Summary of Study ST000358

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 PR000285. The data can be accessed directly via it's Project DOI: 10.21228/M82W27 This work is supported by NIH grant, U2C- DK119886.

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

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 IDST000358
Study TitleBroad Spectrum MS analysis of mouse hippocampus from Anxiety Prone HSV-Latently Infected Obese Mice
Study TypeBroad Spectrum LCMS
Study SummaryBrain tissue samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics.
Institute
RTI International
DepartmentDiscovery Sciences
LaboratorySystem and Translational Sciences
Last NameSumner
First NameSusan
Address3040 Cornwallis Rd, RTP,NC
Emailssumner@rti.org
Phone919-541-7479
Submit Date2016-07-05
Num Groups4
Total Subjects31
Num Males31
Raw Data AvailableYes
Raw Data File Type(s)raw(Waters)
Analysis Type DetailLC-MS
Release Date2017-03-04
Release Version1
Susan Sumner Susan Sumner
https://dx.doi.org/10.21228/M82W27
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000285
Project DOI:doi: 10.21228/M82W27
Project Title:Metabolic Profiling of Anxiety Prone HSV-Latently Infected Obese Mice
Project Type:Broad Spectrum LCMS
Project Summary:The biological factors that lead children from low socioeconomic backgrounds to be at greater risk for the development of anxiety and learning problems are not well understood. While it is clear that there are genetic components to the risk of developing mental health disorders, a role for environmental factors in inducing these problems has been suggested. Many of these factors that affect the brain likely involve exposures that occur early in life. Two such factors are the higher rates of herpes simplex virus (HSV)-1 seropositivity and prevalence of obesity among these children. HSV-1 infection has been associated with impaired cognition during childhood and mental health problems in adulthood. Additionally, obese adults are shown to have higher HSV-1 titers. Similarly, other studies have correlated a diet high in saturated fat and obesity with increased risk of mood disorders and anxiety. A mouse model of obese HSV-1 latent infection was developed. Broad spectrum metabolomics analysis was performed to better understand the metabolomic profile of hippocampus and to compare this metabolomics profile with that of the hypothalamus, microglia, and peripheral blood mononuclear cells. Brain tissue samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics.
Institute:University of North Carolina at Chapel Hill
Department:Gillings School of Public Health
Laboratory:Department of Nutrition
Last Name:Sheridan
First Name:Patricia
Address:2002 Hooker Research Center, CB#7461, Chapel Hill NC, 27599
Email:patricia_sheridan@med.unc.edu
Phone:919-843-6434
Funding Source:NIH, Common Fund, Pilot and Feasibility

Subject:

Subject ID:SU000922
Subject Type:mouse
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57Bl/6
Age Or Age Range:5.5 mo
Weight Or Weight Range:29.1 g-49.2 g
Gender:male
Animal Animal Supplier:JAX
Animal Housing:4/cage
Animal Feed:Purina chow
Species Group:Mammal

Factors:

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

mb_sample_id local_sample_id HSV-1 Diet at Week 2
SA051517Total_Pool_03- -
SA051518Total_Pool_02- -
SA051519Total_Pool_04- -
SA051520Total_Pool_01- -
SA051521Total_Pool_05- -
SA051522Hipp402no HF
SA051523Hipp387no HF
SA051524Hipp401no HF
SA051525Hipp403no HF
SA051526Hipp385no HF
SA051527Hipp386no HF
SA051528Hipp388no HF
SA051529Hipp405no LF
SA051530Hipp407no LF
SA051531Hipp390no LF
SA051532Hipp408no LF
SA051533Hipp391no LF
SA051534Hipp406no LF
SA051535Hipp389no LF
SA051536Hipp392no LF
SA051537Hipp393yes HF
SA051538Hipp395yes HF
SA051539Hipp411yes HF
SA051540Hipp396yes HF
SA051541Hipp412yes HF
SA051542Hipp409yes HF
SA051543Hipp394yes HF
SA051544Hipp410yes HF
SA051545Hipp414yes LF
SA051546Hipp398yes LF
SA051547Hipp400yes LF
SA051548Hipp399yes LF
SA051549Hipp415yes LF
SA051550Hipp397yes LF
SA051551Hipp416yes LF
SA051552Hipp413yes LF
Showing results 1 to 36 of 36

Collection:

Collection ID:CO000916
Collection Summary:Brain tissue samples were homogenized in 50:50 Acetonitrile:Water (10 µL to every 1 mg of tissue) with washed ceramic beads on a MagNA Lyser, using two 30 sec pulses at 2,000 rpm with a 1 min chilling step in between pulses. Samples were vortexed on a multi-tube vortexer for 2 min at 5,000 rpm and centrifuged at room temperature for 4 min at 16,000 rcf. A 50 µL aliquot of each experimental brain tissue homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind eppendorf tube. Analytical pooled samples were created by combining 15 µL aliquot from each experimental sample in a 2.0 mL Lo-Bind eppendorf tube. The QC pooled sample was vortexed for 30 sec and 50 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf tubes. Next, 350 µL of acetonitrile was added to study sample and QC pooled sample tubes. The samples were vortexed on a multi-tube vortexer for 2 min at 5,000 rpm, and centrifuged at room temperature for 4 min at 16,000 rcf. The supernatants were transferred to new labeled tubes and dried on a lyophilizer overnight. Dried samples were reconstituted in 50 µL of 95:5 Water:Methanol, mixed on an analog vortex mixer for 1 min with speed set at 10. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition. UPLC-MS Methods: UPLC-MS spectra were collected for all samples. UPLC was performed on a Waters Acquity UPLC with an Acquity HSS T3 column (2.1x 100mm x 1.8 µm) at 50 °C using the reversed-phase separation. Water with 0.1% formic acid (mobile phase A) and methanol with 0.1% formic acid (mobile phase B) were used as mobile phases and the metabolites were chromatographically separated using a gradient separation: (see the 3. Sheridan-Mice-Hypothalamus_RP-Metadata and Analytical Metadata.xlsx file for the flow gradient). Mass spectroscopy analysis was performed using a Synapt G2 Si ESI-Q-TOF using a 10 µL injection volume. UPLC-MS data were collected over 70-1000 m/z in both positive and negative modes.
Sample Type:brain homogenate

Treatment:

Treatment ID:TR000936
Treatment Summary:3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics.

Sample Preparation:

Sampleprep ID:SP000929
Sampleprep Summary:Brain tissue samples were homogenized in 50:50 Acetonitrile:Water (10 µL to every 1 mg of tissue) with washed ceramic beads on a MagNA Lyser, using two 30 sec pulses at 2,000 rpm with a 1 min chilling step in between pulses. Samples were vortexed on a multi-tube vortexer for 2 min at 5,000 rpm and centrifuged at room temperature for 4 min at 16,000 rcf. A 50 µL aliquot of each experimental brain tissue homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind eppendorf tube. Analytical pooled samples were created by combining 15 µL aliquot from each experimental sample in a 2.0 mL Lo-Bind eppendorf tube. The QC pooled sample was vortexed for 30 sec and 50 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf tubes. Next, 350 µL of acetonitrile was added to study sample and QC pooled sample tubes. The samples were vortexed on a multi-tube vortexer for 2 min at 5,000 rpm, and centrifuged at room temperature for 4 min at 16,000 rcf. The supernatants were transferred to new labeled tubes and dried on a lyophilizer overnight. Dried samples were reconstituted in 50 µL of 95:5 Water:Methanol, mixed on an analog vortex mixer for 1 min with speed set at 10. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition.

Combined analysis:

Analysis ID AN001446 AN001447
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Waters Acquity Waters Acquity
Column Waters ACQUITY UPLC HSS T3 Waters ACQUITY UPLC HSS T3
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Waters Synapt G2 Si QTOF Waters Synapt G2 Si QTOF
Ion Mode POSITIVE NEGATIVE
Units intensity intensity

Chromatography:

Chromatography ID:CH001016
Chromatography Summary:UPLC was performed on a Waters Acquity UPLC with an Acquity HSS T3 column (2.1x 100mm x 1.8 µm) at 50 °C using the reversed-phase separation. Water with 0.1% formic acid (mobile phase A) and methanol with 0.1% formic acid (mobile phase B) were used as mobile phases and the metabolites were chromatographically separated using a gradient separation:
Methods Filename:RTI-RCMRC-RP
Instrument Name:Waters Acquity
Column Name:Waters ACQUITY UPLC HSS T3
Column Temperature:50
Solvent A:100% water; 0.1% formic acid
Solvent B:100% methanol; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS001336
Analysis ID:AN001446
Instrument Name:Waters Synapt G2 Si QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS001337
Analysis ID:AN001447
Instrument Name:Waters Synapt G2 Si QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
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