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MB Sample ID: SA273548
| Local Sample ID: | 5B |
| Subject ID: | SU002819 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002819 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 5B | SA273548 | FL035355 | F | sex |
| 5B | SA273548 | FL035355 | WT | genetic status |
Collection:
| Collection ID: | CO002812 |
| Collection Summary: | Mice were bred and maintained in the animal facility of the University of Lausanne. All animal experiments were authorized by “Service de la consommation et des Affaires vétérinaires” (SCAV) of the Canton de Vaud in Switzerland. Cx3cr1CREERT2;MCT4flox mice were obtained by crossing the B6.129P2(Cg)-Cx3cr1tm2.1(cre/ERT2)Litt/WganJ mice (Cx3cr1CREERT2; No: 021160, The Jackson Laboratory) with C57Bl/6.MCT4tm1flox mice (MCT4flox; kindly donated by Prof. Luc Pellerin). For specific experiments, the Cx3cr1CREERT2;MCT4flox line was further crossed with B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J mice (TdTomatoflox; No: 007914, The Jackson Laboratory). Mice were group-housed and kept in a 12 h day/night cycle, with food and water ad libitum, at 20-22°C. All the experiments were performed during the day cycle. Both male and female mice were analyzed, unless differently specified. For in vivo gene KO induction, tamoxifen (Sigma Aldrich) was prepared in 10% ethanol in corn oil at a concentration of 6mg/ml. The whole litter was injected intraperitoneally (i.p.) at P6 and P8 (75mg/kg). For biochemical and histological assessments, mice at P15 were terminally anesthetized with sodium pentobarbital diluted in saline (150mg/kg, i.p.) and perfused with cold HBSS (Life Technologies, 3ml/min). Upon brain collection, the hippocampus was dissected from the right hemisphere, immediately frozen in dry ice and stored at -80°C. |
| Sample Type: | Brain |
Treatment:
| Treatment ID: | TR002828 |
| Treatment Summary: | For in vivo gene KO induction, tamoxifen (Sigma Aldrich) was prepared in 10% ethanol in corn oil at a concentration of 6mg/ml. The whole litter was injected intraperitoneally (i.p.) at P6 and P8 (75mg/kg). |
Sample Preparation:
| Sampleprep ID: | SP002825 |
| Sampleprep Summary: | Frozen tissue samples were weighed to the nearest 0.1 mg then treated with cold 5:3:2 MeOH:acetonitrile:water to a final concentration of 15 mg/mL. Samples were vortexed 30 min at 4 degrees C then supernatants clarified by centrifugation (10 min, 18,000 g, 4 degrees C) and transferred to autosampler vials. |
| Processing Storage Conditions: | 4℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN004399 | AN004400 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH003301 |
| Chromatography Summary: | Negative C18 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min 100-0% B, 3-5 min hold at 0% B |
| Flow Rate: | 450 uL/min |
| Sample Injection: | 10 uL |
| Solvent A: | 95% water 5% acetonitrile 1 mM ammonium acetate |
| Solvent B: | 95% acetonitrile 5% water 1 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003302 |
| Chromatography Summary: | Positive C18 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min 95-5% B, 3-5 min hold at 5% B |
| Flow Rate: | 450 uL/min |
| Sample Injection: | 10 uL |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100%acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004148 |
| Analysis ID: | AN004399 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004149 |
| Analysis ID: | AN004400 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database. |
| Ion Mode: | POSITIVE |
