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Genomic Changes Driven by Radiation-Induced DNA Damage and Microgravity in Human Cells

The space environment consists of a complex mixture of different types of ionizing radiation and altered gravity that represents a threat to humans during space missions. In particular, individual radiation sensitivity is strictly related to the risk of space radiation carcinogenesis. Therefore, in view of future missions to the Moon and Mars, there is an urgent need to estimate as accurately as possible the individual risk from space exposure to improve the safety of space exploration. In this review, we survey the combined effects from the two main physical components of the space environment, ionizing radiation and microgravity, to alter the genetics and epigenetics of human cells, considering both real and simulated space conditions. Data collected from studies on human cells are discussed for their potential use to estimate individual radiation carcinogenesis risk from space exposure.

space radiation↗

Models and molecular approaches to assessing the effects of the microgravity environment on vertebrate development

The extent to which gravity, and especially the lack thereof, can affect normal development in higher organisms is poorly understood. Underlying this question is the assumption that normal development depends on the embryo's ability to maintain a programmed temporal and spatial coordination of morphogenetic events. There are several reports documenting the apparently normal development of several vertebrate species, including mammals, under conditions of exposure to space flight during various periods of the development process. Evidence to the contrary also exists and it is therefore likely that some alterations in morphology do occur in a microgravity environment. Although subsequent development may appear overtly normal, more subtle abnormalities result. In all studies, the evaluation is restricted by the few numbers of specimens that can be examined and the relatively insensitive techniques for assessing potentially subtle effects. In the present discussion, we summarize some observations of mammalian development made in microgravity and consider which stages might be expected to be differentially sensitive to altered gravity conditions. While we emphasize mammalian development, we discuss the suitability of another model system for examining such effects in a cross-species context. Furthermore, we consider recent developments in our understanding of the molecular genetic program regulating embryogenesis that could serve as markers for assessing perturbations of development.

Cosmos 1129 Project↗

Investigating Neuro-Consequences of Spaceflight Using Drosophila Melanogaster

Research on human acclimation to spaceflight, including the recent NASA's Twin Study, reports complex effects of the spaceflight environment on health, with both acute and prolonged changes in multiple tissues. Spaceflight includes multiple factors such as microgravity, ionizing radiation, physiological stress, and disrupted circadian rhythms, that have been shown to contribute to pathophysiological responses that target immunity, bone and muscle integrity, cardiovascular and nervous systems. In this study, we used a well-established spaceflight model organism, Drosophila melanogaster, to assess spaceflight-associated changes on the nervous system. With 75% disease gene orthology to humans, short generation time, large sample size and ease of genetic, neuronal and behavioral studies, Drosophila is an excellent model to study nervous system dysfunction. Here, we present results from MVP-Fly-01 spaceflight mission that was launched on SpaceX CRS-14. The MVP hardware (developed by Techshot) used in this mission enabled us to have an in-flight 1g centrifuge, to distinguish the changes resulting from gravity versus those induced by other environmental factors associated with spaceflight. We observe behavioral impairments (p<0.001) and synaptic deficits, including decreased synaptic connections (p<0.05), in 3rd instar larvae which were developed in space. Furthermore, space-grown microgravity adults show a decrease in neuronal (p<0.05) and dendritic field (p<0.01) in adult brains coupled with an increased number of apoptotic cells (p<0.001) compared to in-flight 1g controls, suggesting increased neuronal loss under spaceflight conditions. In summary, we observe that altered gravity leads to gross neurological deficits. To better understand the long-term effects of spaceflight on the nervous system, longitudinal and multigenerational changes were also identified. This study will help elucidate the different approaches to prevent nervous system dysfunction in astronauts during spaceflight, while also contributing to a better understanding of the pathways that are related to some CNS disorders on Earth.

Iyer, Janani↗

Customized Science Carrier Modules and Accessories Developed for Microgravity Simulation Devices

Because the access to the true weightless environment of the ISS and other Low Earth Orbit (LEO) vehicles is limited and cost prohibitive, the need for ground-based microgravity simulation analogs to bridge the gravity continuum gap between 1g gravity and costly space flight studies with true weightlessness was realized. As such, the Microgravity Simulation Support Facility (MSSF) was established at NASA John F. Kennedy Space Center (KSC) to fulfill the need of the scientific community. Our facility provides the US science community with a focal point in the field of gravitational research by granting access to an array of two-dimensional(2D) and three-dimensional (3D)clinostats, and Random Positioning Machine (RPM) platforms developed by teams throughout the world. The availability of these capabilities will allow further understanding the role of altered gravity vectors from “functional weightlessness”, partial gravity conditions (1 x 10-3to 0.9g, and Moon and Mars ‘g vectors), to hypergravity (up to3g) on biological processes. We have designed and 3D printed a variety of specialized attachments(and associated “static control” hardware)to allow researchers to mount their samples onto these microgravity simulation devices to meet their unique science requirements. A summary of each attachment module will be presented below. Note: All software or code information should be requested to the author Ye Zhang, Kennedy Space Center ye.zhang-1@nasa.gov

Ye Zhang↗

Plasticity of Neurovestibular Systems Following Micro- and Hyper-Gravity Exposure and Readaptation to Earth's 1G

The gravity-sensing organs sense the sum of inertial force due to head translation and head orientation relative to gravity. Normally gravity is constant, and yet the neural sensors show remarkable plasticity. When the force of gravity changes, such as in spaceflight or during centrifugation, the neurovestibular system responds by regulating its neural output, and this response is similar for the vertebrate utricular nerve afferents and for the statocyst hair cell in invertebrates. First, we examine the response of utricular afferents in toadfish following exposure to G on two orbital missions (STS-90 and 95). Within the first day after landing, magnitude of neural response to an applied acceleration was significantly elevated, and re-adaptation back to control values occurred within approximately 30 hours. Time course of return to normal approximately parallels the decrease in vestibular disorientation in astronauts following return. Next, we use well-controlled hyper-G experiments in the vertebrate model to address: If G leads to adaptation and subsequent re-adaptation neural processes, does the transfer from 1G to hyper-G impart the opposite effects and do the effects accompanying transfer from the hyper-G back to the 1G conditions resemble as an analog the transfer from 1G to the microG Results show a biphasic pattern in reaction to 3G exposures: an initial sensitivity up-regulation (3- and 4-day) followed by a significant decrease after longer exposure. Return to control values is on the order of 4-8 days. Utricular sensitivity is strongly regulated up or down by gravity load and the duration of exposure. Interestingly, we found no correlation of response and hair cell synaptic body counts despite the large gain difference between 4- and 16-Day subjects. Lastly, we examine responses of statocyst receptors in land snail following exposure to G on two unmanned Russian Orbital missions (Foton M-2 and -3). Here, we have the ability to measure the output directly from the hair cells. Similar to afferents in vertebrates the hair cells increased their response sensitivity to vestibular stimulation. Two major pieces of information are needed: the precise vertebrate hair cell response to altered gravity and the impact of longer duration exposures on sensory plasticity.

Boyle, Richard D.↗

Cardiovascular Disease Spaceflight Equivalent Stressor Risk Modeling Project

The risk of cardiovascular disease (CVD) may be exacerbated by spaceflight hazards including radiation, isolation, distance from the Earth, gravity fields, and closed or confined environments. This project aims to use epidemiologic, scientific research evidence of surrogate spaceflight stressors on analog populations in terrestrial cohorts to model CVD risk for spaceflight conditions. To date, Dr. Butler and the multi-disciplinary CVD Spaceflight Equivalent Stressors (SES) team have conducted a formal systematic literature review on the risk of sleep loss as it relates to cardiovascular disease outcomes and are currently working on a meta-analysis for the data gathered from nearly 100 studies and their various CVD outcomes. The research will continue to examine other surrogate stressors in depth, such as altered gravity, radiation exposure, noise, etc. with cardiovascular outcomes to determine an overall combined spaceflight risk for long term future missions such as Artemis Lunar and Mars missions.

Jennifer Butler↗

Reduced receptor aggregation and altered cytoskeleton in cultured myocytes after space-flight

We carried out parallel experiments first on the slow clinostat and then in space-flight to examine the effects of altered gravity on the aggregation of the nicotinic acetylcholine receptors and the structure of the cytoskeleton in cultured Xenopus embryonic muscle cells. By examining the concordance between results from space flight and the clinostat, we tested whether the slow clinostat is a relevant simulation paradigm. Space-flown cells showed marked changes in the distribution and organization of actin filaments and had a reduced incidence of acetylcholine receptor aggregates at the site of contact with polystyrene beads. Similar effects were found after clinostat rotation. The sensitivity of synaptic receptor aggregation and cytoskeletal morphology suggests that in the microgravity of space cell behavior may be importantly altered.

STS-56 Shuttle Project↗

MicroRNA Expression Profiles in Cultured Human Fibroblasts in Space

Microgravity, or an altered gravity environment from the static 1g, has been shown to influence global gene expression patterns and protein levels in living organisms. However, it is unclear how these changes in gene and protein expressions are related to each other or are related to other factors regulating such changes. A different class of RNA, the small non-coding microRNA (miRNA), can have a broad effect on gene expression networks by mainly inhibiting the translation process. Previously, we investigated changes in the expression of miRNA and related genes under simulated microgravity conditions on the ground using the NASA invented bioreactor. In comparison to static 1 g, simulated microgravity altered a number of miRNAs in human lymphoblastoid cells. Pathway analysis with the altered miRNAs and RNA expressions revealed differential involvement of cell communication and catalytic activity, as well as immune response signaling and NGF activation of NF-kB pathways under simulated microgravity condition. The network analysis also identified several projected networks with c- Rel, ETS1 and Ubiquitin C as key factors. In a flight experiment on the International Space Station (ISS), we will investigate the effects of actual spaceflight on miRNA expressions in nondividing human fibroblast cells in mostly G1 phase of the cell cycle. A fibroblast is a type of cell that synthesizes the extracellular matrix and collagen, the structural framework for tissues, and plays a critical role in wound healing and other functions. In addition to miRNA expressions, we will investigate the effects of spaceflight on the cellular response to DNA damages from bleomycin treatment.

Wu, Honglu↗

Spatial orientation of optokinetic nystagmus and ocular pursuit during orbital space flight

On Earth, eye velocity of horizontal optokinetic nystagmus (OKN) orients to gravito-inertial acceleration (GIA), the sum of linear accelerations acting on the head and body. We determined whether adaptation to micro-gravity altered this orientation and whether ocular pursuit exhibited similar properties. Eye movements of four astronauts were recorded with three-dimensional video-oculography. Optokinetic stimuli were stripes moving horizontally, vertically, and obliquely at 30 degrees/s. Ocular pursuit was produced by a spot moving horizontally or vertically at 20 degrees/s. Subjects were either stationary or were centrifuged during OKN with 1 or 0.5 g of interaural or dorsoventral centripetal linear acceleration. Average eye position during OKN (the beating field) moved into the quick-phase direction by 10 degrees during lateral and upward field movement in all conditions. The beating field did not shift up during downward OKN on Earth, but there was a strong upward movement of the beating field (9 degrees) during downward OKN in the absence of gravity; this likely represents an adaptation to the lack of a vertical 1-g bias in-flight. The horizontal OKN velocity axis tilted 9 degrees in the roll plane toward the GIA during interaural centrifugation, both on Earth and in space. During oblique OKN, the velocity vector tilted towards the GIA in the roll plane when there was a disparity between the direction of stripe motion and the GIA, but not when the two were aligned. In contrast, dorsoventral acceleration tilted the horizontal OKN velocity vector 6 degrees in pitch away from the GIA. Roll tilts of the horizontal OKN velocity vector toward the GIA during interaural centrifugation are consistent with the orientation properties of velocity storage, but pitch tilts away from the GIA when centrifuged while supine are not. We speculate that visual suppression during OKN may have caused the velocity vector to tilt away from the GIA during dorsoventral centrifugation. Vertical OKN and ocular pursuit did not exhibit orientation toward the GIA in any condition. Static full-body roll tilts and centrifugation generating an equivalent interaural acceleration produced the same tilts in the horizontal OKN velocity before and after flight. Thus, the magnitude of tilt in OKN velocity was dependent on the magnitude of interaural linear acceleration, rather than the tilt of the GIA with regard to the head. These results favor a 'filter' model of spatial orientation in which orienting eye movements are proportional to the magnitude of low frequency interaural linear acceleration, rather than models that postulate an internal representation of gravity as the basis for spatial orientation.

Flight Experiment↗

Centrifuge Facility for the International Space Station Alpha

The Centrifuge Facility planned for the International Space Station Alpha has under-one considerable redesign over the past year, primarily because the Station is now viewed as a 10 year mission rather than a 30 year mission and because of the need to simply the design to meet budget constraints and a 2000 launch date. The basic elements of the Centrifuge Facility remain the same, i.e., a 2.5 m diameter centrifuge, a micro-g holding unit, plant and animal habitats, a glovebox and a service unit. The centrifuge will still provide the full range of artificial gravity from 0.01 a to 2 - as originally planned; however, the extractor to permit withdrawal of habitats from the centrifuge without stopping the centrifuge has been eliminated. The specimen habitats have also been simplified and are derived from other NASA programs. The Plant Research Unit being developed by the Gravitational Biology Facility will be used to house plants in the Centrifuge Facility. Although not as ambitious as the Centrifuge Facility plant habitat, it will provide much better environmental control and lighting than the current Shuttle based Plant Growth Facility. Similarly, rodents will be housed in the Advanced Animal Habitat being developed for the Shuttle program. The Centrifuge Facility and ISSA will provide the opportunity to perform repeatable, high quality science. The long duration increments available on the Station will permit multigeneration studies on both plants and animals which have not previously been possible. The Centrifuge Facility will accommodate sufficient number of specimens to permit statistically significant sampling of specimens to investigate the time course of adaptation to altered gravity environments. The centrifuge will for the first time permit investigators to use gravity itself as a tool to investigate fundamental processes, to investigate the intensity and duration of gravity to maintain normal structure and function, to separate the effects of micro-g from other 0 environmental factors and to examine artificial gravity as a potential countermeasure for the physical deconditioning observed during spaceflight.

Johnson, Catherine C.↗

Functional Responses in Otolith Structures from Micro- to Hyper-Gravity

Vertebrates and invertebrates sense gravito-inertial acceleration by mechanoreceptors in the otolith and statolith organs, respectively. These structures consist of ciliated sensory hair cells surmounted by biomineral grains of calcium carbonate (CaCO3) called oto-orstato-conia. The grains provide mechanical loading of hair cell cilia, and their high density increases sensitivity to acceleration. A widely considered mechanism by which the animal responds to a chronic change in amplitude of gravity is a change in weight lending otoconia. In G, it is argued, the organism counters the loss of gravity by increasing CaCO3 production, thereby increasing otolith mass, as a means to increase system gain. In hypergravity (HG), the converse is argued. Here, we present the results obtained in 3 species exposed both to G and HG. Adult toadfish, Opsanus tau, were exposed to G in 2 short-duration shuttle missions and to 1.24 1.73G centrifugation for 1-32 days; re-adaptation was studied following 1-8 days of 1G. Results show a biphasic pattern in response to 1.73G: initial hypersensitivity, similar to that observed after G exposure, followed by transition to a significant decrease at 16-32 days. Recovery from HG exposure is 4-8 days. Next, we examined directly the responses of statocyst receptors in the land snail after exposure to G on two unmanned Russian Orbital missions and at 1.24G. Similar to vertebrate afferents snail receptors increased their sensitivity to tilt after G exposure, and decrease it after 16-32 days of HG. Two major pieces of information are still needed: vertebrate hair cell response to altered gravity and impact of longer duration exposures on sensory plasticity. To address the latter, we applied electron microscopic techniques to image otoconia mass obtained from 1) mice subjected to 91-days of weightlessness in the Mouse Drawer System (MDS) flown on International Space Station, 2) mice subjected to 91-days of 1.24G centrifugation on ground, and 3) mice flown on 2 short-duration orbital missions. Images indicate a clear restructuring of individual otoconia, suggesting deposition to the outer shell. Images from their HG counterparts indicate the converse - an ablation of the otoconia mass. For shorter duration exposures to weightlessness on 13-day shuttle missions, mice otoconia appear normal. Despite the permanence of 1G in evolution, the animal senses exposure to a novel, non-1G, environment and adaptive mechanisms are initiated - in the short term, compensation is likely confined to the peripheral sensory receptors, the brain or both. For longer exposures structural modifications of the endorgan may also result. Support Contributed By: NASA 03-OBPR-04 and 11-11_Omni_2-0002

Vertebrates↗

Hypergravity Effects on Rodent Pregnancy and Parturition

No mammal has yet undergone birth, or parturition, in the microgravity of space. Previous studies (Ronco & Alberts, 2000) have shown that mid-pregnant rat dams exposed to spaceflight (0-g) and landed 48-72 hrs before term successfully delivered robust, healthy offspring Microgravity-exposed dams exhibited twice the expected numbers of labor contractions whereas length of pregnancy, duration of labor, fetal wastage, number of neonates born and litter gender ratios were identical to controls. In the present study, we report the results of rodent pregnancy and parturition at the opposite end of the gravity spectrum, in hypergravity. Dams exposed to either: 1.0-g, 1.5-g, 1.75-g or 2.0-g from Gestational day (G) 11 and throughout the births of their litters had comparable pregnancy and labor durations, fetal wastage, numbers of neonates born and litter Tender ratios. During parturition, hypergravity-exposed dams exhibited significantly fewer labor contractions as compared to 1.0-g controls. Dams that underwent birth in hypergravity had significantly fewer offspring surviving the immediate postpartum period (P1: 1.0-g, 11.92 +/- 2.84; 1.5-g, 10.88 +/- 2.17; 1.75-g, 9.22 +/-1.99; 2.0-g, 8.83 +/- 3.31). Within 24 hrs postpartum, neonatal survival was further diminished in hypergravity [P2: 100% (1.0-g); 96% (1.5-g); 96% (1.75-g); 73% (2.0-g)] and continued to decline (P10: 100%(1.0-g.); 90%(1.5-g); 87%(1.75-g), 40%(2.0-g)]. Neonatal losses stabilized by P5 for the 1.5-g andl.75-g conditions but continued until P9 for the 2.0-g condition. Together, these findings show that postnatal, but not prenatal, survival is compromised following birth in hypergravity, Maternal and neonatal factors that contribute to peri-parturitional vulnerability to altered gravity environments will be discussed.

Ronca, A. E.↗

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. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

NASA Program Fundamental Space Biology↗

Effects of static orientation upon human optokinetic afternystagmus

"Normal" human subjects were placed in a series of 5 static orientations with respect to gravity and were asked to view an optokinetic display moving at a constant angular velocity. The axis of rotation coincided with the subject's rostro-caudal axis and produced horizontal optokinetic nystagmus and afternystagmus. Wall (1) previously reported that these optokinetic afternystagmus responses were not well characterized by parametric fits to slow component velocity. The response for nose-up, however, was larger than for nose-down. This suggested that the horizontal eye movements measured during optokinetic stimulation might include an induced linear VOR component as presented in the body of this paper. To investigate this hypothesis, another analysis of these data has been made using cumulative slow component eye position. Some subjects' responses had reversals in afternystagmus direction. These reversals were "filled in" by a zero slow component velocity. This method of analysis gives a much more consistent result across subjects and shows that, on average, responses from the nose-down horizontal (prone) orientation are greatly reduced (p < 0.05) compared to other horizontal and vertical orientations. Average responses are compared to responses predicted by a model previously used to predict successfully the responses to post-rotatory nystagmus after earth horizontal axis rotation. Ten of 11 subjects had larger responses in their supine than their prone orientation. Application of horizontal axis optokinetic afternystagmus for clinical otolith function testing, and implications for altered gravity experiments are discussed.

Non-NASA Center↗

Feasibility of Earthbound Motion in Lunar Gravity

BACKGROUND: Marginal stability of astronaut movement while performing lunar surface tasks has been well documented, and is clearly demonstrated in videos of falls, and near falls, during Apollo Lunar Extravehicular Activities. Referencing mission reports from Apollo 15 and 16 [1, 2], suspected causes for falls include: surface conditions, visibility, and gravitational effects (hypogravity). In this preliminary test, we employ the open-source biomechanical tool OpenSim [3, 4] to analyze the impact of lunar gravity (Lg) on two object-pickup motions performed by a single shirt-sleeved subject. Specifically, we attempt to answer the following questions based on an estimation of the Center of Mass Projection (CoMP) and ground reaction force Center of Pressure (COP) as it relates to the astronaut Base of Support (BOS) for 1g and Lg conditions: 1. Is the task motion, as performed in 1g, dynamically feasible in 1g and Lg? 2. Can we make the motion dynamically feasible in Lg by slowing it down? 3. Is the Lg COP equal to the 1g COP at a theoretically predicted reduction in motion speed? METHODS AND RESULTS: To answer the first question, the gravitational acceleration in the OpenSim model is modified from a nominal 9.81 m/s2 to 1.64 m/s2, and the 1g joint trajectory is input to an OpenSim based method [5] for estimating ground reaction forces and moments. From this method, the position of the CoMP and COP can be estimated and checked to see if they remain within a simulated BOS formed from the footprint of the OpenSim model to determine whether the motion is dynamically feasible. As expected, both of the motions were estimated to be feasible in a 1g environment, however, both motions had periods of infeasibility in Lg. It is well known that crew members make adjustments to motion trajectories in altered gravity fields to maintain balance. As a first step, we considered the simple adjustment of slowing the motion in Lg by a constant factor. This was accomplished by scaling the time stamps in the motion trajectory file by that factor. For the two motions considered, it was found that scale factors of 1.3 and 1.4 kept the COP just within the subject BOS. The CoMP is unchanged by the gravity field. Simple analysis of an inverted pendulum in the Lg environment, which generalizes to a general multibody system, leads to a theoretical prediction that a reduction in speed factor of √1g/Lg, or 2.445, will make the COP trajectory in Lg equivalent to that in 1g. When the above procedure was performed with a factor of 2.445, the estimated COP in Lg, was observed to be very close to that in 1g. In summary, we have developed a method for estimating the CoMP and COP in Lg, for subject motion collected in 1g. We believe this method can prove to be a valuable check and balance for simulated Lg training and testing by exposing potential simulator-induced artifacts that make the simulated task motion seem possible, when in fact, it would violate the above criteria. We also note that a reduction in task speed should tend the task motion towards stability, with a theoretical slowdown factor of √1g/Lg making the motion stability equal to that in 1g according to the CoMP and COP criteria.

R K Huffman↗

Early Development of Gravity-Sensing Organs in Microgravity

Most animals have organs that sense gravity. These organs use dense stones (called otoliths or statoconia), which rest on the sensitive hairs of specialized gravity- and motion-sensing cells. The weight of the stones bends the hairs in the direction of gravitational pull. The cells in turn send a coded representation of the gravity or motion stimulus to the central nervous system. Previous experiments, in which the eggs or larvae of a marine mollusk (Aplysia californica, the sea hare) were raised on a centrifuge, demonstrated that the size of the stones (or test mass) was reduced in a graded manner as the gravity field was increased. This suggests that some control mechanism was acting to normalize the weight of the stones. The experiments described here were designed to test the hypothesis that, during their initial development, the mass of the stones is regulated to achieve a desired weight. If this is the case, we would expect a larger-than-normal otolith would develop in animals reared in the weightlessness of space. To test this, freshwater snails and swordtail fish were studied after spaceflight. The snails mated in space, and the stones (statoconia) in their statocysts developed in microgravity. Pre-mated adult female swordtail fish were flown on the Space Shuttle, and the developing larvae were collected after landing. Juvenile fish, where the larval development had taken place on the ground, were also flown. In snails that developed in space, the total volume of statoconia forming the test mass was 50% greater than in size-matched snails reared in functionally identical equipment on the ground. In the swordtail fish, the size of otoliths was compared between ground- and flight-reared larvae of the same size. For later-stage larvae, the growth of the otolith was significantly greater in the flight-reared fish. However, juvenile fish showed no significant difference in otolith size between flight- and ground-reared fish. Thus, it appears that fish and snails reared in space do produce larger-than-normal otoliths (or their analogs), apparently in an attempt to compensate for the reduced weight of the stones in space. The fish data suggest that there is a critical period during which altered gravity can affect the size of the test mass, since the larval, but not the juvenile, fish showed the changes.

Wiederhold, Michael L.↗

A Brief Description of the Two Primate Experiments to Be Carried Out on SL-4

Two primate experiments to be carried on Spacelab 4 are discussed. One of these investigates thermoregulation of primates during altered gravity. In previous ground-based tests, primate heat distribution was shown to change during centrifugation (a way of subjecting subjects to higher-than-Earth gravity), the inner body cooling off and the skin temperature rising. The adaptive and homeostatic mechanisms triggered by spaceflight are studied. Ways in which to correct any undesirable shifts in the homeostatic capabilities of the thermoregulatory control system are addressed. The other experiment involving primates addressess the changes in fluid distribution and electrolyte content of blood that occur in spaceflight. During previous space missions, fluid shifts from the legs to the chest and head have been noted. Ground-based studies have shown decreases in blood potassium levels and increases in potassium excretion.

Elsea, L.↗

Proliferation and performance of hybridoma cells in microgravity (7-IML-1)

The purpose of this experiment is to study how cell performance (biosynthesis and secretion) is altered by altered gravity conditions. Hybridoma cells are obtained by fusion of an activated B-lymphocyte with a myeloma cell. Activated B-lymphocytes, derived from a human or an animal, carry the information required to produce antibodies of a certain specificity and can survive only a few days in culture. Myeloma cells are tumor cells which can grow indefinitely in culture. Therefore, the product of the fusion is an immortal cell line capable of producing homogeneous antibodies (monoclonal antibodies). Experimental procedures are explained in some detail.

Cogoli, Augusto↗