#METABOLOMICS WORKBENCH amitch_20151112_9216941_mwtab.txt DATATRACK_ID:411 efahy_20151226_121228 STUDY_ID:ST000287 ANALYSIS_ID:AN000456 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 Thermo Hypercarb (50 x 2.1mm, 3um) CH:METHODS_FILENAME SB_NAD-ATP_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 NAD/ATP 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 NMN 2.57 3.26 3.32 3.57 2.49 3.30 5.35 3.63 3.16 3.40 5.17 2.50 2.91 2.67 2.27 2.97 NAD 1208.52 1601.82 1484.41 1837.65 1892.69 1772.16 1732.87 1157.78 1392.64 1710.87 1209.64 852.58 1478.12 1416.72 924.83 2118.29 2313.90 2318.14 2186.24 2341.62 NADP 37.08 42.88 34.52 AMP 0.61 0.34 0.62 0.35 0.47 0.51 0.34 0.36 0.45 0.27 0.33 1.18 0.22 0.23 0.34 0.19 0.32 ADP 4.16 3.50 4.70 3.57 4.01 4.36 4.07 3.08 4.16 3.66 3.33 4.20 3.02 3.13 2.66 2.72 3.30 3.11 3.23 3.64 ATP 17.74 25.06 22.81 24.31 20.41 24.48 26.02 14.27 18.93 23.10 16.90 10.52 18.07 16.88 10.84 23.07 26.83 24.82 27.29 26.14 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Index Moverz Quant PubChem CID NMN 1.20 335.2, 123.1 14180 NAD 1.80 664.2, 136.1 5893 NADP 1.70 744.2, 136.1 5886 AMP 1.50 348.1, 136.1 6083 ADP 1.50 428.0, 136.1 6022 ATP 1.50 508.1, 136.1 5957 METABOLITES_END #END