Summary of Study ST003249
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 PR002017. The data can be accessed directly via it's Project DOI: 10.21228/M8WJ8H 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 | ST003249 |
| Study Title | Mitochondrial respiration impairment in microglia dampens response to demyelinating injury but is not sufficient to induce an aging phenotype |
| Study Summary | Microglia are necessary for CNS function during development and play roles in aging, Alzheimer’s Disease (AD) and the response to demyelinating injury1–5. Mitochondrial respiratory chain (RC) controls macrophage-dependent immune responses6–9. However, whether mitochondrial RC is essential to microglia function is not known. We conditionally deleted the mitochondrial complex III subunit Rieske Iron-Sulfur Protein (RISP) in the microglia of adult mice to assess the requirement of microglial RC for survival, proliferation, and adult CNS function in vivo. Surprisingly, mitochondrial RC function was not required for survival or proliferation of microglia in vivo. RNA-seq analysis showed that loss of RC function in microglia caused changes in gene expression distinct from aged or disease-associated microglia (DAM). Microglia-specific loss of mitochondrial RC function did not affect cognitive decline during aging or in the 5xFAD model of Alzheimer’s disease (AD). However, Abeta plaque coverage decreased and microglial interaction with Abeta plaques increased in the hippocampus of 5xFAD mice with mitochondrial RC-deficient microglia. Microglia-specific loss of mitochondrial RC function did impair remyelination following an acute, reversible demyelinating event. Thus, mitochondrial respiration in microglia is dispensable for maintenance of normal cognitive function but is essential to maintain a proper response to CNS demyelinating injury. |
| Institute | Northwestern University |
| Last Name | Stoolman |
| First Name | Joshua |
| Address | 303 E Superior Street, |
| joshua.stoolman@northwestern.edu | |
| Phone | 7343559440 |
| Submit Date | 2024-01-19 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-06-11 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002017 |
| Project DOI: | doi: 10.21228/M8WJ8H |
| Project Title: | Mitochondrial respiration in microglia is essential for response to demyelinating injury but not proliferation. |
| Project Summary: | Microglia are necessary for CNS function during development and play roles in aging, Alzheimer’s Disease (AD) and the response to demyelinating injury. Mitochondrial respiratory chain (RC) controls macrophage-dependent immune responses. However, whether mitochondrial RC is essential to microglia function is not known. We conditionally deleted the mitochondrial complex III subunit Rieske Iron-Sulfur Protein (RISP) in the microglia of adult mice to assess the requirement of microglial RC for survival, proliferation, and adult CNS function in vivo. Surprisingly, mitochondrial RC function was not required for survival or proliferation of microglia in vivo. RNA-seq analysis showed that loss of RC function in microglia caused changes in gene expression distinct from aged or disease-associated microglia (DAM). Microglia-specific loss of mitochondrial RC function did not affect cognitive decline during aging or in the 5xFAD model of Alzheimer’s disease (AD). However, Abeta plaque coverage decreased and microglial interaction with Abeta plaques increased in the hippocampus of 5xFAD mice with mitochondrial RC-deficient microglia. Microglia-specific loss of mitochondrial RC function did impair remyelination following an acute, reversible demyelinating event. Thus, mitochondrial respiration in microglia is dispensable for maintenance of normal cognitive function but is essential to maintain a proper response to CNS demyelinating injury. |
| Institute: | Northwestern University Feinberg School of Medicine |
| Last Name: | Stoolman |
| First Name: | Joshua |
| Address: | 303 E Superior Street, |
| Email: | joshua.stoolman@northwestern.edu |
| Phone: | 7343559440 |
Subject:
| Subject ID: | SU003368 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 1 year |
| Gender: | Male and female |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Sample source |
|---|---|---|---|
| SA353485 | Nav-Jos-20220209-02 | HET | Microglia |
| SA353486 | Nav-Jos-20220209-04 | HET | Microglia |
| SA353487 | Nav-Jos-20220209-07 | HET | Microglia |
| SA353488 | Nav-Jos-20220209-09 | HET | Microglia |
| SA353489 | Nav-Jos-20220209-19 | HET | Microglia |
| SA353490 | Nav-Jos-20220209-22 | HET | Microglia |
| SA353491 | Nav-Jos-20220209-03 | KO | Microglia |
| SA353492 | Nav-Jos-20220209-05 | KO | Microglia |
| SA353493 | Nav-Jos-20220209-08 | KO | Microglia |
| SA353494 | Nav-Jos-20220209-10 | KO | Microglia |
| SA353495 | Nav-Jos-20220209-20 | KO | Microglia |
| SA353496 | Nav-Jos-20220209-21 | KO | Microglia |
| SA353497 | Nav-Jos-20220209-01 | N/A | Plasma |
| SA353498 | Nav-Jos-20220209-06 | N/A | Plasma |
| SA353499 | Nav-Jos-20220209-16 | N/A | Plasma |
| Showing results 1 to 15 of 15 |
Collection:
| Collection ID: | CO003361 |
| Collection Summary: | Following CNS isolation, 100,000 YFP+TdTomato+ cells were sorted at 4°C on the MACS Tyto, centrifuged at 500xg, resuspended in 30μL (3,333 cells/µL) 80% acetonitrile in HPLC-grade H2O, vortexed vigorously for 1 minute and then quickly frozen on dry ice. The samples were stored at -80°C for a minimum of 12 hours prior to performing 3 additional freeze thaw cycles with a warm water bath and liquid nitrogen. Samples were then centrifuged at 16000xg for 30 minutes at 4°C. Supernatants were transferred to a fresh tube for analysis. |
| Sample Type: | Microglia |
Treatment:
| Treatment ID: | TR003377 |
| Treatment Summary: | Cx3CR1-creERT2 with RISP (Fx/Fx) or RISP (Fx/Wt) genotype were given tamoxifen chow for 3 weeks starting at 8 weeks of age to generate microglia-targeted KO of mitochondrial complex III subunit RISP. At 1 yr of age microglia from these mice were isolated for untargeted metabolomics analysis. |
Sample Preparation:
| Sampleprep ID: | SP003375 |
| Sampleprep Summary: | A 15μl aliquot of the sample was used for high-resolution HPLC-tandem mass spectrometry. High-resolution HPLC-tandem mass spectrometry was performed on a Q-Exactive (ThermoFisher Scientific) in line with an electrospray source and an UltiMate 3000 (ThermoFisher Scientific) series HPLC consisting of a binary pump, degasser and autosampler outfitted with a Xbridge Amide column (Waters; 4.6 mm × 100 mm dimension and a 3.5 μm particle size). Mobile phase A contained water and acetonitrile (95/5, v/v), 10 mM ammonium hydroxide and 10 mM ammonium acetate (pH 9.0). Mobile phase B was 100% acetonitrile. The gradient was set to 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1–18 min, 75% A; 18–25 min, 15% A, with a flow rate of 400 μl min–1. The capillary of the electrospray ionization source was set to 275 °C, with sheath gas at 45 arbitrary units, auxiliary gas at 5 arbitrary units and the spray voltage at 4.0 kV. A mass/charge ratio scan ranging from 70 to 850 was used in positive/negative polarity switching mode. MS1 data were collected at a resolution The automatic gain control (AGC) target was set at 1 × 106, with a maximum injection time of 200 ms. The top five precursor ions were fragmented using the higher-energy collisional dissociation cell with normalized collision energy of 30% in MS2 at a resolution of 17,500. Data were acquired with Xcalibur software (v.4.1; ThermoFisher Scientific). |
Combined analysis:
| Analysis ID | AN005322 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Waters XBridge BEH Amide (100 x 3mm, 3.5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH004026 |
| Chromatography Summary: | Solvent A at pH 9.0 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters XBridge BEH Amide (100 x 3mm, 3.5um) |
| Column Temperature: | 25 |
| Flow Gradient: | 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A |
| Flow Rate: | 400 uL/min |
| Injection Temperature: | 275 |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium hydroxide; 20 mM ammonium acetate |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS005052 |
| Analysis ID: | AN005322 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | A 15μl aliquot of the sample was used for high-resolution HPLC-tandem mass spectrometry. High-resolution HPLC-tandem mass spectrometry was performed on a Q-Exactive (ThermoFisher Scientific) in line with an electrospray source and an UltiMate 3000 (ThermoFisher Scientific) series HPLC consisting of a binary pump, degasser and autosampler outfitted with a Xbridge Amide column (Waters; 4.6 mm × 100 mm dimension and a 3.5 μm particle size). Mobile phase A contained water and acetonitrile (95/5, v/v), 10 mM ammonium hydroxide and 10 mM ammonium acetate (pH 9.0). Mobile phase B was 100% acetonitrile. The gradient was set to 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1–18 min, 75% A; 18–25 min, 15% A, with a flow rate of 400 μl min–1. The capillary of the electrospray ionization source was set to 275 °C, with sheath gas at 45 arbitrary units, auxiliary gas at 5 arbitrary units and the spray voltage at 4.0 kV. A mass/charge ratio scan ranging from 70 to 850 was used in positive/negative polarity switching mode. MS1 data were collected at a resolution The automatic gain control (AGC) target was set at 1 × 106, with a maximum injection time of 200 ms. The top five precursor ions were fragmented using the higher-energy collisional dissociation cell with normalized collision energy of 30% in MS2 at a resolution of 17,500. Data were acquired with Xcalibur software (v.4.1; ThermoFisher Scientific). |
| Ion Mode: | UNSPECIFIED |
