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.

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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.

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Study IDST002714
Study TitleLoss of microglial MCT4 leads to defective synaptic pruning and anxiety-like behavior in mice
Study SummaryMicroglia, 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 NameHaines
First NameJulie
Address12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Emailjulie.haines@cuanschutz.edu
Phone3037243339
Submit Date2023-05-24
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2023-06-22
Release Version1
Julie Haines Julie Haines
https://dx.doi.org/10.21228/M8914J
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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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
SA2735427F KO
SA27354312F KO
SA27354415F KO
SA2735456F KO
SA27354611F KO
SA2735474BF WT
SA2735485BF WT
SA2735499F WT
SA2735502F WT
SA2735513BM KO
SA2735521BM KO
SA27355314M KO
SA27355410M KO
SA2735554M KO
SA2735561M KO
SA2735578M WT
SA2735582BM WT
SA2735593M WT
SA27356013M WT
SA2735615M 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
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