#METABOLOMICS WORKBENCH juliehaines_20230524_112714 DATATRACK_ID:4041 STUDY_ID:ST002714 ANALYSIS_ID:AN004400 PROJECT_ID:PR001682 VERSION 1 CREATED_ON May 30, 2023, 11:59 am #PROJECT PR:PROJECT_TITLE Loss of microglial MCT4 leads to defective synaptic pruning and anxiety-like PR:PROJECT_TITLE behavior in mice PR:PROJECT_SUMMARY Microglia, the innate immune cells of the central nervous system, actively PR:PROJECT_SUMMARY participate in brain development by supporting neuronal maturation and refining PR:PROJECT_SUMMARY synaptic connections. These cells are emerging as highly metabolically flexible, PR:PROJECT_SUMMARY able to oxidize different energetic substrates to meet their energy demand. PR:PROJECT_SUMMARY Lactate is particularly abundant in the brain, but whether microglia use it as a PR:PROJECT_SUMMARY metabolic fuel has been poorly explored. Here we show that microglia can import PR:PROJECT_SUMMARY lactate, and this is coupled with increased lysosomal acidification. In vitro, PR:PROJECT_SUMMARY loss of the monocarboxylate transporter MCT4 in microglia prevents PR:PROJECT_SUMMARY lactate-induced lysosomal modulation and leads to defective cargo degradation. PR:PROJECT_SUMMARY Microglial depletion of MCT4 in vivo leads to impaired synaptic pruning, PR:PROJECT_SUMMARY associated with increased excitation in hippocampal neurons, enhanced E/I ratio, PR:PROJECT_SUMMARY vulnerability to seizures and anxiety-like phenotype. Overall, these findings PR:PROJECT_SUMMARY show that selective disruption of the MCT4 transporter in microglia is PR:PROJECT_SUMMARY sufficient to alter synapse refinement and to induce defects in brain PR:PROJECT_SUMMARY development and adult behavior. PR:INSTITUTE University of Colorado Denver PR:LABORATORY Lab of Angelo D'Alessandro in collaboration with lab of Rosa Paolicelli (Univ of PR:LABORATORY Lausanne) PR:LAST_NAME Haines PR:FIRST_NAME Julie PR:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA PR:EMAIL julie.haines@cuanschutz.edu PR:PHONE 3037243339 #STUDY ST:STUDY_TITLE Loss of microglial MCT4 leads to defective synaptic pruning and anxiety-like ST:STUDY_TITLE behavior in mice ST:STUDY_SUMMARY Microglia, the innate immune cells of the central nervous system, actively ST:STUDY_SUMMARY participate in brain development by supporting neuronal maturation and refining ST:STUDY_SUMMARY synaptic connections. These cells are emerging as highly metabolically flexible, ST:STUDY_SUMMARY able to oxidize different energetic substrates to meet their energy demand. ST:STUDY_SUMMARY Lactate is particularly abundant in the brain, but whether microglia use it as a ST:STUDY_SUMMARY metabolic fuel has been poorly explored. Here we show that microglia can import ST:STUDY_SUMMARY lactate, and this is coupled with increased lysosomal acidification. In vitro, ST:STUDY_SUMMARY loss of the monocarboxylate transporter MCT4 in microglia prevents ST:STUDY_SUMMARY lactate-induced lysosomal modulation and leads to defective cargo degradation. ST:STUDY_SUMMARY Microglial depletion of MCT4 in vivo leads to impaired synaptic pruning, ST:STUDY_SUMMARY associated with increased excitation in hippocampal neurons, enhanced E/I ratio, ST:STUDY_SUMMARY vulnerability to seizures and anxiety-like phenotype. Overall, these findings ST:STUDY_SUMMARY show that selective disruption of the MCT4 transporter in microglia is ST:STUDY_SUMMARY sufficient to alter synapse refinement and to induce defects in brain ST:STUDY_SUMMARY development and adult behavior. ST:INSTITUTE University of Colorado Denver ST:LAST_NAME Haines ST:FIRST_NAME Julie ST:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA ST:EMAIL julie.haines@cuanschutz.edu ST:PHONE 3037243339 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 1 sex:M | genetic status:KO RAW_FILE_NAME=FC8-1 SUBJECT_SAMPLE_FACTORS - 2 sex:F | genetic status:WT RAW_FILE_NAME=FC8-2 SUBJECT_SAMPLE_FACTORS - 3 sex:M | genetic status:WT RAW_FILE_NAME=FC8-3 SUBJECT_SAMPLE_FACTORS - 4 sex:M | genetic status:KO RAW_FILE_NAME=FC8-4 SUBJECT_SAMPLE_FACTORS - 5 sex:M | genetic status:WT RAW_FILE_NAME=FC8-5 SUBJECT_SAMPLE_FACTORS - 6 sex:F | genetic status:KO RAW_FILE_NAME=FC8-6 SUBJECT_SAMPLE_FACTORS - 7 sex:F | genetic status:KO RAW_FILE_NAME=FC8-7 SUBJECT_SAMPLE_FACTORS - 8 sex:M | genetic status:WT RAW_FILE_NAME=FC8-8 SUBJECT_SAMPLE_FACTORS - 9 sex:F | genetic status:WT RAW_FILE_NAME=FC8-9 SUBJECT_SAMPLE_FACTORS - 10 sex:M | genetic status:KO RAW_FILE_NAME=FC8-10 SUBJECT_SAMPLE_FACTORS - 11 sex:F | genetic status:KO RAW_FILE_NAME=FC8-11 SUBJECT_SAMPLE_FACTORS - 12 sex:F | genetic status:KO RAW_FILE_NAME=FC8-12 SUBJECT_SAMPLE_FACTORS - 13 sex:M | genetic status:WT RAW_FILE_NAME=FC8-13 SUBJECT_SAMPLE_FACTORS - 14 sex:M | genetic status:KO RAW_FILE_NAME=FC8-14 SUBJECT_SAMPLE_FACTORS - 15 sex:F | genetic status:KO RAW_FILE_NAME=FC8-15 SUBJECT_SAMPLE_FACTORS - 1B sex:M | genetic status:KO RAW_FILE_NAME=FC8-16 SUBJECT_SAMPLE_FACTORS - 2B sex:M | genetic status:WT RAW_FILE_NAME=FC8-17 SUBJECT_SAMPLE_FACTORS - 3B sex:M | genetic status:KO RAW_FILE_NAME=FC8-18 SUBJECT_SAMPLE_FACTORS - 4B sex:F | genetic status:WT RAW_FILE_NAME=FC8-19 SUBJECT_SAMPLE_FACTORS - 5B sex:F | genetic status:WT RAW_FILE_NAME=FC8-20 #COLLECTION CO:COLLECTION_SUMMARY Mice were bred and maintained in the animal facility of the University of CO:COLLECTION_SUMMARY Lausanne. All animal experiments were authorized by “Service de la CO:COLLECTION_SUMMARY consommation et des Affaires vétérinaires” (SCAV) of the Canton de Vaud in CO:COLLECTION_SUMMARY Switzerland. Cx3cr1CREERT2;MCT4flox mice were obtained by crossing the CO:COLLECTION_SUMMARY B6.129P2(Cg)-Cx3cr1tm2.1(cre/ERT2)Litt/WganJ mice (Cx3cr1CREERT2; No: 021160, CO:COLLECTION_SUMMARY The Jackson Laboratory) with C57Bl/6.MCT4tm1flox mice (MCT4flox; kindly donated CO:COLLECTION_SUMMARY by Prof. Luc Pellerin). For specific experiments, the Cx3cr1CREERT2;MCT4flox CO:COLLECTION_SUMMARY line was further crossed with B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J mice CO:COLLECTION_SUMMARY (TdTomatoflox; No: 007914, The Jackson Laboratory). Mice were group-housed and CO:COLLECTION_SUMMARY kept in a 12 h day/night cycle, with food and water ad libitum, at 20-22°C. All CO:COLLECTION_SUMMARY the experiments were performed during the day cycle. Both male and female mice CO:COLLECTION_SUMMARY were analyzed, unless differently specified. For in vivo gene KO induction, CO:COLLECTION_SUMMARY tamoxifen (Sigma Aldrich) was prepared in 10% ethanol in corn oil at a CO:COLLECTION_SUMMARY concentration of 6mg/ml. The whole litter was injected intraperitoneally (i.p.) CO:COLLECTION_SUMMARY at P6 and P8 (75mg/kg). For biochemical and histological assessments, mice at CO:COLLECTION_SUMMARY P15 were terminally anesthetized with sodium pentobarbital diluted in saline CO:COLLECTION_SUMMARY (150mg/kg, i.p.) and perfused with cold HBSS (Life Technologies, 3ml/min). Upon CO:COLLECTION_SUMMARY brain collection, the hippocampus was dissected from the right hemisphere, CO:COLLECTION_SUMMARY immediately frozen in dry ice and stored at -80°C. CO:SAMPLE_TYPE Brain #TREATMENT TR:TREATMENT_SUMMARY For in vivo gene KO induction, tamoxifen (Sigma Aldrich) was prepared in 10% TR:TREATMENT_SUMMARY ethanol in corn oil at a concentration of 6mg/ml. The whole litter was injected TR:TREATMENT_SUMMARY intraperitoneally (i.p.) at P6 and P8 (75mg/kg). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Frozen tissue samples were weighed to the nearest 0.1 mg then treated with cold SP:SAMPLEPREP_SUMMARY 5:3:2 MeOH:acetonitrile:water to a final concentration of 15 mg/mL. Samples were SP:SAMPLEPREP_SUMMARY vortexed 30 min at 4 degrees C then supernatants clarified by centrifugation (10 SP:SAMPLEPREP_SUMMARY min, 18,000 g, 4 degrees C) and transferred to autosampler vials. SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Positive C18 CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100%acetonitrile; 0.1% formic acid CH:FLOW_GRADIENT 0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min CH:FLOW_GRADIENT 95-5% B, 3-5 min hold at 5% B CH:FLOW_RATE 450 uL/min CH:COLUMN_TEMPERATURE 45 CH:SAMPLE_INJECTION 10 uL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, MS:MS_COMMENTS microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, MS:MS_COMMENTS capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas MS:MS_COMMENTS 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were MS:MS_COMMENTS annotated and integrated using Maven in conjunction with the KEGG database. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples 2 9 4B 5B 1 4 10 14 1B 3B 3 5 8 13 2B 6 11 12 15 Factors sex:F | genetic status:WT sex:F | genetic status:WT sex:F | genetic status:WT sex:F | genetic status:WT sex:M | genetic status:KO sex:M | genetic status:KO sex:M | genetic status:KO sex:M | genetic status:KO sex:M | genetic status:KO sex:M | genetic status:KO sex:M | genetic status:WT sex:M | genetic status:WT sex:M | genetic status:WT sex:M | genetic status:WT sex:M | genetic status:WT sex:F | genetic status:KO sex:F | genetic status:KO sex:F | genetic status:KO sex:F | genetic status:KO L-alanine 18708870 25974210 22744650 26207830 18807370 18556850 22396420 18158610 24038570 30819830 19565430 21903050 17737750 20265470 30185520 17440700 15518890 17915250 23929550 L-arginine 4745954 3783467 4337314 5626634 3651894 3912812 3864312 3727371 3728948 4717536 5534902 5050974 4574801 4913766 6294500 5154906 1743050 2724454 4584048 L-asparagine 737165 1132755 647858 768478 868396 761047 1038758 762314 609036 1101922 898081 813910 1028420 788992 1005813 794605 776314 923143 930405 L-aspartate 11610260 18398580 13680170 14755210 16990420 11933850 18987400 12884160 13166660 15784390 13232080 15897190 20884750 13563870 18623380 15486870 13069950 17604630 16468240 L-glutamate 140950400 173776600 222660300 175127000 194292700 143061700 202275200 159956900 179324000 216971000 150017800 168685400 174791200 166408700 196915300 166765900 144579600 181954500 204548700 L-glutamine 70844950 136087900 137925500 151268200 95520980 74835540 124637400 83194840 118920500 151401600 94265980 94949150 100264400 74998100 136946700 95887020 92438140 130820400 99615780 glycine 3852559 4837328 4252931 4457556 4053850 4649200 5208594 3611669 4276260 5697966 4596568 4181281 3808820 3390356 5052950 3967107 4791984 4731082 4803058 L-histidine 7193650 8595998 8770076 6082570 9045638 7760248 9813711 10226190 7390944 7955874 8885792 8283056 8088200 6567538 7420508 7582140 7498344 9000734 10518130 L-leucine 4418642 6512998 6201420 5807506 5500076 4883474 4944628 6163922 4704005 8204186 6358660 3400859 6226780 7193282 8304928 3714078 4505340 4852124 6741112 L-lysine 13511840 14303020 12874560 9752447 14001560 12889010 13711500 14104260 9731347 15141120 13785930 14431230 10291490 15774750 11727560 10769730 14124880 16431690 18021130 L-methionine 7088648 5959866 5912008 5257072 8491281 5415257 7795810 9636192 5593698 6108702 7118104 7382864 11778320 9725288 7725652 6033906 4980086 6734340 7726876 L-phenylalanine 1823948 1668300 2809258 3043452 2230982 2590284 2644581 2088027 1749979 2250645 2466120 2034593 1944907 3283980 2993546 1967487 1528075 2369322 2593226 L-proline 22776020 28656290 25354280 22244340 24448520 19704750 26363560 23200660 19402600 23942010 28788210 21955630 22178070 28883770 29224540 15697830 17884370 27729800 30808390 L-serine 7159564 10811020 9792367 8524944 8582912 6269976 11166680 7967608 8610015 11984940 8363730 8343594 10005800 7371838 10576800 7633168 8472209 8926496 9147147 L-threonine 9061256 9015699 11125150 8600537 9415589 6477196 8896564 7708218 8438606 14957120 8332952 8555587 10582790 8118832 10210350 8877399 4289399 9868692 8022552 L-tryptophan 7046384 7214788 10601040 6279106 9014250 8766429 10646240 6373966 7638092 7805000 7340804 7678130 6278960 6371742 9809490 6538162 4762164 8214672 10191610 L-tyrosine 9442998 6518416 10320380 6351628 11475250 7917314 9639556 11342740 9001695 8459412 10033210 9691429 11645020 9909675 10467370 8418632 5797820 7547566 8792517 L-valine 14536450 24248780 15054100 10800580 15480920 14296200 16035080 16751640 12484700 17965580 17688880 15452760 20056980 19620910 13572100 13672400 22149050 17800020 18152180 Adenosine 1243016 918982 15366910 1657452 1934819 964453 1717768 1376814 1299720 2470599 1183391 1108965 1481098 2163333 1537941 929027 1446291 1047335 699721 GMP 898672 990085 1428852 1118917 1493024 1076728 1288406 1251542 1320097 1829480 1201769 1525800 1258243 1651598 1024938 1143586 1088262 1248821 1420236 Guanosine 873592 761788 306294 1129086 722324 932786 616214 977356 885981 1359413 1063226 873332 1197248 1528695 1183376 762858 395082 459798 914770 CMP 546509 664207 1236881 663448 997887 511920 968686 886119 584075 656513 786688 884730 890558 868432 337000 715453 877685 1002082 908664 Cytidine 2158904 1898639 1209948 2529579 2192153 3779717 1957228 2839671 2495029 2936930 2623850 2551910 1658222 2625513 4031659 2039850 1202515 1487352 2362833 Cytosine 1711206 1321953 948651 1373951 1410745 1875734 1166365 1838416 1670516 1834888 1907990 1657886 1336774 1765355 2692104 1350282 997730 1066312 1557380 Thymine 16002 18923 42042 57023 17810 115319 6159 14902 36539 114560 24534 18357 29705 31500 19487 7363 9337 13505 68870 Uracil 650116 946770 274624 591429 737950 1162590 800890 869108 798627 1154149 1018466 768593 934592 894683 1346413 596632 425812 659993 888309 IMP 3157185 5163746 1842169 5491350 5282538 4682784 5470730 5863950 5085416 7081502 5440057 6563328 3930327 4378787 4799112 4327730 3211889 5502486 6935082 Inosine 5791252 4345863 3755944 5439698 4458464 4778000 4655834 5659150 5650040 7707632 6114848 4290606 7313392 9580605 8291308 4651290 3034500 2965846 3748685 Hypoxanthine 56760310 56607760 30733910 60714820 52148900 62916270 51650660 69720930 67153860 99787270 80239660 54130270 87276560 101913300 122196600 49067110 31973050 42379960 56471440 Allantoate 678169 654335 1029666 889386 691437 474856 693263 594167 904720 892920 618051 666229 613016 531852 901654 592093 492736 594391 819987 5-Hydroxyisourate 9287737 9307888 10939990 15051280 7794052 10639700 11206030 5420868 12440340 12359370 8776595 11072850 7848614 8049662 14071910 9495309 8819691 10889720 8348154 Nicotinamide 34169140 35380200 27560910 33833170 38253200 29733540 40647190 40620780 41012520 41170270 39653040 39897920 38792380 41307900 39582820 31002320 27328190 28679500 35906340 ADP-D-ribose 189246 220237 284659 361734 158096 189039 251988 126481 284022 202475 198161 154973 168002 138549 238773 226089 198975 285967 136334 Glutathione 75403340 101206700 135705100 85744000 126690300 71667320 119215400 115078900 81837380 108567200 100637400 112741300 99686140 94350100 63131450 82298650 95136040 115956200 109752200 Glutathione disulfide 74138 52887 27845 97564 83514 49007 146203 114284 76992 222873 127495 67439 103014 206180 203756 89950 17259 105436 61593 5-Oxoproline 92003170 146131400 153208800 158743700 104750300 99115110 145268900 96398090 126234900 166420100 109118500 109263000 121805100 91693530 143993300 102048100 109544400 142737200 121755400 Cys-Gly 589305 776647 1088953 770015 826474 559358 919614 905938 669198 756228 816750 890823 886447 760632 499362 641388 774329 816069 938054 gamma-L-Glutamyl-L-cysteine 61626 100504 218253 135751 131715 81505 124568 62115 90926 154022 86960 135314 135615 87831 61469 91845 104138 109625 135126 gamma-Glutamyl-gamma-aminobutyrate 262666 449452 481986 346630 378019 350603 433712 449331 320591 464795 421767 414215 444623 428190 425804 320686 382139 421940 489967 gamma-L-Glutamylputrescine 19273 27728 53227 47901 19023 8949 28771 27998 37458 38743 26854 20923 28564 9449 17864 18357 24371 30292 5771 (5-L-Glutamyl)-L-glutamine 186970 532572 81162 714690 301430 328494 413660 170049 427135 760514 389641 386403 220394 358584 645707 387351 338534 397440 546118 Cystathionine 555057 619974 784389 719493 628091 466563 615015 473188 535698 927004 473140 550870 696332 489011 637872 436015 506885 772887 560324 Dimethylglycine 113287300 128686100 104468100 130633600 115868500 120226200 121127300 124905100 122944600 144618300 106696500 121954400 119205700 118173700 191825500 119964200 97702880 119055800 116225200 Phosphoserine 1224940 1059364 1152427 1352618 1169844 1451207 1308397 1409644 1292148 1294193 956026 1403498 1207268 1496234 1288351 1392249 983341 1004015 1395844 S-Adenosyl-L-homocysteine 259392 188263 117695 258689 221861 200505 181785 262925 180040 271840 244826 271232 212447 260630 172828 182592 108530 113514 222122 S-Adenosyl-L-methionine 303703 379338 399530 328583 348011 346653 427679 358877 352412 380908 312194 373646 337822 317649 268201 335059 415912 371022 464753 5-10-Methenyltetrahydrofolate 273864 236411 518246 184042 302364 232011 264857 188173 140727 199294 236910 299214 210405 183411 276884 226108 130096 250866 152342 Ornithine 359085 437690 566957 325581 433852 322755 364270 425677 366386 305653 444164 379217 504812 508694 418934 352813 294411 437684 492514 L-Citrulline 1050079 1930586 1211046 1500066 1256506 1382960 1857759 1405583 1436949 1947305 1374934 1461582 1814303 1273086 1631068 1334684 1251578 1440096 1968057 Putrescine 1289776 1141063 1673695 1764097 1049549 1529527 1131985 1577793 1416417 1521128 1455008 1114675 1013529 1141081 1512447 1137283 1883313 1144062 1216993 Spermine 764134 440871 175900 359147 1111722 1295531 1182359 378565 349931 400481 4379894 228687 1137752 12373200 780243 298743 828323 389378 346308 5-Guanidino-2-oxopentanoate 197633 293229 239209 176904 217588 197131 272955 258124 194498 211063 210033 255865 256196 214112 216810 167335 161550 216059 298813 Creatinine 6462582 9164164 9719581 7841766 7808914 5951700 9243351 8655919 8771711 10980370 7395336 8931680 6937222 9357712 10792130 5449613 7338946 9267407 9503120 Guanidinoacetate 1326329 3195221 1784282 1484537 1518506 1550422 3320355 1364334 1170127 1871206 1863618 1656589 2125317 1621180 1579364 1622011 783402 2959455 2586904 5-6-Dihydrouracil 491120 385958 479407 378811 504672 488173 481157 522364 513496 640238 432569 350577 381664 497641 632322 218874 395410 500681 486597 Hypotaurine 763642 757914 199654 408322 742599 213045 788661 458592 573806 752849 300278 737879 820720 1209806 820340 614895 501612 936143 657984 3-Sulfocatechol 3774 9318 6872 11675 11198 6215 7084 8899 8761 9139 10253 5042 5353 5988 18450 5042 5145 9081 14428 L-Cysteate 154444 242176 214167 207181 179524 129142 201195 136618 194382 235264 201276 145542 195509 193275 211949 119500 140883 201812 249510 Indole 143248 190902 283825 151766 258718 172179 287426 145327 190627 207044 167557 174002 171448 174659 260877 150241 131936 235213 279677 Indole-3-acetaldehyde 70364 55711 118832 37820 84751 53116 94114 40154 99718 66604 42222 72036 38444 64007 62606 40617 75537 78401 51476 Picolinic acid 1635061 4453314 3435764 2015788 2568606 1949485 3005444 2339703 2526618 2670782 3284941 1848404 1685106 6340012 2068842 1391728 6665804 2557666 3707635 g-Oxalo-crotonate 1541755 833448 252358 944475 852610 1577142 754093 1068696 1184135 1240595 1287525 962293 1479254 1771588 2185660 1015336 573961 624773 889130 2-Aminomuconate 2819438 3045763 4083685 3375253 3790382 2490391 3394339 2998200 3280867 3262671 3358365 3244764 2952720 3298658 3761753 2798367 2753269 3232131 2977177 Dopamine 6760 6351 7350 6348 11934 4834 9498 4817 4938 5634 5215 5802 5877 4583 6330 6859 5951 5406 6579 Serotonin 16009 13486 27125 3910 17627 3553 23934 0 20599 9829 2950 12299 0 7505 9720 5174 16236 13417 0 Sphingosine 56938 179772 306237 54848 224210 62048 233917 72541 64377 208064 248391 35282 173691 64480 271386 47707 89418 362131 268363 Acetylcholine 14699870 14113400 16182170 13217100 12756880 14600150 17418390 15725640 10333460 10199070 13675670 17135140 9454319 14319770 15508790 11485150 12315040 15051600 13024760 acyl-C4-OH (Hydroxybutyrylcarnitine) 992954 1043284 2095049 881013 958844 1308213 1494384 1356316 1475731 1782188 1103140 1025922 781043 1054033 563670 839652 1935375 1616689 1096020 acyl-C5-OH 262991 424158 432757 261589 290729 382219 401516 356983 342105 407289 350980 407010 358692 412475 362081 327502 360699 451728 332808 acyl-C6 (hexanoyl-l-carnitine) 92222 121215 407564 46580 142994 44703 246868 97308 66048 71451 42197 77688 71139 46169 11317 90353 167788 353067 74507 acyl-C16:1 (Hexadecenoyl-carnitine) 2801 5116 19022 4817 5688 6333 7857 10737 4518 31454 12981 3380 3980 5484 5946 5957 4451 5045 10014 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID m/z r.t. L-alanine C00041 90.05515 0.658537 L-arginine C00062 175.1192 0.6319906 L-asparagine C00152 133.0609 0.6477286 L-aspartate C00049 134.0449 0.6519988 L-glutamate C00025 148.0606 0.6551487 L-glutamine C00064 147.0768 0.6503452 glycine C00037 76.03951 0.6509777 L-histidine C00135 156.0771 0.6124547 L-leucine C00123 132.1022 0.7681905 L-lysine C00047 147.1132 0.6105534 L-methionine C00073 150.0586 0.7245071 L-phenylalanine C00079 166.0866 1.209986 L-proline C00148 116.0707 0.6757178 L-serine C00065 106.05 0.6472368 L-threonine C00188 120.0658 0.6528462 L-tryptophan C00078 205.0975 1.716568 L-tyrosine C00082 182.0814 0.7022607 L-valine C00183 118.0863 0.680799 Adenosine C00212 268.1037 0.6995918 GMP C00144 364.0655 0.7235308 Guanosine C00387 284.099 0.6995821 CMP C00055 324.0591 0.6677667 Cytidine C00475 244.093 0.683457 Cytosine C00380 112.0507 0.6830011 Thymine C00178 127.0504 0.7037253 Uracil C00106 113.035 0.7050622 IMP C00130 349.0547 0.7237406 Inosine C00294 269.088 0.7003214 Hypoxanthine C00262 137.046 0.7063929 Allantoate C00499 177.0589 0.8380048 5-Hydroxyisourate C11821 185.0323 0.6449929 Nicotinamide C00153 123.0554 0.7291929 ADP-D-ribose C01882 560.0765 0.6525124 Glutathione C00051 308.0913 0.7097494 Glutathione disulfide C00127 613.1599 0.7003402 5-Oxoproline C01879 130.0501 0.6526915 Cys-Gly C01419 179.0488 0.7053834 gamma-L-Glutamyl-L-cysteine C00669 251.0694 0.6945988 gamma-Glutamyl-gamma-aminobutyrate C15767 233.1133 0.6990371 gamma-L-Glutamylputrescine C15699 218.1502 0.6084161 (5-L-Glutamyl)-L-glutamine C05283 276.1191 0.6665633 Cystathionine C00542 223.0749 0.6454355 Dimethylglycine C01026 104.0707 0.6540116 Phosphoserine C02532 186.0171 0.6603054 S-Adenosyl-L-homocysteine C00021 385.1289 0.6981629 S-Adenosyl-L-methionine C00019 399.1446 0.6404675 5-10-Methenyltetrahydrofolate C00445 457.1662 0.6751891 Ornithine C01602 133.0975 0.6077038 L-Citrulline C00327 176.1034 0.6534141 Putrescine C00134 89.10743 0.5902053 Spermine C00750 203.2234 0.5527307 5-Guanidino-2-oxopentanoate C03771 174.0877 0.6969466 Creatinine C00791 114.0664 0.6676642 Guanidinoacetate C00581 118.0612 0.6537535 5-6-Dihydrouracil C00429 115.0504 0.6832282 Hypotaurine C00519 110.0272 0.6568711 3-Sulfocatechol C06336 191.0012 0.5382885 L-Cysteate C00506 170.0125 0.845418 Indole C00463 118.0654 1.732132 Indole-3-acetaldehyde C00637 160.076 1.134227 Picolinic acid C10164 124.0405 0.5881027 g-Oxalo-crotonate C03453 159.0278 0.6977988 2-Aminomuconate C02220 158.045 0.7024754 Dopamine C03758 154.0866 0.8418379 Serotonin C00780 177.1025 1.131402 Sphingosine C00319 300.2897 1.903268 Acetylcholine C01996 146.1179 0.672938 acyl-C4-OH (Hydroxybutyrylcarnitine) HMDB13127 248.1493 0.7000383 acyl-C5-OH ac107 262.1649 0.7034601 acyl-C6 (hexanoyl-l-carnitine) HMDB00756 260.1856 1.777901 acyl-C16:1 (Hexadecenoyl-carnitine) HMDB06317 398.3272 1.950467 METABOLITES_END #END