Assessing Mitochondrial Function in In Vitro and Ex Vivo Models of Huntington's Disease

Methods Mol Biol. 2018:1780:415-442. doi: 10.1007/978-1-4939-7825-0_19.

Abstract

Mitochondrial dysfunction has gained a preponderant role in the pathogenesis of Huntington's disease (HD). Mutant huntingtin (mHTT) directly interacts with mitochondria in a deleterious manner. As the central hub of the cell, not only mitochondrial bioenergetics is affected but there is also diminished mitochondrial membrane potential (Δψ m) and altered production of reactive oxygen species (ROS). Restoration of mitochondrial function has proven to be a major player in the search and establishment of therapeutics for HD patients. As such, performing an overall study of mitochondrial function is crucial. In this chapter, we describe some methodologies used to study mitochondrial function by determining the oxygen consumption, changes in Δψ m, mitochondrial calcium handling, and levels of mitochondrial ROS. Here we focus on biological samples derived from HD versus control cells and/or animal models, namely functional isolated brain mitochondria, an ex vivo animal model, and cultured cells, including cell lines and primary neural cultures, as in vitro models.

Keywords: Bioenergetics; Cells; Mitochondria; Mitochondrial calcium handling; Mitochondrial function; Mitochondrial membrane potential; Mitochondrial reactive oxygen species; Neurons; Oxygen consumption.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology
  • Brain / metabolism
  • Brain / pathology*
  • Calcium / metabolism
  • Cell Line
  • Disease Models, Animal
  • Humans
  • Huntingtin Protein / genetics
  • Huntingtin Protein / metabolism
  • Huntington Disease / genetics
  • Huntington Disease / pathology*
  • Intravital Microscopy / instrumentation
  • Intravital Microscopy / methods
  • Membrane Potential, Mitochondrial
  • Mice
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Mitochondria / metabolism
  • Mitochondria / pathology*
  • Mutation
  • Neurons / cytology
  • Neurons / pathology*
  • Optical Imaging / instrumentation
  • Optical Imaging / methods
  • Oxygen Consumption
  • Primary Cell Culture / instrumentation
  • Primary Cell Culture / methods*
  • Reactive Oxygen Species / metabolism
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods

Substances

  • Huntingtin Protein
  • Reactive Oxygen Species
  • Calcium