Isolation and functional assessment of mitochondria from small amounts of mouse brain tissue

Christos Chinopoulos, Steven F. Zhang, Bobby Thomas, Vadim Ten, Anatoly A. Starkov

Research output: Chapter in Book/Report/Conference proceedingChapter

27 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationNeurodegeneration
Subtitle of host publicationMethods and Protocols
EditorsGiovanni Manfredi, Hibiki Kawamata
Pages311-324
Number of pages14
DOIs
Publication statusPublished - Oct 24 2011

Publication series

NameMethods in Molecular Biology
Volume793
ISSN (Print)1064-3745

    Fingerprint

Keywords

  • Adenine nucleotide carrier
  • Adenine nucleotide translocase
  • Systems biology

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Chinopoulos, C., Zhang, S. F., Thomas, B., Ten, V., & Starkov, A. A. (2011). Isolation and functional assessment of mitochondria from small amounts of mouse brain tissue. In G. Manfredi, & H. Kawamata (Eds.), Neurodegeneration: Methods and Protocols (pp. 311-324). (Methods in Molecular Biology; Vol. 793). https://doi.org/10.1007/978-1-61779-328-8_20