Summary of Study ST001213

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 IDST001213
Study TitleSerum lipidomic profile of cold-exposed Ucp1cre/12-LOX KO mice
Study SummaryWe aimed to evaluate whether specific deletion of 12-Lipoxygenase (12-LOX) in brown fat can affect the serum concentrations of 12-LOX products under cold exposure.
Institute
Joslin Diabetes Center
Last NameLeiria
First NameLuiz
AddressOne Joslin Place, 02215, Boston, MA-USA
Emailluiz.leiria@joslin.harvard.edu
Phone617-309-1967
Submit Date2019-06-26
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2019-09-23
Release Version1
Luiz Leiria Luiz Leiria
https://dx.doi.org/10.21228/M8VD62
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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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:SU001280
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Male

Factors:

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

mb_sample_id local_sample_id Temperature Genotype
SA0858074621 0222C KO
SA0858084661 0122C KO
SA0858094831 0222C KO
SA0858104801 0322C KO
SA0858114671 0122C KO
SA0858124841 0222C KO
SA0858134601 0422C WT
SA0858144621 0322C WT
SA0858154801 0222C WT
SA0858164831 0122C WT
SA0858174801 0122C WT
SA0858184671 0222C WT
SA0858194761 015C KO
SA0858204761 025C KO
SA0858214601 035C KO
SA0858224801 055C KO
SA0858234831 035C KO
SA0858244621 045C KO
SA0858254841 015C WT
SA0858264621 015C WT
SA0858274801 045C WT
SA0858284601 025C WT
SA0858294661 025C WT
SA0858304671 035C WT
Showing results 1 to 24 of 24

Collection:

Collection ID:CO001274
Collection Summary:Mice were anaesthetised with Isoflurane (under controlled flow-rate), and placed in a hot pad, while the blood was drawn from the tail into a tube. The blood was left at 22C for 30 - 60 minutes, and then centrifuged at 14,000 RPM for 3 minutes. The serum fraction was collected and frost at -20C.
Sample Type:Blood (serum)

Treatment:

Treatment ID:TR001295
Treatment Summary:Wild-type and Ucp1CRE/12-LOX KO mice were exposed to a short-term cold temperature (1 hour at 5C) or kept at room temperature. After this period, the serum was collected as described. Ucp1CRE/12-LOX KO mice were created through CRISPR-Cas9 technology, as described in Leiria et al., 2019, Cell Metabolism.

Sample Preparation:

Sampleprep ID:SP001288
Sampleprep Summary:Aliquots of 100 µL serum or 1mg protein from homogenized tissue (measured by BCA) were taken, depending on the experiment. 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 31 −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 H 2 O (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:H 2 O (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 LCMS/MS mediator lipidomics platform.

Combined analysis:

Analysis ID AN002024
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:CH001465
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:MS001877
Analysis ID:AN002024
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. Fullscan 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).
Ion Mode:NEGATIVE
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