Summary of Study ST000850
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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000850 |
| Study Title | Targeting Myelin Cholesterol of Kallikrein 6 Signals through PAR1 and PAR2 to Enchance Recovery of Function after SCI |
| Study Summary | Targeting mouse myelin Cholesterol 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 Name | Scarisbrick |
| First Name | Isobel |
| Address | 200 First St. SW, Rochester, Minnesota, 55905, USA |
| scarisbrick.isobel@mayo.edu | |
| Phone | 507-284-0124 |
| Submit Date | 2017-08-09 |
| Analysis Type Detail | GC-MS |
| Release Date | 2021-02-17 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
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: | SU000877 |
| 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 |
|---|---|---|---|
| SA047018 | 211-15 | P21d | CTRL |
| SA047019 | 214-15 | P21d | CTRL |
| SA047020 | 212-15 | P21d | CTRL |
| SA047021 | 372-15 | P21d | K6+/+ |
| SA047022 | 373-15 | P21d | K6+/+ |
| SA047023 | 219-15 | P21d | K6+/+ |
| SA047024 | 317-15 | P21d | K6-/- |
| SA047025 | 316-15 | P21d | K6-/- |
| SA047026 | 318-15 | P21d | K6-/- |
| SA047027 | 202-15 | P21d | PAR1 |
| SA047028 | 228-15 | P21d | PAR1 |
| SA047029 | 267-15 | P21d | PAR1 |
| SA047030 | 281-15 | P21d | PAR2 |
| SA047031 | 264-15 | P21d | PAR2 |
| SA047032 | 266-15 | P21d | PAR2 |
| SA047033 | 250-15 | P60d | PAR1-/- |
| SA047034 | 248-15 | P60d | PAR1-/- |
| SA047035 | 249-15 | P60d | PAR1-/- |
| SA047036 | 184-15 | P60d | PAR2-/- |
| SA047037 | 183-15 | P60d | PAR2-/- |
| SA047038 | 185-15 | P60d | PAR2-/- |
| SA047039 | 270-15 | P60d | Wt |
| SA047040 | 271-15 | P60d | Wt |
| SA047041 | 272-15 | P60d | Wt |
| SA047042 | 369-15 | P90d | K6+/+ |
| SA047043 | 352-15 | P90d | K6+/+ |
| SA047044 | 353-15 | P90d | K6+/+ |
| SA047045 | 130-15 | P90d | K6-/- |
| SA047046 | 246-15 | P90d | K6-/- |
| SA047047 | 245-15 | P90d | K6-/- |
| Showing results 1 to 30 of 30 |
Collection:
| Collection ID: | CO000871 |
| 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: | TR000891 |
| 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: | SP000884 |
| Sampleprep Summary: | Cholesterol 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 | AN001374 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890A |
| Column | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Agilent 5975C |
| Ion Mode | POSITIVE |
| Units | ug/ul of protein |
Chromatography:
| Chromatography ID: | CH000959 |
| Instrument Name: | Agilent 7890A |
| Column Name: | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS001266 |
| Analysis ID: | AN001374 |
| Instrument Name: | Agilent 5975C |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
