JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_10.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_12.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_9.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_4.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_11.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_7.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_4.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_1.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_2.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_6.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_1.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_8.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_5.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_3.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_5.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_7.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_2.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_3.rawhttps://storage.jpostdb.org/JPST002135/files/180419QE_SpaceMouse_bone_12.rawhttps://storage.jpostdb.org/JPST002135/files/220107QE_DIA_JAXA_Space_mouse_Pulp_bone_11.rawprimaryOK200ProteomicsYayoi KimuraMus Musculus (mouse)https://repository.jpostdb.org/entry/JPST002135Yokohama City UniversityjPOSTAlthough the microgravity (μ-g) environment that astronauts encounter during spaceflight can cause severe acute bone loss, the molecular mechanism of this bone loss remains unclear. To investigate the gravity-response proteins involved in bone metabolism, it is important to comprehensively determine which proteins exhibit differential abundance associated with mechanical stimuli. However, comprehensive proteomic analysis using small bone samples is difficult because protein extraction in mineralized bone tissue is inefficient. Here, we established a high-sensitivity analysis system for mouse bone proteins using data-independent acquisition mass spectrometry. This system successfully detected 40 proteins in the femoral diaphysis showing differential abundance between mice raised in a μ-g environment, where the bone mass was reduced by gravity unloading, and mice raised in an artificial 1-gravity environment on the International Space Station. Additionally, 22 proteins, including noncollagenous bone matrix proteins, showed similar abundance between the two groups in the mandible, where bone mass was unaltered due to mastication stimuli, suggesting that these proteins are responsive to mechanical stimuli. One of these proteins, SPARCL1, is suggested to promote osteoclastogenesis induced by receptor activator of nuclear factor-κB ligand. We expect these findings to lead to new insights into the mechanisms of bone metabolism induced by mechanical stimuli. SIGNIFICANCE: We aimed to investigate the gravity-response proteins involved in bone metabolism. To this end, we established a comprehensive analysis system for mouse bone proteins using data-independent acquisition mass spectrometry, which is particularly useful in comprehensively analyzing the bone proteome using small sample volumes. In addition, a comprehensive proteomic analysis of the femoral diaphysis and mandible, which exhibit different degrees of bone loss in mice raised on the International Space Station, identified proteins that respond to mechanical stimuli. SPARCL1, a mechanical stimulus-responsive protein, was consequently suggested to be involved in osteoclast differentiation associated with bone remodeling. Our findings represent an important step toward elucidating the molecular mechanism of bone metabolism induced by mechanical stimuli.Identification of gravity-responsive proteins in the femur of spaceflight mice using a quantitative proteomic approach.Egashira Kenji K, Ino Yoko Y, Nakai Yusuke Y, Ohira Takashi T, Akiyama Tomoko T, Moriyama Kayano K, Yamamoto Yu Y, Kimura Mitsuo M, Ryo Akihide A, Saito Tomoyuki T, Inaba Yutaka Y, Hirano Hisashi H, Kumagai Ken K, Kimura Yayoi Y3.4.22.-, thigh, Trochanters, Greater Trochanter, Laboratory, proximal segment of free lower limb, Mus domesticus, Proteins, Lesser Trochanters, CASP14, mouse, number, Gene, mini-ICE, CASP-14, hindlimb stylopodium, hind limb stylopod, os femoris, House Mouse, Trochanter, presence., mesomere 1 of pevlic appendage, upper leg, count in organism, hindlimb stylopod, House, Mus, Protein, Mini-ICE, Gene Products, Mus musculus domesticus, femoral bone, Mice, hindlimb propodium, Greater Trochanters, Mus musculus, hind propodium, Lesser Trochanter, Caspase-14 subunit p10, stylopod of hind limb, Swiss, os longissimum, hind limb stylopodium, mice, os femorale, Swiss Mouse, stylopod of lower limb, Lesser, House Mice, Swiss Mice, Caspase-14 subunit p19, MICE, Laboratory Mice, domesticus, Protein Gene Products, stylopod of hindlimb, Gene Proteins, femur, thigh bone, Mouse, responsive, Greater, Laboratory Mouse, proximal metapterygial mesomere of pelvic appendage, Femurs, proximal metapterygial mesomere of pelvic finbiochemical pathways, Bones and Bone Tissue, Metabolic Process, artificial sequence, determination, Laboratory, mandibular series, Mus domesticus, Metabolic Concepts, Cosmonaut, CASP-14, protein, hevin, mandible, Anabolism., mandibula, House Mouse, osteogenic tissue, Bone and Bone, responsivity, GRP1, Grp1, symptoms, Concepts, Impacts, Analysis, Environmental Impacts, Metabolism Concept, protein aggregate, Phenomenon, Bone Tissues, Ridge, Mass Spectrum Analysis, bone remodelling, Analyses, catabolism, PTPSTEP, Tissue, hypoplasia, Mylohyoid Groove, Swiss Mice, metabolic process resulting in cell growth, Mandibles, DIA, allergic reaction, Environmental Impact, l(2)k08110, sample, body of femur, Mylohyoid Grooves, Condyles, biotransformation, house mouse, Bony Apophyses, Catabolism, GPH, screening, Astronaut, Ecm2, Ligand, Process, metabolism resulting in cell growth, mouse, CASP14, osteoclast cell differentiation, Spectrum Analysis, Acute onset, Spectroscopy, Mylohyoid, mandibles, Ridges, Mini-ICE, 3.1.3.48, secretion, femoral shaft, END, Mus musculus, PIG33, Caspase-14 subunit p10, Step, CG11628, Remodeling, mice, Bone Matrices, portion of bone tissue, Swiss Mouse, Spectrometry, signs, Caspase-14 subunit p19, Striatum-enriched protein-tyrosine phosphatase, Condyle, MICE, domesticus, SYNTHETIC CONSTRUCT sequences, Bony, Specificity and Sensitivity, bone, STEP, chewing, MAST 9, artificial, Mouse, Bony Apophysis, Bone, Proteomes, 3.4.22.-, GRP1/cytohesin 1, Chewing, Processes, Matrix, Mylohyoid Ridge, Gene, mini-ICE, Turnover, Spectrum Analyses, Metabolic Processes, protein-containing complex, Matrices, CG11633, cytohesin/GRP1, reduced, House, Grooves, sensitive, Metabolism, Bone Turnover, Gene Products, Mass, Mus musculus domesticus, Bone mass, lower mandibula, tiny, Metabolism Phenomena, Mice, Mylohyoid Ridges, sensitivity, Mass Spectroscopy, inferior maxillary bone, reactivity, anatomical systems, Tissues, Swiss, ligand, Turnovers, l(2)SH2 0323, Metabolic Concept, Neural-specific protein-tyrosine phosphatase, Sensitivity, Apophysis, femoral diaphysis, HHT1, small, lumen, findings, Apophyses, Edg, Bone Loss, degradation, protein complex, space, Proteins, total expressed protein, artificial gene, stepk, synthetic DNA, Impact, Concept, Metabolic Phenomena, Bones and Bone, Environmental, Metabolism Concepts, MS, native protein, Mus, Protein, chemical analysis, Phenomena, proteomic analysis, synthetic, bones, anatomical spaces, lower jaw, Groove, metabolism, l(2)SH0323, Mass Spectrum Analyses, Metabolic Phenomenon, MAST9, Sc1, CYH1, Cosmonauts, Mass Spectrum, multicellular organism metabolic process, lumen space, osseous tissue, Exhibit, biodegradation, mast9, Metabolic, underdeveloped, Bone Turnovers, Specificity, House Mice, Bones, synthetic constructs, sample population, DmelCG11628, Laboratory Mice, Protein Gene Products, SC1, Gene Proteins, mineralized bone tissue, Bone Tissue, shaft of femur, lower jaw bone, Environments, calcium tissue, assay, response, responsive, ORW1, bone organ, Laboratory Mouse, Severe, Anabolism, corpus femoris3.4.22.-, thigh, Trochanters, Greater Trochanter, Laboratory, proximal segment of free lower limb, Mus domesticus, Proteins, Lesser Trochanters, CASP14, mouse, number, Gene, mini-ICE, CASP-14, hindlimb stylopodium, hind limb stylopod, os femoris, House Mouse, Trochanter, presence., mesomere 1 of pevlic appendage, upper leg, count in organism, hindlimb stylopod, House, Mus, Protein, Mini-ICE, Gene Products, Mus musculus domesticus, femoral bone, Mice, hindlimb propodium, Greater Trochanters, Mus musculus, hind propodium, Lesser Trochanter, Caspase-14 subunit p10, stylopod of hind limb, Swiss, os longissimum, hind limb stylopodium, mice, os femorale, Swiss Mouse, stylopod of lower limb, Lesser, House Mice, Swiss Mice, Caspase-14 subunit p19, MICE, Laboratory Mice, domesticus, Protein Gene Products, stylopod of hindlimb, Gene Proteins, femur, thigh bone, Mouse, responsive, Greater, Laboratory Mouse, proximal metapterygial mesomere of pelvic appendage, Femurs, proximal metapterygial mesomere of pelvic finliquid chromatography tandem mass spectroscopy, Bones and Bone Tissue, CDF, Plantaris, artificial sequence, Anterior, short stature, Activity, determination, Laboratory, mandibular series, Blood, Skeletal, fracture, Mus domesticus, Anterior Tibial Muscle, Osteoporosis, Cosmonaut, mandible, Serum, mandibula, House Mouse, osteogenic tissue, Space, Readability, POF, Bone and Bone, AGAMOUS-like 61, Research Activity, Eastern, Laboratory Research, East, DiA, Bone Tissues, Ridge, Priorities, average, Eastern Asian Peoples, LCMSMS, l(2)k07135, Tissue, Mylohyoid Groove, Swiss Mice, Mandibles, DIASP, Post-Traumatic Osteoporoses, DIA, Dia, Involutional, 4-(4-dihexadecylaminostyryl)N-methylpyridium iodide, set, ISS, Muscles, involutional, Age Related Osteoporosis, body of femur, 38E.16, Mylohyoid Grooves, OSTEOP, Condyles, Age-Related Bone Loss, idiopathic, Research Priority, house mouse, Age-Related, Gastrocnemius, Bony Apophyses, LC-MS2, Senile, Astronaut, Explorations, Post Traumatic, Age-Related Osteoporosis, mouse, LC-MS/MS, Research Priorities, Normalcy, Blood Serum, Spaceflights, Generalized osteopenia, Far East, results, Osteopenia, Senile Osteoporoses, Bone Disease, ms(2)04138, Desmoplastic infantile astrocytoma, Voluntary Muscle, Mylohyoid, mandibles, Voluntary, Ridges, Voluntary Muscles, Individual, femoral shaft, East Asian Peoples, Research and Development, Mus musculus, Yamato people, mice, portion of bone tissue, POF2, susceptibility to, F27C12_24, Swiss Mouse, F27C12.24, Flights, Condyle, Post-Traumatic, domesticus, Activities, Bone Density, SYNTHETIC CONSTRUCT sequences, Health, Bony, People, Exploration, Patient, Metabolic Bone Diseases, bone, liquid chromatography-tandem mass spectroscopy, Eastern Asians, Space Travel, DIANA, Travel, artificial, Mouse, Bony Apophysis, Proteomes, Bone, Gruppe, Age-Related Bone Losses, Flight, Gastrocnemius Muscle, X-linked, Asian, Goal, Mylohyoid Ridge, Age-Related Osteoporoses, Gene, Normalcies, skeletal muscle, LC-MS-MS, Senile Osteoporosis, Grooves, House, Japanese, DmelCG1768, LC-MSMS, Gene Products, HILDA, Age Related, Mus musculus domesticus, Bone mass, lower mandibula, Low, Space Travels, Space Flights, Low Bone Mineral Density, Mylohyoid Ridges, Mice, Bone Losses, Space Explorations, inferior maxillary bone, Asian People, Normalities, study, Research, Tibial Muscle, Tissues, Swiss, Eastern Asian People, Space Exploration, Skeletal Muscles, Far East Asian, Skeletal Muscle, Plantaris Muscle, grupos, Clients, Apophysis, Development and Research, Generalised osteopenia, DIA2, Anterior Tibial, femoral diaphysis, striated muscle, Individual Health, lumen, Spaceflight, Apophyses, grupo, Bone Loss, Metabolic Bone Disease, Low Bone Densities, hip, space, Desmoplastic astrocytoma of infancy, Proteins, total expressed protein, artificial gene, CG1768, East Asian, synthetic DNA, Muscle, DRF2, C57BL6J, Client, Osteoporoses, group, Low Bone Density, Bones and Bone, Soleus, Osteopenias, somatic muscle, LC/MS/MS, Experiment, Priority, Mus, osteoporosis, chemical analysis, Protein, MLPLI, Research Activities, synthetic, bones, anatomical spaces, lower jaw, Dias, Groove, Post-Traumatic Osteoporosis, Cosmonauts, lumen space, osseous tissue, Soleus Muscle, Metabolic, ensemble, Normality, Protein., Rest, House Mice, Travels, Understanding, Bones, synthetic constructs, Laboratory Mice, Protein Gene Products, Gene Proteins, East Asians, Far East Asians, mineralized bone tissue, Bone Tissue, Osteopaenia, shaft of femur, lower jaw bone, liquid chromatography tandem mass spectrometry, calcium tissue, Eastern Asian, assay, bone organ, Laboratory Mouse, groupe, Serums, skeletal muscle system, corpus femorisfalseIdentification of gravity-responsive proteins in the femur of spaceflight mice using a quantitative proteomic approachThis project is “Medical proteome analysis of osteoporosis- and bone mass-related proteins using the Kibo Japanese Experiment Module of International Space Station (ISS)”. Utilizing proteomic technologies, this research aims to identify proteins involved in space flight-induced osteopenia as well as osteoporosis occurring on the ground by searching serum, and skeletal muscle and bone (femoral diaphysis, mandible) proteomes. The goal of this study is to get better understanding to protect the health of both astronauts and osteoporosis patients. Proteome analyses of C57BL/6J space flown mouse (micro-G and artificial 1-G [n = 5/group, in femoral diaphysis and n = 6/group in mandible]) were carried out tryptic digestion followed by LC-MS/MS analysis (DIA). The obtained results were analyzed using Spectronaut Pulsar X against the mouse-bone spectral library for identification and quantification of proteins.Tue Aug 01 00:00:00 BST 2023PXD0417381009037482271