Related Experiment Video
Updated: Jun 30, 2025
11:46
Shotgun Lipidomics of Rodent Tissues
Published on: November 18, 2022
2.0K
A targeted proteomics method for quantifying plasma apolipoprotein kinetics in individual mice using stable isotope
Baohai Shao1, Masami Shimizu-Albergine1, Farah Kramer1
1Division of Metabolism, Endocrinology and Nutrition, Department of Medicine, UW Medicine Diabetes Institute, University of Washington, Seattle, WA, USA.
Journal of Lipid Research
|March 15, 2024
Summary
This study introduces a new mass spectrometry method to measure apolipoprotein kinetics in mice, crucial for understanding dyslipidemia and atherosclerosis. The findings reveal altered protein clearance and production rates in genetic models, aiding disease research.
Area of Science:
- Biochemistry
- Pharmacology
- Genetics
Background:
- Apolipoprotein kinetics are vital in dyslipidemia and atherogenesis.
- Studying mouse apolipoprotein kinetics is challenging due to limited blood volumes.
- Existing methods lack the sensitivity for sequential analysis in small animal models.
Purpose of the Study:
- To develop a sensitive mass spectrometry method for quantifying apolipoprotein kinetics in mice.
- To analyze apolipoprotein fractional clearance rates (FCRs) and production rates (PRs) in wild-type and atherosclerosis-prone mouse models.
- To enable robust kinetic studies using small, serial blood samples.
Main Methods:
- Developed a targeted liquid chromatography tandem mass spectrometry (LC-MS/MS) assay.
- Utilized stable isotope labeling with 13C6,15N2-lysine (Lys8) for peptide quantification.
- Analyzed plasma samples from wild-type, LDL receptor-deficient (Ldlr-/-), and apolipoprotein E-deficient (Apoe-/-) mice.
Main Results:
- The method successfully quantified stable isotope enrichment in multiple apolipoproteins from 15 μl blood samples.
- Ldlr-/- mice exhibited significantly slower FCRs for several key apolipoproteins (APOA1, APOB, APOE, etc.).
- Apoe-/- mice showed distinct alterations in FCRs and PRs compared to wild-type and Ldlr-/- mice.
Conclusions:
- The developed LC-MS/MS method is robust and reproducible for assessing plasma apolipoprotein kinetics in mouse models.
- This technique facilitates the study of dyslipidemia and atherogenesis mechanisms in genetically modified mice.
- The method's adaptability allows for the analysis of numerous proteins in various animal models of human disease.
