In prospective human exploration of outer space the need to maintain a species over several generations under changed gravity conditions may arise. This paper reports the analysis of the third generation of fruit fly Drosophila melanogaster obtained during the 44.5-day space flight (Foton-M4 satellite 2014 Russia) followed by the fourth generation on Earth and the fifth generation under conditions of a 12-day space flight (2014 in the Russian Segment of the ISS). The obtained results show that it is possible to obtain the third-fifth generations of a complex multicellular Earth organism under changed gravity conditions (in the cycle weightlessness - Earth - weightlessness) which preserves fertility and normal development. However there were a number of changes in the expression levels and content of cytoskeletal proteins that are the key components of the spindle apparatus and the contractile ring of cells.

Gene expression levels were determined in 3rd instar and adult Drosophila melanogaster reared during spaceflight, to elucidate the genetic and molecular mechanisms underpinning the effects of microgravity on the immune system. The goal was to validate the Drosophila model for understanding alterations of innate immune responses in humans due to spaceflight. Five containers of flies, with ten female and five male fruit flies in each container, were housed and bred on the space shuttle (average orbit altitude of 330.35 km) for 12 days and 18.5 hours, with a new generation reared in microgravity. RNA was extracted on the day of shuttle landing from whole body animals (3rd instar larvae and adults), hybridized to Drosophila 2.0 Affymetrix genome arrays, and the expression level of all genes was normalized against the gene expression level from the corresponding developmental stage animals raised on ground. Spaceflight altered the expression of larval genes involved in the maturation of plasmatocytes (macrophages) and their phagocytic response, as well as the level of constitutive expression of pattern recognition receptors and opsonins that specifically recognize bacteria, and of lysozymes, antimicrobial peptide pathway and immune stress genes, hallmarks of humoral immunity. Larval microarrays (FL 6 samples) are based on RNA extracted from 6 independent sets of 50 mid 3rd instar larvae reared in microgravity and collected on the day of landing after 12 days and 18.5 hours on the space shuttle and the same number of control larvae raised on ground (GL 6 samples). Adults microarrays (F1 3 samples) are based on RNA from 3 sets of 20 adult females each, that emerged during spaceflight and within 4 hours of landing and the same number of adult females from the corresponding ground control containers (G1 3 samples).

In this study, fruit flies and their parasitic wasps were sent to the ISS to examine changes in host immunity and parasite virulence. Key findings from this work are as follows: (a) Spaceflight spiked the expression of inflammation genes in fruit flies; (b) Spaceflight enhanced tumor growth in flies already suffering from chronic inflammation; (c) Spaceflight did not significantly affect parasitic wasp development, emergence, or the expression of virulence genes; (d) Parasite mutants with visible wing color and shape alterations were obtained. This study contains Leptopilina heterotoma14 data. The drosophila data is deposited in OSD-588, https://osdr.nasa.gov/bio/repo/data/studies/OSD-588, and the Leptopilina boulardi17 data is deposited in OSD-610, https://osdr.nasa.gov/bio/repo/data/studies/OSD-610.

In this study, fruit flies and their parasitic wasps were sent to the ISS to examine changes in host immunity and parasite virulence. Key findings from this work are as follows: (a) Spaceflight spiked the expression of inflammation genes in fruit flies; (b) Spaceflight enhanced tumor growth in flies already suffering from chronic inflammation; (c) Spaceflight did not significantly affect parasitic wasp development, emergence, or the expression of virulence genes; (d) Parasite mutants with visible wing color and shape alterations were obtained. This study contains Leptopilina boulardi 17 data. The drosophila data is deposited in OSD-588, https://osdr.nasa.gov/bio/repo/data/studies/OSD-588, and the Leptopilina heterotoma 14 data is deposited in OSD-609, https://osdr.nasa.gov/bio/repo/data/studies/OSD-609.

In this study, fruit flies and their parasitic wasps were sent to the ISS to examine changes in host immunity and parasite virulence. Key findings from this work are as follows: (a) Spaceflight spiked the expression of inflammation genes in fruit flies; (b) Spaceflight enhanced tumor growth in flies already suffering from chronic inflammation; (c) Spaceflight did not significantly affect parasitic wasp development, emergence, or the expression of virulence genes; (d) Parasite mutants with visible wing color and shape alterations were obtained. This study contains Drosophila melanogaster data. The Leptopilina heterotoma14 data is deposited in OSD-609, https://osdr.nasa.gov/bio/repo/data/studies/OSD-609, and the Leptopilina boulardi17 data is deposited in OSD-610, https://osdr.nasa.gov/bio/repo/data/studies/OSD-610.

Physical forces greatly influence the growth and function of an organism. Altered gravity can perturb normal development and induce corresponding changes in gene expression. Understanding this relationship between the physical and biological realms is important for NASA's space travel goals. We use combined RNA-Seq and qRT-PCR to profile changes in early Drosophila melanogaster pupae exposed to chronic hypergravity (3 g, three times Earth's gravity) to highlight gravity-dependent pathways and gene products. Robust transcriptional response was evident among the pupae developed in a hypergravity environment compared to control. 1,513 genes showed significantly (p less than 0.05) altered gene expression in the 3 g samples. These findings were supported with qRT-PCR data. Major biological processes affected include ion transport, redox homeostasis, immune and humoral stress response, proteolysis, and cuticle development.

Experimentation on the International Space Station has reached the stage where repeated and nuanced transcriptome studies are beginning to illuminate the structural and metabolic differences between plants grown in space compared to plants on the Earth. Genes that are important in establishing the spaceflight responses are being identified, their roles in spaceflight physiological adaptation are increasingly understood, and the fact that different genotypes adapt differently is recognized. However, the basic question of whether these spaceflight responses are actually required for survival has yet to be posed, and the fundamental notion that spaceflight responses may be non-adaptive has yet to be explored. Therefore the experiments presented here were designed to ask if portions of the plant spaceflight response can be genetically removed without causing loss of spaceflight survival and without causing increased stress responses. The CARA experiment compared the spaceflight transcriptome responses in the root tips of two Arabidopsis ecotypes, Col-0 and WS, as well as that of a PhyD mutant of Col-0. When grown with the ambient light of the ISS, phyD plants displayed a significantly reduced spaceflight transcriptome response compared to Col-0, suggesting that altering the activity of a single gene can actually improve spaceflight adaptation by reducing the transcriptome cost of physiological adaptation. The WS genotype showed an even simpler spaceflight transcriptome response in the ambient light of the ISS, more broadly indicating that the plant genotype can be manipulated to reduce the cost of spaceflight adaptation, as measured by transcriptional response. These differential genotypic responses suggest that genetic manipulation could further reduce, or perhaps eliminate the metabolic cost of spaceflight adaptation. When plants were germinated and then left in the dark on the ISS, the WS genotype actually mounted a larger transcriptome response than Col-0, suggesting that the in-space light environment affects physiological adaptation, which implies that manipulating the local habitat can also substantially impact the metabolic cost of spaceflight adaptation.

Life in spaceflight demonstrates remarkable adaptive processes within the specialized environments of space vehicles which are subject to the myriad of attending and unique environmental issues associated with orbital trajectories. To examine the adaptive processes that occur in plants in space, leaves and roots from Arabidopsis seedlings that were grown from seed for 12 days on the International Space Station and preserved on orbit in RNAlater were returned to earth and analyzed using iTRAQ broad scale proteomics procedures.

Space travel presents unlimited opportunities for exploration and discovery but requires a more complete understanding of the immunological consequences of long-term exposure to the conditions of spaceflight. To understand these consequences better and to contribute to design of effective countermeasures we used the Drosophila model to compare innate immune responses to bacteria and fungi in flies that were either raised on earth or in outer space aboard the NASA Space Shuttle Discovery (STS-121). Microarrays were used to characterize changes in gene expression that occur in response to infection by bacteria and fungus in drosophila that were either hatched and raised in outer space (microgravity) or on earth (normal gravity). Whole Oregon R strain drosophila melanogaster fruit flies either raised on earth or in space that were (1) uninfected (2) infected with bacteria (Escherichia coli) or (3) infected with fungus (Beauveria bassiana) were used for RNA extraction and hybridization on Affymetrix microarrays.

Spaceflight poses risks to the central nervous system (CNS), and understanding neurological responses is important for future missions. We report CNS changes in Drosophila aboard the International Space Station in response to spaceflight microgravity (SFμg) and artificially simulated Earth gravity (SF1g) via inflight centrifugation as a countermeasure. While inflight behavioral analyses of SFμg exhibit increased activity, postflight analysis displays significant climbing defects, highlighting the sensitivity of behavior to altered gravity. Multi-omics analysis shows alterations in metabolic, oxidative stress and synaptic transmission pathways in both SFμg and SF1g; however, neurological changes immediately postflight, including neuronal loss, glial cell count alterations, oxidative damage, and apoptosis, are seen only in SFμg. Additionally, progressive neuronal loss and a glial phenotype in SF1g and SFμg brains, with pronounced phenotypes in SFμg, are seen upon acclimation to Earth conditions. Overall, our results indicate that artificial gravity partially protects the CNS from the adverse effects of spaceflight. This study derives results from the in-flight video analysis (video recording assay).