#METABOLOMICS WORKBENCH araskind_20170802_132535 DATATRACK_ID:1181 STUDY_ID:ST000832 ANALYSIS_ID:AN001839 PROJECT_ID:PR000593 VERSION 1 CREATED_ON January 16, 2019, 8:47 am #PROJECT PR:PROJECT_TITLE Regulatory roles of glycolysis and its enzymes in photoreceptor survival PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY The goals specific to this proposal are to understand how the metabolic flux PR:PROJECT_SUMMARY through glycolysis and its associated pathways, such as oxidative PR:PROJECT_SUMMARY phosphorylation and pentose phosphate pathway, is regulated during periods of PR:PROJECT_SUMMARY photoreceptor stress. PR:INSTITUTE University of Michigan PR:DEPARTMENT Ophthamology and Visual Sciences PR:LABORATORY Besirli Lab PR:LAST_NAME Besirli PR:FIRST_NAME Cagri PR:ADDRESS Ann Arbor, MI PR:EMAIL cbesirli@umich.edu PR:PHONE 734-615-6099 #STUDY ST:STUDY_TITLE Rat Retinal Detachment Metabolomics Timecourse ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY Detachments were created in adult male Brown-Norway rats (300?400 g; Charles ST:STUDY_SUMMARY River Laboratories, Wilmington, MA). Briefly, rodents were anesthetized with a ST:STUDY_SUMMARY 50:50 mix of ketamine/xylazine, and pupils were dilated with topical ST:STUDY_SUMMARY phenylephrine (2.5%) and tropicamide (1%). A 20-gauge microvitreoretinal blade ST:STUDY_SUMMARY was used to create a sclerotomy 2 mm posterior to the limbus, carefully avoiding ST:STUDY_SUMMARY lens damage. A subretinal injector (Glaser, 32-gauge tip; BD Ophthalmic Systems, ST:STUDY_SUMMARY Franklin Lakes, NJ) was introduced through the sclerotomy into the vitreous ST:STUDY_SUMMARY cavity and then through a peripheral retinotomy into the subretinal space. ST:STUDY_SUMMARY Sodium hyaluronate (10 mg/mL, Healon O.D Abbott Medical Optics, Uppsala, ST:STUDY_SUMMARY Sweden) was slowly injected to detach the neurosensory retina from the ST:STUDY_SUMMARY underlying RPE. In all experiments, approximately one third to one half of the ST:STUDY_SUMMARY neurosensory retina was detached. Detachments were created in the left eye. The ST:STUDY_SUMMARY right eye served as the control, with all the steps of the procedure performed ST:STUDY_SUMMARY except for introduction of the subretinal injector and injection of the sodium ST:STUDY_SUMMARY hyaluronate. At varying intervals after creation of the detachment, the animals ST:STUDY_SUMMARY were euthanatized, and the eyes were enucleated.The retina was then dissected ST:STUDY_SUMMARY away from the retinal pigment epithelium taking just the detached portion in ST:STUDY_SUMMARY those eyes experimentally detached. All experiments were performed in accordance ST:STUDY_SUMMARY with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research ST:STUDY_SUMMARY and the guidelines established by the University Committee on Use and Care of ST:STUDY_SUMMARY Animals of the University of Michigan. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@med.umich.edu ST:PHONE (734) 232-8175 ST:NUM_GROUPS 30 ST:TOTAL_SUBJECTS 21 #SUBJECT SU:SUBJECT_TYPE RAT SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10114 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0014158 S00021535 Attachment:Detached | Time:5 min Rat #=10.0 SUBJECT_SAMPLE_FACTORS SU0014158 S00021536 Attachment:Attached | Time:5 min Rat #=10.0 SUBJECT_SAMPLE_FACTORS SU0014159 S00021537 Attachment:Detached | Time:5 min Rat #=11.0 SUBJECT_SAMPLE_FACTORS SU0014159 S00021538 Attachment:Attached | Time:5 min Rat #=11.0 SUBJECT_SAMPLE_FACTORS SU0014160 S00021539 Attachment:Detached | Time:5 min Rat #=12.0 SUBJECT_SAMPLE_FACTORS SU0014160 S00021540 Attachment:Attached | Time:5 min Rat #=12.0 SUBJECT_SAMPLE_FACTORS SU0014161 S00021541 Attachment:Detached | Time:15 min Rat #=13.0 SUBJECT_SAMPLE_FACTORS SU0014161 S00021542 Attachment:Attached | Time:15 min Rat #=13.0 SUBJECT_SAMPLE_FACTORS SU0014162 S00021543 Attachment:Detached | Time:15 min Rat #=14.0 SUBJECT_SAMPLE_FACTORS SU0014162 S00021544 Attachment:Attached | Time:15 min Rat #=14.0 SUBJECT_SAMPLE_FACTORS SU0014163 S00021545 Attachment:Detached | Time:15 min Rat #=15.0 SUBJECT_SAMPLE_FACTORS SU0014163 S00021546 Attachment:Attached | Time:15 min Rat #=15.0 SUBJECT_SAMPLE_FACTORS SU0014164 S00021547 Attachment:Detached | Time:30 min Rat #=16.0 SUBJECT_SAMPLE_FACTORS SU0014164 S00021548 Attachment:Attached | Time:30 min Rat #=16.0 SUBJECT_SAMPLE_FACTORS SU0014165 S00021549 Attachment:Detached | Time:30 min Rat #=17.0 SUBJECT_SAMPLE_FACTORS SU0014165 S00021550 Attachment:Attached | Time:30 min Rat #=17.0 SUBJECT_SAMPLE_FACTORS SU0014166 S00021551 Attachment:Detached | Time:30 min Rat #=18.0 SUBJECT_SAMPLE_FACTORS SU0014166 S00021552 Attachment:Attached | Time:30 min Rat #=18.0 SUBJECT_SAMPLE_FACTORS SU0014167 S00021553 Attachment:Detached | Time:45 min Rat #=22.0 SUBJECT_SAMPLE_FACTORS SU0014167 S00021554 Attachment:Attached | Time:45 min Rat #=22.0 SUBJECT_SAMPLE_FACTORS SU0014168 S00021555 Attachment:Detached | Time:45 min Rat #=23.0 SUBJECT_SAMPLE_FACTORS SU0014168 S00021556 Attachment:Attached | Time:45 min Rat #=23.0 SUBJECT_SAMPLE_FACTORS SU0014169 S00021557 Attachment:Detached | Time:45 min Rat #=24.0 SUBJECT_SAMPLE_FACTORS SU0014169 S00021558 Attachment:Attached | Time:45 min Rat #=24.0 SUBJECT_SAMPLE_FACTORS SU0014170 S00021559 Attachment:Detached | Time:90 min Rat #=19.0 SUBJECT_SAMPLE_FACTORS SU0014170 S00021560 Attachment:Attached | Time:90 min Rat #=19.0 SUBJECT_SAMPLE_FACTORS SU0014171 S00021561 Attachment:Detached | Time:90 min Rat #=20.0 SUBJECT_SAMPLE_FACTORS SU0014171 S00021562 Attachment:Attached | Time:90 min Rat #=20.0 SUBJECT_SAMPLE_FACTORS SU0014172 S00021563 Attachment:Detached | Time:90 min Rat #=21.0 SUBJECT_SAMPLE_FACTORS SU0014172 S00021564 Attachment:Attached | Time:90 min Rat #=21.0 SUBJECT_SAMPLE_FACTORS SU0014173 S00021565 Attachment:Detached | Time:3 hr Rat #=7.0 SUBJECT_SAMPLE_FACTORS SU0014173 S00021566 Attachment:Attached | Time:3 hr Rat #=7.0 SUBJECT_SAMPLE_FACTORS SU0014174 S00021567 Attachment:Detached | Time:3 hr Rat #=8.0 SUBJECT_SAMPLE_FACTORS SU0014174 S00021568 Attachment:Attached | Time:3 hr Rat #=8.0 SUBJECT_SAMPLE_FACTORS SU0014175 S00021569 Attachment:Detached | Time:3 hr Rat #=9.0 SUBJECT_SAMPLE_FACTORS SU0014175 S00021570 Attachment:Attached | Time:3 hr Rat #=9.0 SUBJECT_SAMPLE_FACTORS SU0014176 S00021571 Attachment:Detached | Time:5 hr Rat #=1.0 SUBJECT_SAMPLE_FACTORS SU0014176 S00021572 Attachment:Attached | Time:5 hr Rat #=1.0 SUBJECT_SAMPLE_FACTORS SU0014177 S00021573 Attachment:Detached | Time:5 hr Rat #=2.0 SUBJECT_SAMPLE_FACTORS SU0014177 S00021574 Attachment:Attached | Time:5 hr Rat #=2.0 SUBJECT_SAMPLE_FACTORS SU0014178 S00021575 Attachment:Detached | Time:5 hr Rat #=3.0 SUBJECT_SAMPLE_FACTORS SU0014178 S00021576 Attachment:Attached | Time:5 hr Rat #=3.0 #COLLECTION CO:COLLECTION_SUMMARY - CO:SAMPLE_TYPE Retina #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME A011_Glycolysis-TCA-nucleotides.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent GC_7890N CH:COLUMN_NAME Agilent HP5-MS (15m × 0.25mm, 0.25 um) #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE A011 - Glycolysis-TCA-nucleotides.pdf AN:ACQUISITION_PARAMETERS_FILE HEIDIFAMES.M #MS MS:INSTRUMENT_NAME Agilent MS_5975 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS none #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol/ug protein MS_METABOLITE_DATA_START Samples S00021535 S00021536 S00021537 S00021538 S00021539 S00021540 S00021541 S00021542 S00021543 S00021544 S00021545 S00021546 S00021547 S00021548 S00021549 S00021550 S00021551 S00021552 S00021553 S00021554 S00021555 S00021556 S00021557 S00021558 S00021559 S00021560 S00021561 S00021562 S00021563 S00021564 S00021565 S00021566 S00021567 S00021568 S00021569 S00021570 S00021571 S00021572 S00021573 S00021574 S00021575 S00021576 Factors Attachment:Detached | Time:5 min Attachment:Attached | Time:5 min Attachment:Detached | Time:5 min Attachment:Attached | Time:5 min Attachment:Detached | Time:5 min Attachment:Attached | Time:5 min Attachment:Detached | Time:15 min Attachment:Attached | Time:15 min Attachment:Detached | Time:15 min Attachment:Attached | Time:15 min Attachment:Detached | Time:15 min Attachment:Attached | Time:15 min Attachment:Detached | Time:30 min Attachment:Attached | Time:30 min Attachment:Detached | Time:30 min Attachment:Attached | Time:30 min Attachment:Detached | Time:30 min Attachment:Attached | Time:30 min Attachment:Detached | Time:45 min Attachment:Attached | Time:45 min Attachment:Detached | Time:45 min Attachment:Attached | Time:45 min Attachment:Detached | Time:45 min Attachment:Attached | Time:45 min Attachment:Detached | Time:90 min Attachment:Attached | Time:90 min Attachment:Detached | Time:90 min Attachment:Attached | Time:90 min Attachment:Detached | Time:90 min Attachment:Attached | Time:90 min Attachment:Detached | Time:3 hr Attachment:Attached | Time:3 hr Attachment:Detached | Time:3 hr Attachment:Attached | Time:3 hr Attachment:Detached | Time:3 hr Attachment:Attached | Time:3 hr Attachment:Detached | Time:5 hr Attachment:Attached | Time:5 hr Attachment:Detached | Time:5 hr Attachment:Attached | Time:5 hr Attachment:Detached | Time:5 hr Attachment:Attached | Time:5 hr PYRUVIC ACID 1.914960012 5.749664404 6.018999909 3.712134628 3.233896277 7.994760809 2.45814733 3.388307139 2.346863833 3.707007745 3.767880528 5.722722103 3.900903322 6.716524579 1.753887465 11.46572208 3.281163996 3.602056143 2.609178356 6.278992075 2.028007717 11.13737539 1.764516118 5.133485154 1.530608248 5.565228238 1.948324704 4.51216678 2.897409035 1.423542278 3.056264257 6.554697624 8.25841931 9.84186998 0 12.87259958 2.733773994 4.312095619 13.63295968 4.826686773 2.974103287 7.316334626 FUMARIC ACID 2.928037589 4.934183615 5.671400625 10.32169115 6.276423579 6.463732986 3.755339869 13.55063177 4.458397438 7.198796349 4.561678219 8.082123864 6.307513961 8.433229832 4.473076795 8.333050833 9.784582803 5.805891342 4.758073761 8.676303698 3.275210622 8.32445507 2.365422918 7.933511507 8.118226181 15.29953984 7.072342509 6.036885696 3.281864631 5.099946472 5.195030763 6.687546465 6.795161934 9.33728031 3.225273937 7.851276921 4.021701301 8.996187512 9.53725604 10.51370536 3.914503005 10.05754303 LACTIC ACID 1181.189054 6018.953802 3154.777017 8053.081863 1145.998186 7697.42363 2006.209877 6870.435211 2035.402853 3369.220301 3638.203301 2093.122305 2850.16654 7924.013453 2448.227514 8770.468299 3276.357115 2552.816541 2832.550429 5613.947156 2005.549158 6845.513606 1342.177941 5083.680984 5029.723722 10795.14526 3553.082525 5041.68004 2299.525461 2033.591173 2626.362269 6069.345314 4494.384846 8025.490843 2610.490392 6779.067024 2238.486372 5015.601143 7935.837166 6177.271033 2253.274375 6180.379815 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type PYRUVIC ACID 1060 FUMARIC ACID 444972 LACTIC ACID 612 METABOLITES_END #END