Gravitational biology and the mammalian circadian timing system.

Mammals have evolved under the influence of many selective pressures. Two of these pressures have been the static force of gravity and the daily variations in the environment due to the rotation of the earth. It is now clear that each of these pressures has led to specific adaptations which influence how organisms respond to changes in either gravity or daily time cues. However, several unpredicted responses to altered gravitational environments occur within the homeostatic and circadian control systems. These results may be particularly relevant to biological and medical issues related to spaceflight. This paper demonstrates that the homeostatic regulation of rat body temperature, heart rate, and activity become depressed following exposure to a 2 G hyperdynamic field, and recovers within 5-6 days. In addition, the circadian rhythms of these same variables exhibit a depression of rhythm amplitude; however, recovery required a minimum of 7 days.     

Calcium signaling in plant cells in altered gravity.

Changes in the intracellular Ca2+ concentration in altered gravity (microgravity and clinostating) evidence that Ca2+ signaling can play a fundamental role in biological effects of microgravity. Calcium as a second messenger is known to play a crucial role in stimulus-response coupling for many plant cellular signaling pathways. Its messenger functions are realized by transient changes in the cytosolic ion concentration induced by a variety of internal and external stimuli such as light, hormones, temperature, anoxia, salinity, and gravity. Although the first data on the changes in the calcium balance in plant cells under the influence of altered gravity have appeared in 80th, a review highlighting the performed research and the possible significance of such Ca2+ changes in the structural and metabolic rearrangements of plant cells in altered gravity is still lacking. In this paper, an attempt was made to summarize the available experimental results and to consider some hypotheses in this field of research. It is proposed to distinguish between cell gravisensing and cell graviperception; the former is related to cell structure and metabolism stability in the gravitational field and their changes in microgravity (cells not specialized to gravity perception), the latter is related to active use of a gravitational stimulus by cells presumebly specialized to gravity perception for realization of normal space orientation, growth, and vital activity (gravitropism, gravitaxis) in plants. The main experimental data concerning both redistribution of free Ca2+ ions in plant cell organelles and the cell wall, and an increase in the intracellular Ca2+ concentration under the influence of altered gravity are presented. Based on the gravitational decompensation hypothesis, the consequence of events occurring in gravisensing cells not specialized to gravity perception under altered gravity are considered in the following order: changes in the cytoplasmic membrane surface tension --> alterations in the physicochemical properties of the membrane --> changes in membrane permeability, --> ion transport, membrane-bound enzyme activity, etc. --> metabolism rearrangements --> physiological responses. An analysis of data available on biological effects of altered gravity at the cellular level allows one to conclude that microgravity environment appears to affect cytoskeleton, carbohydrate and lipid metabolism, cell wall biogenesis via changes in enzyme activity and protein expression, with involvement of regulatory Ca2+ messenger system. Changes in Ca2+ influx/efflux and possible pathways of Ca2+ signaling in plant cell biochemical regulation in altered gravity are discussed.     

The biological clock of Neurospora in a microgravity environment.

The circadian rhythm of conidiation in Neurospora crassa is thought to be an endogenously derived circadian oscillation; however, several investigators have suggested that circadian rhythms may, instead, be driven by some geophysical time cue(s). An experiment was conducted on space shuttle flight STS-9 in order to test this hypothesis; during the first 7-8 cycles in space, there were several minor alterations observed in the conidiation rhythm, including an increase in the period of the oscillation, an increase in the variability of the growth rate and a diminished rhythm amplitude, which eventually damped out in 25% of the flight tubes. On day seven of flight, the tubes were exposed to light while their growth fronts were marked. Some aspect of the marking process reinstated a robust rhythm in all the tubes which continued throughout the remainder of the flight. These results from the last 86 hours of flight demonstrated that the rhythm can persist in space. Since the aberrant rhythmicity occurred prior to the marking procedure, but not after, it was hypothesized that the damping on STS-9 may have resulted from the hypergravity pulse of launch. To test this hypothesis, we conducted investigations into the effects of altered gravitational forces on conidiation. Exposure to hypergravity (via centrifugation), simulated microgravity (via the use of a clinostat) and altered orientations (via alterations in the vector of a 1 g force) were used to examine the effects of gravity upon the circadian rhythm of conidiation.     

Adaptations of the vestibular system to short and long-term exposures to altered gravity.

Long-term space flight creates unique environmental conditions to which the vestibular system must adapt for optimal survival of a given organism. The development and maintenance of vestibular connections are controlled by environmental gravitational stimulation as well as genetically controlled molecular interactions. This paper describes the effects of hypergravity on axonal growth and dendritic morphology, respectively. Two aspects of this vestibular adaptation are examined: (1) How does long-term exposure to hypergravity affect the development of vestibular axons? (2) How does short-term exposure to extremely rapid changes in gravity, such as those that occur during shuttle launch and landing, affect dendrites of the vestibulocerebellar system? To study the effects of longterm exposures to altered gravity, embryonic rats that developed in hypergravity were compared to microgravity-exposed and control rats. Examination of the vestibular projections from epithelia devoted to linear and angular acceleration revealed that the terminal fields segregate differently in rat embryos that gestated in each of the gravitational environments.To study the effects of short-term exposures to altered gravity, mice were exposed briefly to strong vestibular stimuli and the vestibulocerebellum was examined for any resulting morphological changes. My data show that these stimuli cause intense vestibular excitation of cerebellar Purkinje cells, which induce up-regulation of clathrin-mediated endocytosis and other morphological changes that are comparable to those seen in long-term depression. This system provides a basis for studying how the vestibular environment can modify cerebellar function, allowing animals to adapt to new environments.     

Exposure to altered gravity affects all stages of endochondral cartilage differentiation.

Chondrogenesis has a number of well-defined steps: (1) condensation, which involves cell aggregation, adhesion and communication; (2) activation of cartilage genes, which is accompanied by rounding up of the cells and intracellular differentiation; and (3) production and secretion of cartilage specific matrix molecules. Our studies show that each of these steps is affected by exposure to gravitational changes. Clinorotation and centrifugation affected initial aggregation and condensation. In the CELLS experiment, where cells were exposed to microgravity after some condensation occurred preflight, intracellular differentiation and matrix production were delayed relative to controls. Once cartilage has developed, in rats, further differentiation (hypertrophy, matrix production) was also affected by spaceflight and hind limb suspension. For the process of chondrogenesis to proceed as we know it, loading and other factors present at 1g are required at each step of the process. This requirement means that not only will skeletal development and bone healing, processes involving chondrogenesis, be altered by long term exposure to microgravity, but that continuous intervention will be necessary to correct any defects produced by altered gravity environments.     

Physico-chemical characteristics of biomembranes and cell gravisensitivity.

The resistance of living systems to the action of environmental factors is known to be largely determined by molecular organization of biomembranes constituting the basis of the cell per se and of all intracellular organelles. Gravity as one of the environmental factors, plays a definite role in the vital activity of organisms. Therefore, the problem of altered gravity impact on biological objects should be considered in close relation to the functional state of membranes and contractible elements of cytoskeleton. Moreover, the involvement of membrane structures and cytoskeleton in the processes of reception and realization of gravitational stimulus allows us to evaluate the extent of the direct or indirect influence of gravity on cell functioning in the gravitational field. The results of experimental studies having been conducted up to this time on a variety of cells and cell organelles under altered gravity conditions demonstrated noticeable alterations in the molecular organization of the membranes.     

Neuroplastic reactivity of fish induced by altered gravity conditions: a review of recent results.

A review is being presented concerning behavioural, biochemical, histochemical and electronmicroscopical data on the influence of altered gravitational forces on the swimming performance and on the neuronal differentiation of the brain of cichlid fish larvae and adult swordtail fish that had been exposed to hyper-gravity (3g in laboratory centrifuges), hypo-gravity (>10(-2) g in a fast-rotating clinostat) and to near weightlessness (10(-4) g aboard the Spacelab D-2 mission). After long-term alterations of gravity (and parallel light deprivation), initial disturbances in the swimming behaviour followed by a stepwise regain of normal swimming modes are induced. Parallel, neuroplastic reactivities on different levels of investigation were found, such as adaptive alterations of activities of various enzymes in whole brain as well as in specific neuronal integration centers and an intraneuronal reactivity on ultrastructural level in individual brain parts and in the sensory epithelia of the inner ear. Taken together, these data reveal distinct adaptive neuroplastic reactions of fish to altered gravity conditions.     

Biorhythms on cellular and organismic level (introduction).

The regular change of day and night, of light and darkness during millions of years has strongly affected the development of life on earth. Many organisms adapted themselves to this environmental condition and, finally, evolved an endogenous timer which usually is in phase with the earth's rotation and causes many functions to perform one oscillation per day. Such circadian rhythms (derived from circa dies i.e. about 1 day) were found in almost all classes of plants and animals, and even in protozoans. They persist in a constant environment and, therefore, are independent of any known external trigger signals. Since even unicells perform circadian rhythms which are similar to those observed in highly developed multicellular organisms many scientists favor the existence of a basic mechanism common to all kinds of biological clocks that is located somewhere in the single cell and probably comprises many different biochemical reactions. One purpose of this topical meeting was to discuss how organisms respond to the absence of gravity and terrestrial zeitgeber and how they may react to the imposing of hypergravity fields. Another aim was to develop model-mechanisms appropriate to describe these responses.     

Crickets in space: morphological, physiological and behavioral alterations induced by space flight and hypergravity.

"Crickets in Space" was a Neurolab experiment by which the balance between genetic programs and the gravitational environment for the development of a gravity sensitive neuronal system was studied. The model character of crickets was justified by their external gravity receptors, identified position-sensitive interneurons (PSI) and gravity-related compensatory head response, and by the specific relation of this behavior to neuronal arousal systems activated by locomotion. These advantages allowed to study the impact of modified gravity on cellular processes in a complex organism. Eggs, 1st, 4th and 6th stage larvae of Acheta domesticus were used. Post-flight experiments revealed a low susceptibility of the behavior to micro- and hypergravity while the physiology of the PSI was significantly affected. Immunocytological investigations revealed a stage-dependent sensitivity of thoracic GABAergic motoneurons to 3 g-conditions concerning their soma sizes but not their topographical arrangement. The morphology of neuromuscular junctions was not affected by 3 g-hypergravity. Peptidergic neurons from cerebral sensorimotor centers revealed no significant modifications by microgravity (micro g). The contrary physiological and behavioral results indicate a facilitation of 1 g-readaptation originating from accessory gravity, proprioceptive and visual sense organs. Absence of anatomical modifications point to an effective time window of micro g or 3 g-expo-sure related to the period of neuronal proliferation. The analysis of basic mechanisms of how animals and man adapt to altered gravitational conditions will profit from a continuation of the project "Crickets in Space".     

空间里的时间:微重力等环境下的生物节律研究

生物钟是地球上的生物为适应环境周期性变化经历长期演化而来的内在机制.在分子水平上受生物钟基因及其他相关基因的调节;在组织水平上,生物钟由主生物钟和外周生物钟组成.生物钟对于各种生物的生理、认知和行为等具有重要功能,是生物适应环境的决定因素之一.空间环境下的微重力、辐射、光照条件、社会性因素等与地面存在很大差异,这些因素均可能导致节律紊乱,影响生物的生理及环境适应性.因此,对地外生命的研究也应该考虑生物钟因素.对航天员而言,节律紊乱可引起睡眠障碍,并且对骨肌系统、神经系统、心血管系统及内分泌系统等造成不利影响,导致人的认知和工效水平下降.在未来空间生命探索以及航天员健康保障研究中,生物钟是一个不可忽视的重要因素.      

Preliminary characterization of persisting circadian rhythms during space flight.

In order to evaluate the function of the circadian timing system in space, the circadian rhythm of conidiation of the fungus Neurospora crassa was monitored in constant darkness on the STS 9 flight of the Space Shuttle Columbia. During the first 7 days of spaceflight many tubes showed a marked reduction in the apparent amplitude of the conidiation rhythm, and some cultures appeared arrhythmic. There was more variability in the growth rate and circadian rhythms of individual cultures in space than is usually seen on earth. The results of this experiment indicate that while the circadian rhythm of Neurospora conidiation can persist outside of the Earth's environment, either the timekeeping process or its expression is altered in space.     

Teratogenic effects of gravitational changes.

In investigating the effect of gravitational changes on development, it is instructive to think of altered gravity (delta g) as a teratogen--that is, an environmental factor influencing development. Observed effects on skeletal development include: suppression of morphogenesis in centrifuged mouse limb buds; advanced fusion stages in centrifuged mouse palates; smaller crown rump lengths (CRL) and decreased number of pregnancies in centrifuged rats and mice; altered differentiation of growth plates in young growing rats in space; and decreased length of calcified long bone regions in fetal rats exposed to microgravity in utero. These studies show that delta g is able to alter development in vivo and in vitro and suggest that delta g operates, at least in part, at the cellular level.     

Environmental impacts on the developing CNS: CD15, NCAM-L1, and GFAP expression in rat neonates exposed to hypergravity.

We have previously reported that the developing rat cerebellum is affected by hypergravity exposure. The effect is observed during a period of both granule and glial cell proliferation and neuronal migration in the cerebellum and coincides with changes in thyroid hormone levels. The present study begins to address the molecular mechanisms involved in the cerebellar response to hypergravity. Specifically, the study focuses on the expression of cerebellar proteins that are known to be directly involved in cell-cell interactions [protein expressing 3-fucosyl-N-acetyl-lactosamine antigen (CD15), neuronal cell adhesion molecule (NCAM-L1)] and those that affect cell-cell interactions indirectly [glial fibrillary acidic protein (GFAP)] in rat neonates exposed to centrifuge-produced hypergravity. Cerebellar mass and protein expression in rat neonates exposed to hypergravity (1.5 G) from gestational day (G) 11 to postnatal day (P) 30 were compared at one of six time points between P6 and P30 against rat neonates developing under normal gravity. Proteins were analyzed by quantitative western blots of cerebellar homogenates prepared from male or female neonates. Cerebellar size was most clearly reduced in male neonates on P6 and in female neonates on P9, with a significant gender difference; differences in cerebellar mass remained significant even when change in total body mass was factored in. Densitometric analysis of western blots revealed both quantitative and temporal changes in the expression of selected cerebellar proteins that coincided with changes in cerebellar mass and were gender-specific. In fact, our data indicated certain significant differences even between male and female control animals. A maximal decrease in expression of CD15 was observed in HG females on P9, coinciding with maximal change in their cerebellar mass. A shift in the time-course of NCAM-L1 expression resulted in a significant increase in NCAM-L1 in HG males on P18, an isolated time at which cerebellar mass does not significantly differ between HG and SC neonates. A maximal decrease in expression of GFAP was observed in HG males on P6, coinciding with maximal change in their cerebellar mass. Altered expression of cerebellar proteins is likely to affect a number of developmental processes and contribute to the structural and functional alterations seen in the CNS developing under altered gravity. Our data suggest that both cerebellar development and its response to gravitational manipulations differ in males and females.     

Living plant systems: How robust are they in the absence of gravity?

The following hierarchical levels can be recognised in plant systems: cells, organs, organisms and gamodemes (interbreeding members of a community). Each level in this ‘living hierarchy’ is both defined and supported by a similar set of sub-systems. Within this framework of plant organization, two complementary questions are relevant for interpreting plant-oriented space experiments: 1) What role, if any, does gravity play in enabling the development of each organizational level? and 2) Does abnormal development in an altered gravity environment indicate sub-system inefficiency? Although a few representatives of the various organizational levels in plant systems have already been the subject of microgravity experiments in space laboratories—from cells in culture to gamodemes, the latter being found in some Closed Environment Life Support Systems—it would be of interest to investigate additional systems with respect to their response to microgravity. Recognition of the sub-systems at each level might be relevant not only for a more complete understanding of plant development but also for the successful cultivation and propagation of plants during long-term space flights and the establishment of plants in extra-terrestrial environments.     

Amphibian egg cytoplasm response to altered g-forces and gravity orientation

Elucidation of dorsal/ventral polarity and primary embryonic axis development in amphibian embryos requires an understanding of cytoplasmic rearrangements in fertile eggs at the biophysical, physiological, and biochemical levels. Evidence is presented that amphibian egg cytoplasmic components are compartmentalized. The effects of altered orientation to the gravitational vector (i.e., egg inversion) and alterations in gravity force ranging from hypergravity (centrifugation) to simulated microgravity (i.e., horizontal clinostat rotation) on cytoplasmic compartment rearrangements are reviewed. The behavior of yolk compartments as well as a newly defined (with monoclonal antibody) non-yolk cytoplasmic compartment, in inverted eggs and in eggs rotated on horizontal clinostats at their buoyant density, is discussed.     

Modeling irregular small bodies gravity field via extreme learning machines and Bayesian optimization

Close proximity operations around small bodies are extremely challenging due to their uncertain dynamical environment. Autonomous guidance and navigation around small bodies require fast and accurate modeling of the gravitational field for potential on-board computation. In this paper, we investigate a model-based, data-driven approach to compute and predict the gravitational acceleration around irregular small bodies. More specifically, we employ Extreme Learning Machine (ELM) theories to design, train and validate Single-Layer Feedforward Networks (SLFN) capable of learning the relationship between the spacecraft position and the gravitational acceleration. ELM-base neural networks are trained without iterative tuning therefore dramatically reducing the training time. Analysis of performance in constant density models for asteroid 25143 Itokawa and comet 67/P Churyumov-Gerasimenko show that ELM-based SLFN are able learn the desired functional relationship both globally and in selected localized areas near the surface. The latter results in a robust neural algorithm for on-board, real-time calculation of the gravity field needed for guidance and control in close-proximity operations near the asteroid surface.     

Artificial gravity as a countermeasure of physiological deconditioning in space.

The displacement of humans from our planet to space and back to our planet or to another planet necessarily causes the exposure to transitional gravitational forces which create a need for countermeasures of physiological deconditioning. One of the alternative approaches to counteracting physiological deconditioning is the simulation of a gravitational field. An apparatus named the artificial gravity simulator (AGS) has been designed and constructed. The AGS is a short radius rotating platform capable of producing a +Gz 100% gradient of a variable magnitude up to 3-g. It supports four beds for testing four subjects at a time. The AGS is suitable for short and long duration experiments on various physiological systems. At present it is being used to study the cardiovascular response to various g-levels and exposure times.     

Water wave communication in the genus Bombina (amphibia).

Amphibians were phylogenetically the first vertebrates to leave the aquatic environment and cope with terrestrial conditions including effects of gravity and substrate on movement and communication. Studies of extant primitive amphibians, which have conserved ancestral morphology and behavior, may help us to understand how gravitational adaptation from aquatic to terrestrial environments occurred. The anuran genus Bombina is a candidate for this type of investigation. In particular, a member of this genus, B. orientalis, is known for its low reaction threshold to minor changes of angular acceleration. We hypothesize that a heightened sensitivity to angular and mechanical accelerations evolved with wave communication. Comparisons of such behavior among B. variegata, B. bombina and B. orientalis may shed light on the evolution of reproductive systems based on water wave communication and relevant vestibular sensitivity. This may represent a transition to derived vocalization modes, which is seen in B. bombina to a certain degree.     

Histochemical investigations on the influence of long-term altered gravity on the CNS of developing cichlid fish: results from the 2nd German Spacelab Mission D-2.

The effect of long-term (10 days) altered gravitational conditions upon succinate dehydrogenase (SDH) reactivity in total brains as well as in individual brain nuclei of developing cichlid fish larvae had been investigated by means of semiquantitative histochemical methods (densitometric grey value analysis). Increasing accelerations from near weightlessness (spaceflight) via 1g controls to 3g hyper gravity (centrifuge) resulted in slightly increasing "all over the brain" (total brain) SDH reactivity. When focusing on distinct neuronal integration centers within the same brains in order to find the anatomical substratum of the gross histochemical data, significant effects of altered gravity only within vestibulum related brain parts were obtained.     

Theory and experimental results on gravitational effects on monocellular algae.

The orientation of a body which has an anisotropic distribution of mass and which is suspended in water is biased by gravitational torque, so that the center of gravity lies below the center of buoyancy. Many species of unicellular swimming algae are gravitationally oriented in this manner. Their axis of propulsion is essentially fixed within their bodies, so that when the cells swim, they swim upwards. Gravitaxis is an exotaxis, which requires no sensory processing. Nevertheless, gravity affects the lives of these cells both individually and collectively. For single cells, gravity intervenes in the execution and mechanism of sense-dependent taxes, such as phototaxis, it provides for fail-safe locomotion toward the upper interface of their habitat, the source of light and air, and it may cause up-accumulation. Populations of single cells, swimming in the presence of gravity, are coupled through fluid-mechanical interactions which cause spatial and temporal patterns of fluid convection and cell concentration. These patterns modify the cell's environmental interactions, by facilitating downward migrations of cell populations, by mixing the embedding fluid and its contents, and by providing a collective mechanism for controlling light intensity at the individual cell level. Summarizing, gravity modulates the interaction of algal cells with each other and with their environment.