Measurement of mitochondrial respiration in the murine retina using a Seahorse extracellular flux analyzer

STAR Protoc. 2021 May 12;2(2):100533. doi: 10.1016/j.xpro.2021.100533. eCollection 2021 Jun 18.

Abstract

Mitochondrial metabolism is a critical mechanism that is deregulated in numerous retinal diseases. Here, we elaborate a protocol to quantify oxygen consumption rate as a measure of mitochondrial respiration directly from mouse retinal tissue pieces. Our procedure combines the use of Seahorse extracellular flux technology and ex vivo retinal tissue isolation and is robustly reproducible under different treatment conditions. This protocol allows direct assessment of mitochondrial function in response to drug treatments or genetic manipulation in mouse models. For complete details on the use and execution of this protocol, please refer to Shetty et al. (2020), Sardar Pasha et al. (2021), Kooragayala et al. (2015), and Joyal et al. (2016).

Keywords: Metabolism; Model Organisms; Neuroscience.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Respiration / physiology*
  • Metabolic Flux Analysis / methods*
  • Mice
  • Mitochondria* / metabolism
  • Mitochondria* / physiology
  • Oxygen / analysis
  • Oxygen / metabolism
  • Oxygen Consumption / physiology
  • Retina / cytology*

Substances

  • Oxygen