Summary of Study ST003601

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 PR002229. The data can be accessed directly via it's Project DOI: 10.21228/M8H822 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 IDST003601
Study TitleRapid modulation of gut microbiota composition by hypothalamic circuits in mice
Study TypeMetabolomics
Study SummaryWhether the hypothalamus is able to influence gut microbiota composition remains enigmatic. Here, we present a proof-of-concept study designed to unravel this challenging question. To this aim, we centrally administered leptin or ghrelin to male mice. Subsequently, we conducted microbiota composition analysis throughout the gut using 16S rRNA gene sequencing. Our results showed that these brain interventions significantly changed the gut microbiota in an anatomical and short-term (two to four hours) fashion. Metabolomics analysis was performed on the intestinal content of the duodenum 4h after leptin injection, revealing changed metabolite levels.
Institute
University of Luebeck
DepartmentBioanalytic Core Facility
Last NameInderhees
First NameJulica
AddressRatzeburger Allee 34b, 23568 Luebeck
Emailjulica.inderhees@uni-luebeck.de
Phone+4945131012805
Submit Date2024-11-30
Num Groups2
Total Subjects23
Num Males23
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2025-02-27
Release Version1
Julica Inderhees Julica Inderhees
https://dx.doi.org/10.21228/M8H822
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR002229
Project DOI:doi: 10.21228/M8H822
Project Title:Rapid modulation of gut microbiota composition by hypothalamic circuits in mice
Project Type:Metabolomics
Project Summary:Whether the hypothalamus is able to influence gut microbiota composition remains enigmatic. Here, we present a proof-of-concept study designed to unravel this challenging question. To this aim, we centrally administered leptin or ghrelin to male mice. Subsequently, we conducted microbiota composition analysis throughout the gut using 16S rRNA gene sequencing. Our results showed that these brain interventions significantly changed the gut microbiota in an anatomical and short-term (two to four hours) fashion. Metabolomics analysis was performed on the intestinal content of the duodenum 4h after leptin injection, revealing changed metabolite levels.
Institute:University of Luebeck
Department:Bioanalytic Core Facility
Last Name:Inderhees
First Name:Julica
Address:Ratzeburger Allee 34b, 23568 Luebeck
Email:julica.inderhees@uni-luebeck.de
Phone:+4945131012805
Funding Source:European Research Council ERC-Synergy-Grant-2019-WATCH No. 810331
Contributors:Míriam Toledo, Sara Martínez-Martínez, Rubén Nogueiras, Marc Claret

Subject:

Subject ID:SU003791
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:8 weeks
Gender:Male
Animal Light Cycle:12-hour light/dark cycle
Animal Feed:standard diet (Teklad maintenance diet 14% protein; Envigo)
Animal Water:ad libitum

Factors:

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

mb_sample_id local_sample_id Treatment Group Sample source
SA400459extraction_blank_2Blank Blank
SA400460extraction_blank_1Blank Blank
SA400461solvent_blank_2Blank Blank
SA400462solvent_blank_1Blank Blank
SA400463Duo_23Leptin Intestinal content
SA400464Duo_02Leptin Intestinal content
SA400465Duo_24Leptin Intestinal content
SA400466Duo_01Leptin Intestinal content
SA400467Duo_21Leptin Intestinal content
SA400468Duo_14Leptin Intestinal content
SA400469Duo_17Leptin Intestinal content
SA400470Duo_07Leptin Intestinal content
SA400471Duo_09Leptin Intestinal content
SA400472Duo_11Leptin Intestinal content
SA400473Duo_04Leptin Intestinal content
SA400474QC2_3Quality Control Quality Control
SA400475QC3_2Quality Control Quality Control
SA400476QC5_3bQuality Control Quality Control
SA400477QC5_2bQuality Control Quality Control
SA400478QC5_1bQuality Control Quality Control
SA400479QC5_3Quality Control Quality Control
SA400480QC5_2Quality Control Quality Control
SA400481QC5_1Quality Control Quality Control
SA400482QC4_3Quality Control Quality Control
SA400483QC4_2Quality Control Quality Control
SA400484QC4_1Quality Control Quality Control
SA400485QC2_1Quality Control Quality Control
SA400486QC2_2Quality Control Quality Control
SA400487QC3_3Quality Control Quality Control
SA400488QC3_1Quality Control Quality Control
SA400489Duo_12Vehicle Intestinal content
SA400490Duo_15Vehicle Intestinal content
SA400491Duo_03Vehicle Intestinal content
SA400492Duo_05Vehicle Intestinal content
SA400493Duo_06Vehicle Intestinal content
SA400494Duo_08Vehicle Intestinal content
SA400495Duo_22Vehicle Intestinal content
SA400496Duo_10Vehicle Intestinal content
SA400497Duo_19Vehicle Intestinal content
SA400498Duo_18Vehicle Intestinal content
SA400499Duo_16Vehicle Intestinal content
SA400500Duo_20Vehicle Intestinal content
Showing results 1 to 42 of 42

Collection:

Collection ID:CO003784
Collection Summary:Samples of mucus and luminal content were collected from the duodenum of each mouse at the selected time point for the subsequent analysis of gut metabolomics.
Sample Type:Duodenum
Storage Conditions:-80℃

Treatment:

Treatment ID:TR003800
Treatment Summary:Intracerebroventricular (ICV) leptin (L3772, Sigma) treatment was performed three days after a cannula implantation. One hour after the start of the light period, mice were food-deprived for two hours before treatment. At the time of injection, mice were immobilized, and the cannula was opened to inject leptin (3 µg/mouse) or sterile saline for controls, using a 25 µL syringe (Gastight Syringe, Hamilton). Animals were euthanized four hours after the ICV injection for sample collection.

Sample Preparation:

Sampleprep ID:SP003798
Sampleprep Summary:50 µl of water and 700 µl acetone/acetonitrile/methanol (1:1:1, v/v/v), containing 2.5 µM Metabolomics Amino Acid Mix Standard (Cambridge Isotope Laboratories), were added to each sample (34 mg ± 8.6 mg). After incubation and centrifugation, 600 µl of the supernatant were dried under vacuum. The leftover supernatants of all samples were pooled and used for quality control samples (QCs). Dried supernatants were reconstituted in 70 µl methanol/acetonitrile (1:1, v/v) for LC-MS/MS analysis.
Processing Storage Conditions:On ice
Extract Storage:4℃

Combined analysis:

Analysis ID AN006012
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Dionex Ultimate 3000 RS
Column Merck SeQuant ZIC-HILIC (150 x 2.1 mm, 5 µm)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode UNSPECIFIED
Units relative concentration

Chromatography:

Chromatography ID:CH004568
Chromatography Summary:Metabolites were separated on a SeQuant ZIC-HILIC column (150 × 2.1 mm, 5 μm; Merck) using water with 5 mM ammonium acetate as eluent A and acetonitrile/eluent A (95:5, v/v) as eluent B, both containing 0.1% formic acid. The gradient elution was set as follows: isocratic step of 100% B for 3 min, 100% B to 60% B in 15 min, held for 5 min, returned to initial conditions in 5 min and held for 5 min. Flow rate was 0.5 mL/min.
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:Merck SeQuant ZIC-HILIC (150 x 2.1 mm, 5 µm)
Column Temperature:35°C
Flow Gradient:Isocratic step of 100% B for 3 min, 100% B to 60% B in 15 min, held for 5 min, returned to initial conditions in 5 min and held for 5 min
Flow Rate:0.5 mL/min
Solvent A:100% water; 5mM ammonium acetate; 0.1% formic acid
Solvent B:5% water/95% acetonitrile; 5 mM ammonium acetate; 0.1% formic acid
Chromatography Type:HILIC

MS:

MS ID:MS005723
Analysis ID:AN006012
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Data acquisition with data-dependent MS2 scans (top 10) was performed with polarity switching. Compound Discoverer 3.3 (ThermoFisher Scientific) was used for data processing. Metabolites were identified based on exact mass, retention time, fragmentation spectra and isotopic pattern. We used an in-house fragmentation library, a microbiome-specific library as well as the online library mzCloud. A QC-based normalization was performed. The area under the peak was additionally normalized to the appropriate internal standard and sample weight. QCs at 4 concentrations were used to ensure signal stability and linearity.
Ion Mode:UNSPECIFIED
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