Measurement of oxygen consumption rate in mouse aortic tissue

• Published in: Biology methods and protocols Vol. 10; no. 1; p. bpaf031
• Main Authors: Zhou, Zhen, Sarkar, Ripon, Esparza Pinelo, Jose Emiliano, Richard, Alexis, Dunn, Jay, Ren, Zhao, Kwartler, Callie S, Milewicz, Dianna M
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.01.2025
• Subjects: Aorta; Aortic aneurysms; Cell death; Homeostasis; Metabolism; Methods; Mitochondria; Oligomycin; Oxygen consumption; Respiration; Rotenone; Smooth muscle; Thorax; aortic tissue; Seahorse extracellular flux; oxygen consumption rate; mitochondrial respiration
• ISSN: 2396-8923; 2396-8923
• DOI: 10.1093/biomethods/bpaf031

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.