#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