Sensitization of primary cultures from rat dorsal root ganglia with lipopolysaccharide (LPS) requires a robust inflammatory response

Inflamm Res. 2022 Feb;71(2):187-190. doi: 10.1007/s00011-021-01534-2. Epub 2021 Dec 23.

Abstract

Objective: We investigated whether it is possible to induce a state of "LPS-sensitization" in neurons of primary cultures from rat dorsal root ganglia by pre-treatment with ultra-low doses of LPS.

Methods: DRG primary cultures were pre-treated with low to ultra-low doses of LPS (0.001-0.1 µg/ml) for 18 h, followed by a short-term stimulation with a higher LPS-dose (10 µg/ml for 2 h). TNF-α in the supernatants was measured as a sensitive read out. Using the fura-2 340/380 nm ratio imaging technique, we further investigated the capsaicin-evoked Ca2+-signals in neurons from DRG, which were pre-treated with a wide range of LPS-doses.

Results: Release of TNF-α evoked by stimulation with 10 µg/ml LPS into the supernatant was not significantly modified by pre-exposure to low to ultra-low LPS-doses. Capsaicin-evoked Ca2+-signals were significantly enhanced by pre-treatment with LPS doses being above a certain threshold.

Conclusion: Ultra-low doses of LPS, which per se do not evoke a detectable inflammatory response, are not sufficient to sensitize neurons (Ca2+-responses) and glial elements (TNF-α-responses) of the primary afferent somatosensory system.

Keywords: Ca2+-imaging; Capsaicin; Cytokines; Dorsal root ganglia; Inflammation; LPS sensitization; Mixed neuro-glial cultures.

MeSH terms

  • Animals
  • Calcium Signaling / drug effects
  • Capsaicin / pharmacology
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Ganglia, Spinal / drug effects*
  • Ganglia, Spinal / immunology
  • Lipopolysaccharides / pharmacology*
  • Rats
  • Rats, Wistar
  • Tumor Necrosis Factor-alpha / biosynthesis*

Substances

  • Lipopolysaccharides
  • Tumor Necrosis Factor-alpha
  • Capsaicin