Summary of Study ST001207

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 PR000811. The data can be accessed directly via it's Project DOI: 10.21228/M8VD62 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001207
Study TitleLipidomics in the serum of cold exposed mice treated with 12-LOX inhibitor LOXBlock-1
Study SummaryWe aimed to investigate whether the cold-induced release of 12-LOX products into the circulation were dependent on 12-LOX activation. We pre-treated C57BL6/J mice with the pharmacological inhibitor LOXBlock-1 or its vehicle (DMSO), and after 15 minutes we placed them under cold temperature (5C) for 4 hours. A control group was injected with DMSO and kept at room temperature for the same 4 hours. After this period of time we, collected the blood, and obtained the serum fraction that was immediately frozen and submitted for untargeted lipidomics.
Institute
Joslin Diabetes Center
Last NameLeiria
First NameLuiz
AddressOne Joslin Place, Boston-MA, 02215
Emailluiz.leiria@joslin.harvard.edu
Phone617-309-1967
Submit Date2019-06-26
Num Groups3
Study CommentsJoslin Diabetes Center affiliate of Harvard Medical School
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2019-07-17
Release Version1
Luiz Leiria Luiz Leiria
https://dx.doi.org/10.21228/M8VD62
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR000811
Project DOI:doi: 10.21228/M8VD62
Project Title:12-LOX metabolites in cold adaptation
Project Summary:The goal of this project is to understand the role of the enzyme 12-lipoxygenase in the adaptive thermogenesis. We found this enzyme is activated by cold stimulation, then producing lipid metabolites in adipose tissue and releasing them into the circulation to regulate fuel utilisation and thermogenic pathways required for the cold adaptation.
Institute:Joslin Diabetes Center
Department:Integrative Physiology and Metabolism
Laboratory:Yu-Hua Tseng lab
Last Name:Leiria
First Name:Luiz
Address:One Joslin Place, Boston, MA-USA, 02215
Email:luiz.leiri@joslin.harvard.edu
Phone:1 6173091967

Subject:

Subject ID:SU001274
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL6/J
Age Or Age Range:12 weeks old
Weight Or Weight Range:25-30 g
Gender:Male
Animal Animal Supplier:Jax Laboratories
Animal Housing:Conventional
Animal Light Cycle:regular 12h light /dark cycles
Animal Feed:ad libitum normal chow diet
Animal Water:ad libitum

Factors:

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

mb_sample_id local_sample_id Temperature Treatment
SA084803T522C DMSO
SA084804T622C DMSO
SA084805T122C DMSO
SA084806T222C DMSO
SA084807T322C DMSO
SA084808T722C DMSO
SA084809T422C DMSO
SA084810C45C DMSO
SA084811C35C DMSO
SA084812C55C DMSO
SA084813C65C DMSO
SA084814C15C DMSO
SA084815C75C DMSO
SA084816C25C DMSO
SA084817LB35C LoxBlock-1
SA084818LB15C LoxBlock-1
SA084819LB25C LoxBlock-1
SA084820LB45C LoxBlock-1
SA084821LB55C LoxBlock-1
Showing results 1 to 19 of 19

Collection:

Collection ID:CO001268
Collection Summary:Mice were anaesthetised with Avertine and blood was collected through cardiac puncture. Samples were let in room temperature for 30 minutes, and then centrifuged for 3 minutes at 14,000RPM. Serum fas collected and frozen at -20C for further lipidomics analysis.
Sample Type:Blood (serum)
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001289
Treatment Summary:Mice were injected i.p. 15 minutes before the cold exposure, with 50ul of DMSO (vehicle) or LOXBlock-1. Then, the animas were placed in 5C or 22C temperature for 4 hours, until the samples were collected.

Sample Preparation:

Sampleprep ID:SP001282
Sampleprep Summary:Aliquots of 100 µL serum were taken and a mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H2O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H2O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LC–MS/MS mediator lipidomics platform.

Combined analysis:

Analysis ID AN002009
Analysis type MS
Chromatography type Reversed phase
Chromatography system Ekspert MicroLC 200 system
Column Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
MS Type ESI
MS instrument type Triple TOF
MS instrument name ABI Sciex 5600+ TripleTOF
Ion Mode NEGATIVE
Units Peak Area

Chromatography:

Chromatography ID:CH001453
Instrument Name:Ekspert MicroLC 200 system
Column Name:Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
Chromatography Type:Reversed phase

MS:

MS ID:MS001862
Analysis ID:AN002009
Instrument Name:ABI Sciex 5600+ TripleTOF
Instrument Type:Triple TOF
MS Type:ESI
MS Comments:MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Full-scan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX). C18SPE cartridges were purchased from Biotage. All solvents were of HPLC or LC-MS/MS grade and were acquired from Sigma-Aldrich, Fisher Scientific, or VWR International.
Ion Mode:NEGATIVE
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