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MB Sample ID: SA318230
| Local Sample ID: | bs_r4 |
| Subject ID: | SU003045 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 20 wks |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Lab |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003045 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 20 wks |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Lab |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| bs_r4 | SA318230 | FL037869 | brainstem | Factor |
Collection:
| Collection ID: | CO003038 |
| Collection Summary: | Extraction of fecal metabolites: 50 mg of fresh feces were collected from each mouse and frozen in liquid nitrogen. Samples were spiked with 0.5 mL water, homogenized for 5 min followed by 10 s of sonication, and centrifuged at 14K rpm X 10 min at 4°C. Collected supernatants were evaporated to dryness under a nitrogen gas stream and stored at −80°C. Extraction of brain metabolites: Approximately 20-40 mg of frozen tissues was resuspended in 4 volumes of MPER and homogenized on ice. 130 µL of tissue homogenates were spin filtered, as above, and the flow through was diluted 1:1 with deionized water, followed by sonication for 10 sec using a Sonic Dismembrator on ice, vortexed for 10 s, and centrifuged at 14K rpm X 15 min at 4°C. 100 uL of eluate was evaporated to dryness under a nitrogen gas stream and stored at −80°C. |
| Sample Type: | feces, hippocampus, brainstem |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003054 |
| Treatment Summary: | To evaluate our hypothesis that running exercise reshapes gut microbiota diversity that balances TRP metabolism in the gut, we used an established model of voluntary running exercise in 20-week-old male mice (C57BL/6J, Jackson Lab) housed individually in a standard cage in temperature-controlled (21°C) quarters with a 12-h light/12-h dark cycle. Animals were given water and food (Purina Chow) ad libitum as previously described (Chorna et al., 2013). Briefly, mice were randomized into two groups, sedentary control (n=12) and running experimental (n=12), housed with free access to a wireless running wheel (Med Associates) for six weeks. For the sedentary group, the wheels were locked in the experiment. Therefore, this group of mice could not perform running exercises. Exercise activities of the running group were recorded for each animal for the investigation to ensure that each mouse was physically active. The recording was conducted using an automatic counter and Med Associates software. |
Sample Preparation:
| Sampleprep ID: | SP003051 |
| Sampleprep Summary: | Extraction of fecal metabolites: 50 mg of fresh feces were collected from each mouse and frozen in liquid nitrogen. Samples were spiked with 0.5 mL water, homogenized for 5 min followed by 10 s of sonication, and centrifuged at 14K rpm X 10 min at 4°C. Collected supernatants were evaporated to dryness under a nitrogen gas stream and stored at −80°C. Extraction of brain metabolites: Approximately 20-40 mg of frozen tissues was resuspended in 4 volumes of MPER and homogenized on ice. 130 µL of tissue homogenates were spin filtered, as above, and the flow through was diluted 1:1 with deionized water, followed by sonication for 10 sec using a Sonic Dismembrator on ice, vortexed for 10 s, and centrifuged at 14K rpm X 15 min at 4°C. 100 uL of eluate was evaporated to dryness under a nitrogen gas stream and stored at −80°C. |
| Processing Storage Conditions: | Described in summary |
Combined analysis:
| Analysis ID | AN004810 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Synapt-XS Waters |
| Column | Waters ACQUITY UPLC BEH HILIC (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Ion Mobility TOF |
| MS instrument name | Waters Synapt-XS |
| Ion Mode | POSITIVE |
| Units | peak intensities |
Chromatography:
| Chromatography ID: | CH003635 |
| Chromatography Summary: | The LC/MS experiments were conducted using an i-Class LC system coupled to a Synapt-XS mass spectrometer (Waters). Liquid chromatography was performed on fecal, hippocampus, and brainstem samples collected from both the SED and RUN groups. The chromatographic separation was conducted using a BEH HILIC column with 1.7 µm particle size dimensions and 2.1 x 100 mm (Waters). The column temperature was maintained at 50ºC, and the flow rate was set at 0.4 mL/min. The mobile phase consisted of 0.1% formic acid in water as solvent A and 0.1% formic acid in acetonitrile as solvent B. The initial mobile phase composition was 5% A and 95% B, which was held constant for 2 minutes. Subsequently, a linear gradient from 5% to 50% A was applied over 8 minutes, followed by a 2-minute hold at 50% B before returning to the initial condition for a 5-minute re-equilibration. The total run time for each analysis was 19 minutes. |
| Instrument Name: | Synapt-XS Waters |
| Column Name: | Waters ACQUITY UPLC BEH HILIC (100 x 2.1mm,1.7um) |
| Column Temperature: | 50ºC |
| Flow Gradient: | 5% to 50% A was applied over 8 minutes, followed by a 2-minute hold at 50% B before returning to the initial condition for a 5-minute re-equilibration. |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004556 |
| Analysis ID: | AN004810 |
| Instrument Name: | Waters Synapt-XS |
| Instrument Type: | Ion Mobility TOF |
| MS Type: | ESI |
| MS Comments: | The mass spectrometer operated at a frequency of 5 Hz, covering a mass range of 50-1200 m/z. |
| Ion Mode: | POSITIVE |
