Summary of Study ST001439

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 PR000989. The data can be accessed directly via it's Project DOI: 10.21228/M8VH7G This work is supported by NIH grant, U2C- DK119886.

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Study IDST001439
Study TitleMetabolites in contents of small intestine in wild type and DAOG181R/G181R mice
Study SummaryTo investigate if DAO regulates metabolites in intestinal lumen, we compared metabolites in contents of small intestine in wild type and DAOG181R/G181R mice. All mice have C57BL/6 background and 8 weeks of age.
Institute
Keio University
DepartmentPharmacology
Last NameSuzuki
First NameMasataka
Address35, Shinanomachi, Shinjuku-ku, Tokyo
Emailmasataka.s@keio.jp
Phone+81-3-5363-3750
Submit Date2019-12-17
Analysis Type DetailMS(Dir. Inf.)
Release Date2020-08-04
Release Version1
Masataka Suzuki Masataka Suzuki
https://dx.doi.org/10.21228/M8VH7G
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000989
Project DOI:doi: 10.21228/M8VH7G
Project Title:D-amino acids control IgA production
Project Summary:Intestinal microbiota produces D-amino acids, which are bacteria-specific metabolites, for their growth and their communication. Host intestine release D-amino acid oxidase (DAO) to degrade bacterial D-amino acids that results in regulation of microbiota. However, it is not clarified whether bacterial D-amino acids affect host’s immunity. In the present study, we investigated the metabolites in intestinal contents of DAO null mice compared to wild type animals. We did not find any significant difference in metabolites when we did not distinguish chirality. However, we found that DAO null mice had much higher amount of D-alanine in the intestinal epithelium, suggesting that D-alanine are involved in activated immune response in DAO null mice.
Institute:Keio University
Department:Pharmacology
Last Name:Suzuki
First Name:Masataka
Address:35, Shinanomachi,, Shinjuku-ku, Tokyo, 160-8582, Japan
Email:masataka.s@keio.jp
Phone:+81-3-5363-3750

Subject:

Subject ID:SU001513
Subject Type:Bacteria
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL/6
Age Or Age Range:8 weeks of age
Gender:Male

Factors:

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

mb_sample_id local_sample_id Genotype
SA122100CSI-DAO3DAOG181R
SA122101CSI-DAO4DAOG181R
SA122102CSI-DAO5DAOG181R
SA122103CSI-DAO2DAOG181R
SA122104CSI-DAO1DAOG181R
SA122105CSI-wt2wild type
SA122106CSI-wt3wild type
SA122107CSI-wt4wild type
SA122108CSI-wt5wild type
SA122109CSI-wt1wild type
Showing results 1 to 10 of 10

Collection:

Collection ID:CO001508
Collection Summary:Mice were euthanized and collect half length of small intestine (ileum). Contents of small intestine were mechanically squeezed by pipet tip into 1.5 mL tube and then flash-frozen in liquid nitrogen.
Sample Type:contents of small intestine
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001528
Treatment Summary:Mice were generated by in vitro fertilization and transplantation to surrogate mothers, and pups C-sectioned on E20 were fostered to CD-1 mother with strain-selected microbiota in the animal facility of CIEA.

Sample Preparation:

Sampleprep ID:SP001521
Sampleprep Summary:For extracting ionic metabolites. approximately 20 mg of contents of small intestine were dissolved in H2O containing internal standards (H3304-1002, HMT, Japan) with ratio of 1:9. After centrifugation, the supernatant was the centrifugically filtered through a Milipore 5000-Da cutoff filter to remove macromolecules for subsequent analysis with CE-TOFMS. For extracting non-ionic metabolites, approximately 20 mg of contents of small intestine were dissolved in 1 mL of methanol containing internal standard (H3304-1002, HMT). After centrifugation, 1o uL of the supernatant was transferred into grass vials for evaporation under nitrogen gas and reconstituted with 200 uL of 50% isopropanol for subsequent analysis with LC-TOFMS.

Combined analysis:

Analysis ID AN002403 AN002404
Analysis type MS MS
Chromatography type None (Direct infusion) None (Direct infusion)
Chromatography system none none
Column none none
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Agilent 6210 TOF Agilent 6230 TOF
Ion Mode UNSPECIFIED UNSPECIFIED
Units relative_area relative_area

Chromatography:

Chromatography ID:CH001766
Instrument Name:none
Column Name:none
Chromatography Type:None (Direct infusion)

MS:

MS ID:MS002244
Analysis ID:AN002403
Instrument Name:Agilent 6210 TOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:System control: Agilent G2201AA ChemStation software version B .03.01 Peaks were extracted using automatic integration software MasterHands to obtain peal information including m/z, peak area and migration time. Detected metabolites were plotted on metabolic pathway maps using VANTED software.
Ion Mode:UNSPECIFIED
  
MS ID:MS002245
Analysis ID:AN002404
Instrument Name:Agilent 6230 TOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:System control: Agilent G2201AA ChemStation software version B .03.01 Peaks were extracted using automatic integration software MasterHands to obtain peal information including m/z, peak area and retention time. Detected metabolites were plotted on metabolic pathway maps using VANTED software.
Ion Mode:UNSPECIFIED
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