Summary of Study ST003239

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 PR002013. The data can be accessed directly via it's Project DOI: 10.21228/M8DJ7S This work is supported by NIH grant, U2C- DK119886.

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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 IDST003239
Study TitleLipidomic analysis of adipose from Gclc WT and whole-body Gclc KO mice.
Study SummaryWe found decreased triglycerides in the adipose of whole-body deletion of Gclc mice compared to wildtype mice.
Institute
University of Rochester Medical Center
Last NameHarris
First NameIsaac
Address601 Elmwood Ave, Rochester, New York, 14642-0001, USA
Emailisaac_harris@urmc.rochester.edu
Phone8572348624
Submit Date2024-05-29
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2024-06-10
Release Version1
Isaac Harris Isaac Harris
https://dx.doi.org/10.21228/M8DJ7S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002013
Project DOI:doi: 10.21228/M8DJ7S
Project Title:Glutathione synthesis in the mouse liver supports lipid abundance through NRF2 repression.
Project Summary:Cells rely on antioxidants to survive. The most abundant antioxidant is glutathione (GSH). The synthesis of GSH is non-redundantly controlled by the glutamate-cysteine ligase catalytic subunit (GCLC). GSH imbalance is implicated in many diseases, but the requirement for GSH in adult tissues is unclear. To interrogate this, we have developed a series of in vivo models to induce Gclc deletion in adult animals. We find that GSH is essential to lipid abundance in vivo. GSH levels are highest in liver tissue, which is also a hub for lipid production. While the loss of GSH does not cause liver failure, it decreases lipogenic enzyme expression, circulating triglyceride levels, and fat stores. Mechanistically, we find that GSH promotes lipid abundance by repressing NRF2, a transcription factor induced by oxidative stress. These studies identify GSH as a fulcrum in the liver's balance of redox buffering and triglyceride production.
Institute:University of Rochester Medical Center
Last Name:Harris
First Name:Isaac
Address:601 Elmwood Ave, Rochester, New York, 14642-0001, USA
Email:isaac_harris@urmc.rochester.edu
Phone:8572348624

Subject:

Subject ID:SU003358
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Male and female

Factors:

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

mb_sample_id local_sample_id Sample source Genotype
SA353329Pos_Fat_CreERT2_223adipose tissue Gclc KO
SA353330Pos_Fat_CreERT2_210adipose tissue Gclc KO
SA353331Pos_Fat_CreERT2_232adipose tissue Gclc KO
SA353332Pos_Fat_CreERT2_224adipose tissue Gclc KO
SA353333Pos_Fat_FF_231adipose tissue Gclc WT
SA353334Pos_Fat_FF_226adipose tissue Gclc WT
SA353335Pos_Fat_FF_225adipose tissue Gclc WT
Showing results 1 to 7 of 7

Collection:

Collection ID:CO003351
Collection Summary:Mice were euthanized with CO2 inhalation. Tissues were isolated, snap frozen on dry ice, and stored at -80C.
Sample Type:Adipose tissue

Treatment:

Treatment ID:TR003367
Treatment Summary:Gclc WT (Gclc f/f) and whole-body Gclc KO (Gclc f/f Rosa26-CreERT2) were treated with 160 mg/kg i.p. injection of tamoxifen daily for five consecutive days. After approximately 14 days, mice were euthanized and tissue was isolated and stored.

Sample Preparation:

Sampleprep ID:SP003365
Sampleprep Summary:Tissues were homogenized with a pre-chilled BioPulverizer (59012MS, BioSpec) and then placed on dry ice. The chloroform:methanol extraction solvent (v:v = 1:1) was added to the homogenate to meet 50 mg/mL. The samples were then sonicated in ice-cold water using BiorupterTM UCD-200 sonicator for 5 min (30 s sonication and 30 s rest cycle; high voltage mode). The lipid extracts were cleared by centrifugation (17,000g, 20°C, 10 min), and the lipids in the supernatant were analyzed by LC-MS.

Combined analysis:

Analysis ID AN005305 AN005306
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish
Column Perkin Elmer Brownlee SPP C18 (75 × 2.1mm, 2.7um) Perkin Elmer Brownlee SPP C18 (75 × 2.1mm, 2.7um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive HF hybrid Orbitrap Thermo Q Exactive HF hybrid Orbitrap
Ion Mode POSITIVE NEGATIVE
Units Normalized to the median value of total lipid signals Normalized to the median value of total lipid signals

Chromatography:

Chromatography ID:CH004013
Instrument Name:Thermo Vanquish
Column Name:Perkin Elmer Brownlee SPP C18 (75 × 2.1mm, 2.7um)
Column Temperature:50
Flow Gradient:0–2 min 35% B, 2–8 min from 35 to 80% B, 8–22 min from 80 to 99% B, 22–36 min 99% B, and 36.1–40 min from 99 to 35% B.
Flow Rate:0.400 mL/min
Solvent A:100% water; 0.1% formic acid, 10mM ammonium acetate
Solvent B:50% acetonitrile/50% isopropanol; 0.1% formic acid; 10mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS005035
Analysis ID:AN005305
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:The scan range was from m/z 250–1500, resolution was 60,000 for MS, and 30,000 for DDMS2 (top 10), and the AGC target was 3E6 for full MS and 1E5 for DDMS2, allowing ions to accumulate for up to 200 ms for MS and 50 ms for MS/MS. For MS/MS, the following settings are used: isolation window width 1.2 m/z with an offset of 0.5 m/z, stepped NCE at 10, 15, and 25 a.u., minimum AGC 5E2, and dynamic exclusion of previously sampled peaks for 8 s.
Ion Mode:POSITIVE
  
MS ID:MS005036
Analysis ID:AN005306
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:The scan range was from m/z 250–1500, resolution was 60,000 for MS, and 30,000 for DDMS2 (top 10), and the AGC target was 3E6 for full MS and 1E5 for DDMS2, allowing ions to accumulate for up to 200 ms for MS and 50 ms for MS/MS. For MS/MS, the following settings are used: isolation window width 1.2 m/z with an offset of 0.5 m/z, stepped NCE at 10, 15, and 25 a.u., minimum AGC 5E2, and dynamic exclusion of previously sampled peaks for 8 s.
Ion Mode:NEGATIVE
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