Herbst 2015 J Physiol
|Herbst EA, Holloway GP (2015) Permeabilization of brain tissue in situ enables multi-region analysis of mitochondrial function in a single mouse brain. J Physiol 593:787-801.|
Abstract: Mitochondria function as the core energy providers in the brain and symptoms of neurodegenerative diseases are often attributed to their dysregulation. Assessing mitochondrial function is classically performed in isolated mitochondria, however this process requires significant isolation time, the demand for abundant tissue, and the disruption of the cooperative mitochondrial reticulum, all of which reduce reliability when attempting to assess in vivo mitochondrial bioenergetics. Here we introduce a method that advances the assessment of mitochondrial respiration in the brain by permeabilizing existing brain tissue to grant direct access to the mitochondrial reticulum in situ. The permeabilized brain preparation allows for instant analysis of mitochondrial function with unaltered mitochondrial morphology using significantly small sample sizes (~2 mg), which permits the analysis of mitochondrial function in multiple subregions within a single mouse brain. Here this technique was applied to assess regional variation in brain mitochondrial function with acute ischemia-reperfusion injuries and to determine the role of reactive oxygen species in exacerbating dysfunction through the application of a transgenic mouse model overexpressing catalase within mitochondria. Through creating accessibility to small regions for the investigation of mitochondrial function, the permeabilized brain preparation enhances the capacity for examining regional differences in mitochondrial regulation within the brain, as the majority of genetic models used for unique approaches exist in the mouse model.
• Keywords: Mitochondrial function, Respiration, Ischemia-reperfusion
• O2k-Network Lab: CA Guelph Holloway GP
Labels: MiParea: Respiration, Genetic knockout;overexpression
Stress:Ischemia-reperfusion, Oxidative stress;RONS Organism: Mouse Tissue;cell: Nervous system Preparation: Permeabilized tissue, Isolated mitochondria
Regulation: Cyt c Coupling state: LEAK, OXPHOS Pathway: N, S, NS HRR: Oxygraph-2k