Editor's Highlight: Multiparametric Image Analysis of Rat Dorsal Root Ganglion Cultures to Evaluate Peripheral Neuropathy-Inducing Chemotherapeutics

Toxicol Sci. 2017 Mar 1;156(1):275-288. doi: 10.1093/toxsci/kfw254.


Chemotherapy-induced peripheral neuropathy (CIPN) is a major, dose-limiting adverse effect experienced by cancer patients. Advancements in mechanism-based risk mitigation and effective treatments for CIPN can be aided by suitable in vitro assays. To this end, we developed a multiparametric morphology-centered rat dorsal root ganglion (DRG) assay. Morphologic alterations in subcellular structures of neurons and non-neurons were analyzed with an automated microscopy system. Stains for NeuN (a neuron-specific nuclear protein) and Tuj-1 (β-III tubulin) were used to identify neuronal cell nuclei and neuronal cell bodies/neurites, respectively. Vimentin staining (a component of Schwann cell intermediate filaments) was used to label non-neuronal supporting cells. Nuclei that stained with DAPI, but lacked NeuN represented non-neuronal cells. Images were analyzed following 24 h of continuous exposure to CIPN-inducing agents and 72 h after drug removal to provide a dynamic measure of recovery from initial drug effects. Treatment with bortezomib, cisplatin, eribulin, paclitaxel or vincristine induced a dose-dependent loss of neurite/process areas, mimicking the 'dying back' degeneration of axons, a histopathological hallmark of clinical CIPN in vivo. The IC50 for neurite loss was within 3-fold of the maximal clinical exposure (Cmax) for all five CIPN-inducing drugs, but was >4- or ≥ 28-fold of the Cmax for 2 non-CIPN-inducing agents. Compound-specific effects, eg, neurite fragmentation by cisplatin or bortezomib and enlarged neuronal cell bodies by paclitaxel, were also observed. Collectively, these results support the use of a quantitative, morphologic evaluation and a DRG cell culture model to inform risk and examine mechanisms of CIPN.

Keywords: automated image analysis; early safety evaluation.; chemotherapy-induced peripheral neuropathy (CIPN); high content analysis (HCA); rat dorsal root ganglion cells (DRGs).

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antineoplastic Agents / adverse effects*
  • Biomarkers / metabolism
  • Cell Body / drug effects
  • Cell Body / metabolism
  • Cell Body / pathology
  • Cell Nucleus Shape / drug effects
  • Cell Shape / drug effects
  • Cells, Cultured
  • Drug Evaluation, Preclinical / methods
  • Electrophoresis, Capillary
  • Fluorescent Antibody Technique
  • Ganglia, Spinal / drug effects*
  • Ganglia, Spinal / metabolism
  • Ganglia, Spinal / pathology
  • Image Processing, Computer-Assisted
  • Kinetics
  • Molecular Weight
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / metabolism
  • Neurites / drug effects
  • Neurites / metabolism
  • Neurites / pathology
  • Neurons / drug effects*
  • Neurons / metabolism
  • Neurons / pathology
  • Neurotoxicity Syndromes / etiology
  • Neurotoxicity Syndromes / metabolism
  • Neurotoxicity Syndromes / pathology
  • Organelle Shape / drug effects
  • Organelle Size / drug effects
  • Peripheral Nervous System Diseases / etiology
  • Peripheral Nervous System Diseases / metabolism
  • Peripheral Nervous System Diseases / pathology
  • Rats


  • Antineoplastic Agents
  • Biomarkers
  • Nerve Tissue Proteins