Release of neuropeptides from a neuro-cutaneous co-culture model: A novel in vitro model for studying sensory effects of ciguatoxins

Toxicon. 2016 Jun 15:116:4-10. doi: 10.1016/j.toxicon.2015.11.004. Epub 2015 Nov 10.

Abstract

Ciguatoxins are the major toxins responsible for ciguatera fish poisoning, a disease dominated by muco-cutaneous sensory disorders including paresthesiae, cold dysesthesia and pruritus. While the ciguatoxins are well known to target voltage-gated sodium channels (VGSCs), the ensuing molecular mechanisms underlying these sensory disorders remain poorly understood. In this study, we propose a primary sensory neuron-keratinocyte co-culture as an appropriate model to study the neuro-cutaneous effects of ciguatoxins. Using this model, we show for the first time that nanomolar concentrations of Pacific ciguatoxin-2 (P-CTX-2) induced a VGSC-dependent release of substance P (SP) and calcitonin gene-related peptide (CGRP). As these neuropeptides are known mediators of pain and itch sensations, the ciguatoxin-induced sensory disturbances in ciguatera fish poisoning may involve the release of these neuropeptides. We further determined time- and P-CTX-2 concentration-dependence of the release of SP and CGRP from the co-culture model. Moreover, we highlighted the influence of extracellular calcium on the release of neuropeptides elicited by P-CTX-2. These findings underline the usefulness of this novel in vitro model for studying the cellular and molecular mechanisms of the neuro-cutaneous effects of ciguatoxins, which may assist with identifying potential therapeutics for ciguatera fish poisoning.

Keywords: Ciguatoxins; In vitro model; Neuropeptide release; Sensory disorders.

MeSH terms

  • Animals
  • Calcitonin Gene-Related Peptide / metabolism
  • Calcium / metabolism
  • Ciguatera Poisoning
  • Ciguatoxins / chemistry
  • Ciguatoxins / pharmacology*
  • Coculture Techniques
  • Humans
  • Immunohistochemistry
  • Keratinocytes / cytology
  • Keratinocytes / drug effects*
  • Models, Biological*
  • Neuropeptides / metabolism*
  • Rats
  • Sensory Receptor Cells / cytology
  • Sensory Receptor Cells / drug effects*

Substances

  • Neuropeptides
  • Ciguatoxins
  • ciguatoxin 2
  • Calcitonin Gene-Related Peptide
  • Calcium