Microglia as a surrogate biosensor to determine nanoparticle neurotoxicity

Cayla M. Duffy, Shihab Ahmed, Ce Yuan, Vijayakumar Mavanji, Joshua P Nixon, Tammy Butterick

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Nanoparticles found in air pollutants can alter neurotransmitter profiles, increase neuroinflammation, and alter brain function. Therefore, the assay described here will aid in elucidating the role of microglia in neuroinflammation and neurodegenerative diseases. The use of microglia, resident immune cells of the brain, as a surrogate biosensor provides novel insight into how inflammatory responses mediate neuronal insults. Here, we utilize an immortalized murine microglial cell line, designated BV2, and describe a method for nanoparticle exposure using silver nanoparticles (AgNPs) as a standard. We describe how to expose microglia to nanoparticles, how to remove nanoparticles from supernatant, and how to use supernatant from activated microglia to determine toxicity, using hypothalamic cell survival as a measure. Following AgNP exposure, BV2 microglial activation was validated using a tumor necrosis factor alpha (TNF-α) enzyme linked immunosorbent assay (ELISA). The supernatant was filtered to remove the AgNP and to allow cytokines and other secreted factors to remain in the conditioned media. Hypothalamic cells were then exposed to supernatant from AgNP activated microglia and survival of neurons was determined using a resazurinbased fluorescent assay. This technique is useful for utilizing microglia as a surrogate biomarker of neuroinflammation and determining the effect of neuroinflammation on other cell types.

Original languageEnglish (US)
Article numbere54662
JournalJournal of Visualized Experiments
Volume2016
Issue number116
DOIs
StatePublished - Oct 25 2016

Bibliographical note

Publisher Copyright:
© 2016 Journal of Visualized Experiments.

Keywords

  • Cell death
  • Conditioned media
  • Hypothalamic cells
  • Issue 116
  • Microglia
  • Nanoparticle
  • Neuroinflammation
  • Neuroscience

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