title: Release of neuropeptides from a neuro-cutaneous co-culture model: A novel in vitro model for studying sensory effects of ciguatoxins.
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.
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France. Electronic address: rlegarrec@univ-brest.fr.
authors.forename: Raphaele
authors.initials: R
authors.lastname: Le Garrec
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France.
authors.forename: Killian
authors.initials: K
authors.lastname: L'herondelle
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Le Gall-Ianotto
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Lebonvallet
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France.
authors.forename: Raphael
authors.initials: R
authors.lastname: Leschiera
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Buhe
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France; Department of Pathology, University Hospital of Brest, 2 avenue Foch, 29200 Brest, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Talagas
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Irina
authors.initials: I
authors.lastname: Vetter
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Lewis
authors.affiliation: Laboratory of Neurosciences of Brest (EA4685), University of Western Brittany, 22 avenue Camille Desmoulins, 29200 Brest, France; Department of Dermatology, University Hospital of Brest, 2 avenue Foch, 29200 Brest, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Misery
mesh_terms: D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D036841:Ciguatera Poisoning; D002922:Ciguatoxins / Q000737:chemistry / Q000494:pharmacology*; D018920:Coculture Techniques; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects*; D008954:Models, Biological; D009479:Neuropeptides / Q000378:metabolism*; D051381:Rats; D011984:Sensory Receptor Cells / Q000166:cytology / Q000187:drug effects*
chemical_list: D009479:Neuropeptides; D002922:Ciguatoxins; C082191:ciguatoxin 2; D015740:Calcitonin Gene-Related Peptide; D002118:Calcium
keywords: Ciguatoxins; In vitro model; Neuropeptide release; Sensory disorders