Advanced and high-throughput method for mitochondrial bioenergetics evaluation in neurotrauma

Jignesh D. Pandya, Patrick G. Sullivan, Lai Yee Leung, Frank C. Tortella, Deborah A. Shear, Ying Deng-Bryant

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

15 Scopus citations

Abstract

Mitochondrial dysfunction is one of the key posttraumatic neuropathological events observed in various experimental models of traumatic brain injury (TBI). The extent of mitochondrial dysfunction has been associated with the severity and time course of secondary injury following brain trauma. Critically, several mitochondrial targeting preclinical drugs used in experimental TBI models have shown improved mitochondrial bioenergetics, together with cortical tissue sparing and cognitive behavioral outcome. Mitochondria, being a central regulator of cellular metabolic pathways and energy producer of cells, are of a great interest for researchers aiming to adopt cutting-edge methodology for mitochondrial bioenergetics assessment. The traditional way of mitochondrial bioenergetics analysis utilizing a Clark-type oxygen electrode (aka. oxytherm) is time-consuming and labor-intensive. In the present chapter, we describe an advanced and highthroughput method for mitochondrial bioenergetics assessments utilizing the Seahorse Biosciences XF e 24 Flux Analyzer. This allows for simultaneous measurement of multiple samples with higher efficiency than the oxytherm procedure. This chapter provides helpful guidelines for conducting mitochondrial isolation and studying mitochondrial bioenergetics in brain tissue homogenates following experimental TBI.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages597-610
Number of pages14
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1462
ISSN (Print)1064-3745

Keywords

  • Brain metabolism
  • Mitochondria
  • Mitochondrial bioenergetics
  • Neurotrauma
  • Oxidative phosphorylation
  • Therapeutic drugs preclinical screening
  • Traumatic brain injury

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