# 0.33g mitigates muscle atrophy while 0.67g preserves muscle function and myofiber type composition in mice during spaceflight

**Authors:** Ryosuke Tsuji, Ryo Fujita, Takuto Hayashi, Shunya Sadaki, Tatsuya Matsumoto, Yuri Inoue, Yuka Murakami, Michito Hamada, Masafumi Muratani, Hiroe Kobayashi, Akane Yumoto, Maki Okada, Daisuke Kamimura, Risa Okada, Takafumi Suzuki, Ryo Kurosawa, Akihito Otsuki, Seizo Koshiba, Martha Hotz Vitaterna, Charles A. Fuller, Marie Mortreux, Dong-Min Sung, Jason Ciola, Seward B. Rutkove, Jennifer Coulombe, Takashi Kudo, Masayuki Yamamoto, Mary L. Bouxsein, Dai Shiba, Satoru Takahashi

PMC · DOI: 10.1126/sciadv.aed2258 · 2026-03-13

## TL;DR

This study shows that 0.33g prevents muscle loss in mice during spaceflight, but 0.67g is needed to maintain muscle function and fiber type.

## Contribution

The study identifies 0.67g as the critical gravity threshold for preserving muscle function and myofiber composition in space.

## Key findings

- 0.33g prevents muscle atrophy but does not stop myofiber type transitions.
- 0.67g fully prevents myofiber type transitions and maintains muscle function.
- 11 gravity-dependent metabolites were identified as potential biomarkers for physiological adaptation.

## Abstract

As human space exploration advances, understanding how different gravity levels affect skeletal muscle is critical for long-term health. Among the major organ systems, skeletal muscle is particularly sensitive to gravitational unloading, yet the gravity threshold required to maintain homeostasis remains unclear. Using the Multiple Artificial-gravity Research System aboard the International Space Station, mice were exposed to graded gravity levels, microgravity, 0.33g, 0.67g, and 1g, and their muscles were analyzed postflight. In the gravity-sensitive soleus, the cross-sectional area was preserved at 0.33g, while the slow-to-fast myofiber transition was partially suppressed at 0.33g and fully prevented at 0.67g. Functional measures, including forelimb grip strength and electrical impedance myography, indicated that 0.67g was sufficient to maintain muscle performance. Plasma metabolomics identified 11 metabolites with gravity-dependent changes, suggesting potential biomarkers for monitoring physiological adaptation. Collectively, these results identify 0.67g as a critical threshold for mitigating spaceflight-induced muscle atrophy and myofiber type transitions.

Spaceflight tests reveal that 0.33g mitigates muscle loss, but 0.67g is needed to keep function, impacting future Mars missions.

## Linked entities

- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** Tnnt1 (troponin T1, skeletal, slow) [NCBI Gene 21955] {aka Tnt, sTnT, ssTnT}, Actn3 (actinin alpha 3) [NCBI Gene 11474], MYH2 (myosin heavy chain 2) [NCBI Gene 4620] {aka CMYO6, CMYP6, IBM3, MYH2A, MYHSA2, MYHas8}, Myh7 (myosin, heavy polypeptide 7, cardiac muscle, beta) [NCBI Gene 140781] {aka B-MHC, MYH-beta/slow, MyHC-I, Myhc-b, Myhcb, beta-MHC}, MAF (MAF bZIP transcription factor) [NCBI Gene 4094] {aka AYGRP, CCA4, CTRCT21, c-MAF}, Atp2a1 (ATPase, Ca++ transporting, cardiac muscle, fast twitch 1) [NCBI Gene 11937] {aka SERCA1}, Mfn2 (mitofusin 2) [NCBI Gene 170731] {aka D630023P19Rik, Fzo}, Atp2a2 (ATPase, Ca++ transporting, cardiac muscle, slow twitch 2) [NCBI Gene 11938] {aka 9530097L16Rik, D5Wsu150e, SERCA2, SERCA2B, Serca2a, mKIAA4195}, MAFA (MAF bZIP transcription factor A) [NCBI Gene 389692] {aka INSDM, RIPE3b1, hMafA}, Ercc8 (excision repaiross-complementing rodent repair deficiency, complementation group 8) [NCBI Gene 71991] {aka 2410022P04Rik, 2810431L23Rik, 4631412O06Rik, B130065P18Rik, Ckn1, Csa}, Trim63 (tripartite motif-containing 63) [NCBI Gene 433766] {aka MuRF1, RF1, Rnf28}, AKT1 (AKT serine/threonine kinase 1) [NCBI Gene 207] {aka AKT, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA}, MYH7 (myosin heavy chain 7) [NCBI Gene 4625] {aka CMD1S, CMH1, CMYO7A, CMYO7B, CMYP7A, CMYP7B}, Gcg (glucagon) [NCBI Gene 14526] {aka GLP-1, Glu, PPG}, Ppargc1a (peroxisome proliferative activated receptor, gamma, coactivator 1 alpha) [NCBI Gene 19017] {aka A830037N07Rik, Gm11133, PGC-1, PPARGC-1-alpha, Pgc-1alpha, Pgc1}, Gast (gastrin) [NCBI Gene 14459] {aka GAS}, Cacna1s (calcium channel, voltage-dependent, L type, alpha 1S subunit) [NCBI Gene 12292] {aka Cav1.1, Cchl1a3, DHPR, DHPR alpha1s, fmd, mdg}, MAFB (MAF bZIP transcription factor B) [NCBI Gene 9935] {aka DURS3, KRML, MCTO}, Mfn1 (mitofusin 1) [NCBI Gene 67414] {aka 2310002F04Rik, 6330416C07Rik, D3Ertd265e, HR2, mKIAA4032}, PIK3CB (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta) [NCBI Gene 5291] {aka P110BETA, PI3K, PI3KBETA, PIK3C1}, MYH4 (myosin heavy chain 4) [NCBI Gene 4622] {aka MYH2B, MyHC-2B, MyHC-IIb}, Foxo1 (forkhead box O1) [NCBI Gene 56458] {aka Afxh, FKHR, Fkhr1, Foxo1a}, Atp9b (ATPase, class II, type 9B) [NCBI Gene 50771] {aka Atpc2b, IIb, MMR}, Mafa (MAF bZIP transcription factor A) [NCBI Gene 378435] {aka RIPE3b1}, Myh4 (myosin, heavy polypeptide 4, skeletal muscle) [NCBI Gene 17884] {aka MHC2B, MM, MYH-2B, Minimsc, Minmus, MyHC-IIb}, Mafb (MAF bZIP transcription factor B) [NCBI Gene 16658] {aka Kreisler, Krml, Krml1, kr}, Capn1 (calpain 1) [NCBI Gene 12333] {aka Capa-1, Capa1, mu-calpin}, Maf (MAF bZIP transcription factor) [NCBI Gene 17132] {aka 2810401A20Rik, A230108G15Rik, c-maf}, Ryr1 (ryanodine receptor 1, skeletal muscle) [NCBI Gene 20190] {aka RYR-1, Ryr, skrr}, MTOR (mechanistic target of rapamycin kinase) [NCBI Gene 2475] {aka FRAP, FRAP1, FRAP2, RAFT1, RAPT1, SKS}, Foxo3 (forkhead box O3) [NCBI Gene 56484] {aka 1110048B16Rik, 2010203A17Rik, FKHRL1, Fkhr2, Foxo3a}, Fbxo32 (F-box protein 32) [NCBI Gene 67731] {aka 4833442G10Rik, ATROGIN1, Gm20361, MAFbx}, TRIM63 (tripartite motif containing 63) [NCBI Gene 84676] {aka CMH31, IRF, MURF1, MURF2, RNF28, SMRZ}, FBXO32 (F-box protein 32) [NCBI Gene 114907] {aka Fbx32, MAFbx}, MYH1 (myosin heavy chain 1) [NCBI Gene 4619] {aka HEL71, MYHSA1, MYHa, MyHC-2X/D, MyHC-2x}, PIK3R1 (phosphoinositide-3-kinase regulatory subunit 1) [NCBI Gene 5295] {aka AGM7, GRB1, IMD36, p85, p85-ALPHA, p85alpha}, Myh2 (myosin, heavy polypeptide 2, skeletal muscle, adult) [NCBI Gene 17882] {aka MHC2A, MyHC-2a, MyHC-IIa, Myh2a, Myhs-f, Myhs-f1}
- **Diseases:** Muscle atrophy (MESH:D009133), impaired glucose tolerance (MESH:D018149), muscle weakness (MESH:D018908), SOL (MESH:D019042), EDL (MESH:D009127), cachexia (MESH:D002100), frailty (MESH:D000073496), impaired (MESH:D060825), EIM (MESH:D004556), type IIx (MESH:C562844), sarcopenia (MESH:D055948), atrophy (MESH:D001284), inflammatory (MESH:D007249), amyotrophic lateral sclerosis (MESH:D000690), Duchenne muscular dystrophy (MESH:D020388), muscle mass loss (MESH:C536030), decline of muscle functions (MESH:D009135), neuromuscular and muscular disorders (MESH:D009468), aggressive behavior (MESH:D010554), weight loss (MESH:D015431)
- **Chemicals:** tragacanth (MESH:D014144), eosin (MESH:D004801), ketone bodies (MESH:D007657), xylene (MESH:D014992), lipid (MESH:D008055), TRIzol (MESH:C411644), isopentane (MESH:C067038), glycerol (MESH:D005990), Creatine (MESH:D003401), 3-hydroxybutyrate (MESH:D020155), Alexa Fluor (-), calcium (MESH:D002118), ethanol (MESH:D000431), acetone (MESH:D000096), lactate (MESH:D019344), fatty acid (MESH:D005227), hematoxylin (MESH:D006416), Carbon dioxide (MESH:D002245), betaine (MESH:D001622), steroid (MESH:D013256), NaCl (MESH:D012965), nitrogen (MESH:D009584), glucose (MESH:D005947), Glycine (MESH:D005998), water (MESH:D014867), acetoacetate (MESH:C016635), N-acetylcysteine (MESH:D000111), glutathione (MESH:D005978), glycogen (MESH:D006003), amino acid (MESH:D000596), H&amp;E (MESH:D006371), glutamate (MESH:D018698), methanol (MESH:D000432)
- **Species:** Homo sapiens (human, species) [taxon 9606], Mus musculus (house mouse, species) [taxon 10090], Sus scrofa (pig, species) [taxon 9823]
- **Mutations:** F to I, D to H, (F) to (L), (A) to (D)
- **Cell lines:** /6J — Homo sapiens (Human), Cutaneous melanoma, Cancer cell line (CVCL_W797)

## Figures

5 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12985678/full.md

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Source: https://tomesphere.com/paper/PMC12985678