#METABOLOMICS WORKBENCH anne_miller_20230404_060909 DATATRACK_ID:3837 STUDY_ID:ST002699 ANALYSIS_ID:AN004374 PROJECT_ID:PR001671 VERSION 1 CREATED_ON May 5, 2023, 8:11 am #PROJECT PR:PROJECT_TITLE Spatially resolved metabolomics and isotope tracing reveal dynamic metabolic PR:PROJECT_TITLE responses of dentate granule neurons with acute stimulation PR:PROJECT_TYPE Mass spectometry imaging PR:PROJECT_SUMMARY Metabolomics of dentate gyrus in mouse hippocamal slices PR:INSTITUTE Harvard University PR:LAST_NAME Yellen PR:FIRST_NAME Gary PR:ADDRESS 220 Longwood Ave PR:EMAIL gary_yellen@hms.harvard.edu PR:PHONE 617-432-0137 #STUDY ST:STUDY_TITLE Spatially resolved metabolomics and isotope tracing reveal dynamic metabolic ST:STUDY_TITLE responses of dentate granule neurons with acute stimulation ST:STUDY_SUMMARY To define the metabolic adaptations of the neuron-enriched dentate granule cell ST:STUDY_SUMMARY layer within the native brain environment, we utilize acute hippocampal brain ST:STUDY_SUMMARY slices and perform fast metabolite preservation followed by mass spectrometry ST:STUDY_SUMMARY imaging (MALDI-MSI) to obtain spatially resolved metabolomics and isotope ST:STUDY_SUMMARY tracing data. ST:INSTITUTE Harvard University ST:LAST_NAME Yellen ST:FIRST_NAME Gary ST:ADDRESS 220 Longwood Ave ST:EMAIL gary_yellen@hms.harvard.edu ST:PHONE 617-432-0137 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #FACTORS #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 - Fig2 N1 ctrl Treatment:ctrl RAW_FILE_NAME=n1_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N1 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n1_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N1 1min KCl Treatment:1min KCl RAW_FILE_NAME=n1_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N1 2min KCl Treatment:2min KCl RAW_FILE_NAME=n1_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N1 3min KCl Treatment:3min KCl RAW_FILE_NAME=n1_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N2 ctrl Treatment:ctrl RAW_FILE_NAME=n2_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N2 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n2_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N2 1min KCl Treatment:1min KCl RAW_FILE_NAME=n2_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N2 2min KCl Treatment:2min KCl RAW_FILE_NAME=n2_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N2 3min KCl Treatment:3min KCl RAW_FILE_NAME=n2_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N2 5min KCl Treatment:5min KCl RAW_FILE_NAME=n2_5min_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N3 ctrl Treatment:ctrl RAW_FILE_NAME=n3_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N3 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n3_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N3 1min KCl Treatment:1min KCl RAW_FILE_NAME=n3_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N3 2min KCl Treatment:2min KCl RAW_FILE_NAME=n3_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N3 3min KCl Treatment:3min KCl RAW_FILE_NAME=n3_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N3 5min KCl Treatment:5min KCl RAW_FILE_NAME=n3_5min_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N4 ctrl Treatment:ctrl RAW_FILE_NAME=n4_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N4 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n4_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N4 1min KCl Treatment:1min KCl RAW_FILE_NAME=n4_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N4 2min KCl Treatment:2min KCl RAW_FILE_NAME=n4_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N4 3min KCl Treatment:3min KCl RAW_FILE_NAME=n4_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N4 5min KCl Treatment:5min KCl RAW_FILE_NAME=n4_5min_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N5 ctrl Treatment:ctrl RAW_FILE_NAME=n5_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N5 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n5_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N5 1min KCl Treatment:1min KCl RAW_FILE_NAME=n5_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N5 2min KCl Treatment:2min KCl RAW_FILE_NAME=n5_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N5 3min KCl Treatment:3min KCl RAW_FILE_NAME=n5_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N5 5min KCl Treatment:5min KCl RAW_FILE_NAME=n5_5min_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N6 ctrl Treatment:ctrl RAW_FILE_NAME=n6_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N6 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n6_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N6 1min KCl Treatment:1min KCl RAW_FILE_NAME=n6_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N6 2min KCl Treatment:2min KCl RAW_FILE_NAME=n6_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N6 3min KCl Treatment:3min KCl RAW_FILE_NAME=n6_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N6 5min KCl Treatment:5min KCl RAW_FILE_NAME=n6_5min_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N7 ctrl Treatment:ctrl RAW_FILE_NAME=n7_ctrl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N7 30 sec KCl Treatment:30 sec KCl RAW_FILE_NAME=n7_30sec kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N7 1min KCl Treatment:1min KCl RAW_FILE_NAME=n7_1min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N7 2min KCl Treatment:2min KCl RAW_FILE_NAME=n7_2min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N7 3min KCl Treatment:3min KCl RAW_FILE_NAME=n7_3min kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig2 N7 5min KCl Treatment:5min KCl RAW_FILE_NAME=n7_5min_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 unlabeled Treatment:unlabeled RAW_FILE_NAME=n1_unlabeled_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 5min 13C Gluc Treatment:5min 13C Gluc RAW_FILE_NAME=n1_5min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 10min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n1_10min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 10min 13C Gluc KCl Treatment:10min 13C Gluc KCl RAW_FILE_NAME=n1_10min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 20min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n1_20min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 35min 13C Gluc Treatment:35min 13C Gluc RAW_FILE_NAME=n1_35min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N1 35min 13C Gluc KCl Treatment:35min 13C Gluc KCl RAW_FILE_NAME=n1_35min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 unlabeled Treatment:unlabeled RAW_FILE_NAME=n2_unlabeled_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 5min 13C Gluc Treatment:5min 13C Gluc RAW_FILE_NAME=n2_5min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 10min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n2_10min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 10min 13C Gluc KCl Treatment:10min 13C Gluc KCl RAW_FILE_NAME=n2_10min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 20min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n2_20min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 35min 13C Gluc Treatment:35min 13C Gluc RAW_FILE_NAME=n2_35min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N2 35min 13C Gluc KCl Treatment:35min 13C Gluc KCl RAW_FILE_NAME=n2_35min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 unlabeled Treatment:unlabeled RAW_FILE_NAME=n3_unlabeled_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 5min 13C Gluc Treatment:5min 13C Gluc RAW_FILE_NAME=n3_5min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 10min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n3_10min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 10min 13C Gluc KCl Treatment:10min 13C Gluc KCl RAW_FILE_NAME=n3_10min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 20min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n3_20min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 35min 13C Gluc Treatment:35min 13C Gluc RAW_FILE_NAME=n3_35min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N3 35min 13C Gluc KCl Treatment:35min 13C Gluc KCl RAW_FILE_NAME=n3_35min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 unlabeled Treatment:unlabeled RAW_FILE_NAME=n4_unlabeled_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 5min 13C Gluc Treatment:5min 13C Gluc RAW_FILE_NAME=n4_5min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 10min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n4_10min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 10min 13C Gluc KCl Treatment:10min 13C Gluc KCl RAW_FILE_NAME=n4_10min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 20min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n4_20min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 35min 13C Gluc Treatment:35min 13C Gluc RAW_FILE_NAME=n4_35min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N4 35min 13C Gluc KCl Treatment:35min 13C Gluc KCl RAW_FILE_NAME=n4_35min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 unlabeled Treatment:unlabeled RAW_FILE_NAME=n5_unlabeled_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 5min 13C Gluc Treatment:5min 13C Gluc RAW_FILE_NAME=n5_5min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 10min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n5_10min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 10min 13C Gluc KCl Treatment:10min 13C Gluc KCl RAW_FILE_NAME=n5_10min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 20min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n5_20min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 35min 13C Gluc Treatment:35min 13C Gluc RAW_FILE_NAME=n5_35min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N5 35min 13C Gluc KCl Treatment:35min 13C Gluc KCl RAW_FILE_NAME=n5_35min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 unlabeled Treatment:unlabeled RAW_FILE_NAME=n6_unlabeled_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 5min 13C Gluc Treatment:5min 13C Gluc RAW_FILE_NAME=n6_5min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 10min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n6_10min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 10min 13C Gluc KCl Treatment:10min 13C Gluc KCl RAW_FILE_NAME=n6_10min 13c gluc kcl_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 20min 13C Gluc Treatment:10min 13C Gluc RAW_FILE_NAME=n6_20min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 35min 13C Gluc Treatment:35min 13C Gluc RAW_FILE_NAME=n6_35min 13c gluc_dg-total ion count SUBJECT_SAMPLE_FACTORS - Fig3 N6 35min 13C Gluc KCl Treatment:35min 13C Gluc KCl RAW_FILE_NAME=n6_35min 13c gluc kcl_dg-total ion count #COLLECTION CO:COLLECTION_SUMMARY Hippocampal slices were stimulated with 50mM potassium chloride over a time CO:COLLECTION_SUMMARY course of 5 minutes. The experiment was terminated with thermal preservation at CO:COLLECTION_SUMMARY the indicated time points. CO:SAMPLE_TYPE Brain #TREATMENT TR:TREATMENT_SUMMARY 50mM potassium chloride #SAMPLEPREP SP:SAMPLEPREP_SUMMARY At the specific time chosen for thermal preservation, the slices were flash SP:SAMPLEPREP_SUMMARY heated to 80°C for 2 seconds and then cooled to 4°C; the slices were then SP:SAMPLEPREP_SUMMARY flash-frozen to -20 °C. The brain slices were stored at -80°C for subsequent SP:SAMPLEPREP_SUMMARY thin sectioning in a cryomicrotome. The thin sections were mounted onto SP:SAMPLEPREP_SUMMARY conductive glass slides (ITO slides) and sprayed with matrix SP:SAMPLEPREP_SUMMARY (1,5-diaminonaphthalene). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME none CH:COLUMN_NAME none CH:SOLVENT_A none CH:SOLVENT_B none CH:FLOW_GRADIENT none CH:FLOW_RATE none CH:COLUMN_TEMPERATURE none #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Bruker timsTOF fleX MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE MALDI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Mass spectra were acquired on a timsTOF fleX in negative ion mode. Hippocampal MS:MS_COMMENTS sections were outlined as individual ROIs with a pixel size of 20 µm. Most MS:MS_COMMENTS metabolites were acquired in a single imaging run (m/z range 50 - 1000). Each MS:MS_COMMENTS MSI pixel consisted of 1000 laser shots. The laser Smartbeam parameter was set MS:MS_COMMENTS to single with a frequency of 10 kHz. MS:MS_RESULTS_FILE ST002699_AN004374_Results.txt UNITS:a.u. Has m/z:Yes Has RT:No RT units:No RT data #END