Quantitative analysis of neuronal mitochondrial movement reveals patterns resulting from neurotoxicity of rotenone and 6-hydroxydopamine

FASEB J. 2021 Dec;35(12):e22024. doi: 10.1096/fj.202100899R.


Alterations in mitochondrial dynamics, including their intracellular trafficking, are common early manifestations of neuronal degeneration. However, current methodologies used to study mitochondrial trafficking events rely on parameters that are primarily altered in later stages of neurodegeneration. Our objective was to establish a reliable applied statistical analysis to detect early alterations in neuronal mitochondrial trafficking. We propose a novel quantitative analysis of mitochondria trajectories based on innovative movement descriptors, including straightness, efficiency, anisotropy, and kurtosis. We evaluated time- and dose-dependent alterations in trajectory descriptors using biological data from differentiated SH-SY5Y cells treated with the mitochondrial toxicants 6-hydroxydopamine and rotenone. MitoTracker Red CMXRos-labelled mitochondria movement was analyzed by total internal reflection fluorescence microscopy followed by computational modelling to describe the process. Based on the aforementioned trajectory descriptors, this innovative analysis of mitochondria trajectories provides insights into mitochondrial movement characteristics and can be a consistent and sensitive method to detect alterations in mitochondrial trafficking occurring in the earliest time points of neurodegeneration.

Keywords: exploratory data analysis; live cell imaging; mitochondria movement; neurotoxicants; principal component analysis; trajectory descriptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenergic Agents / adverse effects
  • Cell Differentiation
  • Humans
  • Mitochondria / drug effects
  • Mitochondria / pathology*
  • Mitochondrial Dynamics*
  • Neuroblastoma / chemically induced
  • Neuroblastoma / pathology*
  • Neurons / drug effects
  • Neurons / pathology*
  • Oxidopamine / adverse effects*
  • Rotenone / adverse effects*
  • Uncoupling Agents / adverse effects


  • Adrenergic Agents
  • Uncoupling Agents
  • Rotenone
  • Oxidopamine