#METABOLOMICS WORKBENCH kameiyasutomi_20190506_194029 DATATRACK_ID:1719 STUDY_ID:ST001179 ANALYSIS_ID:AN001957 PROJECT_ID:PR000791 VERSION 1 CREATED_ON May 7, 2019, 12:25 pm #PROJECT PR:PROJECT_TITLE Metabolomic analysis of skeletal muscle in young and aged mice PR:PROJECT_SUMMARY To better understand the metabolic changes in skeletal muscle during aging, we PR:PROJECT_SUMMARY performed a metabolomic analysis of skeletal muscle in young (8-week-old) and PR:PROJECT_SUMMARY aged (28-month-old) mice using CE-TOFMS. PR:INSTITUTE Kyoto Prefectural University PR:LAST_NAME Kamei PR:FIRST_NAME Yasutomi PR:ADDRESS 1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan PR:EMAIL ran960116@yahoo.co.jp PR:PHONE +81-75-703-5661 #STUDY ST:STUDY_TITLE Metabolomic analysis of skeletal muscle in young and aged mice ST:STUDY_SUMMARY Sarcopenia is the age-induced, progressive loss of skeletal muscle mass and ST:STUDY_SUMMARY function, which results in poor muscle performance. To better understand changes ST:STUDY_SUMMARY in skeletal muscles during sarcopenia, we performed a metabolomic analysis of ST:STUDY_SUMMARY skeletal muscle in young (8-week-old) and aged (28-month-old) mice using ST:STUDY_SUMMARY CE-TOFMS. Our data shows that the metabolites including glucose and polyamine ST:STUDY_SUMMARY metabolism were decreased in aged mice compared with young mice. In addition, ST:STUDY_SUMMARY neurotransmitter levels were higher in aged mice. ST:INSTITUTE Kyoto Prefectural University ST:LAST_NAME Kamei ST:FIRST_NAME Yasutomi ST:ADDRESS 1-5 Hangi-cho, Shimogamo, Sakyo-ku Kyoto, Kyoto, Kyoto, 606-8522, Japan ST:EMAIL ran960116@yahoo.co.jp ST:PHONE +81-75-703-5661 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN wild type SU:AGE_OR_AGE_RANGE 8-week-old and 28-month-old SU:WEIGHT_OR_WEIGHT_RANGE 25.4 ± 0.2 g for young and 32.0 ± 0.2 g for aged SU:GENDER Male SU:ANIMAL_ANIMAL_SUPPLIER Shimizu Laboratory Supplies (Kyoto, Japan) SU:ANIMAL_LIGHT_CYCLE 12-h light-dark cycle SU:ANIMAL_FEED CE2 SU:SPECIES_GROUP C57BL/6J #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - Young-1 Age:8w SUBJECT_SAMPLE_FACTORS - Young-2 Age:8w SUBJECT_SAMPLE_FACTORS - Young-3 Age:8w SUBJECT_SAMPLE_FACTORS - Young-4 Age:8w SUBJECT_SAMPLE_FACTORS - Young-5 Age:8w SUBJECT_SAMPLE_FACTORS - Old-1 Age:28m SUBJECT_SAMPLE_FACTORS - Old-2 Age:28m SUBJECT_SAMPLE_FACTORS - Old-3 Age:28m SUBJECT_SAMPLE_FACTORS - Old-4 Age:28m SUBJECT_SAMPLE_FACTORS - Old-5 Age:28m #COLLECTION CO:COLLECTION_SUMMARY Gastrocnemius muscles were collected and flash-frozen in liquid N2. CO:SAMPLE_TYPE Muscle CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY No specific treatment to mention. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Gastrocnemius muscles of 8-week-old male mice (N=5, young group) and SP:SAMPLEPREP_SUMMARY 28-month-old male mice (N=5, aged group) were used for the metabolomic analysis SP:SAMPLEPREP_SUMMARY (Human Metabolome Technologies Inc., Tsuruoka, Japan). Frozen muscle specimens SP:SAMPLEPREP_SUMMARY were transferred into 500 μL of methanol containing 50 mM external standard. SP:SAMPLEPREP_SUMMARY After homogenization by BMSM10N21 (BMS, Tokyo) at 1,500 rpm for 120 s performed SP:SAMPLEPREP_SUMMARY 5 times, 500 μL of chloroform and 200 μL of ultrapure water were added to the SP:SAMPLEPREP_SUMMARY homogenate. The solution was mixed well and centrifuged at 2,300 × g for 5 min SP:SAMPLEPREP_SUMMARY at 4 °C. The resultant water phase was ultrafiltered using a Millipore SP:SAMPLEPREP_SUMMARY Ultrafree-MC PLHCC HMT Centrifugal Filter Device, 5 kDa (Millipore, Billerica, SP:SAMPLEPREP_SUMMARY MA). The filtrate was dried and then dissolved in 50 μL of ultrapure water. The SP:SAMPLEPREP_SUMMARY samples obtained were then subjected to CE-TOFMS analysis using the Agilent SP:SAMPLEPREP_SUMMARY CE-TOFMS system (Agilent Technologies, Santa Clara, CA) at 4 °C. The detected SP:SAMPLEPREP_SUMMARY peaks were aligned according to their m/z values and normalized migration times. SP:SAMPLEPREP_SUMMARY The peaks were mean-centered and scaled using their standard deviations on a per SP:SAMPLEPREP_SUMMARY peak basis as a pretreatment. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME none CH:COLUMN_NAME none #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent CE-TOFMS MS:INSTRUMENT_TYPE Other MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS - #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS fold MS_METABOLITE_DATA_START Samples Young-1 Young-2 Young-3 Young-4 Young-5 Old-1 Old-2 Old-3 Old-4 Old-5 Factors Age:8w Age:8w Age:8w Age:8w Age:8w Age:28m Age:28m Age:28m Age:28m Age:28m Lactic acid 0.225585325 0.217333933 0.169365125 0.231525651 0.224748872 0.243354395 0.228333419 0.186296691 0.195701049 0.219346315 2-Hydroxyisobutyric acid 0 0 0 0 0 0 0 0 0 0.000141745 3-Hydroxybutyric acid 0.001455232 0.001443166 0.001182724 0.00096359 0.001122551 0.002181388 0.00126583 0.00104279 0.001133621 0.001810175 Fumaric acid 0.002153938 0.002058432 0.001637814 0.001668941 0.001699128 0.002474962 0.002300558 0.001874621 0.001578476 0.002008705 2-Oxoisovaleric acid 0 0 0 0 0 0 0.000299125 0 0 0 Succinic acid 0.012058389 0.009878876 0.010095181 0.010450312 0.012025181 0.013137761 0.009494816 0.010798795 0.011698267 0.012327049 2-Hydroxyvaleric acid 0.000229645 0.000216186 0.000216507 0.000189291 0.000180069 0.000214485 0.000173109 0.000166795 0.000173198 0.000173822 Isethionic acid 0.000417758 0.00035158 0.000327733 0.000354347 0.000369896 0.000469804 0.00047267 0.000412485 0.000442618 0.000482453 5-Oxoproline 0.000310265 0.000193547 0.000175328 0.000206424 0.000211559 0.000217494 0.000306974 0.000225966 0.000340131 0.000170354 4-Methyl-2-oxovaleric acid 3-Methyl-2-oxovaleric acid 0.000434045 0 0 0.000286235 0 0.000616377 0.000554645 0 0.000398759 0.000468582 Malic acid 0.017727848 0.01576252 0.011477006 0.014119976 0.013168134 0.018906942 0.017698511 0.014813452 0.012163261 0.015713325 Threonic acid 0.000178341 0.000164692 0.000160895 0.000203499 0.000196923 0.000256179 0.000245492 0.000244833 0.000177674 0.000208933 p-Toluic acid m-Toluic acid o-Toluic acid 0.000337138 0.000191771 0.000161744 0.000231078 0 0.000350741 0.000282555 0.000252551 0.000320887 0.000293026 Ethanolamine phosphate 0.000699928 0.000981051 0.000730606 0.000796027 0.000659072 0.000966688 0.000923101 0.000996911 0.001052168 0.000793685 Octanoic acid 0 0 0 0 0 0.000186976 0 0 0.000212582 0 2-Hydroxyglutaric acid 0.000736981 0.000638793 0.00051707 0.000546564 0.000516258 0.00075908 0.000759585 0.000580567 0.00062969 0.000984846 Pelargonic acid 0.000372969 0.000278778 0.000261082 0.000254478 0.000340624 0.000341285 0.000319183 0.000288997 0.000385333 0.000357614 Terephthalic acid 0.000288278 0.000326277 0.00028061 0.000290832 0.000322883 0.000333118 0.00033488 0.000337878 0.000301642 0.000301696 Uric acid 0.000338767 0.000243709 0.000244101 0.000218124 0.000233292 0.000385988 0.000287788 0.000251693 0.000365193 0.000408763 Dihydroxyacetone phosphate 0.013590983 0.015495727 0.00708106 0.017098914 0.018284147 0.00870879 0.007580591 0.008341464 0.007360255 0.008304005 Glycerol 3-phosphate 0.025406291 0.021746489 0.022915806 0.021738019 0.021795502 0.016775844 0.021202109 0.017541343 0.015937372 0.020217956 N-Acetylaspartic acid 0.000333881 0.000722693 0.000368487 0.000478034 0.000369896 0.000448313 0.000404647 0.000662463 0.000736204 0.000637202 Ascorbic acid 0.001221108 0.002718533 0.002770444 0.001291612 0.001496439 0.001142059 0.00182876 0.001770428 0.000696821 0.000783282 XA0017 0.000149839 0 0.000111225 0 0 0 0 0 0.00016917 0 Citric acid 0.008261513 0.008912917 0.006617056 0.007330554 0.007209875 0.01005029 0.010011089 0.009433563 0.009356286 0.012050061 Lauric acid 0.00047802 0.000475433 0.000489052 0.000491406 0.000429328 0.000590587 0.000668452 0.000535974 0.000519147 0.000418733 Phosphocreatine 0.088448682 0.08149118 0.129866987 0.075470479 0.088833236 0.065998281 0.091409239 0.083877289 0.089831266 0.086803876 Pantothenic acid 0.000401064 0.000336487 0.000339619 0.000402819 0.000411144 0.000690307 0.000586041 0.000696337 0.000992645 0.000622031 XA0027 0.000194221 0 0 0.000180099 0 0.000180529 0 0.000163794 0.000238092 0 Ribulose 5-phosphate 0.000530138 0.000577533 0.000374006 0.000511463 0.000427554 0.000560499 0.000445199 0.000380327 0.000369669 0.000430003 Ribose 5-phosphate 0.000232495 0 0 0.000201827 0.000276313 0.000193424 0 0 0 0 XA0033 0.000252854 0.000340926 0.000337921 0.000353929 0.000243049 0 0.000218457 0 0.000240777 0.00022974 Glucose 6-phosphate 0.087976363 0.032653915 0.067297243 0.031383528 0.044494456 0.069229744 0.054406711 0.052367618 0.045195346 0.09791894 Fructose 6-phosphate 0.022432309 0.008459681 0.019656735 0.00784327 0.01139272 0.018116054 0.014878191 0.014790726 0.012242476 0.025137419 Glucose 1-phosphate 0.005138507 0.002078852 0.004361135 0.001928433 0.002686849 0.004082527 0.003131654 0.003007026 0.002734474 0.005721818 myo-Inositol 1-phosphate myo-Inositol 3-phosphate 0.001447902 0.001340178 0.001321543 0.001037134 0.00108707 0.001839244 0.001892859 0.002132746 0.00227843 0.0019931 Fructose 1,6-diphosphate 0.026137164 0.038289854 0.009885466 0.042076617 0.039311471 0.011992263 0.011471394 0.011568025 0.009896468 0.012643483 IMP 0.007275751 0.004517719 0.003149968 0.005409224 0.003117508 0.012191704 0.00489152 0.005119191 0.005167753 0.010182235 CoA_divalent 0.000395364 0.000497184 0.000435986 0.000333036 0.000301594 0.000368794 0.000384153 0.000361461 0.000452016 0.000417866 PRPP 0.000344874 0.000449686 0.00036976 0.000320918 0.000490091 0.00040533 0.000353194 0.000321584 0.000212582 0.00026355 FAD_divalent 0 0.000185112 0.00015198 0 0.000179182 0.000180959 0.000204068 0.000170225 0.000211687 0.000190727 Cholic acid 0 0 0 0 0 0 0 0 0.001294288 0 ADP 0.002171854 0.002389592 0.001716351 0.002580298 0.002139101 0.002123361 0.001812627 0.002714599 0.002161174 0.001739519 XA0065 0.000181191 0.000195766 0.00016047 0.000195142 0.000178739 0.000217924 0.000190114 0.000282136 0.000252413 0.000192895 CTP 0.000933237 0.000940211 0.000931406 0.000911357 0.001022759 0.001000215 0.000860747 0.001034643 0.000898214 0.000948001 UTP 0.004389311 0.004212306 0.004349673 0.004306904 0.004488431 0.003095207 0.003043137 0.003816132 0.003560634 0.003434391 ATP 0.21578063 0.20119231 0.212911948 0.203129001 0.204649837 0.188138835 0.19016099 0.197490392 0.195325563 0.18737956 Taurocholic acid 0 0 0 0 0 0 0 0 0.000349082 0 GTP 0.002940594 0.003053245 0.003245486 0.002791318 0.00274362 0.002594025 0.002452737 0.003125369 0.002907672 0.002524535 ADP-ribose 0.001783819 0.001976308 0.001456117 0.002273169 0.002646489 0.001652267 0.001823964 0.001421831 0.001312637 0.001618581 UDP-glucose UDP-galactose 0.000603428 0.00050695 0.000634663 0.000609243 0.000571255 0.000587148 0.000611331 0.000732354 0.000704877 0.000770712 UDP-N-acetylgalactosamine UDP-N-acetylglucosamine 0.000571262 0.000589963 0.000559098 0.000625957 0.000514484 0.000623254 0.000495344 0.000666751 0.000874047 0.000554409 CMP-N-acetylneuraminate 0.000205622 0.00017579 0.000203347 0.000169652 0.000228413 0.000247582 0.000224126 0.000246548 0.000226903 0.000224105 NAD+ 0.011533138 0.010754275 0.011795824 0.011792484 0.011074272 0.008474103 0.008346717 0.008676769 0.009205912 0.008907396 NADH 0.000174677 0.000262354 0.000173206 0.000157534 0.000230187 0.000131528 0 0 0 0 NADP+ 0.000267512 0.000297423 0.000288676 0.000261582 0.000228413 0.000299592 0.000289532 0.000243975 0.00032939 0.00027222 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID HMDB ID m/z MT/RT Lactic acid C00186,C00256,C01432 HMDB00190,HMDB01311 89.0247784 10.987768 2-Hydroxyisobutyric acid HMDB00729 103.04019 10.085938 3-Hydroxybutyric acid C01089,C03197 HMDB00011,HMDB00357,HMDB00442 103.0405668 9.68519015 Fumaric acid C00122 HMDB00134 115.003679 25.6369806 2-Oxoisovaleric acid C00141 HMDB00019 115.04068 10.350212 Succinic acid C00042 HMDB00254 117.0195006 21.1901742 2-Hydroxyvaleric acid HMDB01863 117.0565299 9.3734367 Isethionic acid C05123 HMDB03903 124.9922302 11.8360354 5-Oxoproline C01879 HMDB00267 128.03589 9.64513775 4-Methyl-2-oxovaleric acid 3-Methyl-2-oxovaleric acid C00233, C00671,C03465 HMDB00695, HMDB00491 129.0555567 9.70501975 Malic acid C00149,C00497,C00711 HMDB00156,HMDB00744 133.014858 21.5838626 Threonic acid C01620 HMDB00943 135.030485 9.3867846 p-Toluic acid m-Toluic acid o-Toluic acid C01454, C07211, C07215 135.0456489 9.295916889 Ethanolamine phosphate C00346 HMDB00224 140.011914 8.1697449 Octanoic acid C06423 HMDB00482 143.10783 8.4872195 2-Hydroxyglutaric acid C02630,C01087,C03196 HMDB00606,HMDB00694 147.030441 17.0522023 Pelargonic acid C01601 HMDB00847 157.124406 8.2744281 Terephthalic acid C06337 HMDB02428 165.019325 16.7940757 Uric acid C00366 HMDB00289 167.021505 9.0649833 Dihydroxyacetone phosphate C00111 HMDB01473 168.990849 12.83340805 Glycerol 3-phosphate C00093 HMDB00126 171.006854 12.32361765 N-Acetylaspartic acid C01042 HMDB00812 174.040382 14.7599331 Ascorbic acid C00072 HMDB00044 175.02504 8.6184882 XA0017 186.11286 8.003117133 Citric acid C00158 HMDB00094 191.020598 26.6184488 Lauric acid C02679 HMDB00638 199.17045 7.79895608 Phosphocreatine C02305 HMDB01511 210.028591 12.46162905 Pantothenic acid C00864 HMDB00210 218.103245 7.81677746 XA0027 227.20155 7.60193386 Ribulose 5-phosphate C00199,C01101 HMDB00618 229.012029 11.1458278 Ribose 5-phosphate C00117 HMDB01548 229.0127075 10.73758338 XA0033 242.07977 7.8030808 Glucose 6-phosphate C00668,C01172,C00092 HMDB01401 259.022507 10.0380613 Fructose 6-phosphate C05345,C00085 HMDB00124 259.022628 10.1305196 Glucose 1-phosphate C00103 HMDB01586 259.022714 10.30869055 myo-Inositol 1-phosphate myo-Inositol 3-phosphate C01177, C04006 HMDB00213, HMDB06814 259.022878 10.440029 Fructose 1,6-diphosphate C00354 HMDB01058 338.989064 14.7710764 IMP C00130 HMDB00175 347.040347 9.7497771 CoA_divalent C00010 HMDB01423 382.550225 10.8008063 PRPP C00119 HMDB00280 388.946735 17.0088161 FAD_divalent C00016 HMDB01248 391.5717563 8.02999275 Cholic acid C00695 HMDB00619 407.2817 7.034793 ADP C00008 HMDB01341 426.023398 11.05458215 XA0065 445.052524 7.11626824 CTP C00063 HMDB00082 481.978473 12.5406645 UTP C00075 HMDB00285 482.962009 12.746615 ATP C00002 HMDB00538 505.988969 12.0286459 Taurocholic acid C05122 HMDB00036 514.2893 6.901324 GTP C00044 HMDB01273 521.984277 11.637898 ADP-ribose C00301 HMDB01178 558.065307 8.6329599 UDP-glucose UDP-galactose C00029, C00052 HMDB00286, HMDB00302 565.049469 8.77997875 UDP-N-acetylgalactosamine UDP-N-acetylglucosamine C00203, C00043 HMDB00304, HMDB00290 606.075897 8.5917757 CMP-N-acetylneuraminate C00128 HMDB01176 613.141508 8.3201072 NAD+ C00003 HMDB00902 662.103054 6.73651017 NADH C00004 HMDB01487 664.1195267 8.329429417 NADP+ C00006 HMDB00217 742.068148 9.5449303 METABOLITES_END #END