Summary of Study ST000885

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 PR000614. The data can be accessed directly via it's Project DOI: 10.21228/M8KD9Q This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST000885
Study Title	Gut Microbiota Modulate Brain Insulin Sensitivity and Neurobehavior
Study Type	Metabolite profiling of cecal contents and brains of mice under diet-induced obesity (DIO) with and without antibiotic treatments.
Study Summary	C57BL/6J mice were purchased from Jackson Laboratory and maintained on either a normal chow containing 22% of calories from fat, 23% from protein, and 55% from carbohydrates (Mouse diet 9F 5020; PharmaServ) or a high-fat diet (Open Source Diet, D12492; Research Diets) containing 60% of calories from fat, 20% from protein, and 20% from carbohydrates for 6 weeks. During the last 2 weeks, some of the HFD mice were treated with vancomycin or metronidazole (1 g/L in the drinking water). All mice were housed at 22°C on a 12 h light/dark cycle. All animal studies were approved by the IACUC of Joslin Diabetes Center (# 97-05) and Harvard Medical School (# 05131) and were in accordance with NIH guidelines. The metabolite profiling was conducted on plasma, hypothalamus and nucleus accumbens.
Institute	Broad Institute of MIT and Harvard
Last Name	Avila-Pacheco
First Name	Julian
Address	415 Main Street
Email	jravilap@broadinstitute.org
Phone	617-714-8264
Submit Date	2017-10-30
Num Groups	12
Total Subjects	24
Raw Data Available	Yes
Raw Data File Type(s)	d
Analysis Type Detail	LC-MS
Release Date	2017-10-30
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP000926
Sampleprep Summary:	Plasma: LC-MS samples were prepared from plasma (10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly. Nucleus accumbens and Hypothalamus: Frozen brain tissue samples were homogenized in 4 volumes of HPLC water (J. T. Baker, Center Valley Pa.) using a TissueLyser II (Qiagen, Hilden, Germany) with 3mm tungsten beads at 20 Hz in two 2-minute cycles. Homogenates were then aliquoted for profiling. Samples were prepared from lysates(10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly.

Sampleprep ID:

SP000926

Sampleprep Summary:

Plasma: LC-MS samples were prepared from plasma (10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly. Nucleus accumbens and Hypothalamus: Frozen brain tissue samples were homogenized in 4 volumes of HPLC water (J. T. Baker, Center Valley Pa.) using a TissueLyser II (Qiagen, Hilden, Germany) with 3mm tungsten beads at 20 Hz in two 2-minute cycles. Homogenates were then aliquoted for profiling. Samples were prepared from lysates(10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly.