#METABOLOMICS WORKBENCH hormel101_20170623_152446 DATATRACK_ID:1102 STUDY_ID:ST000653 ANALYSIS_ID:AN000991 PROJECT_ID:PR000460 VERSION 1 CREATED_ON June 26, 2017, 8:42 am #PROJECT PR:PROJECT_TITLE The dystrophic muscle metabolome: effects of exercise and NO donor therapy PR:PROJECT_SUMMARY "In Duchenne and Becker muscular dystrophy (DMD, BMD), loss of the cytoskeletal PR:PROJECT_SUMMARY protein dystrophin weakens the sarcolemma and disrupts cellular signaling, PR:PROJECT_SUMMARY rendering the diseased muscles susceptible to contractioninduced damage. We and PR:PROJECT_SUMMARY others have shown that loss of neuronal nitric oxide synthase (nNOSμ) from the PR:PROJECT_SUMMARY sarcolemma of dystrophin-deficient muscle causes functional muscle ischemia PR:PROJECT_SUMMARY during exercise due to unopposed sympathetic vasoconstriction, thereby PR:PROJECT_SUMMARY exacerbating fatigue and injury of the diseased muscles. Genetic and PR:PROJECT_SUMMARY pharmacologic strategies targeting nNOSμ-NO signaling ameliorate functional PR:PROJECT_SUMMARY muscle ischemia, as well as many other features of the dystrophic phenotype in PR:PROJECT_SUMMARY the mdx mouse model of DMD/BMD. These findings suggest that the therapeutic PR:PROJECT_SUMMARY benefit of NO likely extends beyond its vascular effects. A growing body of PR:PROJECT_SUMMARY evidence indicates that NO directly influences muscle metabolism through effects PR:PROJECT_SUMMARY on glucose transport as well as mitochondrial biogenesis and function. Both PR:PROJECT_SUMMARY nNOS-/- mice and mdx mice exhibit muscle mitochondrial dysfunction, decreased PR:PROJECT_SUMMARY resistance to fatigue, and exercise-induced muscle injury, suggesting a causal PR:PROJECT_SUMMARY role of nNOSμ-NO deficiency. However, the specific metabolic changes resulting PR:PROJECT_SUMMARY from reduced NO signaling that might render dystrophic muscle susceptible to PR:PROJECT_SUMMARY fatigue and use-dependent injury remain poorly defined. Therefore, the goal of PR:PROJECT_SUMMARY this pilot metabolomics study is to identify the unique biochemical profiles of PR:PROJECT_SUMMARY skeletal and cardiac muscles of mdx mice to gain further mechanistic insight PR:PROJECT_SUMMARY into the pathophysiological role of NO deficiency in muscular dystrophy. In Aim PR:PROJECT_SUMMARY 1, we will characterize the skeletal and cardiac muscle metabolomes of mdx and PR:PROJECT_SUMMARY nNOS-/- mice at rest and following a single bout of treadmill exercise with the PR:PROJECT_SUMMARY goal of discovering common metabolic signatures caused by loss of NO signaling. PR:PROJECT_SUMMARY In Aim 2, we will evaluate the potential of a NO donor drug that is under PR:PROJECT_SUMMARY development as a therapeutic for DMD/BMD to improve the skeletal and cardiac PR:PROJECT_SUMMARY muscle metabolomes in mdx mice. As a result of this pilot study, we hope to gain PR:PROJECT_SUMMARY new understanding of the metabolic derangements in dystrophin-deficient muscle, PR:PROJECT_SUMMARY insight into the therapeutic effects of NO replacement, and to identify new PR:PROJECT_SUMMARY pathogenic mechanisms and putative therapeutic targets that will form the basis PR:PROJECT_SUMMARY of future grant applications." PR:INSTITUTE Mayo Clinic PR:LAST_NAME Thomas PR:FIRST_NAME Gail PR:ADDRESS Penn State Hershey Heart and Vascular Institute Penn State College of Medicine PR:ADDRESS 500 University Drive, MC H047 Hershey, PA 17033 PR:EMAIL gthomas4@hmc.psu.edu PR:PHONE 717-531-0003, ext. 287087 #STUDY ST:STUDY_TITLE Effects of Exercise on Dystrophic Mouse Heart Meabolome (part IX) ST:STUDY_SUMMARY We will use male C57BL10, mdx, and nNOS-/- mice (n = 10 each group) to ST:STUDY_SUMMARY characterize the skeletal and cardiac muscle metabolomes. Half of the mice in ST:STUDY_SUMMARY each group will remain sedentary while the other half will be subjected to a ST:STUDY_SUMMARY single bout of treadmill exercise to exhaustion. Mice will be euthanized ST:STUDY_SUMMARY immediately postexercise and blood, hearts, and hindlimb muscles will be ST:STUDY_SUMMARY harvested and frozen as detailed in the General Methods. The heart and ST:STUDY_SUMMARY gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to ST:STUDY_SUMMARY the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will ST:STUDY_SUMMARY be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius ST:STUDY_SUMMARY and soleus muscles will be used for targeted analyses of amino acids plus amino ST:STUDY_SUMMARY metabolites, non-esterified fatty acids, and citric acid cycle intermediates. ST:INSTITUTE Mayo Clinic ST:LAST_NAME Thomas ST:FIRST_NAME Gail ST:ADDRESS Penn State Hershey Heart and Vascular Institute Penn State College of Medicine ST:ADDRESS 500 University Drive, MC H047 Hershey, PA 17033 ST:EMAIL gthomas4@hmc.psu.edu ST:PHONE 717-531-0003, ext. 287087 #SUBJECT SU:SUBJECT_TYPE Mouse SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS ms5940-1 nC18-18may16-001-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-1 nC18-18may16-001-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-2 nC18-18may16-002-r001 Group:SED-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-2 nC18-18may16-002-r002 Group:SED-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-3 nC18-18may16-003-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-3 nC18-18may16-003-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-4 nC18-18may16-004-r001 Group:RUN-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-4 nC18-18may16-004-r002 Group:RUN-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-5 nC18-18may16-005-r001 Group:SED-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-5 nC18-18may16-005-r002 Group:SED-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-6 nC18-18may16-006-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-6 nC18-18may16-006-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-7 nC18-18may16-007-r001 Group:SED-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-7 nC18-18may16-007-r002 Group:SED-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-8 nC18-18may16-008-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-8 nC18-18may16-008-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-9 nC18-18may16-009-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-9 nC18-18may16-009-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-10 nC18-18may16-010-r001 Group:RUN-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-10 nC18-18may16-010-r002 Group:RUN-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-11 nC18-18may16-011-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-11 nC18-18may16-011-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-12 nC18-18may16-012-r001 Group:RUN-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-12 nC18-18may16-012-r002 Group:RUN-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-13 nC18-18may16-013-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-13 nC18-18may16-013-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-14 nC18-18may16-014-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-14 nC18-18may16-014-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-15 nC18-18may16-015-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-15 nC18-18may16-015-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-16 nC18-18may16-016-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-16 nC18-18may16-016-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-17 nC18-18may16-017-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-17 nC18-18may16-017-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-18 nC18-18may16-018-r001 Group:SED-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-18 nC18-18may16-018-r002 Group:SED-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-19 nC18-18may16-019-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-19 nC18-18may16-019-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-20 nC18-18may16-020-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-20 nC18-18may16-020-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-21 nC18-18may16-021-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-21 nC18-18may16-021-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-22 nC18-18may16-022-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-22 nC18-18may16-022-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-23 nC18-18may16-023-r001 Group:RUN-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-23 nC18-18may16-023-r002 Group:RUN-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-24 nC18-18may16-024-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-24 nC18-18may16-024-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-25 nC18-18may16-025-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-25 nC18-18may16-025-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-26 nC18-18may16-026-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-26 nC18-18may16-026-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-27 nC18-18may16-027-r001 Group:RUN-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-27 nC18-18may16-027-r002 Group:RUN-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-28 nC18-18may16-028-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-28 nC18-18may16-028-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-29 nC18-18may16-029-r001 Group:SED-MDX | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-29 nC18-18may16-029-r002 Group:SED-MDX | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-30 nC18-18may16-030-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-30 nC18-18may16-030-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NC18 SUBJECT_SAMPLE_FACTORS ms5940-1 pC18-16may16-001-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-1 pC18-16may16-001-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-2 pC18-16may16-002-r001 Group:SED-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-2 pC18-16may16-002-r002 Group:SED-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-3 pC18-16may16-003-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-3 pC18-16may16-003-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-4 pC18-16may16-004-r001 Group:RUN-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-4 pC18-16may16-004-r002 Group:RUN-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-5 pC18-16may16-005-r001 Group:SED-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-5 pC18-16may16-005-r002 Group:SED-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-6 pC18-16may16-006-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-6 pC18-16may16-006-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-7 pC18-16may16-007-r001 Group:SED-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-7 pC18-16may16-007-r002 Group:SED-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-8 pC18-16may16-008-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-8 pC18-16may16-008-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-9 pC18-16may16-009-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-9 pC18-16may16-009-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-10 pC18-16may16-010-r001 Group:RUN-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-10 pC18-16may16-010-r002 Group:RUN-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-11 pC18-16may16-011-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-11 pC18-16may16-011-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-12 pC18-16may16-012-r001 Group:RUN-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-12 pC18-16may16-012-r002 Group:RUN-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-13 pC18-16may16-013-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-13 pC18-16may16-013-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-14 pC18-16may16-014-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-14 pC18-16may16-014-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-15 pC18-16may16-015-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-15 pC18-16may16-015-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-16 pC18-16may16-016-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-16 pC18-16may16-016-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-17 pC18-16may16-017-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-17 pC18-16may16-017-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-18 pC18-16may16-018-r001 Group:SED-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-18 pC18-16may16-018-r002 Group:SED-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-19 pC18-16may16-019-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-19 pC18-16may16-019-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-20 pC18-16may16-020-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-20 pC18-16may16-020-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-21 pC18-16may16-021-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-21 pC18-16may16-021-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-22 pC18-16may16-022-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-22 pC18-16may16-022-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-23 pC18-16may16-023-r001 Group:RUN-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-23 pC18-16may16-023-r002 Group:RUN-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-24 pC18-16may16-024-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-24 pC18-16may16-024-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-25 pC18-16may16-025-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-25 pC18-16may16-025-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-26 pC18-16may16-026-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-26 pC18-16may16-026-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-27 pC18-16may16-027-r001 Group:RUN-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-27 pC18-16may16-027-r002 Group:RUN-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-28 pC18-16may16-028-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-28 pC18-16may16-028-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-29 pC18-16may16-029-r001 Group:SED-MDX | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-29 pC18-16may16-029-r002 Group:SED-MDX | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-30 pC18-16may16-030-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-30 pC18-16may16-030-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PC18 SUBJECT_SAMPLE_FACTORS ms5940-1 nhilic-18may16-001-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-1 nhilic-18may16-001-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-2 nhilic-18may16-002-r001 Group:SED-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-2 nhilic-18may16-002-r002 Group:SED-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-3 nhilic-18may16-003-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-3 nhilic-18may16-003-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-4 nhilic-18may16-004-r001 Group:RUN-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-4 nhilic-18may16-004-r002 Group:RUN-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-5 nhilic-18may16-005-r001 Group:SED-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-5 nhilic-18may16-005-r002 Group:SED-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-6 nhilic-18may16-006-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-6 nhilic-18may16-006-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-7 nhilic-18may16-007-r001 Group:SED-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-7 nhilic-18may16-007-r002 Group:SED-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-8 nhilic-18may16-008-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-8 nhilic-18may16-008-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-9 nhilic-18may16-009-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-9 nhilic-18may16-009-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-10 nhilic-18may16-010-r001 Group:RUN-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-10 nhilic-18may16-010-r002 Group:RUN-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-11 nhilic-18may16-011-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-11 nhilic-18may16-011-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-12 nhilic-18may16-012-r001 Group:RUN-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-12 nhilic-18may16-012-r002 Group:RUN-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-13 nhilic-18may16-013-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-13 nhilic-18may16-013-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-14 nhilic-18may16-014-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-14 nhilic-18may16-014-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-15 nhilic-18may16-015-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-15 nhilic-18may16-015-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-16 nhilic-18may16-016-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-16 nhilic-18may16-016-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-17 nhilic-18may16-017-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-17 nhilic-18may16-017-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-18 nhilic-18may16-018-r001 Group:SED-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-18 nhilic-18may16-018-r002 Group:SED-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-19 nhilic-18may16-019-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-19 nhilic-18may16-019-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-20 nhilic-18may16-020-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-20 nhilic-18may16-020-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-21 nhilic-18may16-021-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-21 nhilic-18may16-021-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-22 nhilic-18may16-022-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-22 nhilic-18may16-022-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-23 nhilic-18may16-023-r001 Group:RUN-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-23 nhilic-18may16-023-r002 Group:RUN-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-24 nhilic-18may16-024-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-24 nhilic-18may16-024-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-25 nhilic-18may16-025-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-25 nhilic-18may16-025-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-26 nhilic-18may16-026-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-26 nhilic-18may16-026-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-27 nhilic-18may16-027-r001 Group:RUN-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-27 nhilic-18may16-027-r002 Group:RUN-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-28 nhilic-18may16-028-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-28 nhilic-18may16-028-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-29 nhilic-18may16-029-r001 Group:SED-MDX | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-29 nhilic-18may16-029-r002 Group:SED-MDX | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-30 nhilic-18may16-030-r001 Group:SED-BL10 Control | rep:1 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-30 nhilic-18may16-030-r002 Group:SED-BL10 Control | rep:2 chromatograhy=NHILIC SUBJECT_SAMPLE_FACTORS ms5940-1 philic-16may16-001-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-1 philic-16may16-001-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-2 philic-16may16-002-r001 Group:SED-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-2 philic-16may16-002-r002 Group:SED-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-3 philic-16may16-003-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-3 philic-16may16-003-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-4 philic-16may16-004-r001 Group:RUN-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-4 philic-16may16-004-r002 Group:RUN-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-5 philic-16may16-005-r001 Group:SED-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-5 philic-16may16-005-r002 Group:SED-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-6 philic-16may16-006-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-6 philic-16may16-006-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-7 philic-16may16-007-r001 Group:SED-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-7 philic-16may16-007-r002 Group:SED-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-8 philic-16may16-008-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-8 philic-16may16-008-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-9 philic-16may16-009-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-9 philic-16may16-009-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-10 philic-16may16-010-r001 Group:RUN-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-10 philic-16may16-010-r002 Group:RUN-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-11 philic-16may16-011-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-11 philic-16may16-011-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-12 philic-16may16-012-r001 Group:RUN-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-12 philic-16may16-012-r002 Group:RUN-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-13 philic-16may16-013-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-13 philic-16may16-013-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-14 philic-16may16-014-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-14 philic-16may16-014-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-15 philic-16may16-015-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-15 philic-16may16-015-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-16 philic-16may16-016-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-16 philic-16may16-016-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-17 philic-16may16-017-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-17 philic-16may16-017-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-18 philic-16may16-018-r001 Group:SED-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-18 philic-16may16-018-r002 Group:SED-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-19 philic-16may16-019-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-19 philic-16may16-019-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-20 philic-16may16-020-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-20 philic-16may16-020-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-21 philic-16may16-021-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-21 philic-16may16-021-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-22 philic-16may16-022-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-22 philic-16may16-022-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-23 philic-16may16-023-r001 Group:RUN-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-23 philic-16may16-023-r002 Group:RUN-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-24 philic-16may16-024-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-24 philic-16may16-024-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-25 philic-16may16-025-r001 Group:RUN-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-25 philic-16may16-025-r002 Group:RUN-BL10 Control | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-26 philic-16may16-026-r001 Group:RUN-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-26 philic-16may16-026-r002 Group:RUN-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-27 philic-16may16-027-r001 Group:RUN-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-27 philic-16may16-027-r002 Group:RUN-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-28 philic-16may16-028-r001 Group:SED-nNOS-/- | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-28 philic-16may16-028-r002 Group:SED-nNOS-/- | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-29 philic-16may16-029-r001 Group:SED-MDX | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-29 philic-16may16-029-r002 Group:SED-MDX | rep:2 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-30 philic-16may16-030-r001 Group:SED-BL10 Control | rep:1 chromatograhy=PHILIC SUBJECT_SAMPLE_FACTORS ms5940-30 philic-16may16-030-r002 Group:SED-BL10 Control | rep:2 chromatograhy=PHILIC #COLLECTION CO:COLLECTION_SUMMARY Blood and Tissue Harvesting: Mice will be euthanized and blood will be taken by CO:COLLECTION_SUMMARY cardiac puncture, centrifuged, and the plasma will be stored at -80ºC. The CO:COLLECTION_SUMMARY heart, diaphragm, and muscles of both hindlimbs (gastrocnemius, soleus, CO:COLLECTION_SUMMARY quadriceps) will be dissected and snap frozen in liquid nitrogen. Tibialis CO:COLLECTION_SUMMARY anterior and plantaris muscles from both hindlimbs also will be dissected and CO:COLLECTION_SUMMARY one set will be snap frozen in liquid nitrogen while the other set will be CO:COLLECTION_SUMMARY mounted in OCT and frozen in isopentane cooled by liquid nitrogen. Cryosections CO:COLLECTION_SUMMARY will be used to evaluate disease activity and exercise-induced muscle injury by CO:COLLECTION_SUMMARY staining with: (a) hematoxylin and eosin to assess gross morphology, cellular CO:COLLECTION_SUMMARY infiltration, and necrosis, (b) anti-F4/80 to label macrophages, and (c) CO:COLLECTION_SUMMARY anti-IgG or IgM to label damaged muscle fibers. #TREATMENT TR:TREATMENT_SUMMARY "Studies will be performed in 12-16 week old male mdx mice, C57BL10 control TR:TREATMENT_SUMMARY mice, and nNOS-/- mice obtained from Jackson Laboratory. All protocols will be TR:TREATMENT_SUMMARY approved by the Penn State College of Medicine Institutional Animal Care and Use TR:TREATMENT_SUMMARY Committee. Treadmill Exercise: Mice will run on a horizontal treadmill (Columbus TR:TREATMENT_SUMMARY Instruments) to assess fatigue by a single bout of exercise beginning at 5 m/min TR:TREATMENT_SUMMARY for 5 min followed by 1 m/min increases every minute until exhaustion. Electric TR:TREATMENT_SUMMARY shocks will not be used to stimulate running due to adverse effects in mdx TR:TREATMENT_SUMMARY mice." #SAMPLEPREP SP:SAMPLEPREP_SUMMARY mouse heart untarged metabolomics profiling #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Waters Acquity BEH Amide (150 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6550 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST000653_AN000991_Results.txt UNITS:intensity #END