Protocol for real-time measurement of mitochondrial respiration in the mouse ocular posterior pole using a Seahorse XFe24 analyzer

Tanu Parmar; Vipul M Parmar; Goldis Malek

doi:10.1016/j.xpro.2024.103150

Protocol for real-time measurement of mitochondrial respiration in the mouse ocular posterior pole using a Seahorse XFe24 analyzer

STAR Protoc. 2024 Jul 11;5(3):103150. doi: 10.1016/j.xpro.2024.103150. Online ahead of print.

Authors

Tanu Parmar¹, Vipul M Parmar², Goldis Malek³

Affiliations

¹ Department of Ophthalmology, Albert Eye Research Institute, Duke University, Durham, NC 27710, USA. Electronic address: tanu.parmar@duke.edu.
² Department of Ophthalmology, Albert Eye Research Institute, Duke University, Durham, NC 27710, USA.
³ Department of Ophthalmology, Albert Eye Research Institute, Duke University, Durham, NC 27710, USA; Department of Pathology, Duke University, Durham, NC 27710, USA. Electronic address: gmalek@duke.edu.

Abstract

During aging and in retinal degenerative diseases, vulnerable retinal pigment epithelial (RPE) cells are subject to mitochondrial dysfunction, creating a need for accessibility to tools which can facilitate assessment of the ocular posterior pole bioenergetics. Here, we present a protocol for quantifying mitochondrial respiration in the posterior eye cup (RPE-choroid-sclera) of young and old mice. We describe steps for eye cup dissection, optimization of tissue size, drug concentrations, and cycle conditions using the XF Cell Mito Stress Test.

Keywords: Cell Biology; Metabolism; Model Organisms.

Abstract

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