Summary of Study ST000357

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.

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Study IDST000357
Study TitleBroad Spectrum MS analysis of mouse hypothalmus 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
University of North Carolina
DepartmentDiscovery Sciences
LaboratorySumner Lab
Last NameSumner
First NameSusan
AddressEastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Emailsusan_sumner @unc.edu
Phone704-250-5066
Submit Date2016-03-03
Num Groups4
Total Subjects31
Num Males31
Raw Data AvailableYes
Raw Data File Type(s)raw(Waters)
Analysis Type DetailLC-MS
Release Date2017-03-03
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:SU000378
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
SA015881Total_Pool_04- -
SA015882Total_Pool_02- -
SA015883Total_Pool_01- -
SA015884Total_Pool_05- -
SA015885Total_Pool_03- -
SA015886Hypo388no HF
SA015887Hypo386no HF
SA015888Hypo403no HF
SA015889Hypo401no HF
SA015890Hypo387no HF
SA015891Hypo402no HF
SA015892Hypo385no HF
SA015893Hypo390no LF
SA015894Hypo407no LF
SA015895Hypo392no LF
SA015896Hypo389no LF
SA015897Hypo391no LF
SA015898Hypo405no LF
SA015899Hypo408no LF
SA015900Hypo406no LF
SA015901Hypo411yes HF
SA015902Hypo394yes HF
SA015903Hypo393yes HF
SA015904Hypo396yes HF
SA015905Hypo409yes HF
SA015906Hypo410yes HF
SA015907Hypo412yes HF
SA015908Hypo395yes HF
SA015909Hypo416yes LF
SA015910Hypo399yes LF
SA015911Hypo398yes LF
SA015912Hypo415yes LF
SA015913Hypo413yes LF
SA015914Hypo414yes LF
SA015915Hypo397yes LF
SA015916Hypo400yes LF
Showing results 1 to 36 of 36

Collection:

Collection ID:CO000372
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:TR000392
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:SP000385
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 AN000584 AN000585
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Synapt G2 Si ESI-Q-TOF Synapt G2 Si ESI-Q-TOF
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 Waters Synapt-G2-Si
Ion Mode POSITIVE NEGATIVE
Units intensity intensity

Chromatography:

Chromatography ID:CH000419
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:Synapt G2 Si ESI-Q-TOF
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
  
Chromatography ID:CH000420
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:Synapt G2 Si ESI-Q-TOF
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:MS000520
Analysis ID:AN000584
Instrument Name:Waters Synapt-G2-Si
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000521
Analysis ID:AN000585
Instrument Name:Waters Synapt-G2-Si
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
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