Dias 2018 Anal Biochem
|Dias C, Lourenco CF, Barbosa RM, Laranjinha J, Ledo A (2018) Analysis of respiratory capacity in brain tissue preparations: high-resolution respirometry for intact hippocampal slices. Anal Biochem 551:43-50.|
Abstract: The evaluation of mitochondrial function provides the basis for the study of brain bioenergetics. However, analysis of brain mitochondrial respiration has been hindered by the low yield associated with mitochondria isolation procedures. Furthermore, isolating mitochondria or cells results in loss of the inherent complexity of the central nervous system. High-resolution respirometry (HRR), is a valuable tool to study mitochondrial function and has been used in diverse biological preparations ranging from isolated mitochondria to tissue homogenates and permeabilized tissue biopsies. Here we describe a novel methodology for evaluation of mitochondrial respiration using tissue preparations from the central nervous system, namely acute hippocampal slices from rodents, with HRR. By using acute intact hippocampal slices, tissue cytoarchitecture, intercellular communication and connectivity are preserved. Mitochondrial respiration was evaluated by using an adapted substrate-uncoupler-inhibitor titration (SUIT) protocol and the expected responses were observed. This methodology can be used to detect differences in mitochondrial function at the oxidative phosphorylation level and for studies with different brain oxidative substrates in physiological and neuropathological settings, by using a system that better represents the in vivo conditions than isolated mitochondria and/or cells.
• Keywords: High-resolution respirometry, Oxygen consumption rate, Whole hippocampal slices, Bioenergetics, Oxidative phosphorylation • Bioblast editor: Krumschnabel G • O2k-Network Lab: PT Coimbra Laranjinha J
MitoEAGLE Short-Term Scientific Mission
- Candida Dias was a visiting scientist from the Center for Neuroscience and Cell Biology, University of Coimbra, PT, at Oroboros Instruments within the MitoEAGLE Short-Term Scientific Mission (2019, September 23 to December 21).
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse, Rat Tissue;cell: Nervous system Preparation: Intact cells
Coupling state: LEAK, ROUTINE, ET Pathway: ROX HRR: Oxygraph-2k
Tissue slice, 2018-07