Summary of Study ST003246
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 PR002016. The data can be accessed directly via it's Project DOI: 10.21228/M81C00 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 | ST003246 |
| Study Title | Effects of mitoregulin loss on cardiac and mitochondrial lipids in aged male mice |
| Study Summary | Cardiac lipidome analysis in aged (21 to 23-months old) male wildtype and mtln knockout mice |
| Institute | University of Iowa |
| Last Name | Boudreau |
| First Name | Ryan |
| Address | 4334 PBDB, 169 Newton Rd, Iowa City, IA 52242 |
| ryan-boudreau@uiowa.edu | |
| Phone | 3193535573 |
| Submit Date | 2024-06-02 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-12-31 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002016 |
| Project DOI: | doi: 10.21228/M81C00 |
| Project Title: | Effects of mitoregulin loss on cardiac and mitochondrial lipids in mice |
| Project Summary: | We and others discovered a highly-conserved mitochondrial transmembrane microprotein, named Mitoregulin (Mtln), that supports lipid metabolism. We reported that Mtln strongly binds cardiolipin (CL), increases mitochondrial respiration and Ca2+ retention capacities, and reduces reactive oxygen species (ROS). Here we extend our observation of Mtln-CL binding and examine Mtln influence on cristae structure and mitochondrial membrane integrity during stress. We demonstrate that mitochondria from constitutive- and inducible Mtln-knockout (KO) mice are susceptible to membrane freeze-damage and that this can be rescued by acute Mtln re-expression. In mitochondrial-simulated lipid monolayers, we show that synthetic Mtln decreases lipid packing and monolayer elasticity. Lipidomics revealed that Mtln-KO heart tissues show broad decreases in 22:6-containing lipids and increased cardiolipin damage/remodeling. Lastly, we demonstrate that Mtln-KO mice suffer worse myocardial ischemia-reperfusion injury, hinting at a translationally-relevant role for Mtln in cardioprotection. Our work supports a model in which Mtln binds cardiolipin and stabilizes mitochondrial membranes to broadly influence diverse mitochondrial functions, including lipid metabolism, while also protecting against stress. |
| Institute: | University of Iowa |
| Last Name: | Boudreau |
| First Name: | Ryan |
| Address: | 4334 PBDB, 169 Newton Rd, Iowa City, IA 52242 |
| Email: | ryan-boudreau@uiowa.edu |
| Phone: | 3193535573 |
Subject:
| Subject ID: | SU003365 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammals |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype |
|---|---|---|
| SA353453 | 1134-2 | KO |
| SA353454 | 1134-3 | KO |
| SA353455 | 1134-4 | KO |
| SA353456 | 1134-5 | KO |
| SA353457 | 1153-1 | KO |
| SA353458 | 1102-3 | WT |
| SA353459 | 1116-1 | WT |
| SA353460 | 1116-4 | WT |
| SA353461 | 1134-1 | WT |
| SA353462 | 2191-1 | WT |
| Showing results 1 to 10 of 10 |
Collection:
| Collection ID: | CO003358 |
| Collection Summary: | Mtln-KO and WT mice were fasted for 4-6 h, and while under deep ketamine/xylazine anesthesia, the heart apex was snipped off and immediately snap-frozen by LN2 immersion. |
| Sample Type: | Heart |
Treatment:
| Treatment ID: | TR003374 |
| Treatment Summary: | No treatment. |
Sample Preparation:
| Sampleprep ID: | SP003372 |
| Sampleprep Summary: | Lipid extraction. Lipid extraction, based on Matyash et al.72, was performed as follows. All solutions were pre-chilled on ice. Tissues or mitochondrial pellets were transferred to labeled bead-mill tubes (1.4 mm, MoBio Cat# 13113-50) where lipids were extracted in a solution of 250 µL PBS, 225 µL MeOH containing internal standards, and 750 µL MTBE (methyl tert-butyl ether). Internal standards were Avanti SPLASH LipidoMix (Lot#12) at 10 µL per sample and Cambridge Isotope laboratories NSK-B and NSK-B-G1 (deuterated carnitines) at 10 µL per sample. The samples were homogenized in one 30 s cycle using the Omni Bead Ruptor followed by a rest on ice for 1 h. An addition of 188 µL PBS was made to induce phase separation. After centrifugation at 16,000 g for 5 minutes at 4 °C, the upper phases were collected and evaporated to dryness under a gentle nitrogen stream at room temperature. Lipid samples were reconstituted in 500 µL IPA (isopropyl alcohol) and transferred to an LC-MS vial with insert (Agilent 5182-0554 and 5183-2086) for analysis. Concurrently, a process blank sample and pooled quality control (QC) sample was prepared by taking equal volumes (~50 µL) from each sample after final resuspension. |
Chromatography:
| Chromatography ID: | CH004020 |
| Chromatography Summary: | Positive Mode RP LCMS |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 65 |
| Flow Gradient: | The chromatography gradient for both positive and negative modes started at 15% mobile phase B then increased to 30% B over 2.4 min. It sequentially increased to 48% B from 2.4 – 3.0 min, 82% B from 3 – 13.2 min, and 99% B from 13.2 – 13.8 min where it’s held until 16.7 min and returned to the initial conditions and equilibrated for 5 min. |
| Flow Rate: | 0.4 mL min |
| Solvent A: | 40% water/60% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/9% acetonitrile/1% water; 0.1% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH004021 |
| Chromatography Summary: | Negative Mode RP LCMS |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 65 |
| Flow Gradient: | The chromatography gradient for both positive and negative modes started at 15% mobile phase B then increased to 30% B over 2.4 min. It sequentially increased to 48% B from 2.4 – 3.0 min, 82% B from 3 – 13.2 min, and 99% B from 13.2 – 13.8 min where it’s held until 16.7 min and returned to the initial conditions and equilibrated for 5 min. |
| Flow Rate: | 0.4 mL min |
| Solvent A: | 40% water/60% acetonitrile; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/9% acetonitrile/1% water; 10 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
Analysis:
| Analysis ID: | AN005316 |
| Analysis Type: | MS |
| Chromatography ID: | CH004020 |
| Num Factors: | 2 |
| Num Metabolites: | 285 |
| Rt Units: | Minutes |
| Units: | counts normalized to total lipid signal |
| Analysis ID: | AN005317 |
| Analysis Type: | MS |
| Chromatography ID: | CH004021 |
| Num Factors: | 2 |
| Num Metabolites: | 220 |
| Rt Units: | Minutes |
| Units: | counts normalized to total lipid signal |
