#METABOLOMICS WORKBENCH sven_haange_20200603_022221_mwtab.txt DATATRACK_ID:2041 STUDY_ID:ST001397 ANALYSIS_ID:AN002336 PROJECT_ID:000000 VERSION 1 CREATED_ON June 8, 2020, 5:52 pm #PROJECT PR:PROJECT_TITLE Enteric microbiota and liver metabolomics during the postnatal period PR:PROJECT_SUMMARY Following birth, the neonatal intestine is exposed to maternal and environmental PR:PROJECT_SUMMARY bacteria that successively form a dense and highly dynamic intestinal PR:PROJECT_SUMMARY microbiota. Whereas the effect of exogenous factors has been extensively PR:PROJECT_SUMMARY investigated, endogenous, host-mediated mechanisms have remained largely PR:PROJECT_SUMMARY unexplored. Concomitantly with microbial colonization, the liver undergoes PR:PROJECT_SUMMARY functional transition from a hematopoietic organ to a central organ of metabolic PR:PROJECT_SUMMARY regulation and immune surveillance. The aim of the present study was to analyze PR:PROJECT_SUMMARY the influence of the developing hepatic function and liver metabolism on the PR:PROJECT_SUMMARY early intestinal microbiota. Using metabolomic and microbial profiling in PR:PROJECT_SUMMARY combination with multivariate analysis we characterized the colonization PR:PROJECT_SUMMARY dynamics and liver metabolism in the murine gastrointestinal tract (n=6-10 per PR:PROJECT_SUMMARY age group). We observed major age-dependent microbial and metabolic changes and PR:PROJECT_SUMMARY identified bile acids as potent drivers of the early intestinal microbiota PR:PROJECT_SUMMARY maturation. Consistently, oral administration of tauro-cholic acid or PR:PROJECT_SUMMARY β-tauro-murocholic acid to newborn mice (n= 7-14 per group) accelerated PR:PROJECT_SUMMARY postnatal microbiota maturation. PR:INSTITUTE Helmholtz Centre for Environmental Research - UFZ PR:LAST_NAME Haange PR:FIRST_NAME Sven PR:ADDRESS Permoserstrasse 1, Leipzig, Saxony, 04318, Germany PR:EMAIL sven.haange@ufz.de PR:PHONE 0049 341 2351099 #STUDY ST:STUDY_TITLE Quantitative Hexose study on total murine liver tissue from mice at different ST:STUDY_TITLE age ST:STUDY_TYPE Liver tissue/Primary tissue ST:STUDY_SUMMARY Following birth, the neonatal intestine is exposed to maternal and environmental ST:STUDY_SUMMARY bacteria that successively form a dense and highly dynamic intestinal ST:STUDY_SUMMARY microbiota. Whereas the effect of exogenous factors has been extensively ST:STUDY_SUMMARY investigated, endogenous, host-mediated mechanisms have remained largely ST:STUDY_SUMMARY unexplored. Concomitantly with microbial colonization, the liver undergoes ST:STUDY_SUMMARY functional transition from a hematopoietic organ to a central organ of metabolic ST:STUDY_SUMMARY regulation and immune surveillance. The aim of the present study was to analyze ST:STUDY_SUMMARY the influence of the developing hepatic function and liver metabolism on the ST:STUDY_SUMMARY early intestinal microbiota. Using metabolomic and microbial profiling in ST:STUDY_SUMMARY combination with multivariate analysis we characterized the colonization ST:STUDY_SUMMARY dynamics and liver metabolism in the murine gastrointestinal tract (n=6-10 per ST:STUDY_SUMMARY age group). We observed major age-dependent microbial and metabolic changes and ST:STUDY_SUMMARY identified bile acids as potent drivers of the early intestinal microbiota ST:STUDY_SUMMARY maturation. Consistently, oral administration of tauro-cholic acid or ST:STUDY_SUMMARY β-tauro-murocholic acid to newborn mice (n= 7-14 per group) accelerated ST:STUDY_SUMMARY postnatal microbiota maturation.Summed hexoses in total liver tissue from ST:STUDY_SUMMARY healthy C57BL/6 mice at 1, 7, 14, 21, 28 and 56 day after birth was analyzed. ST:INSTITUTE Helmholtz Centre for Environmental Research - UFZ ST:DEPARTMENT Department of Molecular Systems biology ST:LABORATORY Functional Metabolomics ST:LAST_NAME Rolle-Kampczyk ST:FIRST_NAME Ulrike ST:ADDRESS Permoserstrasse 15, 04318 Leipzig, Germany ST:EMAIL ulrike.rolle-kampczyk@ufz.de ST:PHONE 0049 341 235 1537 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #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 - L_K9D1_neg Litter:9 | Day after birth:1 RAW_FILE_NAME=L_K9D1_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K10D1_neg Litter:10 | Day after birth:1 RAW_FILE_NAME=L_K10D1_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K7D7_neg Litter:7 | Day after birth:7 RAW_FILE_NAME=L_K7D7_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K8D7_neg Litter:8 | Day after birth:7 RAW_FILE_NAME=L_K8D7_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K9D7_neg Litter:9 | Day after birth:7 RAW_FILE_NAME=L_K9D7_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K10D7_neg Litter:10 | Day after birth:7 RAW_FILE_NAME=L_K10D7_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K11D7_neg Litter:11 | Day after birth:7 RAW_FILE_NAME=L_K11D7_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K7D14_neg Litter:7 | Day after birth:14 RAW_FILE_NAME=L_K7D14_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K8D14_neg Litter:8 | Day after birth:14 RAW_FILE_NAME=L_K8D14_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K9D14_neg Litter:9 | Day after birth:14 RAW_FILE_NAME=L_K9D14_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K10D14_neg Litter:10 | Day after birth:14 RAW_FILE_NAME=L_K10D14_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K11D14_neg Litter:11 | Day after birth:14 RAW_FILE_NAME=L_K11D14_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K7D21_neg Litter:7 | Day after birth:21 RAW_FILE_NAME=L_K7D21_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K8D21_neg Litter:8 | Day after birth:21 RAW_FILE_NAME=L_K8D21_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K9D21_neg Litter:9 | Day after birth:21 RAW_FILE_NAME=L_K9D21_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K10D21_neg Litter:10 | Day after birth:21 RAW_FILE_NAME=L_K10D21_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K11D21_neg Litter:11 | Day after birth:21 RAW_FILE_NAME=L_K11D21_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K7D28_neg Litter:7 | Day after birth:28 RAW_FILE_NAME=L_K7D28_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K8D28_neg Litter:8 | Day after birth:28 RAW_FILE_NAME=L_K8D28_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K9D28_neg Litter:9 | Day after birth:28 RAW_FILE_NAME=L_K9D28_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K10D28_neg Litter:10 | Day after birth:28 RAW_FILE_NAME=L_K10D28_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K11D28_neg Litter:11 | Day after birth:28 RAW_FILE_NAME=L_K11D28_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K7D56_neg Litter:7 | Day after birth:56 RAW_FILE_NAME=L_K7D56_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K8D56_neg Litter:8 | Day after birth:56 RAW_FILE_NAME=L_K8D56_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K9D56_neg Litter:9 | Day after birth:56 RAW_FILE_NAME=L_K9D56_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K10D56_neg Litter:10 | Day after birth:56 RAW_FILE_NAME=L_K10D56_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K11D56_neg Litter:11 | Day after birth:56 RAW_FILE_NAME=L_K11D56_neg.wiff SUBJECT_SAMPLE_FACTORS - L_K11D1_neg Litter:11 | Day after birth:1 RAW_FILE_NAME=L_K11D1_neg.wiff #COLLECTION CO:COLLECTION_SUMMARY Animals were sacrificed and the liver tissue was removed, snap frozen in liquid CO:COLLECTION_SUMMARY nitrogen and stored at -80°C until further analysis CO:SAMPLE_TYPE Liver #TREATMENT TR:TREATMENT_SUMMARY All C57BL6J wildtype mice were bred locally and held under specific TR:TREATMENT_SUMMARY pathogen-free conditions at the Institute of Laboratory Animal Science at RWTH TR:TREATMENT_SUMMARY Aachen University Hospital. The day of birth was considered day 0, i.e. animals TR:TREATMENT_SUMMARY screened at day 1 were approximately 24 h old and verified to have ingested TR:TREATMENT_SUMMARY breast milk (abdominal milk spot). Mice were weaned at PND21. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Sample preparation for MetIDQ p180 Kit measurement Solvents: Acetonitril (Merck SP:SAMPLEPREP_SUMMARY KGaA, Darmstadt, Germany hypergrade for LC-MS) Water MiliQ, Extracting agent - SP:SAMPLEPREP_SUMMARY ACN / H2O (1:1) Equipment 4 steel balls size M Eppendorf Tubes 2mL Tissue slicer SP:SAMPLEPREP_SUMMARY (Rettberg, Germany) Centrifuge (Sigma) Work steps Approximately 100mg of sample SP:SAMPLEPREP_SUMMARY and 4 steel balls of size M into each tube. Per mg of sample 5µL of extracting SP:SAMPLEPREP_SUMMARY agent was added. Shake the samples for 10 minutes at 30 Hz in the tissue slicer SP:SAMPLEPREP_SUMMARY and centrifuge for 2 minutes at 14000 rpm. The supernatant was used for the SP:SAMPLEPREP_SUMMARY targeted analytics. Kit reparation The analysis was performed using the SP:SAMPLEPREP_SUMMARY validated MetIDQ p180 Kit and described in Siskos et al. [1]. Data processing SP:SAMPLEPREP_SUMMARY was carried out with the provided quantitation method Kit (Biocrates Life SP:SAMPLEPREP_SUMMARY Sciences AG, Innsbruck, Austria). 1 Siskos AP, Jain P, Römisch-Margl W, Bennett SP:SAMPLEPREP_SUMMARY M, Achaintre D, Asad Y, et al. Interlaboratory Reproducibility of a Targeted SP:SAMPLEPREP_SUMMARY Metabolomics Platform for Analysis of Human Serum and Plasma. Anal Chem SP:SAMPLEPREP_SUMMARY 2017;89:656-65. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME UPLC (Waters Acquity, Waters Corporation, Milford, USA) CH:COLUMN_NAME Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 5500 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Analyst version 1.6 Software from SCIEX. For Validation METIDQ Software version MS:MS_COMMENTS Boron from Biocrates #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS µM MS_METABOLITE_DATA_START Samples L_K9D1_neg L_K10D1_neg L_K11D1_neg L_K7D7_neg L_K8D7_neg L_K9D7_neg L_K10D7_neg L_K11D7_neg L_K7D14_neg L_K8D14_neg L_K9D14_neg L_K10D14_neg L_K11D14_neg L_K7D21_neg L_K8D21_neg L_K9D21_neg L_K10D21_neg L_K11D21_neg L_K7D28_neg L_K8D28_neg L_K9D28_neg L_K10D28_neg L_K11D28_neg L_K7D56_neg L_K8D56_neg L_K9D56_neg L_K10D56_neg L_K11D56_neg Factors Litter:9 | Day after birth:1 Litter:10 | Day after birth:1 Litter:11 | Day after birth:1 Litter:7 | Day after birth:7 Litter:8 | Day after birth:7 Litter:9 | Day after birth:7 Litter:10 | Day after birth:7 Litter:11 | Day after birth:7 Litter:7 | Day after birth:14 Litter:8 | Day after birth:14 Litter:9 | Day after birth:14 Litter:10 | Day after birth:14 Litter:11 | Day after birth:14 Litter:7 | Day after birth:21 Litter:8 | Day after birth:21 Litter:9 | Day after birth:21 Litter:10 | Day after birth:21 Litter:11 | Day after birth:21 Litter:7 | Day after birth:28 Litter:8 | Day after birth:28 Litter:9 | Day after birth:28 Litter:10 | Day after birth:28 Litter:11 | Day after birth:28 Litter:7 | Day after birth:56 Litter:8 | Day after birth:56 Litter:9 | Day after birth:56 Litter:10 | Day after birth:56 Litter:11 | Day after birth:56 Hexose 275.3631285 57.74294671 67.27642276 189.6210873 101.1612903 110.5146036 103.8087774 111.655481 134.1391106 87.96913079 218.8829787 114.4590164 85.91355599 86.23958333 97.73645058 98.01617251 127.9773692 74.48109966 96.31456491 115.8071279 84.96465696 145.1473477 108.7637841 137.7829699 101.0494959 110.7439825 157.7946768 120.7485191 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Description IDs Hexose Summed sugars C00031; C00159; C00221; C00247; C00267; C00738; C00764; C00795; C00936; C00962; C00984; C01452; C01487; C01582; C01719; C01825; C02336; C06464; C06465; C06466; C06467; C06468; C10906; C15923 METABOLITES_END #END