Summary of Study ST002929

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001822. The data can be accessed directly via it's Project DOI: 10.21228/M86H8Z This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002929
Study TitleHypoxia-inducible factor-1a (HIF1a) physiology in mouse skeletal muscle
Study SummaryHypoxia-inducible factor (HIF)-1a is continuously synthesized and degraded in normoxia, whereas during hypoxia, HIF1a stabilization restricts cellular oxygen utilization. Less is known about HIF1a function(s) and sex-specific effects during normoxia in the basal state. Since skeletal muscle is the largest protein store in mammals and protein homeostasis has high energy demands, we determined HIF1a function at baseline during normoxia in skeletal muscle with the use of untargeted metabolomics. Skeletal muscle from mice with and without skeletal muscle HIF1a deletion were harvested and analyzed using untargeted metabolomics (hydrophilic interaction (HILIC) and reverse phase liquid chromatography (RPLC) methods). We identified differentially expressed metabolites in the glycolytic and TCA cycle pathways.
Institute
Cleveland Clinic
DepartmentInflammation and Immunity
Last NameDasarathy
First NameSrinivasan
Address9500 Euclid Avenue, NE4-208
Emaildasaras@ccf.org
Phone2163799846
Submit Date2023-04-27
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailLC-MS
Release Date2024-10-28
Release Version1
Srinivasan Dasarathy Srinivasan Dasarathy
https://dx.doi.org/10.21228/M86H8Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001822
Project DOI:doi: 10.21228/M86H8Z
Project Title:Role of hypoxia inducible factor-1a (HIF1a) in Skeletal Muscle Physiology
Project Summary:Hypoxia-inducible factor (HIF)-1a is continuously synthesized and degraded in normoxia, whereas during hypoxia, HIF1a stabilization restricts cellular oxygen utilization. Less is known about HIF1a function(s) and sex-specific effects during normoxia in the basal state. Since skeletal muscle is the largest protein store in mammals and protein homeostasis has high energy demands, we determined HIF1a function at baseline during normoxia in skeletal muscle with the use of untargeted metabolomics.
Institute:Cleveland Clinic
Department:Inflammation and Immunity
Last Name:Dasarathy
First Name:Srinivasan
Address:9500 Euclid Avenue
Email:dasaras@ccf.org
Phone:2163799846
Funding Source:NIH

Subject:

Subject ID:SU003042
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:12-14 weeks
Gender:Male and female

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Genotype Treatment Sex
SA318000HIFFloxFPBS1-HILICNegFloxed Control PBS Female
SA318001HIFFloxFPBS7-HILICPosFloxed Control PBS Female
SA318002HIFFloxFPBS6-HILICPosFloxed Control PBS Female
SA318003HIFFloxFPBS2-HILICNegFloxed Control PBS Female
SA318004HIFFloxFPBS5-HILICPosFloxed Control PBS Female
SA318005HIFFloxFPBS6-HILICNegFloxed Control PBS Female
SA318006HIFFloxFPBS1-C18PosFloxed Control PBS Female
SA318007HIFFloxFPBS8-HILICNegFloxed Control PBS Female
SA318008HIFFloxFPBS7-HILICNegFloxed Control PBS Female
SA318009HIFFloxFPBS2-HILICPosFloxed Control PBS Female
SA318010HIFFloxFPBS5-HILICNegFloxed Control PBS Female
SA318011HIFFloxFPBS8-HILICPosFloxed Control PBS Female
SA318012HIFFloxFPBS7-C18PosFloxed Control PBS Female
SA318013HIFFloxFPBS8-C18PosFloxed Control PBS Female
SA318014HIFFloxFPBS6-C18PosFloxed Control PBS Female
SA318015HIFFloxFPBS1-HILICPosFloxed Control PBS Female
SA318016HIFFloxFPBS2-C18PosFloxed Control PBS Female
SA318017HIFFloxFPBS1-C18NegFloxed Control PBS Female
SA318018HIFFloxFPBS5-C18PosFloxed Control PBS Female
SA318019HIFFloxFPBS8-C18NegFloxed Control PBS Female
SA318020HIFFloxFPBS2-C18NegFloxed Control PBS Female
SA318021HIFFloxFPBS6-C18NegFloxed Control PBS Female
SA318022HIFFloxFPBS7-C18NegFloxed Control PBS Female
SA318023HIFFloxFPBS5-C18NegFloxed Control PBS Female
SA318024HIFFloxMPBS7-HILICPosFloxed Control PBS Male
SA318025HIFFloxMPBS8-HILICPosFloxed Control PBS Male
SA318026HIFFloxMPBS4-HILICPosFloxed Control PBS Male
SA318027HIFFloxMPBS3-HILICPosFloxed Control PBS Male
SA318028HIFFloxMPBS1-HILICNegFloxed Control PBS Male
SA318029HIFFloxMPBS7-HILICNegFloxed Control PBS Male
SA318030HIFFloxMPBS2-HILICPosFloxed Control PBS Male
SA318031HIFFloxMPBS8-HILICNegFloxed Control PBS Male
SA318032HIFFloxMPBS4-HILICNegFloxed Control PBS Male
SA318033HIFFloxMPBS3-HILICNegFloxed Control PBS Male
SA318034HIFFloxMPBS2-HILICNegFloxed Control PBS Male
SA318035HIFFloxMPBS8-C18PosFloxed Control PBS Male
SA318036HIFFloxMPBS4-C18PosFloxed Control PBS Male
SA318037HIFFloxMPBS7-C18PosFloxed Control PBS Male
SA318038HIFFloxMPBS3-C18PosFloxed Control PBS Male
SA318039HIFFloxMPBS1-HILICPosFloxed Control PBS Male
SA318040HIFFloxMPBS1-C18PosFloxed Control PBS Male
SA318041HIFFloxMPBS1-C18NegFloxed Control PBS Male
SA318042HIFFloxMPBS2-C18PosFloxed Control PBS Male
SA318043HIFFloxMPBS8-C18NegFloxed Control PBS Male
SA318044HIFFloxMPBS4-C18NegFloxed Control PBS Male
SA318045HIFFloxMPBS7-C18NegFloxed Control PBS Male
SA318046HIFFloxMPBS3-C18NegFloxed Control PBS Male
SA318047HIFFloxMPBS2-C18NegFloxed Control PBS Male
SA318048HIFmsdFPBS6-HILICPosMuscle specific deletion PBS Female
SA318049HIFmsdFPBS5-HILICPosMuscle specific deletion PBS Female
SA318050HIFmsdFPBS3-HILICPosMuscle specific deletion PBS Female
SA318051HIFmsdFPBS1-HILICNegMuscle specific deletion PBS Female
SA318052HIFmsdFPBS4-HILICPosMuscle specific deletion PBS Female
SA318053HIFmsdFPBS5-HILICNegMuscle specific deletion PBS Female
SA318054HIFmsdFPBS6-HILICNegMuscle specific deletion PBS Female
SA318055HIFmsdFPBS2-HILICPosMuscle specific deletion PBS Female
SA318056HIFmsdFPBS4-HILICNegMuscle specific deletion PBS Female
SA318057HIFmsdFPBS3-HILICNegMuscle specific deletion PBS Female
SA318058HIFmsdFPBS2-HILICNegMuscle specific deletion PBS Female
SA318059HIFmsdFPBS1-C18PosMuscle specific deletion PBS Female
SA318060HIFmsdFPBS5-C18PosMuscle specific deletion PBS Female
SA318061HIFmsdFPBS6-C18PosMuscle specific deletion PBS Female
SA318062HIFmsdFPBS4-C18PosMuscle specific deletion PBS Female
SA318063HIFmsdFPBS3-C18PosMuscle specific deletion PBS Female
SA318064HIFmsdFPBS1-HILICPosMuscle specific deletion PBS Female
SA318065HIFmsdFPBS1-C18NegMuscle specific deletion PBS Female
SA318066HIFmsdFPBS2-C18PosMuscle specific deletion PBS Female
SA318067HIFmsdFPBS6-C18NegMuscle specific deletion PBS Female
SA318068HIFmsdFPBS2-C18NegMuscle specific deletion PBS Female
SA318069HIFmsdFPBS5-C18NegMuscle specific deletion PBS Female
SA318070HIFmsdFPBS4-C18NegMuscle specific deletion PBS Female
SA318071HIFmsdFPBS3-C18NegMuscle specific deletion PBS Female
SA318072HIFmsdMPBS8-HILICPosMuscle specific deletion PBS Male
SA318073HIFmsdMPBS1-HILICNegMuscle specific deletion PBS Male
SA318074HIFmsdMPBS4-HILICPosMuscle specific deletion PBS Male
SA318075HIFmsdMPBS5-HILICPosMuscle specific deletion PBS Male
SA318076HIFmsdMPBS6-HILICPosMuscle specific deletion PBS Male
SA318077HIFmsdMPBS5-HILICNegMuscle specific deletion PBS Male
SA318078HIFmsdMPBS3-HILICPosMuscle specific deletion PBS Male
SA318079HIFmsdMPBS8-HILICNegMuscle specific deletion PBS Male
SA318080HIFmsdMPBS6-HILICNegMuscle specific deletion PBS Male
SA318081HIFmsdMPBS4-HILICNegMuscle specific deletion PBS Male
SA318082HIFmsdMPBS3-HILICNegMuscle specific deletion PBS Male
SA318083HIFmsdMPBS1-C18NegMuscle specific deletion PBS Male
SA318084HIFmsdMPBS5-C18PosMuscle specific deletion PBS Male
SA318085HIFmsdMPBS6-C18PosMuscle specific deletion PBS Male
SA318086HIFmsdMPBS4-C18PosMuscle specific deletion PBS Male
SA318087HIFmsdMPBS3-C18PosMuscle specific deletion PBS Male
SA318088HIFmsdMPBS1-C18PosMuscle specific deletion PBS Male
SA318089HIFmsdMPBS8-C18PosMuscle specific deletion PBS Male
SA318090HIFmsdMPBS3-C18NegMuscle specific deletion PBS Male
SA318091HIFmsdMPBS8-C18NegMuscle specific deletion PBS Male
SA318092HIFmsdMPBS6-C18NegMuscle specific deletion PBS Male
SA318093HIFmsdMPBS5-C18NegMuscle specific deletion PBS Male
SA318094HIFmsdMPBS4-C18NegMuscle specific deletion PBS Male
SA318095HIFmsdMPBS1-HILICPosMuscle specific deletion PBS Male
Showing results 1 to 96 of 96

Collection:

Collection ID:CO003035
Collection Summary:Mice were sacrificed and gastrocnemius muscle collected, flash frozen, and stored at -80deg until metabolite extraction. Metabolites were extracted from myotubes and skeletal muscle using chilled methanol containing 6 spiked heavy labeled internal standards (Choline-d13, Carnitine-d9, Betaine-d9, Creatinine-d3, Picolinic acid-d4, Hippuric acid-d5). After centrifugation at 14,000g for 20 minutes to precipitate the protein pellet, the supernatant was removed, dried, and reconstituted in 300 uL 98% water / 2% acetonitrile. LC/MS (liquid chromatography/mass spectrometry) analysis was then performed, with 2 chromatography methods (C18 and HILIC) including on pooled quality control samples.
Sample Type:Muscle
Storage Conditions:-80℃

Treatment:

Treatment ID:TR003051
Treatment Summary:Osmotic pumps were filled with phosphate buffered saline (PBS) and surgically implanted in mice aged 10-12 weeks. Mice were sacrificed 28 days after implantation. These mice were in the untreated arm of a larger study comparing hyperammonemic to untreated animals. The ammonia-treated samples are not included in this data submission and will be included as part of a later submission. Only the untreated samples are included in this data submission.

Sample Preparation:

Sampleprep ID:SP003048
Sampleprep Summary:Metabolites were extracted from myotubes and skeletal muscle using chilled methanol containing 6 spiked heavy labeled internal standards (Choline-d13, Carnitine-d9, Betaine-d9, Creatinine-d3, Picolinic acid-d4, Hippuric acid-d5). After centrifugation at 14,000g for 20 minutes to precipitate the protein pellet, the supernatant was removed, dried, and reconstituted in 300 uL 98% water / 2% acetonitrile. LC/MS (liquid chromatography/mass spectrometry) analysis was then performed, with 2 chromatography methods (C18 and HILIC) including on pooled quality control samples.

Combined analysis:

Analysis ID AN004803 AN004804 AN004805 AN004806
Analysis type MS MS MS MS
Chromatography type HILIC HILIC Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish Thermo Vanquish Thermo Vanquish
Column Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) Thermo Accucore C18 (100 x 2.1mm,2.6um) Thermo Accucore C18 (100 x 2.1mm,2.6um)
MS Type ESI ESI ESI ESI
MS instrument type QTRAP QTRAP QTRAP QTRAP
MS instrument name Thermo Q Exactive Focus Thermo Q Exactive Focus Thermo Q Exactive Focus Thermo Q Exactive Focus
Ion Mode POSITIVE NEGATIVE POSITIVE NEGATIVE
Units relative abundance relative abundance relative abundance relative abundance

Chromatography:

Chromatography ID:CH003631
Chromatography Summary:HILIC
Instrument Name:Thermo Vanquish
Column Name:Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um)
Column Temperature:40
Flow Gradient:Time (min.) Flow Rate (mL/min) %B 0.0 0.2 100 2.0 0.2 70 4.0 0.2 60 5.5 0.2 60 10.5 0.2 50 11.5 0.2 50 12.0 0.2 100 16.0 0.2 100
Flow Rate:0.2 mL/min
Solvent A:100% water;10 mM ammonium acetate; 0.125% acetic acid
Solvent B:5% water/95% acetonitrile;10 mM ammonium acetate; 0.125% acetic acid
Chromatography Type:HILIC
  
Chromatography ID:CH003632
Chromatography Summary:C18-RPLC
Instrument Name:Thermo Vanquish
Column Name:Thermo Accucore C18 (100 x 2.1mm,2.6um)
Column Temperature:60
Flow Gradient:Time (min.) Flow Rate (mL/min) %B 0.2 0.3 5 1.0 0.3 15 8.0 0.3 95 10.0 0.3 95 12.0 0.3 5 15.0 0.3 5
Flow Rate:0.3 mL/min
Solvent A:100% water; 0.1% formic acid
Solvent B:100% acetonitrile; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS004549
Analysis ID:AN004803
Instrument Name:Thermo Q Exactive Focus
Instrument Type:QTRAP
MS Type:ESI
MS Comments:Data dependent acquisitions (DDA) on the pooled representative QC samples include MS full scans at a resolution of 120,000 and HCD MS/MS scans taken on the top 5 most abundant ions at a resolution of 30,000 with dynamic exclusion. The resolution of the MS2 scans were taken at a stepped NCE energy of 10.0, 20.0 and 30.0.
Ion Mode:POSITIVE
  
MS ID:MS004550
Analysis ID:AN004804
Instrument Name:Thermo Q Exactive Focus
Instrument Type:QTRAP
MS Type:ESI
MS Comments:Data dependent acquisitions (DDA) on the pooled representative QC samples include MS full scans at a resolution of 120,000 and HCD MS/MS scans taken on the top 5 most abundant ions at a resolution of 30,000 with dynamic exclusion. The resolution of the MS2 scans were taken at a stepped NCE energy of 10.0, 20.0 and 30.0.
Ion Mode:NEGATIVE
  
MS ID:MS004551
Analysis ID:AN004805
Instrument Name:Thermo Q Exactive Focus
Instrument Type:QTRAP
MS Type:ESI
MS Comments:Data dependent acquisitions (DDA) on the pooled representative QC samples include MS full scans at a resolution of 120,000 and HCD MS/MS scans taken on the top 5 most abundant ions at a resolution of 30,000 with dynamic exclusion. The resolution of the MS2 scans were taken at a stepped NCE energy of 10.0, 20.0 and 30.0.
Ion Mode:POSITIVE
  
MS ID:MS004552
Analysis ID:AN004806
Instrument Name:Thermo Q Exactive Focus
Instrument Type:QTRAP
MS Type:ESI
MS Comments:Data dependent acquisitions (DDA) on the pooled representative QC samples include MS full scans at a resolution of 120,000 and HCD MS/MS scans taken on the top 5 most abundant ions at a resolution of 30,000 with dynamic exclusion. The resolution of the MS2 scans were taken at a stepped NCE energy of 10.0, 20.0 and 30.0.
Ion Mode:NEGATIVE
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