Transcriptome analysis of gravitational effects on mouse skeletal muscles under microgravity and artificial 1 g onboard environment

Okada, Risa; Fujita, Shin‐ichiro; Suzuki, Rumiko; Hayashi, Takuto; Tsubouchi, Hirona; Kato, Chiho; Sadaki, Shunya; Kanai, Maho; Fuseya, Sayaka; Inoue, Yuzo; Jeon, Hyojung; Hamada, Michito; Kuno, Akihiro; Ishii, Akiko; Tamaoka, Akira; Tanihata, Jun; Ito, Naoki; Shiba, Dai; Shimizu, Masaki; Muratani, Masafumi; Kudo, Takashi; Takahashi, Satoru

doi:10.1038/s41598-021-88392-4

Cited by 34 publications

(30 citation statements)

References 62 publications

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“…Mammals (i.e., humans and rodents) have evolved with the never-ending downward pull of gravity on Earth, and therefore postural muscles such as the soleus are known to be most affected with spaceflight. Similar to unloading models on Earth, spaceflight and microgravity exposure in rodents unloads the postural soleus causing extensive muscle atrophy and a fiber-type shift from slow-oxidative to fast-glycolytic [ 4 , 5 , 6 , 7 , 8 , 9 ]. Similar changes have also been observed in human soleus muscles after 17 days of spaceflight [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Characterizing SERCA Function in Murine Skeletal Muscles after 35–37 Days of Spaceflight

Braun

Geromella

Hamstra

et al. 2021

IJMS

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It is well established that microgravity exposure causes significant muscle weakness and atrophy via muscle unloading. On Earth, muscle unloading leads to a disproportionate loss in muscle force and size with the loss in muscle force occurring at a faster rate. Although the exact mechanisms are unknown, a role for Ca2+ dysregulation has been suggested. The sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) pump actively brings cytosolic Ca2+ into the SR, eliciting muscle relaxation and maintaining low intracellular Ca2+ ([Ca2+]i). SERCA dysfunction contributes to elevations in [Ca2+]i, leading to cellular damage, and may contribute to the muscle weakness and atrophy observed with spaceflight. Here, we investigated SERCA function, SERCA regulatory protein content, and reactive oxygen/nitrogen species (RONS) protein adduction in murine skeletal muscle after 35–37 days of spaceflight. In male and female soleus muscles, spaceflight led to drastic impairments in Ca2+ uptake despite significant increases in SERCA1a protein content. We attribute this impairment to an increase in RONS production and elevated total protein tyrosine (T) nitration and cysteine (S) nitrosylation. Contrarily, in the tibialis anterior (TA), we observed an enhancement in Ca2+ uptake, which we attribute to a shift towards a faster muscle fiber type (i.e., increased myosin heavy chain IIb and SERCA1a) without elevated total protein T-nitration and S-nitrosylation. Thus, spaceflight affects SERCA function differently between the soleus and TA.

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Section: Introductionmentioning

confidence: 99%

Characterizing SERCA Function in Murine Skeletal Muscles after 35–37 Days of Spaceflight

Braun

Geromella

Hamstra

et al. 2021

IJMS

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“…Skeletal muscle tissue alterations such as atrophy, muscle force decrease, and shift in muscle fiber composition occur during aging, pathology onset, radiation exposure, and mechanical unloading. − A fast deterioration of skeletal muscle tissue, in particular, affects astronauts who are exposed both to gravitational unloading (hereafter termed microgravity or “μ g ”) and to cosmic radiation during spaceflight. For this reason, a variety of strategies for muscular maintenance in vitro and in vivo has also been devised for terrestrial benefit, including physical exercise, mechanical stimulation in the form of vibrations and pressure application, electrical stimulation, exposure to hypergravity, − administration of soluble factors (such as activin type IIB receptor, recombinant myokine irisin, and myostatin antibody YN41), and even genetic transduction finalized to the overexpression of nucleic acids (such as a long noncoding RNA termed lncMUMA) . A common approach against skeletal muscle waste due to mechanical unloading also consists of the supply of antioxidant compounds, like for instance (-)-epicatechin, lecithin, N -acetylcysteine, complex mixtures of polyphenols associated with other antioxidants (such as vitamin E, selenium, and omega-3 fatty acids), or even seed extracts (from Oenothera odorata) .…”

Section: Introductionmentioning

confidence: 99%

Cerium Oxide Nanoparticle Administration to Skeletal Muscle Cells under Different Gravity and Radiation Conditions

Genchi

Degl’Innocenti

Martinelli

et al. 2021

ACS Appl. Mater. Interfaces

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For their remarkable biomimetic properties implying strong modulation of the intracellular and extracellular redox state, cerium oxide nanoparticles (also termed "nanoceria") were hypothesized to exert a protective role against oxidative stress associated with the harsh environmental conditions of spaceflight, characterized by microgravity and highly energetic radiations. Nanoparticles were supplied to proliferating C2C12 mouse skeletal muscle cells under different gravity and radiation levels. Biological responses were thus investigated at a transcriptional level by RNA next-generation sequencing. Lists of differentially expressed genes (DEGs) were generated and intersected by taking into consideration relevant comparisons, which led to the observation of prevailing effects of the space environment over those induced by nanoceria. In space, upregulation of transcription was slightly preponderant over downregulation, implying involvement of intracellular compartments, with the majority of DEGs consistently over-or under-expressed whenever present. Cosmic radiations regulated a higher number of DEGs than microgravity and seemed to promote increased cellular catabolism. By taking into consideration space physical stressors alone, microgravity and cosmic radiations appeared to have opposite effects at transcriptional levels despite partial sharing of molecular pathways. Interestingly, gene ontology denoted some enrichment in terms related to vision, when only effects of radiations were assessed. The transcriptional regulation of mitochondrial uncoupling protein 2 in space-relevant samples suggests perturbation of the intracellular redox homeostasis, and leaves open opportunities for antioxidant treatment for oxidative stress reduction in harsh environments.

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“…4 ). Therefore, while we identified regulation of mitochondrial function and lipid metabolism via DGE (see Additional file 5 ), as has been described previously in skeletal muscle following spaceflight [ 19 – 21 ], we characterized DAS as a novel means of modifying the transcriptome in response to microgravity. Further, we identified coordinate changes in splicing and microgravity-induced physiological adaptations of hind limb muscle, which included muscle atrophy (see Figs.…”

Section: Discussionmentioning

confidence: 99%

“…Most interest has been paid to the transcriptomic effects at the level of differential gene expression (DGE). Recent transcriptome analyses of skeletal muscle in mice [ 19 – 21 ] have annotated microgravity-induced DGE of gene networks related to contractile machinery, calcium homeostasis, muscle development, cellular metabolism, inflammatory/oxidative stress response, and mitochondrial function. One landmark study, the National Aeronautics and Space Administration (NASA) Twins Study [ 22 ], identified DGE between monozygotic twins that were exposed to spaceflight or ground control conditions for one year.…”

Section: Introductionmentioning

confidence: 99%

Alternative splicing diversifies the skeletal muscle transcriptome during prolonged spaceflight

Henrich

Wang

et al. 2022

Skeletal Muscle

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Background As the interest in manned spaceflight increases, so does the requirement to understand the transcriptomic mechanisms that underlay the detrimental physiological adaptations of skeletal muscle to microgravity. While microgravity-induced differential gene expression (DGE) has been extensively investigated, the contribution of differential alternative splicing (DAS) to the plasticity and functional status of the skeletal muscle transcriptome has not been studied in an animal model. Therefore, by evaluating both DGE and DAS across spaceflight, we set out to provide the first comprehensive characterization of the transcriptomic landscape of skeletal muscle during exposure to microgravity. Methods RNA-sequencing, immunohistochemistry, and morphological analyses were conducted utilizing total RNA and tissue sections isolated from the gastrocnemius and quadriceps muscles of 30-week-old female BALB/c mice exposed to microgravity or ground control conditions for 9 weeks. Results In response to microgravity, the skeletal muscle transcriptome was remodeled via both DGE and DAS. Importantly, while DGE showed variable gene network enrichment, DAS was enriched in structural and functional gene networks of skeletal muscle, resulting in the expression of alternatively spliced transcript isoforms that have been associated with the physiological changes to skeletal muscle in microgravity, including muscle atrophy and altered fiber type function. Finally, RNA-binding proteins, which are required for regulation of pre-mRNA splicing, were themselves differentially spliced but not differentially expressed, an upstream event that is speculated to account for the downstream splicing changes identified in target skeletal muscle genes. Conclusions Our work serves as the first investigation of coordinate changes in DGE and DAS in large limb muscles across spaceflight. It opens up a new opportunity to understand (i) the molecular mechanisms by which splice variants of skeletal muscle genes regulate the physiological adaptations of skeletal muscle to microgravity and (ii) how small molecule splicing regulator therapies might thwart muscle atrophy and alterations to fiber type function during prolonged spaceflight.

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Transcriptome analysis of gravitational effects on mouse skeletal muscles under microgravity and artificial 1 g onboard environment

Cited by 34 publications

References 62 publications

Characterizing SERCA Function in Murine Skeletal Muscles after 35–37 Days of Spaceflight

Characterizing SERCA Function in Murine Skeletal Muscles after 35–37 Days of Spaceflight

Cerium Oxide Nanoparticle Administration to Skeletal Muscle Cells under Different Gravity and Radiation Conditions

Alternative splicing diversifies the skeletal muscle transcriptome during prolonged spaceflight

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