Morphometric assessment of toxicant induced neuronal degeneration in full and restricted contact co-cultures of embryonic cortical rat neurons and astrocytes: using m-Dinitrobezene as a model neurotoxicant

Toxicol In Vitro. 2015 Apr;29(3):564-74. doi: 10.1016/j.tiv.2014.12.015. Epub 2014 Dec 30.


With m-Dinitrobenzene (m-DNB) as a selected model neurotoxicant, we demonstrate how to assess neurotoxicity, using morphology based measurement of neurite degeneration, in a conventional "full-contact" and a modern "restricted-contact" co-culture of rat cortical neurons and astrocytes. In the "full-contact" co-culture, neurons and astrocytes in complete physical contact are "globally" exposed to m-DNB. A newly emergent "restricted-contact" co-culture is attained with a microfluidic device that polarizes neuron somas and neurites into separate compartments, and the neurite compartment is "selectively" exposed to m-DNB. Morphometric analysis of the neuronal area revealed that m-DNB exposure produced no significant change in mean neuronal cell area in "full-contact" co-cultures, whereas a significant decrease was observed for neuron monocultures. Neurite elaboration into a neurite exclusive compartment in a compartmentalized microfluidic device, for both monocultures (no astrocytes) and "restricted" co-cultures (astrocytes touching neurites), decreased with exposure to increasing concentrations of m-DNB, but the average neurite area was higher in co-cultures. By using co-culture systems that more closely approach biological and architectural complexities, and the directionality of exposure found in the brain, this study provides a methodological foundation for unraveling the role of physical contact between astrocytes and neurons in mitigating the toxic effects of chemicals such as m-DNB.

Keywords: Co-culture; Cortical astrocytes; Cortical neurons; Microfluidic; Morphology; m-Dinitrobezene (m-DNB).

Publication types

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

MeSH terms

  • Animals
  • Astrocytes / drug effects*
  • Axons / drug effects
  • Cerebral Cortex / cytology*
  • Cerebral Cortex / embryology
  • Coculture Techniques
  • Dinitrobenzenes / toxicity*
  • Immunohistochemistry
  • Microfluidic Analytical Techniques
  • Nerve Degeneration / chemically induced*
  • Nerve Degeneration / pathology
  • Neurites / drug effects
  • Neurons / drug effects*
  • Rats


  • Dinitrobenzenes
  • 3-dinitrobenzene