Chinopoulos 2011 Methods Mol Biol

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Chinopoulos C, Zhang SF, Thomas B, Ten V, Starkov AA (2011) Isolation and functional assessment of mitochondria from small amounts of mouse brain tissue. Methods Mol Biol 793:311-24.

» PMID:21913109

Chinopoulos C, Zhang SF, Thomas B, Ten V, Starkov AA (2011) Methods Mol Biol

Abstract: Recent discoveries have brought mitochondria functions in focus of the neuroscience research community and greatly stimulated the demand for approaches to study mitochondria dysfunction in neurodegenerative diseases. Many mouse disease models have been generated, but studying mitochondria isolated from individual mouse brain regions is a challenge because of small amount of the available brain tissue. Conventional techniques for isolation and purification of mitochondria from mouse brain subregions, such as ventral midbrain, hippocampus, or striatum, require pooling brain tissue from six to nine animals for a single mitochondrial preparation. Working with pooled tissue significantly decreases the quality of data because of the time required to dissect several brains. It also greatly increases the labor intensity and the cost of experiments as several animals are required per single data point. We describe a method for isolation of brain mitochondria from mouse striata or other 7-12 mg brain samples. The method utilizes a refrigerated table-top microtube centrifuge, and produces research grade quality mitochondria in amounts sufficient for performing multiple enzymatic and functional assays, thereby eliminating the necessity for pooling mouse brain tissue. We also include a method of measuring ADP-ATP exchange rate as a function of mitochondrial membrane potential (ΔΨm) in small amounts of isolated mitochondria, adapted to a plate reader format.

Keywords: Adenine nucleotide translocase; Adenine nucleotide carrier; Systems biology Bioblast editor: Sumbalova Z O2k-Network Lab: HU Budapest Chinopoulos C


Labels: MiParea: Respiration, Instruments;methods 


Organism: Rat  Tissue;cell: Nervous system  Preparation: Isolated mitochondria  Enzyme: Adenine nucleotide translocase, Complex V;ATP synthase  Regulation: ATP production, mt-Membrane potential 


HRR: Oxygraph-2k, O2k-Fluorometer, Theory