Journal Article Epublish
PubMed: 40330847 DOI: 10.1093/biomethods/bpaf031
Measurement of oxygen consumption rate in mouse aortic tissue.
Biol Methods Protoc 10(1):bpaf031
Authors: Zhen Zhou, Ripon Sarkar, Jose Emiliano Esparza Pinelo, Alexis Richard, Jay Dunn, Zhao Ren, Callie S Kwartler, Dianna M Milewicz
Affiliations
- Division of Medical Genetics, Department of Internal Medicine, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, 77030, United States.
- Department of Anesthesiology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, 77030, United States.
- Cell Analysis Division, Agilent Technologies, 5301 Stevens Creek Blvd, Santa Clara, CA, 95051, United States.
Abstract
Thoracic aortic aneurysm and dissection (TAD) is a life-threatening vascular disorder, and smooth muscle cell mitochondrial dysfunction leads to cell death, contributing to TAD. Accurate measurements of metabolic processes are essential for understanding cellular homeostasis in both healthy and diseased states. While assays for evaluating mitochondrial respiration have been well established for cultured cells and isolated mitochondria, no optimized application has been developed for aortic tissue. In this study, we generate an optimized protocol using the Agilent Seahorse XFe24 analyzer to measure mitochondrial respiration in mouse aortic tissue. This method allows for precise measurement of mitochondrial oxygen consumption in mouse aorta, providing a reliable assay for bioenergetic analysis of aortic tissue. The protocol offers a reproducible approach for assessing mitochondrial function in aortic tissues, capturing both baseline OCR and responses to mitochondrial inhibitors, such as oligomycin, FCCP, and rotenone/antimycin A. This method establishes a critical foundation for studying metabolic shifts in aortic tissues and offers valuable insights into the cellular mechanisms of aortic diseases, contributing to a better understanding of TAD progression.
MeSH
Cells, Research Design, Respiration, Mitochondria, Tissues