#METABOLOMICS WORKBENCH amitch_20151112_9216941_mwtab.txt DATATRACK_ID:411 efahy_20151226_121228 STUDY_ID:ST000287 ANALYSIS_ID:AN000457 PROJECT_ID:PR000229 VERSION 1 CREATED_ON December 26, 2015, 12:12 pm #PROJECT PR:PROJECT_TITLE Energetics in dystrophic muscle PR:PROJECT_TYPE Cross-sectional PR:PROJECT_SUMMARY This project utilizes mouse models to establish whether lack of PR:PROJECT_SUMMARY sarcolemma-localized nNOS impairs muscle metabolism and perfusion. PR:INSTITUTE University of Florida PR:DEPARTMENT Physical Therapy PR:LABORATORY Muscle Physiology Laboratory PR:LAST_NAME Forbes PR:FIRST_NAME Sean PR:ADDRESS University of Florida, College of Public Health & Health Professions, Department PR:ADDRESS of Physical Therapy, Box 100154, UFHSC PR:EMAIL scforbes@ufl.edu PR:PHONE 352-273-6111 PR:FUNDING_SOURCE Start-up funds #STUDY ST:STUDY_TITLE Energetics in dystrophic muscle ST:STUDY_TYPE Cross-sectional ST:STUDY_SUMMARY This project utilizes mouse models to establish whether lack of ST:STUDY_SUMMARY sarcolemma-localized nNOS impairs muscle metabolism and perfusion. ST:INSTITUTE University of Florida ST:DEPARTMENT SECIM ST:LABORATORY Muscle Physiology Laboratory ST:LAST_NAME Forbes ST:FIRST_NAME Sean ST:ADDRESS University of Florida, College of Public Health & Health Professions, Department ST:ADDRESS of Physical Therapy, Box 100154, UFHSC ST:EMAIL scforbes@ufl.edu ST:PHONE 352-273-6111 ST:NUM_GROUPS 4 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:GENDER Male SU:ANIMAL_ANIMAL_SUPPLIER Jackson Lab SU:ANIMAL_HOUSING McKnight Brain Institute, UF SU:ANIMAL_LIGHT_CYCLE 12 hr light/12 hr dark SU:ANIMAL_FEED ad libtum SU:ANIMAL_WATER ad libtum SU:ANIMAL_INCLUSION_CRITERIA males only #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - scf_01-M1 Strain:nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_01-M2 Strain:nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_01-M3 Strain:nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_01-M4 Strain:nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_01-M5 Strain:nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_02-M1 Strain:mdx-nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_02-M2 Strain:mdx-nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_02-M3 Strain:mdx-nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_02-M4 Strain:mdx-nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_02-M5 Strain:mdx-nNOS-/- SUBJECT_SAMPLE_FACTORS - scf_03-M1 Strain:mdx SUBJECT_SAMPLE_FACTORS - scf_03-M2 Strain:mdx SUBJECT_SAMPLE_FACTORS - scf_03-M3 Strain:mdx SUBJECT_SAMPLE_FACTORS - scf_03-M4 Strain:mdx SUBJECT_SAMPLE_FACTORS - scf_03-M5 Strain:mdx SUBJECT_SAMPLE_FACTORS - scf_04-M1 Strain:C57Bl/10 SUBJECT_SAMPLE_FACTORS - scf_04-M2 Strain:C57Bl/10 SUBJECT_SAMPLE_FACTORS - scf_04-M3 Strain:C57Bl/10 SUBJECT_SAMPLE_FACTORS - scf_04-M4 Strain:C57Bl/10 SUBJECT_SAMPLE_FACTORS - scf_04-M5 Strain:C57Bl/10 #COLLECTION CO:COLLECTION_SUMMARY Muscles extracted under isoflurane, snap frozen with liquid nitrogen, stored at CO:COLLECTION_SUMMARY -80C CO:COLLECTION_PROTOCOL_COMMENTS L3-190 CO:COLLECTION_METHOD muscle extraction, frozen CO:COLLECTION_LOCATION UF, musle physiology laboratory CO:COLLECTION_FREQUENCY once/mouse CO:COLLECTION_DURATION Total ~5 min CO:COLLECTION_TIME no dose CO:VOLUMEORAMOUNT_COLLECTED entire muscles extracted CO:STORAGE_CONDITIONS -80C freezer CO:COLLECTION_VIALS 2 ml vials CO:STORAGE_VIALS 2 ml vials CO:COLLECTION_TUBE_TEMP -80C CO:ADDITIVES none CO:BLOOD_SERUM_OR_PLASMA none CO:TISSUE_CELL_IDENTIFICATION none CO:TISSUE_CELL_QUANTITY_TAKEN none #TREATMENT TR:TREATMENT_SUMMARY No treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY NAD/ATP or NADH SP:SAMPLEPREP_PROTOCOL_FILENAME SB_NAD-ATP_Assay.pdf, SB_NADH_Assay_Agilent.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY NAD-ATP method CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Waters Acquity HSS T3 (100 x 2.1mm, 1.8um) CH:METHODS_FILENAME SB_NADH_Assay_Agilent.pdf #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS NADH MS:INSTRUMENT_NAME Agilent 6490 QQQ MS:INSTRUMENT_NAME Agilent 6490 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmol/mg dry powder MS_METABOLITE_DATA_START Samples scf_01-M1 scf_01-M2 scf_01-M3 scf_01-M4 scf_01-M5 scf_02-M1 scf_02-M2 scf_02-M3 scf_02-M4 scf_02-M5 scf_03-M1 scf_03-M2 scf_03-M3 scf_03-M4 scf_03-M5 scf_04-M1 scf_04-M2 scf_04-M3 scf_04-M4 scf_04-M5 Factors Strain:nNOS-/- Strain:nNOS-/- Strain:nNOS-/- Strain:nNOS-/- Strain:nNOS-/- Strain:mdx-nNOS-/- Strain:mdx-nNOS-/- Strain:mdx-nNOS-/- Strain:mdx-nNOS-/- Strain:mdx-nNOS-/- Strain:mdx Strain:mdx Strain:mdx Strain:mdx Strain:mdx Strain:C57Bl/10 Strain:C57Bl/10 Strain:C57Bl/10 Strain:C57Bl/10 Strain:C57Bl/10 NADH 90.08 128.62 94.34 65.15 155.71 99.68 125.30 34.37 55.37 76.05 56.18 64.50 115.93 17.44 184.07 189.04 198.01 139.52 150.27 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Index Moverz Quant PubChem CID NADH 1.80 666.2, 649.2 439153 METABOLITES_END #END