Return to study ST000792 main page
MB Sample ID: SA043667
| Local Sample ID: | nC18-2jun16-019-r001 |
| Subject ID: | SU000817 |
| Subject Type: | Mouse |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000817 |
| Subject Type: | Mouse |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| nC18-2jun16-019-r001 | SA043667 | FL008963 | HFHS | Group |
| nC18-2jun16-019-r001 | SA043667 | FL008963 | 1 | rep |
Collection:
| Collection ID: | CO000811 |
| Collection Summary: | Uninjured or SCI mice will be randomly assigned to one of four groups, LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet). All diets will be obtained from Research Diets, NJ USA43,45. Dietary fat supplementation will be initiated at 1 week after SCI. This time point for intervention was chosen to provide a meaningful timeframe for clinical translation. Also, a 1 week period will allow time for mice to recover prior to providing access to wheel running (Aim 2). The impact of dietary fat supplementation on myelin metabolism will be examined after a period of 7 weeks, including determination of (i) the lipid profile of the myelin membrane using LC/MS/MS; and (ii) metabolic markers of spinal cord metabolism by NMR. Results will be correlated with (iii) cellular and molecular markers of spinal cord pathophysiology including the appearance of OPCs, oligodendroglia and myelin, axon health, astrogliosis and inflammation; and (iv) the extent of sensorimotor recovery. (v) In addition, to gauge the impact of the dietary fat on systemic metabolic status, Insulin and Glucose Resistance Tests will be performed at 7 weeks. Food intake (g/day) and body weight gain (% initial weight) will be measured daily until the endpoint of each experiment. Mice will be housed individually in a temperature-controlled facility with a 12:12-h light-dark cycle and ad libitum access to each diet and water. A Power Analysis was performed based on histological outcomes in mice with contusion compression injury. To detect a difference between groups of 20%, which would very meaningful, a group size of 8 will be needed to achieve a power of 0.85. An additional 2 mice per group has been added to account for mortality. The Mayo Clinic Institutional Animal Care and Use Committee has approved of the proposed studies. |
| Sample Type: | Spinal cord |
Treatment:
| Treatment ID: | TR000831 |
| Treatment Summary: | To test the hypothesis that optimizing dietary fat will facilitate myelin repair after SCI, the diet of uninjured adult female C57BL6/J mice (12 week, 22-25g, Jackson), or those with experimental contusion-compression SCI of the lumbosacral spinal cord (L2-L3) (Fejota Clip 3g Force, applied for 30s)32,47 will be supplemented with saturated fat. The 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, and to be at a level compatible with examination of exercise training by wheel running (Aim 2). |
Sample Preparation:
| Sampleprep ID: | SP000824 |
| Sampleprep Summary: | large scale profiling 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 | AN001260 | AN001261 | AN001262 | AN001263 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | HILIC | HILIC |
| Chromatography system | Agilent 1290 Infinity | Agilent 1290 Infinity | Agilent 1290 Infinity | Agilent 1290 Infinity |
| Column | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) | Waters Acquity HSS C18 (150 x 2.1mm,1.8um) | Waters Acquity HSS C18 (150 x 2.1mm,1.8um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | QTOF | QTOF | QTOF | QTOF |
| MS instrument name | Agilent 6550 QTOF | Agilent 6550 QTOF | Agilent 6550 QTOF | Agilent 6550 QTOF |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | intensity | intensity | intensity | intensity |
Chromatography:
| Chromatography ID: | CH000878 |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH000879 |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Waters Acquity HSS C18 (150 x 2.1mm,1.8um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS001153 |
| Analysis ID: | AN001260 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| MS ID: | MS001154 |
| Analysis ID: | AN001261 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| MS ID: | MS001155 |
| Analysis ID: | AN001262 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| MS ID: | MS001156 |
| Analysis ID: | AN001263 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
