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Amphibian fertilization and development in microgravity

During the year before launch, female frogs will be tested every 3 months for the quantity and quality of eggs produced. Two weeks or more prior to launch, male and female frogs will be transported to the John F. Kennedy Space Center (KSC). During the few weeks before launch, groups will be periodically tested for egg quality to assure that the frogs have adapted to the KSC laboratory environment. About 27 hours before launch, four females will be placed in a damp foam-lined box, called the Adult Frog Box (AFB), through which 100 cc/min of air wil be circulated. The AFB will be lowered into the Spacelab and loaded into the Frog Environmental Unit (FEU) during the final pre-launch preparations. A sperm suspension, for use in flight to fertilize the eggs, will also be prepared and loaded during the pre-launch period. The sperm suspension, together with a kit of syringes containing Human Chorionic Gonadotropin (HCG), will be stored in a refrigerator aboard the shuttle until needed in flight. On the first day of flight, the AFB will be transferred from the FEU to the General Purpose Work Station (GPWS), which is a type of glove box specially designed to allow the crew to use chemicals and biological materials during the flight without contaminating the shuttle/Spacelab environment. Inside the GPWS the four adult frogs will be injected with the HCG hormone and returned to the FEU. Approximately 16 hours after injection, ovulation should have taken place and 15 to 20 eggs from each frog will be placed on each of two egg baskets and covered with sperm for 10 minutes. The egg baskets are inserted into acrylic egg chambers and 50 ml of 'pond water' (20 percent strength Modified Ringers solution (is added. One of the chambers from each frog will be placed on a centrifuge within the FEU and rotated to simulate normal terrestrial gravity (1 g). The remaining chambers are incubated under microgravity conditions within the FEU. Forty minutes after fertilization, the four chambers exposed to 1 g will be removed and observed within the GPWS using a dissecting microscope and camera system. On the basis of egg rotation and egg appearance, the 'best' and 'second best' frogs will be selected to contribute additional eggs. Using the two best frogs, 22 egg chambers will be loaded with eggs and fertilized. Eleven chambers wil be incubated at microgravity and eleven will be incubated on the centrifuge. At various times during the flight, chambers will be removed from the FEU and transferred to the GPWS where a formaldehyde-based fixative will be injected in order to preserve important developmental stages for in depth study following the flight. Five of the 22 chambers plus the eight fertilization test chambers will be returned to Earth with live tadpoles. The swimming behavior of these free swimming tadpoles will be examined within several hours of landing and some will be fixed for a detailed analysis of their inner ear, the otolity, the animals 'balance system.' Lychakov and Vinikov reported an increase in otolith size in tadpoles that developed (but were not fertilized) in space. Live tadpoles from the SL-J flight will be raised through metamorphsis for studies of maturation including an analysis of the effects of space flight on their ability to produce normal progency. Following the flight, the fixed embryos will be serially sectioned and stained using procedures that were developed for this experiment. The embroys fixed at the 2-4 cell stage will be examined for the distribution of cytoplasmic contents, including the various classes of yolk platelets, and the location and shape of the cleavage furrows. The gastrulae will be assessed for the normality of the complex cellular rearrangements that constitute blastopore formation and involution, i.e., the differentiation of the embryo into its specialized body parts. Using a new procedure, the gastrulae will be bleached which will enable the initial sperm entry point (SEP) to be located. The correlation between the SEP and the dorsal lip of the blastopore will be determined. Under normal terrestrial conditions it was shown that the SEP typically is located on the side of the egg opposite the future dorsal side of the embryo. The neurulae will be examined for the normality and completeness of the neural plate and archenteron expansion. The tadpole stages will be used to study the allometry and morphology of the various organ systems.

Souza, Kenneth A.

Formation of otoconia in the Japanese red-bellied newt, Cynops pyrrhogaster

Pre-mated adult female newts and fertilized eggs will be flown on the International Microgravity Laboratory-2 flight, in 1994. One objective of the flight will be to observe the influence of microgravity on the development of the gravity-sensing organs in the inner ear. These organs contain sensory hair cells covered by a layer of dense stones (otoconia). Gravity and linear acceleration exert forces on these masses, leading to excitation of the nerve fibers innervating the hair cells. If the production of the otoliths is regulated to reach an optimal weight, their development might be abnormal in microgravity. Ground-based control experiments are reported describing the developmental sequence in which both the otoliths and their associated sensory epithelium and the semicircular canals appear and develop. Three-dimensional reconstruction of serial sections through the otic vesicle of newt embryos at stages 31 through 58 demonstrate the first appearance, relative position and growth of the otoliths. Reports of experiments in which fertilized frog eggs were flown on a Russian Cosmos mission conclude that the utricular otolith is increased in volume, whereas the saccular otolith maintains normal size, suggesting that at least in the utricle, the weight of the otolith might be regulated.

Wiederhold, M. L.

Development of the gravity-sensing organs in the Japanese red-bellied newt, Cynops pyrrhogaster

Pre-mated adult female newts and fertilized eggs will be flown on the International Microgravity Laboratory-2 flight, schedule for 1994. One objective of the flight will be to observe the influence of microgravity on the development of the gravity-sensing organs in the inner ear. These organs contain sensory hair cells covered by a layer of dense stones (otoliths). Gravity and linear acceleration exert forces on these masses, leading to excitation of the nerve fibers innervating the hair cells. If the production of the otoliths is regulated to reach an optimal weight, their development would be abnormal in microgravity. Ground-based control experiments are reported describing the developmental sequence in which the otoliths and their associated sensory epithelium appear and increase in size. Three-dimensional reconstruction of serial sections through the otic vesicle of newt embryos at stages 31 through 40 demonstrate the first appearance, relative position and growth of the otoliths. In adult newts, the otoconia in the utricle appear similar to mammalian otoconia, which are composed of calcite. The newt saccular otoconia are at least 99% aragonite, as is found in most aquatic species. Reports of experiments in which fertilized frog eggs were flown on a Russian Cosmos mission conclude that the utricular otolith is increased in volume, whereas the saccular otolith maintains normal size, suggesting that at least the utricular weight might be regulated.

Wiederhold, Michael L.

Trabecular Bone Mechanical Properties and Fractal Dimension

Countermeasures for reducing bone loss and muscle atrophy due to extended exposure to the microgravity environment of space are continuing to be developed and improved. An important component of this effort is finite element modeling of the lower extremity and spinal column. These models will permit analysis and evaluation specific to each individual and thereby provide more efficient and effective exercise protocols. Inflight countermeasures and post-flight rehabilitation can then be customized and targeted on a case-by-case basis. Recent Summer Faculty Fellowship participants have focused upon finite element mesh generation, muscle force estimation, and fractal calculations of trabecular bone microstructure. Methods have been developed for generating the three-dimensional geometry of the femur from serial section magnetic resonance images (MRI). The use of MRI as an imaging modality avoids excessive exposure to radiation associated with X-ray based methods. These images can also detect trabecular bone microstructure and architecture. The goal of the current research is to determine the degree to which the fractal dimension of trabecular architecture can be used to predict the mechanical properties of trabecular bone tissue. The elastic modulus and the ultimate strength (or strain) can then be estimated from non-invasive, non-radiating imaging and incorporated into the finite element models to more accurately represent the bone tissue of each individual of interest. Trabecular bone specimens from the proximal tibia are being studied in this first phase of the work. Detailed protocols and procedures have been developed for carrying test specimens through all of the steps of a multi-faceted test program. The test program begins with MRI and X-ray imaging of the whole bones before excising a smaller workpiece from the proximal tibia region. High resolution MRI scans are then made and the piece further cut into slabs (roughly 1 cm thick). The slabs are X-rayed again and also scanned using dual-energy X-ray absorptiometry (DEXA). Cube specimens are then cut from the slabs and tested mechanically in compression. Correlations between mechanical properties and fractal dimension will then be examined to assess and quantify the predictive capability of the fractal calculations.

Hogan, Harry A.

Morphological Changes in Rat Vestibular System Following Weightlessness

Mammalian gravity receptors (maculas) are morphologically organized for weighted, parallel distributed processing of information. There are two basic circuits: 1) highly channeled, type I cell to calyx; and 2) distributed modifying, type II cells to calyces and processes. The latter circuit should be the more adaptable since it modifies final output. To test this hypothesis, rats were flown in microgravity for 9 days aboard a space shuttle and euthanized shortly after landing. Hair cells and ribbon synapses from maculas of 3 flight and 3 ground control rats were studied ultrastructurally in blocks of 50 serial sections. Synapses increased by approximately 41% in type I cells and by approximately 55% in type II cells in flight animals. There was a shift toward the spherular form of ribbon synapse in both types of hair cells in flight animals. Current findings tend to support the stated hypothesis and indicate that mature utricular hair cells retain synaptic plasticity, permitting adaptation to an altered gravitational environment.

Ross, Muriel D.

Mammalian Vestibular Macular Synaptic Plasticity: Results from SLS-2 Spaceflight

The effects of exposure to microgravity were studied in rat utricular maculas collected inflight (IF, day 13), post-flight on day of orbiter landing (day 14, R+O) and after 14 days (R+ML). Controls were collected at corresponding times. The objectives were 1) to learn whether hair cell ribbon synapses counts would be higher in tissues collected in space than in tissues collected postflight during or after readaptation to Earth's gravity; and 2) to compare results with those of SLS-1. Maculas were fixed by immersion, micro-dissected, dehydrated and prepared for ultrastructural study by usual methods. Synapses were counted in 100 serial sections 150 nm thick and were located to specific hair cells in montages of every 7th section. Counts were analyzed for statistical significance using analysis of variance. Results in maculas of IF dissected rats, one 13 day control (IFC), and one R + 0 rat have been analyzed. Study of an R+ML macula is nearly completed. For type I cells, IF mean is 2.3 +/-1.6; IFC mean is 1.6 +/-1.0; R+O mean is 2.3 +/- 1.6. For type II cells, IF mean is 11.4 +/- 17.1; IFC mean is 5.5 +/-3.5; R+O mean is 10.1 +/- 7.4. The difference between IF and IFC means for type I cells is statistically significant (p less than 0.0464). For type It cells, IF compared to IFC means, p less than 0.0003; and for IFC to R+O means, p less than 0.0139. Shifts toward spheres (p less than 0.0001) and pairs (p less than 0.0139) were significant in type II cells of IF rats. The results are largely replicating findings from SLS-1 and indicate that spaceflight affects synaptic number, form and distribution, particularly in type II hair cells. The increases in synaptic number and in sphere-like ribbons are interpreted to improve synaptic efficacy, to help return afferent discharges to a more normal state. Findings indicate that a great capacity for synaptic plasticity exists in mammalian gravity sensors, and that this plasticity is more dominant in the local circuitry. The local circuit includes type II cells and is interpreted to be responsible for shaping the final output of the system.

Ross, Muriel D.D.

Computational Approaches to Vestibular Research

The Biocomputation Center at NASA Ames Research Center is dedicated to a union between computational, experimental and theoretical approaches to the study of neuroscience and of life sciences in general. The current emphasis is on computer reconstruction and visualization of vestibular macular architecture in three-dimensions (3-D), and on mathematical modeling and computer simulation of neural activity in the functioning system. Our methods are being used to interpret the influence of spaceflight on mammalian vestibular maculas in a model system, that of the adult Sprague-Dawley rat. More than twenty 3-D reconstructions of type I and type II hair cells and their afferents have been completed by digitization of contours traced from serial sections photographed in a transmission electron microscope. This labor-intensive method has now been replace d by a semiautomated method developed in the Biocomputation Center in which conventional photography is eliminated. All viewing, storage and manipulation of original data is done using Silicon Graphics workstations. Recent improvements to the software include a new mesh generation method for connecting contours. This method will permit the investigator to describe any surface, regardless of complexity, including highly branched structures such as are routinely found in neurons. This same mesh can be used for 3-D, finite volume simulation of synapse activation and voltage spread on neuronal surfaces visualized via the reconstruction process. These simulations help the investigator interpret the relationship between neuroarchitecture and physiology, and are of assistance in determining which experiments will best test theoretical interpretations. Data are also used to develop abstract, 3-D models that dynamically display neuronal activity ongoing in the system. Finally, the same data can be used to visualize the neural tissue in a virtual environment. Our exhibit will depict capabilities of our computational approaches and some of our findings from their application. For example, our research has demonstrated that maculas of adult mammals retain the property of synaptic plasticity. Ribbon synapses increase numerically and undergo changes in type and distribution (p<0.0001) in type II hair cells after exposure to microgravity for as few as nine days. The finding of macular synaptic plasticity is pertinent to the clinic, and may help explain some. balance disorders in humans. The software used in our investigations will be demonstrated for those interested in applying it in their own research.

Ross, Muriel D.

New Computer Simulations of Macular Neural Functioning

We use high performance graphics workstations and supercomputers to study the functional significance of the three-dimensional (3-D) organization of gravity sensors. These sensors have a prototypic architecture foreshadowing more complex systems. Scaled-down simulations run on a Silicon Graphics workstation and scaled-up, 3-D versions run on a Cray Y-MP supercomputer. A semi-automated method of reconstruction of neural tissue from serial sections studied in a transmission electron microscope has been developed to eliminate tedious conventional photography. The reconstructions use a mesh as a step in generating a neural surface for visualization. Two meshes are required to model calyx surfaces. The meshes are connected and the resulting prisms represent the cytoplasm and the bounding membranes. A finite volume analysis method is employed to simulate voltage changes along the calyx in response to synapse activation on the calyx or on calyceal processes. The finite volume method insures that charge is conserved at the calyx-process junction. These and other models indicate that efferent processes act as voltage followers, and that the morphology of some afferent processes affects their functioning. In a final application, morphological information is symbolically represented in three dimensions in a computer. The possible functioning of the connectivities is tested using mathematical interpretations of physiological parameters taken from the literature. Symbolic, 3-D simulations are in progress to probe the functional significance of the connectivities. This research is expected to advance computer-based studies of macular functioning and of synaptic plasticity.

Ross, Muriel D.

Synaptic Plasticity In Mammalian Gravity Sensors: Preliminary Results From SLS-2

Sensory conflict is the prevalent theoretical explanation for space adaptation syndrome. This ultrastructural study tests the hypothesis that peripheral gravity sensors (maculae) play a role. Results were obtained from the medial part of utricular maculae of adult rats exposed to microgravity for 14 days, and from controls. Means and statistical significance of synapse counts were calculated using SUPERANOVA(Trademark) and Scheffe's procedure for post-hoc comparisons. Preliminary findings are from 2 sets of 100 serial sections for each dataset. Synapses were doubled numerically in type II hair cells of utricular maculae collected on day 13 inflight compared to controls (11.4 +/- 7.1 vs. 5.3 +/- 3.8; p < 0.0001). Flight mean synaptic number declined rapidly postflight and became comparable to means of controls. Synapses also increased numerically in type I cells inflight (2.4 +/- 1.6 vs. 1.7 +/- 1.0; p < 0.0341). Postflight there were no significant differences in counts. Results concerning shifts in ribbon type and distribution are also largely replicating previous findings from flight studies. Results indicate that mammalian maculae are adaptive endorgans that retain the property of synaptic plasticity into the adult stage. Macular plasticity has clinical implications for balance disorders of peripheral origin.

Ross, M. D.

The Evolution of Dendrite Morphology during Isothermal Coarsening

Dendrite coarsening is a common phenomenon in casting processes. From the time dendrites are formed until the inter-dendritic liquid is completely solidified dendrites are changing shape driven by variations in interfacial curvature along the dendrite and resulting in a reduction of total interfacial area. During this process the typical length-scale of the dendrite can change by orders of magnitude and the final microstructure is in large part determined by the coarsening parameters. Dendrite coarsening is thus crucial in setting the materials parameters of ingots and of great commercial interest. This coarsening process is being studied in the Pb-Sn system with Sn-dendrites undergoing isothermal coarsening in a Pb-Sn liquid. Results are presented for samples of approximately 60% dendritic phase, which have been coarsened for different lengths of times. Presented are three-dimensional microstructures obtained by serial-sectioning and an analysis of these microstructures with regard to interface orientation and interfacial curvatures. These graphs reflect the evolution of not only the microstructure itself, but also of the underlying driving forces of the coarsening process. As a visualization of the link between the microstructure and the driving forces a three-dimensional microstructure with the interfaces colored according to the local interfacial mean curvature is shown.

Alkemper, Jens

Modulation of statolith mass and grouping in white clover (Trifolium repens) growth in 1-g, microgravity and on the clinostat

Current models of gravity perception in higher plants focus on the buoyant weight of starch-filled amyloplasts as the initial gravity signal susceptor (statolith). However, no tests have yet determined if statolith mass is regulated to increase or decrease gravity stimulus to the plant. To this end, the root caps of white clover (Trifolium repens) grown in three gravity environments with three different levels of gravity stimulation have been examined: (i) 1-g control with normal static gravistimulation, (ii) on a slow clinostat with constant gravistimulation, and (iii) in the stimulus-free microgravity aboard the Space Shuttle. Seedlings were germinated and grown in the BioServe Fluid Processing Apparatus and root cap structure was examined at both light and electron microscopic levels, including three-dimensional cell reconstruction from serial sections. Quantitative analysis of the electron micrographs demonstrated that the starch content of amyloplasts varied with seedling age but not gravity condition. It was also discovered that, unlike in starch storage amyloplasts, all of the starch granules of statolith amyloplasts were encompassed by a fine filamentous, ribosome-excluding matrix. From light micrographic 3-D cell reconstructions, the absolute volume, number, and positional relationships between amyloplasts showed (i) that individual amyloplast volume increased in microgravity but remained constant in seedlings grown for up to three days on the clinostat, (ii) the number of amyloplasts per cell remained unchanged in microgravity but decreased on the clinostat, and (iii) the three-dimensional positions of amyloplasts were not random. Instead amyloplasts in microgravity were grouped near the cell centers while those from the clinostat appeared more dispersed. Taken together, these observations suggest that changing gravity stimulation can elicit feedback control over statolith mass by changing the size, number, and grouping of amyloplasts. These results support the starch-statolith theory of graviperception in higher plants and add to current models with a new feedback control loop as a mechanism for modulation of statolith responsiveness to inertial acceleration.

Flight Experiment

Atrial development in the human heart: an immunohistochemical study with emphasis on the role of mesenchymal tissues

The development of the atrial chambers in the human heart was investigated immunohistochemically using a set of previously described antibodies. This set included the monoclonal antibody 249-9G9, which enabled us to discriminate the endocardial cushion-derived mesenchymal tissues from those derived from extracardiac splanchnic mesoderm, and a monoclonal antibody recognizing the B isoform of creatine kinase, which allowed us to distinguish the right atrial myocardium from the left. The expression patterns obtained with these antibodies, combined with additional histological information derived from the serial sections, permitted us to describe in detail the morphogenetic events involved in the development of the primary atrial septum (septum primum) and the pulmonary vein in human embryos from Carnegie stage 14 onward. The level of expression of creatine kinase B (CK-B) was found to be consistently higher in the left atrial myocardium than in the right, with a sharp boundary between high and low expression located between the primary septum and the left venous valve indicating that the primary septum is part of the left atrial gene-expression domain. This expression pattern of CK-B is reminiscent of that of the homeobox gene Pitx2, which has recently been shown to be important for atrial septation in the mouse. This study also demonstrates a poorly appreciated role of the dorsal mesocardium in cardiac development. From the earliest stage investigated onward, the mesenchyme of the dorsal mesocardium protrudes into the dorsal wall of the primary atrial segment. This dorsal mesenchymal protrusion is continuous with a mesenchymal cap on the leading edge of the primary atrial septum. Neither the mesenchymal tissues of the dorsal protrusion nor the mesenchymal cap on the edge of the primary septum expressed the endocardial tissue antigen recognized by 249-9G9 at any of the stages investigated. The developing pulmonary vein uses the dorsal mesocardium as a conduit to reach the primary atrial segment. Initially, the pulmonary pit, which will becomes the portal of entry for the pulmonary vein, is located along the midline, flanked by two myocardial ridges. As development progresses, tissue remodeling results in the incorporation of the portal of entry of the pulmonary vein in left atrial myocardium, which is recognized because of its high level of creatine. Closure of the primary atrial foramen by the primary atrial septum occurs as a consequence of the fusion of these mesenchymal structures. Copyright 2000 Wiley-Liss, Inc.

NASA Discipline Cardiopulmonary

Production of otoconia in the endolymphatic sac in the Japanese red-bellied newt, Cynops pyrrhogaster: light and transmission electron microscopic study

The formation of otoconia in the endolymphatic sac (ES) of the larval newt, Cynops pyrrhogaster, has been studied by light and transmission electron microscopy. Some of the epithelial cells of the ES contain an abundance of swollen vesicles, Golgi complexes, rough endoplasmic reticula and ribosomes at the late larval stages 50 and 51, approximately 26-30 days after eggs are laid. Five days later, at stage 52, crystals are present in the vacuoles between the epithelial cells. Serial sections indicate that these vacuoles actually form small canals which lie in the wall and join the lumen of the ES. Reconstruction of the ES shows that several canals are contained in the ES wall. At stage 56, about 72 days after eggs are laid, a large number of otoconia are present in the ES lumen, while the otoconia disappear from the canals. It appears that the otoconia are first produced in the canals and then released to the lumen. Some epithelial cells of the ES are thought to expel the organic and inorganic material to the canals to form the otoconia in situ. The process of formation of the otoconia in the ES is discussed.

Non-NASA Center

Morphological evidence for local microcircuits in rat vestibular maculae

Previous studies suggested that intramacular, unmyelinated segments of vestibular afferent nerve fibers and their large afferent endings (calyces) on type I hair cells branch. Many of the branches (processes) contain vesicles and are presynaptic to type II hair cells, other processes, intramacular nerve fibers, and calyces. This study used serial section transmission electron microscopy and three-dimensional reconstruction methods to document the origins and distributions of presynaptic processes of afferents in the medial part of the adult rat utricular macula. The ultrastructural research focused on presynaptic processes whose origin and termination could be observed in a single micrograph. Results showed that calyces had 1) vesiculated, spine-like processes that invaginated type I cells and 2) other, elongate processes that ended on type II cells pre- as well as postsynaptically. Intramacular, unmyelinated segments of afferent nerve fibers gave origin to branches that were presynaptic to type II cells, calyces, calyceal processes, and other nerve fibers in the macula. Synapses with type II cells occurred opposite subsynaptic cisternae (C synapses); all other synapses were asymmetric. Vesicles were pleomorphic but were differentially distributed according to process origin. Small, clear-centered vesicles, approximately 40-60 nm in diameter, predominated in processes originating from afferent nerve fibers and basal parts of calyces. Larger vesicles approximately 70-120 nm in diameter having approximately 40-80 nm electron-opaque cores were dominant in processes originating from the necks of calyces. Results are interpreted to indicate the existence of a complex system of intrinsic feedforward (postsynaptic)-feedback (presynaptic) connections in a network of direct and local microcircuits. The morphological findings support the concept that maculae dynamically preprocess linear acceleratory information before its transmission to the central nervous system.

NASA Center ARC

Evidence of a sensory processing unit in the mammalian macula

We cut serial sections through the medial part of the rat vestibular macula for transmission electron microscopic (TEM) examination, computer-assisted 3-D reconstruction, and compartmental modeling. The ultrastructural research showed that many primary vestibular neurons have an unmyelinated segment, often branched, that extends between the heminode (putative site of the spike initiation zone) and the expanded terminal(s) (calyx, calyces). These segments, termed the neuron branches, and the calyces frequently have spine-like processes of various dimensions with bouton endings that morphologically are afferent, efferent, or reciprocal to other macular neural elements. The major questions posed by this study were whether small details of morphology, such as the size and location of neuronal processes or synapses, could influence the output of a vestibular afferent, and whether a knowledge of morphological details could guide the selection of values for simulation parameters. The conclusions from our simulations are (1) values of 5.0 k omega cm2 for membrane resistivity and 1.0 nS for synaptic conductance yield simulations that best match published physiological results; (2) process morphology has little effect on orthodromic spread of depolarization from the head (bouton) to the spike initiation zone (SIZ); (3) process morphology has no effect on antidromic spread of depolarization to the process head; (4) synapses do not sum linearly; (5) synapses are electrically close to the SIZ; and (6) all whole-cell simulations should be run with an active SIZ.

NASA Center ARC

Development of the otolith organs and semicircular canals in the Japanese red-bellied newt, Cynops pyrrhogaster

The sequence in which the otoliths and semicircular canals and their associated sensory epithelia appear and develop in the newt are described. Three-dimensional reconstruction of serial sections through the otic vesicle of newt embryos from stages 31 through 58 demonstrate the first appearance, relative position and growth of the otoliths. A single otolith is first seen in stage 33 embryos (approximately 9 days old); this splits into separate utricular and saccular otoliths at stage 40 (13 days). The lateral semicircular canal is the first to appear, at stage 41 (14 days). The anterior and posterior canals appear approximately one week later and the vestibular apparatus is essentially fully formed at stage 58 (approximately 5 weeks). The data reported here will serve as ground-based controls for fertilized newt eggs flown on the International Microgravity Laboratory-2 Space Shuttle flight, to investigate the influence of microgravity on the development of the gravity-sensing organs.

NASA Discipline Neuroscience

A spaceflight study of synaptic plasticity in adult rat vestibular maculas

Behavioral signs of vestibular perturbation in altered gravity have not been well correlated with structural modifications in neurovestibular centers. This ultrastructural research investigated synaptic plasticity in hair cells of adult rat utricular maculas exposed to microgravity for nine days on a space shuttle. The hypothesis was that synaptic plasticity would be more evident in type II hair cells because they are part of a distributed modifying macular circuitry. All rats were shared with other investigators and were subjected to treatments unrelated to this experiment. Maculas were obtained from flight and control rats after shuttle return (R + 0) and nine days post-flight (R + 9). R + 9 rats had chromodacryorrhea, a sign of acute stress. Tissues were prepared for ultrastructural study by conventional methods. Ribbon synapses were counted in fifty serial sections from medial utricular macular regions of three rats of each flight and control group. Counts in fifty additional consecutive sections from one sample in each group established method reliability. All synapses were photographed and located to specific cells on mosaics of entire sections. Pooled data were analyzed statistically. Flown rats showed abnormal posture and movement at R + 0. They had statistically significant increases in total ribbon synapses and in sphere-like ribbons in both kinds of hair cells; in type II cells, pairs of synapses nearly doubled and clusters of 3 to 6 synapses increased twelve-fold. At R + 9, behavioral signs were normal. However, synapse counts remained high in both kinds of hair cells of flight maculas and were elevated in control type II cells. Only counts in type I cells showed statistically significant differences at R + 9. High synaptic counts at R + 9 may have resulted from stress due to experimental treatments. The results nevertheless demonstrate that adult maculas retain the potential for synaptic plasticity. Type II cells exhibited more synaptic plasticity, but space flight induced synaptic plasticity in type I cells.

Weightlessness

NASA Tech Briefs, August 2003

Topics covered include: Stable, Thermally Conductive Fillers for Bolted Joints; Connecting to Thermocouples with Fewer Lead Wires; Zipper Connectors for Flexible Electronic Circuits; Safety Interlock for Angularly Misdirected Power Tool; Modular, Parallel Pulse-Shaping Filter Architectures; High-Fidelity Piezoelectric Audio Device; Photovoltaic Power Station with Ultracapacitors for Storage; Time Analyzer for Time Synchronization and Monitor of the Deep Space Network; Program for Computing Albedo; Integrated Software for Analyzing Designs of Launch Vehicles; Abstract-Reasoning Software for Coordinating Multiple Agents; Software Searches for Better Spacecraft-Navigation Models; Software for Partly Automated Recognition of Targets; Antistatic Polycarbonate/Copper Oxide Composite; Better VPS Fabrication of Crucibles and Furnace Cartridges; Burn-Resistant, Strong Metal-Matrix Composites; Self-Deployable Spring-Strip Booms; Explosion Welding for Hermetic Containerization; Improved Process for Fabricating Carbon Nanotube Probes; Automated Serial Sectioning for 3D Reconstruction; and Parallel Subconvolution Filtering Architectures.

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