Summary of study ST000848

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 IDST000848
Study TitleTargeting Myelin NEFA of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI
Study SummaryTargeting mouse myelin NEFA of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI. The samples submitted are purified myelin preparations from the postnatal day 21, 60, or 90 mouse spinal cord (SC). There are 9 samples total in Project 1, n=3 for each genotype (control, PAR1 or PAR2) at 21 days. There are 9 samples total in Project 2, n=3 for each genotype (WT, PAR1-/- or PAR2-/-) at 60 days. There are 12 samples total in Project 3, n=3 for K6+/+ or K6-/- at either P21 or P90.
Institute
Mayo Clinic
Last NameScarisbrick
First NameIsobel
Address200 First St. SW, Rochester, Minnesota, 55905, USA
Emailscarisbrick.isobel@mayo.edu
Phone507-284-0124
Submit Date2017-08-09
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:SU000875
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 pt grouping
SA046958ms5959-1P21d CTRL
SA046959ms5959-3P21d CTRL
SA046960ms5959-2P21d CTRL
SA046964ms5959-23P21d K6-/-
SA046965ms5959-22P21d K6-/-
SA046966ms5959-24P21d K6-/-
SA046961ms5959-20P21d K6+/+
SA046962ms5959-21P21d K6+/+
SA046963ms5959-19P21d K6+/+
SA046967ms5959-4P21d PAR1
SA046968ms5959-5P21d PAR1
SA046969ms5959-6P21d PAR1
SA046970ms5959-9P21d PAR2
SA046971ms5959-7P21d PAR2
SA046972ms5959-8P21d PAR2
SA046973ms5959-18P60d PAR1-/-
SA046974ms5959-16P60d PAR1-/-
SA046975ms5959-17P60d PAR1-/-
SA046976ms5959-14P60d PAR2-/-
SA046977ms5959-13P60d PAR2-/-
SA046978ms5959-15P60d PAR2-/-
SA046979ms5959-10P60d Wt
SA046980ms5959-11P60d Wt
SA046981ms5959-12P60d Wt
SA046985ms5959-30P90d K6-/-
SA046986ms5959-29P90d K6-/-
SA046987ms5959-28P90d K6-/-
SA046982ms5959-27P90d K6+/+
SA046983ms5959-25P90d K6+/+
SA046984ms5959-26P90d K6+/+
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Collection:

Collection ID:CO000869
Collection Summary:The samples submitted are purified myelin preparations from the postnatal day 21 or 90 mouse spinal cord (SC). There are two projects but these can be run together since the down stream assays are identical. There are 9 samples total in Project 1, n=3 for each genotype (WT, PAR1-/- or PAR2-/-). There are 12 samples total in Project 2, n=3 for K6+/+ or K6-/- at either P21 or P90. We are submitting these samples for analysis of 1) free fatty acid panel; 2) free fatty acid composition of lipids; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. A total of approximately 130 to 150 ul for each sample is submitted with the protein concentrations already measured by BCA assay and indicated below. Concentrations are provided in ug/ul and the total volume is also indicated. One tube for each sample is submitted but this can be shared across the assays.
Sample Type:Spinal cord

Treatment:

Treatment ID:TR000889
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:SP000882
Sampleprep Summary:"NEFA 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 AN001372
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity
Column Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6460 QQQ
Ion Mode NEGATIVE
Units ng/ug of protein

Chromatography:

Chromatography ID:CH000957
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)
Chromatography Type:Reversed phase

MS:

MS ID:MS001264
Analysis ID:AN001372
Instrument Name:Agilent 6460 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
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