title: Suitability of the Neuro-2a cell line for the detection of palytoxin and analogues (neurotoxic phycotoxins).
abstract: Palytoxin and related compounds are neurotoxic phycotoxins produced by benthic microalgae belonging to the genus Ostreopsis. For several years this family of phycotoxins has been posing a threat to human health since they can bioaccumulate in shellfish. With the aim of replacing current biological assays, such as the mouse or hemolytic assays, we investigated using the Neuro-2a neuroblastoma cell line to detect palytoxin and related compounds. Cell death induced by the effects of PlTX and analogues on Na+, K+-ATPase were measured using the 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay for mitochondrial reductase activity as a surrogate for cell number. The specificity of the Neuro-2a cell-based assay for palytoxin detection was confirmed by using ouabain, which also acts on Na+, K+-ATPase. Pre-treatment of the Neuro-2a cells with ouabain minimizes the effects of palytoxin. The specificity of the Neuro-2a assay was confirmed by the finding that cell death was not detected when Neuro-2a cells were exposed to other phycotoxins with unrelated cellular targets. When the Neuro-2a assay was used to detect palytoxin in mussel extracts spiked with levels of palytoxin around the proposed regulatory value of 250 microg palytoxin/kg shellfish, a good correlation was observed between the levels found and the expected values. We conclude by proposing an experimental design for functional assays using the Neuro-2a cell line for the specific detection of four neurotoxic phycotoxin families: saxitoxins, brevetoxins, ciguatoxins and palytoxins.
authors.affiliation: Unité Ecosystèmes et interactions toxiques, USM 0505/EA 4105 MNHN, Muséum National d'Histoire Naturelle, Case 39, 12 rue Buffon, F-75231 Paris Cedex 5, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Ledreux
authors.forename: Sophie
authors.initials: S
authors.lastname: Krys
authors.forename: Cécile
authors.initials: C
authors.lastname: Bernard
mesh_terms: D000178:Acrylamides / Q000633:toxicity*; D000818:Animals; D001681:Biological Assay; D049872:Bivalvia / Q000737:chemistry; D045744:Cell Line, Tumor; D002470:Cell Survival; D003064:Cnidarian Venoms; D008387:Marine Toxins / Q000633:toxicity*; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism*; D000095702:Polyether Toxins
chemical_list: D000178:Acrylamides; D003064:Cnidarian Venoms; D008387:Marine Toxins; C010272:palytoxin; D000095702:Polyether Toxins