Summary of Study ST002714
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 PR001682. The data can be accessed directly via it's Project DOI: 10.21228/M8914J This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002714 |
| Study Title | Loss of microglial MCT4 leads to defective synaptic pruning and anxiety-like behavior in mice |
| Study Summary | Microglia, the innate immune cells of the central nervous system, actively participate in brain development by supporting neuronal maturation and refining synaptic connections. These cells are emerging as highly metabolically flexible, able to oxidize different energetic substrates to meet their energy demand. Lactate is particularly abundant in the brain, but whether microglia use it as a metabolic fuel has been poorly explored. Here we show that microglia can import lactate, and this is coupled with increased lysosomal acidification. In vitro, loss of the monocarboxylate transporter MCT4 in microglia prevents lactate-induced lysosomal modulation and leads to defective cargo degradation. Microglial depletion of MCT4 in vivo leads to impaired synaptic pruning, associated with increased excitation in hippocampal neurons, enhanced E/I ratio, vulnerability to seizures and anxiety-like phenotype. Overall, these findings show that selective disruption of the MCT4 transporter in microglia is sufficient to alter synapse refinement and to induce defects in brain development and adult behavior. |
| Institute | University of Colorado Denver |
| Last Name | Haines |
| First Name | Julie |
| Address | 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA |
| julie.haines@cuanschutz.edu | |
| Phone | 3037243339 |
| Submit Date | 2023-05-24 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-06-22 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001682 |
| Project DOI: | doi: 10.21228/M8914J |
| Project Title: | Loss of microglial MCT4 leads to defective synaptic pruning and anxiety-like behavior in mice |
| Project Summary: | Microglia, the innate immune cells of the central nervous system, actively participate in brain development by supporting neuronal maturation and refining synaptic connections. These cells are emerging as highly metabolically flexible, able to oxidize different energetic substrates to meet their energy demand. Lactate is particularly abundant in the brain, but whether microglia use it as a metabolic fuel has been poorly explored. Here we show that microglia can import lactate, and this is coupled with increased lysosomal acidification. In vitro, loss of the monocarboxylate transporter MCT4 in microglia prevents lactate-induced lysosomal modulation and leads to defective cargo degradation. Microglial depletion of MCT4 in vivo leads to impaired synaptic pruning, associated with increased excitation in hippocampal neurons, enhanced E/I ratio, vulnerability to seizures and anxiety-like phenotype. Overall, these findings show that selective disruption of the MCT4 transporter in microglia is sufficient to alter synapse refinement and to induce defects in brain development and adult behavior. |
| Institute: | University of Colorado Denver |
| Laboratory: | Lab of Angelo D'Alessandro in collaboration with lab of Rosa Paolicelli (Univ of Lausanne) |
| Last Name: | Haines |
| First Name: | Julie |
| Address: | 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA |
| Email: | julie.haines@cuanschutz.edu |
| Phone: | 3037243339 |
Subject:
| Subject ID: | SU002819 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | sex | genetic status |
|---|---|---|---|
| SA273542 | 7 | F | KO |
| SA273543 | 12 | F | KO |
| SA273544 | 15 | F | KO |
| SA273545 | 6 | F | KO |
| SA273546 | 11 | F | KO |
| SA273547 | 4B | F | WT |
| SA273548 | 5B | F | WT |
| SA273549 | 9 | F | WT |
| SA273550 | 2 | F | WT |
| SA273551 | 3B | M | KO |
| SA273552 | 1B | M | KO |
| SA273553 | 14 | M | KO |
| SA273554 | 10 | M | KO |
| SA273555 | 4 | M | KO |
| SA273556 | 1 | M | KO |
| SA273557 | 8 | M | WT |
| SA273558 | 2B | M | WT |
| SA273559 | 3 | M | WT |
| SA273560 | 13 | M | WT |
| SA273561 | 5 | M | WT |
| Showing results 1 to 20 of 20 |
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 |
