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All sourcesNASA NTRSDOE OSTI
NASA NTRS1996

APS Conference on Understanding the Biological Clock: From Genetics to Physiology

The Conference was designed to take advantage of the fusion of two intellectually dominant but heretofore separate lines of clock research, vertebrate physiology and invertebrate and microbial genetics. The APS Conference attracted 251 scientists, 68 of whom were students. In addition to the excellent speaker program organized by Dunlap and Loros, the attendees also submitted 93 volunteer abstracts that were programmed in poster sessions. Thirty-four percent of the submitted abstracts were first authorized by a female student or scientist.

Frank, Martin↗
NASA NTRS1959

Human Tolerance to Rapidly Applied Accelerations: A Summary of the Literature

The literature is surveyed to determine human tolerance to rapidly applied accelerations. Pertinent human and animal experiments applicable to space flight and to crash impact forces are analyzed and discussed. These data are compared and presented on the basis of a trapezoidal pulse. The effects of body restraint and of acceleration direction, onset rate, and plateau duration on the maximum tolerable and survivable rapidly applied accelerations are shown. Results of the survey indicate that adequate torso and extremity restraint is the primary variable in tolerance to rapidly applied accelerations. The harness, or restraint system, must be arranged to transmit the major portion of the accelerating force directly to the pelvic structure and not via the vertebral column. When the conditions of adequate restraint have been met, then the other variables, direction, magnitude, and onset rate of rapidly applied accelerations, govern maximum tolerance and injury limits. The results also indicate that adequately stressed aft-faced passenger seats offer maximum complete body support with minimum objectionable harnessing. Such a seat, whether designed for 20-, 30-, or 40-G dynamic loading, would include lap strap, chest (axillary) strap, and winged-back seat to increase headward and lateral G protection, full-height integral head rest, arm rests (load-bearing) with recessed hand-holds and provisions to prevent arms from slipping either laterally or beyond the seat back, and leg support to keep the legs from being wedged under the seat. For crew members and others whose duties require forward-facing seats, maximum complete body support requires lap, shoulder, and thigh straps, lap-belt tie-down strap, and full-height seat back with integral head support.

Eiband, A. Martin↗
NASA NTRS1998

Effect of Microgravity on Afferent Innervation

Presentations and publications are: (1) an audiovisual summary web presentation on results from SLM-MIR avian experiments. A color presentation summarizing results from the SLM-MIR and STS-29 avian experiments; (2) color threshold and ratio of S 100B MAP5, NF68/200, GABA and GAD; (3) chicken (Gallus domesticus) inner ear afferents; (4) microgravity in the STS-29 Space Shuttle Discovery affected the vestibular system of chick embryos; (5) expression of S 100B in sensory and secretory cells of the vertebrate inner ear; (6) otoconia biogenesis, phylogeny, composition and functional attributes;(7) the glycan keratin sulfate in inner ear crystals; (8) elliptical-P cells in the avian perilymphatic interface of the tegmentum vasculosum; and (9) LAMP2c and S100B upregulation in brain stem after VIIIth nerve deafferentation.

Source record↗
NASA NTRS1999

The Effect of Spaceflight on Bone Cell Cultures

Understanding the response of bone to mechanical loading (unloading) is extremely important in defining the means of adaptation of the body to a variety of environmental conditions such as during heightened physical activity or in extended explorations of space or the sea floor. The mechanisms of the adaptive response of bone are not well defined, but undoubtedly they involve changes occurring at the cellular level of bone structure. This proposal has intended to examine the hypothesis that the loading (unloading) response of bone is mediated by specific cells through modifications of their activity cytoskeletal elements, and/or elaboration of their extracellular matrices. For this purpose, this laboratory has utilized the results of a number of previous studies defining molecular biological, biochemical, morphological, and ultrastructural events of the reproducible mineralization of a primary bone cell (osteoblast) culture system under normal loading (1G gravity level). These data and the culture system then were examined following the use of the cultures in two NASA shuttle flights, STS-59 and STS-63. The cells collected from each of the flights were compared to respective synchronous ground (1G) control cells examined as the flight samples were simultaneously analyzed and to other control cells maintained at 1G until the time of shuttle launch, at which point they were terminated and studied (defined as basal cells). Each of the cell cultures was assayed in terms of metabolic markers- gene expression; synthesis and secretion of collagen and non-collagenous proteins, including certain cytoskeletal components; assembly of collagen into macrostructural arrays- formation of mineral; and interaction of collagen and mineral crystals during calcification of the cultures. The work has utilized a combination of biochemical techniques (radiolabeling, electrophoresis, fluorography, Western and Northern Blotting, and light microscopic immunofluorescence) and structural methods (conventional and high voltage electron microscopy, inununocytochemistry, stereomicroscopy, and 3D image reconstruction). The studies have provided new knowledge of aspects of bone cell development and structural regulation, extracellular matrix assembly, and mineralization during spaceflight and under normal gravity. The information has contributed to insights into the means in general by which cells respond and adapt to different conditions of gravity (loading). The data may as well have suggested an underlying basis for the observed loss of bone by vertebrates, including man, in microgravity; and these scientific results may have implications for understanding bone loss following fracture healing and extended periods of inactivity such as during long-term bedrest.

Landis, William J.↗
NASA NTRS1999

A Novel Technique for Performing Space Based Radiation Dosimetry Using DNA: Results from GRaDEx-I and the Design of GRaDEx-II

Because of the large amounts of cosmic radiation in the space environment relative to that on earth, the effects of radiation on the physiology of astronauts is of major concern. Doses of radiation which can cause acute or chronic biological effects are to be avoided, therefore determination of the amount of radiation exposure encountered during space flight and assessment of its impact on biological systems is critical. Quantifying the radiation dosage and damage to biological systems, especially to humans during repetitive high altitude flight and during long duration space flight is important for several reasons. Radiation can cause altered biosynthesis and long term genotoxicity resulting in cancer and birth defects, etc. Radiation damage to biological systems depends in a complex way on incident radiation species and their energy spectra. Typically non-biological, i.e. film or electronic monitoring systems with narrow energy band sensitivity are used to perform dosimetry and then results are extrapolated to biological models. For this reason it may be desirable to perform radiation dosimetry by using biological molecules e.g. DNA or RNA strands as passive sensors. A lightweight genotoxicology experiment was constructed to determine the degree to which in-vitro naked DNA extracted from tissues of a variety of vertebrate organisms is damaged by exposure to radiation in a space environment. The DNA is assayed by means of agarose gel electrophoresis to determine damage such as strand breakage caused by high momentum particles and photons, and base oxidation caused by free radicals. The length distribution of DNA fragments is directly correlated with the radiation dose. It is hoped that a low mass, low cost, passive biological system to determine dose-response relationship (increase in strand breaks with increase in exposure) can be developed to perform radiation dosimetry in support of long duration space flight, and to predict negative effects on biological systems (e.g. astronauts and greenhouses) in space. The payload was flown in a 2.5 cubic foot Get Away Special (GAS) container through NASA's GAS program. It was subjected to the environment of the space shuttle cargo bay for the duration of the STS-91 mission (9 days). Results of the genotoxicology and radiation dosimetry experiment (GRaDEx-I) as well as the design of an improved follow on payload are presented.

Ritter, Joe↗
NASA NTRS1999

A Novel Technique for Performing Space Based Radiation Dosimetry Using DNA-Results from GRaDEx-I and the Design of GRaDEx-II

Because of the large amounts of cosmic radiation in the space environment relative to that on earth, the effects of radiation on the physiology of astronauts is of major concern. Doses of radiation which can cause acute or chronic biological effects are to be avoided, therefore determination of the amount of radiation exposure encountered during space flight and assessment of its impact on biological systems is critical. Quantifying the radiation dosage and damage to biological systems, especially to humans during repetitive high altitude flight and during long duration space flight is important for several reasons. Radiation can cause altered biosynthesis and long term genotoxicity resulting in cancer and birth defects etc. Radiation damage to biological systems depends in a complex way on incident radiation species and their energy spectra. Typically non-biological, i.e. film or electronic monitoring systems with narrow energy band sensitivity are used to perform dosimetry and then results are extrapolated to biological models. For this reason it may be desirable to perform radiation dosimetry by using biological molecules e.g. DNA or RNA strands as passive sensors. A lightweight genotoxicology experiment was constructed to determine the degree to which in vitro naked DNA extracted from tissues of a variety of vertebrate organisms is damaged by exposure to radiation in a space environment. The DNA is assayed by means of agarose gel electrophoresis to determine damage such as strand breakage caused by high momentum particles and photons, and base oxidation caused by free radicals. The length distribution of DNA fragments is directly correlated with the radiation dose. It is hoped that a low mass, low cost, passive biological system to determine dose response relationship (increase in strand breaks with increase in exposure) can be developed to perform radiation dosimetry in support of long duration space flight, and to predict negative effects on biological systems (e.g. astronauts and greenhouses) in space. The payload was flown in a 2.5 cubic foot Get Away Special (GAS) container through NASA's GAS program. It was subjected to the environment of the space shuttle cargo bay for the duration of the STS-91 mission (9 days). Results of the genotoxicology and radiation dosimetry experiment (GRaDEx-1) as well as the design of an improved follow on payload are presented.

Ritter, Joe↗
NASA NTRS1997

Development of Gravity-Sensing Organs in Altered Gravity Conditions: Opposite Conclusions From an Amphibian and a Molluscan Preparation

Several components of the systems animals use to orient to gravity might develop differently in micrograms. If the growth of the "test masses" on which gravity acts (otoliths, in vertebrates, statoliths or statoconia in most invertebrates) is controlled on the basis of their weight, larger otoliths (or their analogs) would be expected to develop in micrograms. The vestibular systems in animals reared in altered gravity have been studied in several species, with varied results being reported. Early Russian reports of Xenopus larvae reared in space indicated no qualitative differences in the vestibular organs, compared to ground-reared controls. A similar lack of differences in Xenopus were reported. The ultricular otolith was 30% larger in space-reared Xenopus. No differences in saccular otolith volume between centrifuged and control adult rats were found. A delay in otoconial development in chick embryos reared at 2 grams on a centrifuge was reported but in a later report, no differences in otolith weight between 2 grams and control chicks were found. Increased optokinetic responses in flight-reared Xenopus tadpoles, suggesting that the animals reared in the absence of gravity made greater relative use of their visual system, rather than the vestibular system, in orienting to a moving stimulus was reported. To test early Japanese newt, CYnops pyrrhogaster, were maintained in orbit for 15 days on the IML-2 mission in 1994. All specimens reached orbit before any otoconia were formed and all major components of the inner ear were formed by the end of the flight. In ground-based studies of he Aplysia statocyst, the volume of the statolith in embryos and the number statoconia in post-metamorphic animals were compared between 1-gram controls and specimens reared at 2 to 5.7 grams.

Wiederhold, Michael L.↗
NASA NTRS1999

Quantifying Biomechanical Characteristics of Jumping Exercises in 1G and in Simulated and True Microgravity

Exercise in microgravity is one of the most promising countermeasures to the dual problems of space flight-induced bone loss and muscle atrophy. Although exercise in microgravity has been studied extensively from a metabolic standpoint, little research has focused on the efficacy of different forms of exercise for maintaining musculoskeletal integrity. Exercise protocols have not been effective in preventing muscle atrophy and bone loss during space flight, especially in the lower extremities. In 1-G, however, animal experiments have clearly indicated that: (1) certain bone strains and strain rates do stimulate bone deposition, and (2) repetitive loading of the lower extremity can increase osteonal bone formation even as proximally as the vertebral column. Such studies have also indicated that a relatively small number of appropriate loading cycles may lead to bone deposition. This suggests that an optimal exercise regimen might be able to maintain bone and muscle integrity during space flight. Since there is evidence that the bones and muscles of the lower limbs are particularly affected by space flight, the present study addressed two major aims: (1) quantify externally applied impact loads and rates of loading under the feet during tethered jumping exercises, and (2) determine the amount of eccentric and concentric whole-muscle activity during these jumping exercises in true and in simulated zero-gravity.

Davis, B. L.↗
NASA NTRS1999

The Effects of Gravity on the Circadian Timing System

All vertebrates have a physiological control system that regulates the timing of the rhythms of their daily life. Dysfunction of this system, the circadian timing system (CTS), adversely affects an organism's ability to respond to environmental challenges and has been linked to physiological and psychological disorders. Exposure to altered gravitational environments (the microgravity of space and hyperdynamic environments produced via centrifugation) produces changes in both the functioning of the CTS and the rhythmic variables it controls. The earliest record of primate rhythms in a spaceflight environment come from Biosatellite III. The subject, a pig-tailed macaque, showed a loss of synchronization of the body temperature rhythm and a fragmented sleep-wake cycle. Alterations in the rhythm of body temperature were also seen in rhesus macaques flown on COSMOS 1514. Squirrel monkeys exposed to chronic centrifugation showed an initial decrease in the amplitude and mean of their body temperature and activity rhythms. In a microgravity environment, Squirrel monkeys on Spacelab-3 showed a reduction in the mean and amplitude of their feeding rhythms. Since 1992 we have had the opportunity to participate on three US/Russian sponsored biosatellite missions on which a total of six juvenile male rhesus macaques were flown. These animals uniformly exhibited delays in the phasing of their temperature rhythms, but not their heart rate or activity rhythms during spaceflight. There was also a tendency for changes in waveform mean and amplitude. These data suggest that the spaceflight environment may have a differential effect on the different oscillators controlling different rhythmic variables. Ongoing studies are examining the effects of +G on the CTS. The long-term presence of humans in space highlights the need for effective countermeasures to gravitational effects on the CTS.

Fuller, Charles A.↗
NASA NTRS1994

Dino Fest

This document and video represent the proceedings of the first Dino Fest conference, which was unprecedented in bringing together exhibits of dinosaurs and other fossils and attracting many of the world's leading paleontologists and science educators, students and the public. This first Dino Fest consisted of scores of exhibits that included live and fossil plants, invertebrates and vertebrates. Lasting three weeks, the event concluded with a three-day symposium, providing dinosaur experts from around the country a forum to discuss their research and ideas with the public and other scientists. The document presents the talks of many of the scientists. The video is from an interactive television broadcast relayed by a NASA satellite that enabled children at remote locations to ask questions of a panel of dinosaur experts, literally reaching an audience around the world.

Rosenberg, Gary D.↗
NASA NTRS1995

Adaptation to Space: An Introduction

The cardiovascular and musculoskeletal systems are normally exposed to gradients of blood pressure and weight on Earth. These gradients increase blood pressure and tissue weight in dependent tissues of the body. Exposure to actual and simulated microgravity causes blood and tissue fluid to shift from the legs to the head. Studies of humans in space have documented facial edema, space motion sickness, decreased plasma volume, muscle atrophy, and loss of bone strength. Return of astronauts to Earth is accompanied by orthostatic intolerance, decreased neuromuscular coordination, and reduced exercise capacity. These factors decrease performance during descent from orbit and increase risk during emergency egress from the spacecraft. Models of simulated microgravity include 6 deg head-down tilt, immersion, and prolonged horizontal bedrest. Head-down tilt is the most accepted model and studies using this model of up to one year have been performed in Russia. Animal models which offer clear insights into the role of gravity on vertebrates include the developing giraffe and snakes from various habitats. Finally, possible countermeasures to speed readaptation of astronauts to gravity after prolonged space flight will be discussed.

Hargens, Alan R.↗
NASA NTRS1998

Cardiovascular Responses of Snakes to Gravitational Gradients

Snakes are useful vertebrates for studies of gravitational adaptation, owing to their elongate body and behavioral diversification. Scansorial species have evolved specializations for regulating hemodynamics during exposure to gravitational stress, whereas, such adaptations are less well developed in aquatic and non-climbing species. We examined responses of the amphibious snake,\italicize (Nerodia rhombifera), to increments of Gz (head-to-tail) acceleration force on both a short- and long-arm centrifuge (1.5 vs. 3.7 m radius, from the hub to tail end of snake). We recorded heart rate, dorsal aortic pressure, and carotid arterial blood flow during stepwise 0.25 G increments of Gz force (referenced at the tail) in conscious animals. The Benz tolerance of a snake was determined as the Gz level at which carotid blood flow ceased and was found to be significantly greater at the short- than long-arm centrifuge radius (1.57 Gz vs. 2.0 Gz, respectively; P=0.016). A similar pattern of response was demonstrated in semi-arboreal rat snakes,\italicize{Elaphe obsoleta}, which are generally more tolerant of Gz force (2.6 Gz at 1.5m radius) than are water snakes. The tolerance differences of the two species reflected cardiovascular responses, which differed quantitatively but not qualitatively: heart rates increased while arterial pressure and blood flow decreased in response to increasing levels of Gz. Thus, in both species of snakes, a reduced gradient of Gz force (associated with greater centrifuge radius) significantly decreases the Gz level that can be tolerated.

Hsieh, Shi-Tong T.↗
NASA NTRS2002

Hemichordates and the Origin of Chordates

At the start of the period of the NASA grant three years ago, we had no information on the organization and development of the body axis of the hemichordate, Saccoglossus kowalevskii. Now we have substantial findings about the anteroposterior axis and dorsoventral axis, and based on this information, we have new insights about the origin of chordates from ancestral deuterostomes. We found ways to obtain and preserve large numbers of embryos and hatched juveniles. We can now collect about 40,000 embryos in the month of September, the time of S. kowalevskii spawning at Woods Hole. Excellent cDNA libraries were prepared from three developmental stages. From these libraries, we directly isolated about 30 gene ortholog sequences by screening and pcr techniques, all of these sequences of interest in the inquiry about the animal's organization and development. We also performed a mid-sized EST project (60,000 randomly picked clones, many of these arrayed). About half of these have been analyzed so far by blastx and are suitable for direct use of clones. We have obtained about 50 interesting sequences from this set. The rest still await analysis. Thus, at this time we have isolated orthologs of 80 genes that are known to be expressed in chordates in conserved domains and known to have interesting roles in chordate organization and development. The orthology of the S. kowalevskii sequences has been verified by neighbor joining and parsimony methods, with bootstrap estimates of validity. The S. kowalevskii sequences cluster with other deuterostome sequences, namely, other hemichordates, echinoderms, ascidians, amphioxus, or vertebrates, depending on what sequences are available in the database for comparison. We have used these sequences to do high quality in situ hybridization on S. kowalevskii embryos, and the results can be divided into three sections-those concerning the anteroposterior axis of S. kowalevskii in comparison to the same axis of chordates, those concerning the dorsoventral axis of S. kowalevskii in comparison to the same axis of chordates, and those concerning the signals and transcription factors found in the endoderm, of S. kowalevskii compared to the signals and transcription factors in the endo-mesodermal cells of Spemann's organizer of chordates.

Gerhart, John↗
NASA NTRS2002

Tensile Properties of Hydrogels and of Snake Skin

Stimulus-responsive or 'smart' gels are of potential interest as sensors and actuators, in industrial separations, and as permeable delivery systems. In most applications, a certain degree of mechanical strength and toughness will be required, yet the large-strain behavior of gels has not been widely reported. Some exceptions include work on gelatin and other food gels, some characterization of soft gels applicable for in-vitro cell growth studies, and toughness determinations on commercial contact lens materials. In general, it can be anticipated that the gel stiffness will increase with increasing degree of crosslinking, but the tensile strength may go through a maximum. Gel properties can be tailored by varying not only the degree of crosslinking, but also the polymer concentration and the nature of the polymer backbone (e.g. its stiffness or solubility). Polypeptides provide an especially interesting case, where secondary structure affects trends in moduli and conformational transitions may accompany phase changes. A few papers on the tensile properties of responsive gels have begun to appear. The responsive hydrogel chosen for the present study, crosslinked hydroxypropylcellulose, shrinks over a rather narrow temperature range near 44 C. Some vertebrate skin is also subject to substantial strain. Among reptiles, the morphologies of the skin and scales show wide variations. Bauer et al. described the mechanical properties and histology of gecko skin; longitudinal tensile properties of snake skin were examined by Jayne with reference to locomotion. The present measurements focus on adaptations related to feeding, including the response of the skin to circumferential tension. Tensile properties will be related to interspecific and regional variation in skin structure and folding.

Hinkley, Jeffrey A.↗
NASA NTRS2003

The Zebrafish G12 Gene is required for Nuclear Positioning and Cell Migrations during Early Development

After fertilization Zebrafish embryos undergo synchronous cleavage to form a blastula of cells sitting upon a single multinucleate yolk cell. At the beginning of gastrulation these cells undergo extensive cell migrations to form the major body axes. We have discovered a gene, G12, which is required for cell migrations and positioning of nuclei in the large syncytial yolk cell. Overexpression of a G12-GFP fusion protein is not toxic and shows that the protein localizes inside the yolk cell to the yolk nuclei, microtubules, and to the margin between the blastomeres and the large yolk cell. Morpholino (MO) injection into the 1-cell embryo or into just the yolk syncytium conipletely inhibits cell migrations, doming of the yolk cell, and positioning of nuclei around the margin. This effect can be partially rescued by injection of G12-GFP encoding RNA. Given the known role of microtubules in nuclear positioning of yolk nuclei in Zebrafish, we investigated the microtubules in morpholiiio injected and rescued embryos. We find that microtubules are sparse and disorganized in MO-injected embryos and are restored to normal organization upon G12-GFP rescue. G12 plays a pivotal role in organization of inicrotubules during early development. G12 is highly conserved in vertebrates and two homologues exist in the human genome. One of the human hoinologues is amplified in aggressive breast tumors.

Reinsch, S. S.↗
NASA NTRS2002

Worms on the International Space Station

C. elegans was proposed as a model system for space biology studies in 1991 and has since flown on STS-42, STS-76, and STS-95. Data obtained from these flights have confirmed that C. elegans requires adequate in flight oxygenation and displays an increased rate of mutation, much like other organisms in space. Unlike vertebrates, C. elegans has been observed to successfully complete two continuous full life cycles in space without gross developmental abnormalities. These observations, could with the utility of C. elegans as a terrestrial, fully sequenced, model system make C. elegans a good candidate for long term research onboard the International Space Station (ISS). We are currently working on technology to support biological studies aboard the ISS. A component of this effort is in the development of the Space Station Biological Research Program (SSBRP) Incubator which will be able to house organisms at a constant temperature setpoint ranging from 4deg C to 45deg C. The SSBRP Incubator provides air exchange, power, data and video ports and, when placed in the collaborative NASA/NASDA 2.5M centrifuge rotor, will be capable of providing a lg gravity control. Current plans for validation of the Incubator include video monitoring and periodic sampling of C. elegans in the Incubator onboard the ISS. Once returned to earth, samples will be distributed for analysis via a specimen sharing plan and analyzed for gene expression and other parameters of growth and development in space flight. These data should provide the C. elegans research community with a baseline from which to propose studies for future flights. We have also been developing an appropriate method of culturing C. elegans in liquid media in order to remove the need for the crew to assure that strains are properly fed. Currently, we are growing strains in the chemically defined, axenic, media developed by Dr. Nancy Lu. Wild-type animals complete multiple generations and appear generally healthy after being grown in unchanged media for a minimum of three to four weeks. To assure proper oxygenation we have chosen to grow animals in ten milliliter OptiCells(TradeMark). The choice of the OptiCell(TradeMark) also allows for automation of culturing as demonstrated for cell cultures grown in the OptiCell(TradeMark). Arrangements have been made for the liquid media to be commercially produced by Mediatech, Inc. and OptiCells(TradeMark) are available from BioCrystal Ltd.

Szewczyk, Nate↗
NASA NTRS2003

Morphology of the Vestibular Utricule in Toadfish, Opsanus Tau

The uticle is an otolith organ in the vertebrate inner ear that provides gravitoinertial acceleration information into the vestibular reflex pathways. The aim of the present study was to provide an anatomical description of this structure in the adult oyster toadfish, and establish a morphological basis for interpretation of subsequent functional studies. Light, scanning electron and transmission electron microscopy were applied to visualize the sensory epithelium and its neural innervation. Electrophysiological techniques were used to identify utricular afferents by their response to translation stimuli. Similar to nerve afferents supplying the semicircular canals and lagena, utricular afferents commonly exhibit a short-latency increase of firing rate in response to electrical activation of the central efferent pathway. Afferents were labeled with biocytin either intraaxonally or with extracellular bulk deposits. Light microscope images of serial thick sections were used to make three-dimensional reconstructions of individual labeled afferents to identify the dendritic morphology with respect to epithelial location. Scanning electron microscopy was used to visualize the surface of the otolith mass facing the otolith membrane, and the hair cell polarization patterns of strioler and extrastriolar regions. Transmission electron micrographs of serial thin sections were compiled to create a three-dimensional reconstruction of the labeled afferent over a segment of its dendritic field and to examine the hair cell-afferent synaptic contacts.

Bass, L.↗
NASA NTRS2004

Cellular Mechanisms Underlying Bone-Forming Cell Proliferative Response to Hypergravity

Life on Earth has evolved under the continuous influence of gravity (1-g). As humans explore and develop space, however, we must learn to adapt to an environment with little or no gravity. Studies indicate that lack of weightbearing for vertebrates occurring with immobilization, paralysis, or in a microgravity environment may cause muscle and bone atrophy through cellular and subcellular level mechanisms. We hypothesize that gravity is needed for the efficient transduction of cell growth and survival signals from the extra-cellular matrix (ECM) (consisting of molecules such as collagen, fibronectin, and laminin) in mechanosensitive tissues. We test for the presence of gravity-sensitive pathways in bone-forming cells (osteoblasts) using hypergravity applied by a cell culture centrifuge. Stimulation of 50 times gravity (50-g) increased proliferation in primary rat osteoblasts for cells grown on collagen Type I and fibronectin, but not on laminin or uncoated surfaces. Survival was also enhanced during hypergravity stimulation by the presence of ECM. Bromodeoxyuridine incorporation in proliferating cells showed an increase in the number of actively dividing cells from about 60% at 1-g to over 90% at 25-g. Reverse transcription-polymerase chain reaction was used to test for all possible integrins. Our combined results indicate that beta1 and/or beta3 integrin subunits may be involved. These data indicate that gravity mechanostimulation of osteoblast proliferation involves specific matrix-integrin signalling pathways which are sensitive to g-level. Further research to define the mechanisms involved will provide direction so that we may better adapt and counteract bone atrophy caused by the lack of weightbearing.

Vercoutere, W.↗