title: Exploration of new functional endpoints in neuro-2a cells for the detection of the marine biotoxins saxitoxin, palytoxin and tetrodotoxin.
abstract: Marine neurotoxins accumulate in seafood and therewith represent a threat for consumers. At the European level, the use of in vivo bioassays is banned from 2015 onwards, except for the control of production areas. Cytotoxicity in the neuro-2a assay has been shown a promising in vitro alternative. However, given that cytotoxicity may be sensitive to confounding factors the current study investigates the suitability of functional endpoints as alternatives to cytotoxicity for the detection of marine neurotoxins. Microarray analyses were performed following exposure of neuro-2a cells to three marine neurotoxins (palytoxin (PlTx), saxitoxin (STX) and tetrodotoxin (TTX)) to identify genes up- or down-regulated that can be used as biomarkers for screening purposes. In addition to microarrays, the voltage dependent fluorescent probe bisoxonol was used to assess changes in cellular membrane potential. Biomarkers based on mRNA expression were detected for PlTx but not for STX and TTX. STX and TTX decreased the fluorescence of bisoxonol while PlTx showed no effect. When using cytotoxicity as the read out the neuro-2a assay detects these three neurotoxins at similar concentrations. Therefore it is concluded that the newly investigated endpoints in the neuro-2a assay are not preferred over cytotoxicity in a suitable broad and sensitive bioassay for the detection of marine neurotoxins in real practice.
authors.affiliation: Division of Toxicology, Wageningen University, The Netherlands; RIKILT, Institute of Food Safety, Wageningen, The Netherlands. Electronic address: jonathan.nicolas@wur.nl.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Nicolas
authors.affiliation: RIKILT, Institute of Food Safety, Wageningen, The Netherlands.
authors.forename: Toine F H
authors.initials: TF
authors.lastname: Bovee
authors.affiliation: Division of Toxicology, Wageningen University, The Netherlands.
authors.forename: Lenny
authors.initials: L
authors.lastname: Kamelia
authors.affiliation: Division of Toxicology, Wageningen University, The Netherlands.
authors.forename: Ivonne M C M
authors.initials: IM
authors.lastname: Rietjens
authors.affiliation: RIKILT, Institute of Food Safety, Wageningen, The Netherlands.
authors.forename: Peter J M
authors.initials: PJ
authors.lastname: Hendriksen
mesh_terms: D000178:Acrylamides / Q000633:toxicity*; D000818:Animals; D001681:Biological Assay; D015415:Biomarkers; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003064:Cnidarian Venoms; D023381:Endpoint Determination; D005453:Fluorescence; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D051379:Mice; D009447:Neuroblastoma / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D012530:Saxitoxin / Q000633:toxicity*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013779:Tetrodotoxin / Q000633:toxicity*; D000095702:Polyether Toxins
chemical_list: D000178:Acrylamides; D015415:Biomarkers; D003064:Cnidarian Venoms; D012530:Saxitoxin; D000254:Sodium-Potassium-Exchanging ATPase; D013779:Tetrodotoxin; C010272:palytoxin; D000095702:Polyether Toxins
keywords: Biomarker; Bis-oxonol; Food safety; Marine neurotoxins; Seafood; Toxicogenomics