Summary of Study ST000862

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 PR000575. The data can be accessed directly via it's Project DOI: 10.21228/M8BM2Z This work is supported by NIH grant, U2C- DK119886.

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Study IDST000862
Study TitleTargeting Myelin FFA Compostion in PAR1 and PAR2 Mice after SCI
Study SummaryTargeting Myelin FFA composition in PAR1 and PAR2 Mice after SCI. The samples submitted are purified myelin preparations from the postnatal day 21 or 60 mouse spinal cord (SC). There are 30 total sampes, n=5 for each group. wild type control, PAR1 KO and PAR2 KO at P21 and P60 days.
Institute
Mayo Clinic
Last NameScarisbrick
First NameIsobel
Address200 First Street SW, Rochester, MN 55905
Emailscarisbrick.isobel@mayo.edu
Phone507-284-0124
Submit Date2017-08-10
Analysis Type DetailLC-MS
Release Date2021-02-17
Release Version1
Isobel Scarisbrick Isobel Scarisbrick
https://dx.doi.org/10.21228/M8BM2Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000575
Project DOI:doi: 10.21228/M8BM2Z
Project Title:Mayo Pilot and Feasibility: Targeting Myelin Metabolism to Enhance Recovery of Function after SCI
Project Summary:The loss of myelin, degeneration of the myelin producing oligodendroglia and impaired remyelination are essential features of traumatic spinal cord injury (SCI) that significantly limit patient recovery of function. The lipid rich composition of myelin, including exceptionally high levels of saturated fatty acids, underlie its essential physiological roles, including its structural and signaling properties and electrical insulation of axons to facilitate the conduction of nerve impulses. The myelin sheaths also provide metabolic support to the axons they wrap, and myelin health is therefore essential to the maintenance of axon integrity and function in the brain and spinal cord. The primary goal of this Pilot Proposal to the Mayo Clinic Metabolomics Core is to integrate highly sensitive metabolomics liquid chromatography-tandem mass spectrometry (LC/MS/MS) assays to quantify the lipid composition of the myelin membrane, with our conventional neurobehavioral approaches, enabling us to explore the metabolic basis of new interventions capable of promoting myelin regeneration and restoration of patient function. Metabolomics Core expertise in Magnetic Resonance Spectroscopy (NMR) based evaluation of key metabolites involved in CNS injury and repair (N-acetyl-L-aspartate, choline, myo-inositol, glucose/ glutamine and lactate) will also be applied to strengthen our mechanistic understanding of myelin injury and repair. Specifically, utilizing these innovative approaches we will test a novel hypothesis driven by new preliminary findings that the levels of dietary fatty acids can be optimized alone, or in combination with exercise training, to facilitate myelin regeneration and recovery of neurobehavioral function after injury to the adult spinal cord. In Aim 1, we will determine whether alterations in dietary fat, including saturated fat or omega-3 fatty acids, facilitate restoration of the myelin membrane and metabolite signatures of central nervous system repair after experimental SCI in adult mice. In Aim 2, we will determine whether exercise training alone or in combination with dietary fatty acid supplementation fosters myelin regeneration and recovery of function after experimental SCI. The proposed studies will leverage the expertise of the Mayo Metabolomics Core with that of Dr. Scarisbrick (Mayo) in myelin biology and Dr. Gomez Pinilla (UCLA) in central nervous system plasticity to investigate whether two highly targetable lifestyle variables, that is diet and exercise, can be modulated to improve myelin metabolism and functional recovery after SCI.
Institute:Mayo Clinic
Last Name:Scarisbrick
First Name:Isobel
Address:200 First St. SW, Rochester, Minnesota, 55905, USA
Email:scarisbrick.isobel@mayo.edu
Phone:507-284-0124

Subject:

Subject ID:SU000889
Subject Type:Mouse
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammal

Factors:

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

mb_sample_id local_sample_id time group
SA047678ms6119-18P21 CTRL
SA047679ms6119-19P21 CTRL
SA047680ms6119-20P21 CTRL
SA047681ms6119-17P21 CTRL
SA047682ms6119-16P21 CTRL
SA047683ms6119-24P21 PAR1
SA047684ms6119-25P21 PAR1
SA047685ms6119-23P21 PAR1
SA047686ms6119-22P21 PAR1
SA047687ms6119-21P21 PAR1
SA047688ms6119-26P21 PAR2
SA047689ms6119-27P21 PAR2
SA047690ms6119-30P21 PAR2
SA047691ms6119-29P21 PAR2
SA047692ms6119-28P21 PAR2
SA047693ms6119-2P60 CTRL
SA047694ms6119-3P60 CTRL
SA047695ms6119-4P60 CTRL
SA047696ms6119-5P60 CTRL
SA047697ms6119-1P60 CTRL
SA047698ms6119-9P60 PAR1
SA047699ms6119-10P60 PAR1
SA047700ms6119-7P60 PAR1
SA047701ms6119-8P60 PAR1
SA047702ms6119-6P60 PAR1
SA047703ms6119-11P60 PAR2
SA047704ms6119-14P60 PAR2
SA047705ms6119-15P60 PAR2
SA047706ms6119-13P60 PAR2
SA047707ms6119-12P60 PAR2
Showing results 1 to 30 of 30

Collection:

Collection ID:CO000883
Collection Summary:This prior project involved frozen whole mouse spinal cord (SC) samples for Untargeted Profiling (unbiased metabolomics assay). There were 9 samples total, n=3 for each genotype that includes wild type control, PAR1 KO and PAR2 KO at P60 days. Your team prepared the samples and ran the profiling and provided us with the results. Since the outcome of this preliminary study using 3 animals for each genotype looked so promising, we discussed bringing the total number to 5 and re-running to improve the power. Therefore, we are submitting 2 more samples for each genotype at P60. In addition, we are providing 5 samples for each genotype at P21 (an earlier age) for additional comparisons. To bring n=5 for each P60 genotype we provide 2 more samples per genotype. In addition we provide n=5 for P21 for each genotype (wild type, PAR1 and PAR2).
Sample Type:Spinal cord

Treatment:

Treatment ID:TR000903
Treatment Summary:A 3g Clip produces moderate SCI including demyelination and clinical impairment and we recently published a detailed methodology. At 1 week after injury, the 3g injured mice are expected to have an average Basso Mouse Scale score (BMS)=5 on a 9 point scale such that they have frequent plantar stepping with no or some coordination. This level of impairment was chosen to provide a sufficient window to observe recovery.

Sample Preparation:

Sampleprep ID:SP000896
Sampleprep Summary:FFA composition of mouse spinal cord Lipids will be quantified in myelin isolated in high yield and purity by subcellular fractionation from the lumbosacral spinal cord. While there are no absolutely ‘myelin-specific’ lipids, galactocerebroside is the most typical of myelin in the adult nervous system being directly proportional to the amount of myelin. Sulfatide is another galactolipid enriched in myelin. Together with cholesterol, these form 78% of the total amount of lipid in the myelin membrane and each will be quantified using LC/MS/MS. A highly sensitive assay for galactocerebroside was recently established by the Mayo Metabolomics Core and can be implemented immediately. The LC/MS/MS panel for free fatty acids, including the very long chain fatty acids found in myelin is also routinely performed by the Core. Cholesterol will be quantified using an NMR-based approach by the Mayo Dept. of Laboratory Medicine Clinical Core. Additionally, we have a plan in place with the Metabolomics Core to develop LC/MS/MS assays for sulfatide and sphingomyelin during the Pilot proposal. Having quantitative assays for each of these key myelin lipids will facilitate our goal to comprehensively profile myelin lipid metabolism and will form foundational assays for a future NIH grant focused on myelin metabolism.

Combined analysis:

Analysis ID AN001387
Analysis type MS
Chromatography type HILIC
Chromatography system Cohesive TX2
Column Altma HP HILIC (150 x 2.1mm,5um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex 6500 QTrap
Ion Mode NEGATIVE
Units % composition by nmol

Chromatography:

Chromatography ID:CH000971
Instrument Name:Cohesive TX2
Column Name:Altma HP HILIC (150 x 2.1mm,5um)
Chromatography Type:HILIC

MS:

MS ID:MS001279
Analysis ID:AN001387
Instrument Name:ABI Sciex 6500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
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