward and rising flight.
• (3). 

  When the insects change from level, forward flight to near vertical, rising flight, the following parameters are altered in most of the sequences analysed:

• (a). 

  the stroke plane angle becomes steeply, negatively inclined, associated with an increase in body angle;
• (b). 

  the stroke amplitude is reduced;
• (c). 

  wing beat frequency is lowered, associated with a drop in mean angular velocity;
• (d). 

  the speed of stroke reversal (rotational velocity) is increased. This may be associated with increased wing torsion and tip flexion which in turn could improve any beneficial unsteady aerodynamic effects generated at stroke reversal.

The reasons for this change in flight performance and the deviations from that seen in other insects are discussed.
It is shown that Heteroptera may make use of wing drag in flight, particularly during rising flight.     

The cost of chronic stress: impacts of a nonhabituating stress response on metabolic variables and swimming performance in sturgeon

Metabolic scope for activity (MSA) and critical swimming velocity (U(crit)) were measured in green sturgeon exposed to two stressors daily for 28 consecutive days. The results were compared with unstressed fish in an effort to measure the "cost" of chronic stress. Chronic stress was simulated by exposing fish to a randomized order of acute stressors: a 5-min chasing stressor, a 10-min water depth reduction stressor, or a 5-min confinement stressor. The acute cortisol response to each stressor was initially determined, and the maintenance of that response was verified in 7-d intervals during the chronic stress regime. Exposure to the chronic stress regime resulted in a 25% reduction of MSA caused by significantly increased maintenance metabolic rate (0.27+/-0.01 vs. 0.19+/-0.02 mg O(2) h(-1) g(-1), chronic and control fish, respectively) but did not affect the U(crit) of sturgeon. In addition, a 50% reduction in liver glycogen levels and a twofold increase of resting plasma glucose levels were measured in chronically stressed fish. We conclude that our chronic stress regime resulted in a significant maintenance cost to green sturgeon, possibly because of their inability to habituate to the stressors, but did not decrease their swimming performance.     

亚成体巨须裂腹鱼游泳能力及活动代谢研究

以野生雅鲁藏布江巨须裂腹鱼(Schizothorax macropogon)为对象,通过自制的鱼类游泳实验装置,测定了4个温度(5、10、15和18℃)梯度下亚成体巨须裂腹鱼的临界游泳速度(Ucrit)及流速变化对耗氧率的影响,并通过摄像记录分析了不同游泳速度下的游泳行为。野生亚成体巨须裂腹鱼的临界游速随着温度的变化呈近似线性的递增趋势(P<0.001),4个温度下的绝对临界游速(Ucrit-a)分别为(0.88±0.07)、(1.09±0.07)、(1.24±0.15)和(1.49±0.15)m/s;若以单位时间内游过的体长倍数(BL/s)表示,相对临界游速(Ucrit-r)分别为(3.96±0.21)、(4.4±0.16)、(4.9±0.18)和(5.35±0.14)BL/s。根据不同温度及流速下耗氧率的变化情况,采用非线性拟合得到了4个温度梯度下耗氧率与游泳速度关系的幂函数模型(P<0.05)。模型表明耗氧率随游泳速度的增大而增加,且温度越高耗氧率随游泳速度的变化越显著。4个温度下的速度指数分别为2.4、2.6,2.8及3.1,表明有氧运动的效率随温度升高有所降低。在自然水温条件下(5—9℃),摆尾频率(TBF)与流速的关系呈线性正相关(P<0.001),而运动步长(Ls)的变化与流速没有显著关系,出现由高至低再升高的三个阶段。录像分析表明在流速逐渐增加的过程中,巨须裂腹鱼采用了三种不同的游泳方式,以实现降低能量消耗的目的。研究可为鱼道等过鱼设施的设计提供参考,对数量日益减少的巨须裂腹鱼保护具有较大的意义。     

The effect of clinorotation on vestibular compensation in upside-down swimming catfish.

Upside-down swimming catfish Synodontis nigriventris can keep upside-down swimming posture stably under pseudo-microgravity generated by clinostat. When the vestibular organ is unilaterally ablated, the operated S. nigriventris shows disturbed swimming postures under the clinorotation condition. However, about 1 month after the operation, unilateral vestibular organ-ablated S. nigriventris shows stable upside-down swimming posture under the condition (vestibular compensation). In contrast, a closely related upside-up swimming catfish Synodontis multipunctatus belonging to same Synodontis family can not keep stable swimming postures under the clinorotation conditions. In this study, we examined the effect of continuous clinorotation on vestibular compensation in intact and unilateral vestibular organ-ablated Synodontis nigriventris and Synodontis multipunctatus. After the exposure to continuous clinorotation, the postures of the catfish were observed under microgravity provided by parabolic flights of an aircraft. Unilateral vestibular organ-ablated S. nigriventris which had been exposed to continuous clinorotation showed stable swimming postures and did not show dorsal light reaction (DLR) under microgravity. This postural control pattern of the operated catfish was similar to that of intact catfish. Intact and unilateral vestibular organ-ablated S. multipunctatus showed DLR during microgravity. Our results confirmed that S. nigriventris has a novel balance sensation which is not affected by microgravity. DLR seems not to play an important role in postural control. It remains unclear that the continuous clinorotation effects on vestibular compensation because we could not keep used unilateral vestibular organ-ablated fish alive under continuous clinorotation for uninterrupted 25 days. This study suggests that space flight experiments are required to explore whether gravity information is essential for vestibular compensation.