Summary of Study ST003240

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.

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.

Study ID	ST003240
Study Title	Lipidomic analysis of liver from Gclc WT and whole-body Gclc KO mice.
Study Summary	We found decreased triglycerides in the liver of whole-body deletion of Gclc mice compared to wildtype mice.
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
Submit Date	2024-05-29
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2024-06-07
Release Version	1

Select appropriate tab below to view additional metadata details:

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:	SU003359
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Male and female
Species Group:	Mammals

Factors:

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

mb_sample_id	local_sample_id	Sample source	Genotype
SA353336	Liver_CreERT2_210	liver	Gclc KO
SA353337	Liver_CreERT2_232	liver	Gclc KO
SA353338	Liver_CreERT2_223	liver	Gclc KO
SA353339	Liver_CreERT2_216	liver	Gclc KO
SA353340	Liver_FF_226	liver	Gclc WT
SA353341	Liver_FF_218	liver	Gclc WT
SA353342	Liver_FF_219	liver	Gclc WT
SA353343	Liver_FF_225	liver	Gclc WT

Showing results 1 to 8 of 8

Showing results 1 to 8 of 8

Collection:

Collection ID:	CO003352
Collection Summary:	Mice were euthanized with CO2 inhalation. Tissues were isolated, snap frozen on dry ice, and stored at -80C.
Sample Type:	Liver

Treatment:

Treatment ID:	TR003368
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:	SP003366
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.

Sampleprep ID:

SP003366

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	AN005307	AN005308
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:	CH004014
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:	MS005037
Analysis ID:	AN005307
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:	MS005038
Analysis ID:	AN005308
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