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Arabidopsis thaliana in altered gravity: the 14-3-3κ:GFP overexpression line on the F-104 Starfighter, first flight

ABSTRACT: The increasing availability of flights on suborbital rockets creates new avenues for the study of spaceflight effects on biological systems, in particular the transitions between hypergravity and microgravity. This paper presents an initial comparison of the responses of Arabidopsis thaliana to suborbital and atmospheric parabolic flights as an important step toward characterizing these emerging suborbital platforms and their effects on biology. Transcriptomic profiling of the response of the Arabidopsis ecotype Wassilewskija (WS) to the aggregate spaceflight experiences in the Blue Origin New Shepard and Virgin Galactic SpaceShipTwo rockets revealed that the transcriptomic load induced by flight differed greatly between the two flights, yet was biologically related to traditional parabolic flight responses. The sku5 skewing mutant and 14-3-3κ:GFP regulatory protein overexpression lines each showed altered intra-platform responses compared to WS in the Blue Origin and parabolic flights, respectively. An additional parabolic flight using the F-104 Starfighter showed that the response of 14-3-3κ:GFP to flight was modulated in a similar manner to the WS line. Despite the differing genotypes, experimental workflows, flight profiles and platforms, alteration of gene expression remodeling central metabolic processes was commonly observed as a response to the flights. The processes included carbon and nitrogen metabolism, branched-chain amino acid degradation, and hypoxic responses. The timing and directionality of differentially-expressed genes involved in the conserved pathways differed among the platforms. The data presented herein highlight the potential for various suborbital platforms to contribute insights into biological responses to spaceflight, and further suggest that in-flight fixation during suborbital experiments will provide insights into responses to each phase of flight.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE159863 | GEO | 2021/04/01

REPOSITORIES: GEO

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Project description:BACKGROUND: Understanding physiologic reactions to weightlessness is an indispensable requirement for safe human space missions. While adaptations of human organ systems in response to weightlessness have been described in former studies, their molecular background needs further elucidation. OBJECTIVE: The study aims to analyse changes in the expression of circulating miRNAs in serum in response to gravitational changes induced by parabolic flight as a spaceflight analogue. METHODS: Eight healthy volunteers (age: 25.4 years, male: 4, female: 4) were included. Each subject underwent 31 short-term phases of weightlessness and hypergravity induced by parabolic flight. At different time points (baseline, 1 hour after parabolic flight, and 24 hours parabolic flight), venous blood was withdrawn. Analysis of circulating miRNAs in serum was conducted by means of next generation sequencing. RESULTS: In total, 213 miRNAs were robustly detected by small RNA sequencing in all 24 samples. 4 miRNAs (mir-941, mir-24-3p, mir-486-5p, mir-223-3p) evidenced a significant change in expression after adjusting for multiple testing. mir-941 and mir-24-3p showed a significant decrease 24 hours after parabolic flight compared to 1 hour after parabolic flight. Contrary, mir-486-5p showed a significant increase 24 hours after parabolic flight compared to 1 hour after parabolic flight. Of note, mir-223-3p showed a significant decrease 24 hours after parabolic flight compared to baseline values and values at 1 hour after parabolic flight. A target network analysis identified genes of the p53 signaling pathway and the cell cycle highly enriched among the targets of the four microRNAs. CONCLUSIONS: Our findings suggest cellular adaption to gravitational changes by means of weightlessness and hypergravity already at the transcriptional level. Based on our results, we suggest a change in cell cycle regulation as potential explanation for adaptational changes observed in space missions.

Project description: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 setting up the spaceflight responses are being identified; their role 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 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 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 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 and 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 transcriptome cost of plant physiological adaptation to spaceflight and suggesting that genetic manipulation might 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 further implies that manipulating the local habitat can also substantially impact the metabolic cost of spaceflight adaptation.

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Arabidopsis thaliana in altered gravity: the 14-3-3κ:GFP overexpression line on the F-104 Starfighter, first flight

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