Simultaneous quantitative respirometry and fluorometric assays in dissected hippocampal tissue from mice

Sreemathi Logan; Rojina Ranjit; Hadyn Rose; Anne Bredegaard; Carlos Manlio Díaz-García

doi:10.1016/j.xpro.2024.102988

Simultaneous quantitative respirometry and fluorometric assays in dissected hippocampal tissue from mice

STAR Protoc. 2024 Apr 16;5(2):102988. doi: 10.1016/j.xpro.2024.102988. Online ahead of print.

Authors

Sreemathi Logan¹, Rojina Ranjit², Hadyn Rose², Anne Bredegaard², Carlos Manlio Díaz-García³

Affiliations

¹ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Electronic address: sreemathi-logan@ouhsc.edu.
² Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
³ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Harold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Electronic address: carlosmanlio-diazgarcia@ouhsc.edu.

Abstract

Respirometry is a technique for studying mitochondrial function that has proven compatibility with ≥0.5 mg of brain tissue. Here, we present a protocol for assessing oxygen consumption and H₂O₂ production rates in hippocampal tissue using the Oroboros O2k system. We describe steps for brain harvesting, tissue preparation, hippocampal microdissection, and respirometry assays. This approach has been valuable to study the metabolism of dentate granule cells of the hippocampus and could be applicable to other brain subregions. For complete details on the use and execution of this protocol, please refer to Rose et al.¹.

Keywords: Metabolism; Molecular Biology; Neuroscience.

Abstract

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