#METABOLOMICS WORKBENCH jkrycer_20250216_220502 DATATRACK_ID:5631 STUDY_ID:ST003784 ANALYSIS_ID:AN006218 PROJECT_ID:PR002361
VERSION 1
CREATED_ON March 10, 2025, 9:47 pm
#PROJECT
PR:PROJECT_TITLE Muscle-specific Ryanodine Receptor 1 Properties Underlie Limb-Girdle Muscular
PR:PROJECT_TITLE Dystrophy 2B/R2 Progression
PR:PROJECT_SUMMARY Ryanodine receptor 1 Ca2+ leak is a signal in skeletal muscle, but chronic leak
PR:PROJECT_SUMMARY can underlie pathology. Here we show that in healthy male mouse, limb-girdle
PR:PROJECT_SUMMARY muscle presents higher sympathetic input, elevated ryanodine receptor 1 basal
PR:PROJECT_SUMMARY phosphorylation, Ca2+ leak and mitochondrial Ca2+ content compared to distal leg
PR:PROJECT_SUMMARY muscles. These regional differences are consistent with heat generation in
PR:PROJECT_SUMMARY resting muscle to maintain core temperature. The dysferlin-null mouse develops
PR:PROJECT_SUMMARY severe pathology in the limb-girdle but not leg muscles. Absence of dysferlin
PR:PROJECT_SUMMARY disrupts dihydropyridine receptors’ inhibitory control over ryanodine receptor
PR:PROJECT_SUMMARY 1 leak, synergistically increasing leak through the already phosphorylated
PR:PROJECT_SUMMARY receptor of limb-girdle muscle, altering Ca2+ handling and distribution leading
PR:PROJECT_SUMMARY to reactive oxygen species production prior to disease onset. With age,
PR:PROJECT_SUMMARY oxidation of Ca2+ -handling proteins in dysferlin-null limb-girdle muscle
PR:PROJECT_SUMMARY exacerbated basal Ca2+ movements. Our results show that muscle-specific
PR:PROJECT_SUMMARY pathology in dysferlin-null mice is triggered by increased ryanodine receptor 1
PR:PROJECT_SUMMARY Ca2+ leak.
PR:INSTITUTE University of Queensland
PR:DEPARTMENT School of Biomedical Sciences
PR:LAST_NAME Launikonis
PR:FIRST_NAME Bradley
PR:ADDRESS School of Biomedical Sciences, The University of Queensland, Brisbane, QLD,
PR:ADDRESS 4072, Australia
PR:EMAIL b.launikonis@uq.edu.au
PR:PHONE 61 7 3365 4301
#STUDY
ST:STUDY_TITLE Muscle-specific Ryanodine Receptor 1 Properties Underlie Limb-Girdle Muscular
ST:STUDY_TITLE Dystrophy 2B/R2 Progression
ST:STUDY_SUMMARY Ryanodine receptor 1 Ca2+ leak is a signal in skeletal muscle, but chronic leak
ST:STUDY_SUMMARY can underlie pathology. Here we show that in healthy male mouse, limb-girdle
ST:STUDY_SUMMARY muscle presents higher sympathetic input, elevated ryanodine receptor 1 basal
ST:STUDY_SUMMARY phosphorylation, Ca2+ leak and mitochondrial Ca2+ content compared to distal leg
ST:STUDY_SUMMARY muscles. These regional differences are consistent with heat generation in
ST:STUDY_SUMMARY resting muscle to maintain core temperature. The dysferlin-null mouse develops
ST:STUDY_SUMMARY severe pathology in the limb-girdle but not leg muscles. Absence of dysferlin
ST:STUDY_SUMMARY disrupts dihydropyridine receptors’ inhibitory control over ryanodine receptor
ST:STUDY_SUMMARY 1 leak, synergistically increasing leak through the already phosphorylated
ST:STUDY_SUMMARY receptor of limb-girdle muscle, altering Ca2+ handling and distribution leading
ST:STUDY_SUMMARY to reactive oxygen species production prior to disease onset. With age,
ST:STUDY_SUMMARY oxidation of Ca2+ -handling proteins in dysferlin-null limb-girdle muscle
ST:STUDY_SUMMARY exacerbated basal Ca2+ movements. Our results show that muscle-specific
ST:STUDY_SUMMARY pathology in dysferlin-null mice is triggered by increased ryanodine receptor 1
ST:STUDY_SUMMARY Ca2+ leak.
ST:INSTITUTE University of Queensland
ST:DEPARTMENT School of Biomedical Sciences
ST:LAST_NAME Launikonis
ST:FIRST_NAME Bradley
ST:ADDRESS School of Biomedical Sciences, The University of Queensland, Brisbane, QLD,
ST:ADDRESS 4072, Australia
ST:EMAIL b.launikonis@uq.edu.au
ST:PHONE 61 7 3365 4301
#SUBJECT
SU:SUBJECT_TYPE Mammal
SU:SUBJECT_SPECIES Mus musculus
SU:TAXONOMY_ID 10090
SU:GENOTYPE_STRAIN Wildtype
#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 1.1 RunA_01 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=1; Sample weight (mg)=92.1; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_01.d
SUBJECT_SAMPLE_FACTORS 2.1 RunA_02 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=2; Sample weight (mg)=78.8; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_02.d
SUBJECT_SAMPLE_FACTORS 3.1 RunA_03 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=3; Sample weight (mg)=45.5; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_03.d
SUBJECT_SAMPLE_FACTORS 4.1 RunA_04 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=4; Sample weight (mg)=20.3; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_04.d
SUBJECT_SAMPLE_FACTORS 5.1 RunA_05 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=5; Sample weight (mg)=90.6; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_05.d
SUBJECT_SAMPLE_FACTORS 1.1 RunA_11 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=11; Sample weight (mg)=56.9; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_11.d
SUBJECT_SAMPLE_FACTORS 2.1 RunA_12 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=12; Sample weight (mg)=31.3; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_12.d
SUBJECT_SAMPLE_FACTORS 3.1 RunA_13 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=13; Sample weight (mg)=55.6; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_13.d
SUBJECT_SAMPLE_FACTORS 4.1 RunA_14 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=14; Sample weight (mg)=52.6; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_14.d
SUBJECT_SAMPLE_FACTORS 5.1 RunA_15 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=15; Sample weight (mg)=49.5; Dilution before injection=Neat; RAW_FILE_NAME(Raw file name)=RunA_15.d
SUBJECT_SAMPLE_FACTORS - RunA_21 Sample source:Naïve lysis buffer | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=21; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_21.d
SUBJECT_SAMPLE_FACTORS - RunA_22 Sample source:Naïve lysis buffer | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=22; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_22.d
SUBJECT_SAMPLE_FACTORS - RunA_23 Sample source:STD-1 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=23; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_23.d
SUBJECT_SAMPLE_FACTORS - RunA_23rep Sample source:STD-1 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=23; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_23rep.d
SUBJECT_SAMPLE_FACTORS - RunA_24 Sample source:STD-2 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=24; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_24.d
SUBJECT_SAMPLE_FACTORS - RunA_24rep Sample source:STD-2 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=24; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_24rep.d
SUBJECT_SAMPLE_FACTORS - RunA_25 Sample source:STD-3 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=25; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_25.d
SUBJECT_SAMPLE_FACTORS - RunA_25rep Sample source:STD-3 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=25; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_25rep.d
SUBJECT_SAMPLE_FACTORS - RunA_26 Sample source:STD-4 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=26; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_26.d
SUBJECT_SAMPLE_FACTORS - RunA_26rep Sample source:STD-4 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=26; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_26rep.d
SUBJECT_SAMPLE_FACTORS - RunA_27 Sample source:STD-5 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=27; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_27.d
SUBJECT_SAMPLE_FACTORS - RunA_27rep Sample source:STD-5 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=27; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_27rep.d
SUBJECT_SAMPLE_FACTORS - RunA_28 Sample source:STD-6 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=28; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_28.d
SUBJECT_SAMPLE_FACTORS - RunA_28rep Sample source:STD-6 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=28; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_28rep.d
SUBJECT_SAMPLE_FACTORS - RunA_29 Sample source:STD-7 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=29; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_29.d
SUBJECT_SAMPLE_FACTORS - RunA_29rep Sample source:STD-7 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=29; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_29rep.d
SUBJECT_SAMPLE_FACTORS - RunA_30 Sample source:STD-8 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=30; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_30.d
SUBJECT_SAMPLE_FACTORS - RunA_30rep Sample source:STD-8 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=30; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_30rep.d
SUBJECT_SAMPLE_FACTORS - RunA_31 Sample source:STD-9 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=31; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_31.d
SUBJECT_SAMPLE_FACTORS - RunA_31rep Sample source:STD-9 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=31; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_31rep.d
SUBJECT_SAMPLE_FACTORS - RunA_32 Sample source:STD-10 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=32; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_32.d
SUBJECT_SAMPLE_FACTORS - RunA_32rep Sample source:STD-10 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=32; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunA_32rep.d
SUBJECT_SAMPLE_FACTORS 1.1 RunA_dil01 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=1; Sample weight (mg)=92.1; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil01.d
SUBJECT_SAMPLE_FACTORS 2.1 RunA_dil02 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=2; Sample weight (mg)=78.8; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil02.d
SUBJECT_SAMPLE_FACTORS 3.1 RunA_dil03 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=3; Sample weight (mg)=45.5; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil03.d
SUBJECT_SAMPLE_FACTORS 4.1 RunA_dil04 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=4; Sample weight (mg)=20.3; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil04.d
SUBJECT_SAMPLE_FACTORS 5.1 RunA_dil05 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunA; Sample Number=5; Sample weight (mg)=90.6; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil05.d
SUBJECT_SAMPLE_FACTORS 1.1 RunA_dil11 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=11; Sample weight (mg)=56.9; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil11.d
SUBJECT_SAMPLE_FACTORS 2.1 RunA_dil12 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=12; Sample weight (mg)=31.3; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil12.d
SUBJECT_SAMPLE_FACTORS 3.1 RunA_dil13 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=13; Sample weight (mg)=55.6; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil13.d
SUBJECT_SAMPLE_FACTORS 4.1 RunA_dil14 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=14; Sample weight (mg)=52.6; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil14.d
SUBJECT_SAMPLE_FACTORS 5.1 RunA_dil15 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunA; Sample Number=15; Sample weight (mg)=49.5; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil15.d
SUBJECT_SAMPLE_FACTORS - RunA_dil21 Sample source:Naïve lysis buffer | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=21; Sample weight (mg)=-; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil21.d
SUBJECT_SAMPLE_FACTORS - RunA_dil22 Sample source:Naïve lysis buffer | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunA; Sample Number=22; Sample weight (mg)=-; Dilution before injection=25; RAW_FILE_NAME(Raw file name)=RunA_dil22.d
SUBJECT_SAMPLE_FACTORS 1.1 RunB_Dil50-01 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunB; Sample Number=1; Sample weight (mg)=92.1; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-01.d
SUBJECT_SAMPLE_FACTORS 2.1 RunB_Dil50-02 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunB; Sample Number=2; Sample weight (mg)=78.8; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-02.d
SUBJECT_SAMPLE_FACTORS 3.1 RunB_Dil50-03 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunB; Sample Number=3; Sample weight (mg)=45.5; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-03.d
SUBJECT_SAMPLE_FACTORS 4.1 RunB_Dil50-04 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunB; Sample Number=4; Sample weight (mg)=20.3; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-04.d
SUBJECT_SAMPLE_FACTORS 5.1 RunB_Dil50-05 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Run A or Run B=RunB; Sample Number=5; Sample weight (mg)=90.6; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-05.d
SUBJECT_SAMPLE_FACTORS 1.1 RunB_Dil50-11 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunB; Sample Number=11; Sample weight (mg)=56.9; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-11.d
SUBJECT_SAMPLE_FACTORS 2.1 RunB_Dil50-12 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunB; Sample Number=12; Sample weight (mg)=31.3; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-12.d
SUBJECT_SAMPLE_FACTORS 3.1 RunB_Dil50-13 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunB; Sample Number=13; Sample weight (mg)=55.6; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-13.d
SUBJECT_SAMPLE_FACTORS 4.1 RunB_Dil50-14 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunB; Sample Number=14; Sample weight (mg)=52.6; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-14.d
SUBJECT_SAMPLE_FACTORS 5.1 RunB_Dil50-15 Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Run A or Run B=RunB; Sample Number=15; Sample weight (mg)=49.5; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-15.d
SUBJECT_SAMPLE_FACTORS - RunB_Dil50-21 Sample source:Naïve lysis buffer | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=21; Sample weight (mg)=-; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-21.d
SUBJECT_SAMPLE_FACTORS - RunB_Dil50-22 Sample source:Naïve lysis buffer | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=22; Sample weight (mg)=-; Dilution before injection=50; RAW_FILE_NAME(Raw file name)=RunB_Dil50-22.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA01 Sample source:STDA01 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA01.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA01-rep Sample source:STDA01 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA01-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA01-rep2 Sample source:STDA01 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA01-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA02 Sample source:STDA02 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA02.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA02-rep Sample source:STDA02 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA02-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA02-rep2 Sample source:STDA02 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA02-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA03 Sample source:STDA03 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA03.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA03-rep Sample source:STDA03 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA03-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA03-rep2 Sample source:STDA03 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA03-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA04 Sample source:STDA04 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA04.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA04-rep Sample source:STDA04 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA04-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA04-rep2 Sample source:STDA04 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA04-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA05 Sample source:STDA05 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA05.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA05-rep Sample source:STDA05 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA05-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA05-rep2 Sample source:STDA05 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA05-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA06 Sample source:STDA06 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA06.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA06-rep Sample source:STDA06 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA06-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA06-rep2 Sample source:STDA06 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA06-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA07 Sample source:STDA07 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA07.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA07-rep Sample source:STDA07 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA07-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA07-rep2 Sample source:STDA07 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA07-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA08 Sample source:STDA08 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA08.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA08-rep Sample source:STDA08 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA08-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA08-rep2 Sample source:STDA08 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA08-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA09 Sample source:STDA09 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA09.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA09-rep Sample source:STDA09 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA09-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDA09-rep2 Sample source:STDA09 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDA09-rep2.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB01 Sample source:STDB01 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB01.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB01-rep Sample source:STDB01 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB01-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB02 Sample source:STDB02 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB02.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB02-rep Sample source:STDB02 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB02-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB03 Sample source:STDB03 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB03.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB03-rep Sample source:STDB03 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB03-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB04 Sample source:STDB04 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB04.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB04-rep Sample source:STDB04 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB04-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB05 Sample source:STDB05 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB05.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB05-rep Sample source:STDB05 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB05-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB06 Sample source:STDB06 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB06.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB06-rep Sample source:STDB06 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB06-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB07 Sample source:STDB07 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB07.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB07-rep Sample source:STDB07 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB07-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB08 Sample source:STDB08 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB08.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB08-rep Sample source:STDB08 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB08-rep.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB09 Sample source:STDB09 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB09.d
SUBJECT_SAMPLE_FACTORS - RunB_STDB09-rep Sample source:STDB09 | Muscle type (Ps psoas TA tibialis anterior):- Run A or Run B=RunB; Sample Number=-; Sample weight (mg)=-; Dilution before injection=-; RAW_FILE_NAME(Raw file name)=RunB_STDB09-rep.d
#COLLECTION
CO:COLLECTION_SUMMARY Male C57Bl/6J mice were housed with standard chow food and water ad libitum.
CO:COLLECTION_SUMMARY Mice were euthanised by cervical dislocation and skeletal muscle samples (psoas
CO:COLLECTION_SUMMARY and tibialis anterior) were rapidly excised and frozen.
CO:SAMPLE_TYPE Muscle
#TREATMENT
TR:TREATMENT_SUMMARY No treatment. Skeletal muscle obtained from two distinct sites of male C57BL/6J
TR:TREATMENT_SUMMARY mice (Ps=psoas; TA=tibialis anterior).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY ~50 mg tissue samples were cryoground, homogenised with zirconium oxide beads in
SP:SAMPLEPREP_SUMMARY MeOH/water (1:1), extracted with CHCl3, and aqueous phase lyophilised (GeneVac
SP:SAMPLEPREP_SUMMARY MiVac Duo, max temp 35 *C). Samples were stored at -80 *C prior to analysis.
SP:SAMPLEPREP_SUMMARY Samples were resuspended in 60 uL of ACN/H2O (1:1) and clarified prior to LC/MS.
SP:SAMPLEPREP_SUMMARY Samples were ran either as is ('neat'), or further diluted in ACN/H2O, with the
SP:SAMPLEPREP_SUMMARY fold-dilution specified in the Study Design table.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY Agilent InfinityLab Poroshell 120 HILIC-Z column (100 x 2.1
CH:CHROMATOGRAPHY_SUMMARY mm,2.7µm,stainless-steel lined); Variable flow rate was used as specified in
CH:CHROMATOGRAPHY_SUMMARY the flow gradient.
CH:CHROMATOGRAPHY_TYPE HILIC
CH:INSTRUMENT_NAME Agilent 1290 Infinity II
CH:COLUMN_NAME Agilent InfinityLab Poroshell 120 HILIC-Z (100 x 2.1mm,2.7um)
CH:SOLVENT_A 80% acetonitrile/20% water; 10mM ammonium acetate (pH 9); 5µM medronic acid
CH:SOLVENT_B 10% acetonitrile/90% water; 10mM ammonium acetate (pH 9); 5µM medronic acid
CH:FLOW_GRADIENT 0 min, 0% B, 250 μL/min; 2 min, 0% B, 250 μL/min; 12 min, 40% B, 250 μL/min;
CH:FLOW_GRADIENT 14 min, 40% B, 250 μL/min; 14.5 min, 0% B, 500 μL/min; 18.4 min, 0% B, 500
CH:FLOW_GRADIENT μL/min; 18.5 min, 0% B, 250 μL/min
CH:FLOW_RATE 250-500uL/min
CH:COLUMN_TEMPERATURE 30
#ANALYSIS
AN:ANALYSIS_TYPE MS
#MS
MS:INSTRUMENT_NAME Agilent 6470 QQQ
MS:INSTRUMENT_TYPE Triple quadrupole
MS:MS_TYPE ESI
MS:ION_MODE NEGATIVE
MS:MS_COMMENTS ESI = Jet Stream Technology Ion Source MS acquisition was performed in dMRM
MS:MS_COMMENTS mode, with acquisition parameters (e.g. collision energy, fragmentation voltage)
MS:MS_COMMENTS for each precursor/product transition optimised using neat metabolite standards.
MS:MS_COMMENTS Run A was dMRM analysis, with glutamate and alpha-ketoglutarate being
MS:MS_COMMENTS quantified. Run B was a standard MRM-mode analysis, with lactate and pyruvate
MS:MS_COMMENTS being quantified. Metabolites were identified by their transition and retention
MS:MS_COMMENTS time. For dMRM mode, retention times updated using a neat standard mix injected
MS:MS_COMMENTS immediately prior to performing the dMRM analysis. For MRM mode, retention times
MS:MS_COMMENTS were confirmed using neat standards. Data processing, peak identification, and
MS:MS_COMMENTS quantification was performed using Skyline, with the MRM parameters inputted
MS:MS_COMMENTS prior to file import.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS pmol/mg tissue
MS_METABOLITE_DATA_START
Samples RunA_01 RunA_02 RunA_03 RunA_04 RunA_05 RunA_11 RunA_12 RunA_13 RunA_14 RunA_15 RunB_Dil50-01 RunB_Dil50-02 RunB_Dil50-03 RunB_Dil50-04 RunB_Dil50-05 RunB_Dil50-11 RunB_Dil50-12 RunB_Dil50-13 RunB_Dil50-14 RunB_Dil50-15
Factors Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):Ps Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA Sample source:Mouse tissue | Muscle type (Ps psoas TA tibialis anterior):TA
alpha-ketoglutarate 2.749323882 1.279909872 6.918271257 5.845323673 1.457166777 0.566093945 1.195233735 0.675880013 1.021146173 1.371778272
lactate 52186.68241 65002.87916 67271.22607 190868.799 67248.5792 68294.42465 75963.21526 65773.00664 56919.25148 64172.17952
pyruvate 97.92733578 116.3439665 183.1683434 264.4341672 172.1229661 47.39713517 110.1860061 69.32475476 67.72783139 119.5897472
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name KEGG PubChem Run Precursor Product ion Ret Time (min) Delta Ret Time (min) Fragmentor voltage (V) Collision energy (V) Cell accelerator voltage (V) Dwell time (ms) Polarity
alpha-ketoglutarate C00026 3328 A 145 101.1 6.1 2 60 5 5 Negative
lactate C00186 3486 B 89 43.1 41 8 5 10 Negative
pyruvate C00022 3324 B 87 43.1 61 4 5 10 Negative
METABOLITES_END
#END