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All sourcesNASA NTRSDOE OSTI
NASA NTRSDate not supplied

Multi-Omics Analysis of Mouse Retina Following Low Dose Radiation and/or Hindlimb Unloading

Rodent models have been used as analogs for studying the effects of spaceflight. NASA’s GeneLab provides access to omics datasets generated from spaceflight and ground-based experiments allowing for additional retrospective analysis. We used GeneLab’s GLDS-203, a dataset generated by researchers at Loma Linda University to study the impact of prolonged unloading and/or low-dose radiation on mouse retina. The purpose of this study was to understand the effect of gamma radiation and/or hindlimb unloading on mice retinas through a multi-omics analysis. In the experiment that generated the omics data, mice were irradiated with gamma-ray and/or subjected to hindlimb unloading for 21 days and multi-omics analysis was performed at 7 days, 1 month, or 4 months post exposure. In the current study, for each of the three timepoints, we compared epigenomic profiles for retinas from exposed mice against timepoint-matched controls. We identified a total of 5,271 differentially methylated loci (DML) and 321 differentially methylated regions (DMR; using a sliding window and step size of 500 bp) with methylation difference > 10% and q-value < 0.05 (sliding linear model corrected p-value) across the nine exposure groups. Highest correlation in methylation difference was seen for significant DMLs (q-value < 0.05) across different conditions at same post exposure timepoint (Figure 1).The location of DMLs and DMRs were characterized with respect to CpG islands and shores, putative promoters, gene body, and intergenic regions (Table 1). We analyzed RNA-seq counts and performed gene set enrichment analysis using differential expression results from comparing each exposure group to its timepoint-matched control group. Significant pathways (adjusted p-value <0.05) enriched in all three microgravity-only groups were related to morphogenesis of a branching epithelium, skeletal muscle cell differentiation, and response to fibroblast growth factor. Common processes across all timepoints in the radiation-only groups were retina homeostasis, synaptic vesicle exocytosis-endocytosis, and chemotaxis. In the combination groups, regulation of trans-synaptic signaling, and Rho protein signal transduction were enriched at all three timepoints. Processes related to purine nucleotide metabolism were enriched in all nine exposure groups, with activation at 1 month, and suppression at 7 days and 4 months. A total of 14 genes contained at least one DML and were differentially expressed at adjusted p-value < 0.05, including genes implicated in cataract development (Sipa1l3, Crybb3) and those involved in cytoskeletal organization (Plec, Flnb, Eef1a1). This analysis is part of a larger effort to understand the molecular mechanisms following spaceflight exposures that can help translate effects observed in animal models to human impacts.

Prachi Kothiyal↗
NASA NTRS1991

Performance of the Research Animal Holding Facility (RAHF) and General Purpose Work Station (GPWS) and other hardware in the microgravity environment

This paper discusses the performance of the Research Animal Holding Facility (RAHF) and General Purpose Work Station (GPWS) plus other associated hardware during the recent flight of Spacelab Life Sciences 1 (SLS-1). The RAHF was developed to provide proper housing (food, water, temperature control, lighting and waste management) for up to 24 rodents during flights on the Spacelab. The GPWS was designed to contain particulates and toxic chemicals generated during plant and animal handling and dissection/fixation activities during space flights. A history of the hardware development involves as well as the redesign activities prior to the actual flight are discussed.

Hogan, Robert P.↗
NASA NTRSDate not supplied

Transcriptomic Analysis of Irradiated Mouse Retina Following Readaptation

Rodent models are used as analogs for studying the effects of spaceflight. NASA GeneLab provides access to relevant omics datasets generated from spaceflight and ground-based experiments allowing for additional retrospective analysis. In this study, we used GeneLab’s GLDS-203, a dataset generated by researchers at Loma Linda University to study the impact of prolonged unloading and/or low-dose radiation on mouse retina. We analyzed transcriptomics data from retina of mice irradiated with gamma-rays for 21 days followed by 7 days, 1 month, or 4 months of readaptation. We obtained raw gene counts from GeneLab and performed differential gene expression analysis after data normalization. For each of the three timepoints, we performed differential expression analysis to compare transcriptional profiles for retina from irradiated vs. non-irradiated (controls) mice, all exposed to gravity. We observed the highest number of differentially expressed genes at 7 days, followed by 1 month and 4 months. Enrichment analysis showed top pathways (adjusted p-value < 0.05) were related to transport along microtubule and photoreceptor cell development in the 7-day readaptation group. Fewer significantly enriched pathways were observed for the 1-month group and included mRNA metabolic processes and neuron differentiation. No significantly enriched pathways were found in the 4-month group. The Gene Ontology biological processes common between the 7 days and 1-month groups include visual perception, synapse organization, and perception of light stimulus. This analysis is part of a larger effort to characterize the molecular mechanisms involved in retinal readaptation following radiation exposure. Future analyses will include other related retina datasets in GeneLab repository to assess whether gene expression patterns are consistent across different study cohorts.

Prachi Kothiyal↗
NASA NTRS1994

Physiological Anatomical Rodent Experiment (PARE) .04 feasibility test 2

The objective of this feasibility study was to subject pregnant rats of the same age, strain, and size that will be utilized in a shuttle flight experiment to all flight conditions except the unique microgravity of space flight and determine the feasibility of the proposed experimental design to meet the experimental objectives. The study utilized facilities at NASA, Ames Research Center, Moffett Field, CA to subject the rats to the gravitational stresses of a simulated shuttle launch and simulated shuttle landing. One hundred pregnant rats were received on gestation day (G) 2 (day 1 = day of vaginal sperm) and on G7, eighty rats were laparotomized to determine the condition of pregnancy and allow assignment to test groups. The five test groups (N=10 each group) were as follows: Group 1, Nominal Flight; Group 2, Laparotomy Control; Group 3, Hysterectomy Control; Group 4, Vivarium Control; Group 5, Delayed Recovery. On G9, animals in groups 1,2,3, and 5 were subjected to a shuttle launch simulation. On G18, groups 1,2, and 3 were subjected to a shuttle landing simulation and on this same day groups 1 and 2 were subjected to unilateral hysterectomy to obtain fetuses and placentas for evaluation. Fetal crown-rump length and fetal weight of the Nominal Flight group was significantly less than the Laparotomy Control group, but placentas were similar. On G20, group 5 was subjected to a shuttle landing simulation and on this day this group received a unilateral hysterectomy and fetuses and placentas were weighed. Animals in all groups were allowed to go to term and all animals delivered between 06:00 hours G22 and 18:00 hours G23. After delivery, a blood sample was taken from each experimental dam, and they were euthanized and the thymus and adrenal glands weighed. The thymus weight from all experimental group dams was decreased relative to the Vivarium Control group but adrenal glands and hormone values in dam plasma was similar in all groups. Pups from experimental groups were tattooed for identification, the anogenital distance of male pups was measured, and all pups placed with foster dams and litter sizes were standardized to 10. There was no difference in anogenital distances between male pups from different test groups. Pups delivered from Delayed Recovery animals were smaller than pups delivered from Nominal Flight animals. On neonatal day 7, all pups were euthanized and pup adrenal glands and thymus weighed. There was no difference in weights of thymus and adrenal glands in pups euthanized at neonatal day 7. Collectively, these data confirm the feasibility of the experimental design to meet objectives of the studies proposed for shuttle flight.

Burden, Hubert W.↗
NASA NTRS1985

Conceptual design and programmatics studies of space station accommodations for Life Sciences Research Facilities (LSRF)

Conceptual designs and programmatics of the space station accommodations for the Life Sciences Research Facilities (LSRF) are presented. The animal ECLSS system for the LSRF provides temperature-humidity control, air circulation, and life support functions for experimental subjects. Three ECLSS were studied. All configurations presented satisfy the science requirements for: animal holding facilities with bioisolation; facilities interchangeable to hold rodents, small primates, and plants; metabolic cages interchangeable with standard holding cages; holding facilities adaptable to restrained large primates and rodent breeding/nesting cages; volume for the specified instruments; enclosed ferm-free workbench for manipulation of animals and chemical procedures; freezers for specimen storage until return; and centrifuge to maintain animals and plants at fractional g to 1 g or more, with potential for accommodating humans for short time intervals.

Source record↗
NASA NTRS1995

Spacelab Life Sciences-1

This report provides an historical overview of the Spacelab Life Sciences-1 (SLS-1) mission along with the resultant biomaintenance data and investigators' findings. Only the nonhuman elements, developed by Ames Research Center (ARC) researchers, are addressed herein. The STS-40 flight of SLS-1, in June 1991, was the first spacelab flown after 'return to orbit', it was also the first spacelab mission specifically designated as a Life Sciences Spacelab. The experiments performed provided baseline data for both hardware and rodents used in succeeding missions.

Dalton, Bonnie P.↗
NASA NTRS2000

Modeling Heat-Transfer in Animal Habitats in the Shuttle Orbiter Middeck

A mathematical model has been developed to evaluate the heat transfer characteristics of an Animal Enclosure Module (AEM) in the microgravity environment. The AEM is a spaceflight habitat that provides life support for up to six rodents in the Space Shuttle Middeck. Currently, temperatures within the AEM are recorded in real time using a solid state data recorder; however, the data are only available for analysis post-flight. This temperature information is useful for characterizing the thermal environment of the AEM for researchers, but is unavailable during flight operations. Because animal health in microgravity is directly linked to the thermal environment, the ability to predict internal AEM temperatures is extremely useful to life science researchers. NASA flight crews typically carry hand-held temperature measurement devices which allow them to provide ground researchers with near real time readings of AEM inlet temperature; however, higher priority operations limit the frequency at which these measurements can be made and subsequently downlinked. The mathematical model developed allows users to predict internal cage volume temperatures based on knowledge of the ambient air temperature entering the AEM air intake ports. Additionally, an average convective heat transfer coefficient for the AEM has been determined to provide engineers with the requisite information to facilitate future design improvements and product upgrades. The model has been validated using empirical data from a series of three Space Shuttle missions.

Eodice, Michael T.↗
NASA NTRS1994

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.↗
DOE OSTI2020

Aristolochic acid-induced genotoxicity and toxicogenomic changes in rodents

Aristolochic acid (AA) is a group of structurally related nitrophenanthrene carboxylic acids found in many plants that are widely used by many cultures as traditional herbal medicines. AA is a causative agent for Chinese herbs nephropathy, a term replaced later by AA nephropathy. Evidence indicates that AA is nephrotoxic, genotoxic, and carcinogenic in humans; and it also induces tumors in the forestomach, kidney, renal pelvis, urinary bladder, and lung of rats and mice. Therefore, plants containing AA have been classified as carcinogenic to humans (Group 1) by the International Agency for Research on Cancer. In our laboratories, we have conducted a series of genotoxicity and toxicogenomic studies in the rats exposed to AA of 0.1–10 mg/kg for 12 weeks. Our results demonstrated that AA treatments induced DNA adducts and mutations in the kidney, liver, and spleen of rats, as well as significant alteration of gene expression in both its target and nontarget tissues. AA treatments altered mutagenesis- or carcinogenesis-related microRNA expression in rat kidney and resulted in significant changes in protein expression profiling. We also applied benchmark dose (BMD) modeling to the 3-month AA-induced genotoxicity data. The obtained BMDL10 (the lower 95% confidence interval of the BMD 10 that is a 10% increase over the background level) for AA-induced mutations in the kidney of rats was about 7 μg/kg body weight per day. This review constitutes an overview of our investigations on AA-induced genotoxicity and toxicogenomic changes including gene expression, microRNA expression, and proteomics; and presents updated information focused on AA-induced genotoxicity in rodents.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2004

Effects of heavy ions on visual function and electrophysiology of rodents: the ALTEA-MICE project

ALTEA-MICE will supplement the ALTEA project on astronauts and provide information on the functional visual impairment possibly induced by heavy ions during prolonged operations in microgravity. Goals of ALTEA-MICE are: (1) to investigate the effects of heavy ions on the visual system of normal and mutant mice with retinal defects; (2) to define reliable experimental conditions for space research; and (3) to develop animal models to study the physiological consequences of space travels on humans. Remotely controlled mouse setup, applied electrophysiological recording methods, remote particle monitoring, and experimental procedures were developed and tested. The project has proved feasible under laboratory-controlled conditions comparable in important aspects to those of astronauts' exposure to particle in space. Experiments are performed at the Brookhaven National Laboratories [BNL] (Upton, NY, USA) and the Gesellschaft fur Schwerionenforschung mbH [GSI]/Biophysik (Darmstadt, FRG) to identify possible electrophysiological changes and/or activation of protective mechanisms in response to pulsed radiation. Offline data analyses are in progress and observations are still anecdotal. Electrophysiological changes after pulsed radiation are within the limits of spontaneous variability under anesthesia, with only indirect evidence of possible retinal/cortical responses. Immunostaining showed changes (e.g. increased expression of FGF2 protein in the outer nuclear layer) suggesting a retinal stress reaction to high-energy particles of potential relevance in space. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

NASA Center JSC↗
NASA NTRS2000

In vivo regulation of the beta-myosin heavy chain gene in soleus muscle of suspended and weight-bearing rats

In the weight-bearing hindlimb soleus muscle of the rat, approximately 90% of muscle fibers express the beta-myosin heavy chain (beta-MHC) isoform protein. Hindlimb suspension (HS) causes the MHC isoform population to shift from beta toward the fast MHC isoforms. Our aim was to establish a model to test the hypothesis that this shift in expression is transcriptionally regulated through specific cis elements of the beta-MHC promoter. With the use of a direct gene transfer approach, we determined the activity of different length beta-MHC promoter fragments, linked to a firefly luciferase reporter gene, in soleus muscle of control and HS rats. In weight-bearing rats, the relative luciferase activity of the longest beta-promoter fragment (-3500 bp) was threefold higher than the shorter promoter constructs, which suggests that an enhancer sequence is present in the upstream promoter region. After 1 wk of HS, the reporter activities of the -3500-, -914-, and -408-bp promoter constructs were significantly reduced ( approximately 40%), compared with the control muscles. However, using the -215-bp construct, no differences in promoter activity were observed between HS and control muscles, which indicates that the response to HS in the rodent appears to be regulated within the -408 and -215 bp of the promoter.

Non-NASA Center↗
NASA NTRS1991

Facilities for animal research in space

The animal facilities used aboard or designed for various spacecraft research missions are described. Consideration is given to the configurations used in Cosmos-1514 (1983) and Cosmos-1887 (1987) missions; the reusable Biosatellite capsule flown three times by NASA between 1966 and 1969; the NASA's Lifesat spacecraft that is being currently designed; the Animal Enclosure Module flown on Shuttle missions in 1983 and 1984; the Research Animal Holding Facility developed for Shuttle-Spacelab missions; the Rhesus Research Facility developed for a Spacelab mission; and the Japanese Animal Holding Facility for the Space Station Freedom. Special attention is given to the designs of NASA's animal facilities developed for Space Station Freedom and the details of various subsystems of these facilities. The main characteristics of the rodent and the primate habitats provided by these various facilities are discussed.

Bonting, Sjoerd L.↗
NASA NTRS2000

Simian virus 40, poliovirus vaccines, and human cancer: research progress versus media and public interests

From 1955 through early 1963, millions of people were inadvertently exposed to simian virus 40 (SV40) as a contaminant of poliovirus vaccines; the virus had been present in the monkey kidney cultures used to prepare the vaccines and had escaped detection. SV40 was discovered in 1960 and subsequently eliminated from poliovirus vaccines. This article reviews current knowledge about SV40 and considers public responses to reports in the media. SV40 is a potent tumour virus with broad tissue tropism that induces tumours in rodents and transforms cultured cells from many species. It is also an important laboratory model for basic studies of molecular processes in eukaryotic cells and mechanisms of neoplastic transformation. SV40 neutralizing antibodies have been detected in individuals not exposed to contaminated poliovirus vaccines. There have been many reports of detection of SV40 DNA in human tumours, especially mesotheliomas, brain tumours and osteosarcomas; and DNA sequence analyses have ruled out the possibility that the viral DNA in tumours was due to laboratory contamination or that the virus had been misidentified. However, additional studies are necessary to prove that SV40 is the cause of certain human cancers. A recently published review article evaluated the status of the field and received much media attention. The public response emphasized that there is great interest in the possibility of health risks today from vaccinations received in the past.

Review, Tutorial↗
NASA NTRSDate not supplied

Somatic Mutation in Mice on the International Space Station (ISS): Guanine Substitution Suggests Link to Cancer Risk

We conducted comprehensive analysis of single nucleotide somatic mutations in mice exposed to microgravity and other factors aboard the International Space Station (ISS), using data archived in GeneLab. Animals in the experimental cohort consisted of mice that spent 37 days on the ISS within the Rodent Habitat. Ground control animals consisted of mice of identical age, sex, strain, in a terrestrial Rodent Habitat controlled for temperature, humidity and carbon dioxide levels, to match ISS conditions as closely as possible. RNA extracted from eye, liver, skeletal muscle, and kidney tissue specimens was subjected to next-generation sequencing to acquire primary data. Our analysis employed cutting-edge software developed at NASA Ames Research Center, executed on the NASA Ames Supercomputer and on another high-performance computer, for accurate variant calling of single point mutations. ISS-flown mice exhibited a notably heightened level of somatic mutation compared to control mice. The degree of somatic mutation correlated with the degree of gene expression across the four tissue types, i.e., the greatest rate of mutation accumulation was seen in highly expressed genes. We discovered that guanine substitutions were the most common type of somatic mutation. This observation is consistent with the hypothesis that DNA mutation events stem from reactive oxygen/nitrogen/chlorine species-mediated guanine oxidation induced by the spaceflight environment. Since guanine oxidation is a prominent feature of the DNA mutation landscape that accompanies malignant transformation, our findings suggest a possible link between the spaceflight environment and cancer risk that is independent of radiation carcinogenesis.

ISS↗
NASA NTRS1987

Animal research on the Space Station

The need for in-depth, long- and short-term animal experimentation in space to qualify man for long-duration space missions, and to study the effects of the absence and presence of Earth's gravity and of heavy particle radiation on the development and functioning of vertebrates is described. The major facilities required for these investigations and to be installed on the Space Station are: modular habitats for holding rodents and small primates in full bioisolation; a habitat holding facility; 1.8 and 4.0 m dia centrifuges; a multipurpose workbench; and a cage cleaner/disposal system. The design concepts, functions, and characteristics of these facilities are described.

Bonting, S. L.↗
NASA NTRS2010

Neurolab: Final Report for the Ames Research Center Payload

Neurolab, the final Spacelab mission, launched on STS-90 on April 17, 1998, was dedicated to studying the nervous system. NASA cooperated with domestic and international partners to conduct the mission. ARC's (Ames Research Center's) Payload included 15 experiments designed to study the adaptation and development of the nervous system in microgravity. The payload had the largest number of Principal and Co-Investigators, largest complement of habitats and experiment unique equipment flown to date, and most diverse distribution of live specimens ever undertaken by ARC, including rodents, toadfish, swordtail fish, water snails, hornweed and crickets To facilitate tissue sharing and optimization of science objectives, investigators were grouped into four science discipline teams: Neuronal Plasticity, Mammalian Development, Aquatic, and Neurobiology. Several payload development challenges were experienced and required an extraordinary effort, by all involved, to meet the launch schedule. With respect to hardware and the total amount of recovered science, Neurolab was regarded as an overall success. However, a high mortality rate in one rodent group and several hardware anomalies occurred inflight that warranted postflight investigations. Hardware, science, and operations lessons were learned that should be taken into consideration by payload teams developing payloads for future Shuttle missions and the International Space Station.

Maese, A. Christopher↗
NASA NTRS1997

Monitoring Physiological Variables with Membrane Probes

This project has demonstrated the possibility of using membrane probes in rodents to monitor physiological variables for extended periods of time. The utility of these probes in physiological studies of microgravity has been demonstrated. The feasibility of developing on-line sensors has also been demonstrated and allows for the possibility of developing real-time automated monitoring systems which can be used in ground-base physiological research as well as in research and medical monitoring in space. In addition to space applications these techniques can be extended to medical monitoring in critical care situations on earth as well as facilitating research in many human and animal diseases.

Janle, Elsa M.↗
NASA NTRS1998

AAH Cage Out-Link and In-Link Antenna Characterization

This final report encapsulates the accomplishments of the third year of work on an Advanced Biotelemetry System (ABTS). Overall MU/ABTS project objectives are to provide a biotelemetry system that can collect data from and send commands to an implanted biotransceiver. This system will provide for studies of rodent development in space. The system must be capable of operating in a metal animal cage environment. An important goal is the development of a small, "smart", micropower, -channel data output and single channel command implantable biotransceiver with eight input capabilities with the flexibility for easy customization for a variety of physiologic investigations. The NASA Ames/Marquette University Joint Research work has been devoted to the system design of such a new state of the art biotelemetry system, having multiple physiologic inputs, and bi-directional data transfer capabilities. This work has provided a successful prototype system that connects, by two-way radio links, an addressable biotelemetry system that provides communication between an animal biotelemeter prototype and a personal computer. The operational features of the prototype system are listed below: Two-Way PCM Communication with Implanted Biotelemeter Microcontroller Based Biotelemeter Out-Link: Wideband FSK (60 kbaud) In-Link: OOK (2.4 kbaud) Septum Antenna Arrays (In/Out-Links) Personal Computer Data Interface The important requirement of this third year's work, to demonstrate two-way communication with transmit and receive antennas inside the metal animal cage, has been successfully accomplished. The advances discussed in this report demonstrate that the AAH cage antenna system can provide Out-link and In-link capability for the ABTS bi-directional telemetry system, and can serve as a benchmark for project status.

Jeutter, Dean C.↗