Summary of Study ST000359

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 IDST000359
Study TitleBroad Spectrum MS analysis of mouse microglia cells from Anxiety Prone HSV-Latently Infected Obese Mice
Study TypeBroad Spectrum LCMS
Study SummaryA mouse model of obese HSV-1 latent infection was developed. Broad spectrum metabolomics analysis was performed to better understand the metabolomic profile of microglia cells and to compare this metabolomics profile with that of the hippocampus, hypothalamus, and peripheral blood mononuclear cells. Microglia cell 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 microglia cell 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 Date2016-03-05
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

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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:SU000380
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
SA015953Total_Pool_04- -
SA015954Total_Pool_05- -
SA015955Total_Pool_03- -
SA015956Total_Pool_01- -
SA015957Total_Pool_02- -
SA015962Micro418no HF
SA015963Micro420no HF
SA015964Micro419no HF
SA015965Micro417no HF
SA015958Micro446No HF
SA015959Micro445No HF
SA015960Micro443No HF
SA015961Micro444No HF
SA015966Micro421no LF
SA015967Micro434no LF
SA015968Micro431no LF
SA015969Micro423no LF
SA015970Micro422no LF
SA015971Micro432no LF
SA015972Micro424no LF
SA015973Micro433no LF
SA015974Micro426yes HF
SA015975Micro442yes HF
SA015976Micro425yes HF
SA015977Micro439yes HF
SA015978Micro441yes HF
SA015979Micro427yes HF
SA015980Micro440yes HF
SA015981Micro438yes LF
SA015982Micro435yes LF
SA015983Micro430yes LF
SA015984Micro436yes LF
SA015985Micro429yes LF
SA015986Micro437yes LF
SA015987Micro428yes LF
Showing results 1 to 35 of 35

Collection:

Collection ID:CO000374
Collection Summary:For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser ceramic beads were added to the tubes, and a MagNA lyser was used to beat the samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for 5 min in between pulses. Sonication for 5 min was performed to the beaten cell samples, followed by centrifugation at room temperature and at 16,000rcf for 4 min. A 350 µL aliquot of each experimental cell sample homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube
Sample Type:brain homogenate

Treatment:

Treatment ID:TR000394
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:SP000387
Sampleprep Summary:For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser ceramic beads were added to the tubes, and a MagNA lyser was used to beat the samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for 5 min in between pulses. Sonication for 5 min was performed to the beaten cell samples, followed by centrifugation at room temperature and at 16,000rcf for 4 min. A 350 µL aliquot of each experimental cell sample homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube Analytical pooled samples were created by combining a 125 µL aliquot from each experimental sample in a labeled 20 mL glass vial. The QC pooled sample was vortexed for 30 sec and 350 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf tubes. Next, the supernatants of the study and QC pooled samples were dried overnight using a lyophilizer. The residue was reconstituted in 50 µL of reconstitution buffer (95:5 Water:Methanol, v/v) and 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 AN000588 AN000589
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:CH000422
Chromatography Summary: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-Microglia Cells_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.
Methods Filename:RTI-RCMRC-RP
Instrument Name:Synapt G2 Si ESI-Q-TOF
Column Name:Waters ACQUITY UPLC HSS T3
Flow Rate:.4ml/min
Injection Temperature:8 C
Solvent A:100% water; 0.1% formic acid
Solvent B:100% methanol; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS000524
Analysis ID:AN000588
Instrument Name:Waters Synapt-G2-Si
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
Analysis Protocol File:RTI-RCMRC-RP-POS
  
MS ID:MS000525
Analysis ID:AN000589
Instrument Name:Waters Synapt-G2-Si
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
Analysis Protocol File:RTI-RCMRC-RP-NEG
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