title: Saxitoxins induce cytotoxicity, genotoxicity and oxidative stress in teleost neurons in vitro.
abstract: The aim of this study was establish a protocol for isolation and primary culture of neurons from tropical freshwater fish species Hoplias malabaricus for assessment of the effects of neurotoxic substances as saxitoxins (STXs). Cells from brain of H. malabaricus were treated with different concentrations of trypsin, dispase and papain for tissue dissociation. Cells type was separated by cellular gradient and basic fibroblast growth factor (bFGF) supplement nutrition media were added. The dissociated cells were plated with medium and different STXs concentrations and the toxic cellular effects such as oxidative stress, neurotoxicity, and genotoxicity and apoptosis process were evaluated. Cultures treated with bFGF showed the greatest adherence, survival and cellular development. STXs increased specific activity of glutathione peroxidase and lipoperoxidation levels, were cytotoxic and genotoxic indicated by the comet assay. Although the STXs effects due the blockage of sodium channels is reported to be reversible, the time exposure and concentration of STXs suggested cellular injuries which can lead to neuropathology. The establishment of primary neuronal culture protocol enables new applications for neurotoxicological assessments.
authors.affiliation: Ecology and Conservation Post-graduate Program, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Cesar Aparecido
authors.initials: CA
authors.lastname: da Silva
authors.affiliation: Department of Basic Pathology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Elizabeth Cunha Penna
authors.initials: EC
authors.lastname: de Morais
authors.affiliation: Department of Basic Pathology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Michele Dietrich Moura
authors.initials: MD
authors.lastname: Costa
authors.affiliation: Department of Pharmacology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: João Luiz Coelho
authors.initials: JL
authors.lastname: Ribas
authors.affiliation: Department of Pharmacology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Izonete Cristina
authors.initials: IC
authors.lastname: Guiloski
authors.affiliation: Department of Genetics, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Wanessa A
authors.initials: WA
authors.lastname: Ramsdorf
authors.affiliation: Department of Basic Pathology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Silvio Marques
authors.initials: SM
authors.lastname: Zanata
authors.affiliation: Department of Genetics, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Marta M
authors.initials: MM
authors.lastname: Cestari
authors.affiliation: Department of Cellular Biology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil.
authors.forename: Ciro Alberto Oliveira
authors.initials: CA
authors.lastname: Ribeiro
authors.affiliation: Federal University of Rio de Janeiro, Institute of Biophysics Carlos Chagas Filho, 21941-902 Rio de Janeiro, RJ, Brazil.
authors.forename: Valéria F
authors.initials: VF
authors.lastname: Magalhães
authors.affiliation: Department of Biology, University of Ottawa, K1N 6N5 Ottawa, Ontario, Canada.
authors.forename: Vance L
authors.initials: VL
authors.lastname: Trudeau
authors.affiliation: Department of Pharmacology, Federal University of Paraná, 81531-990, P.O. Box 19031, Curitiba, Paraná, Brazil. Electronic address: helassis@ufpr.br.
authors.forename: Helena C Silva
authors.initials: HC
authors.lastname: de Assis
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000187:drug effects; D018929:Cell Culture Techniques; D005412:Flatfishes; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D018384:Oxidative Stress; D012530:Saxitoxin / Q000633:toxicity*
chemical_list: D012530:Saxitoxin
keywords: Environmental neuroscience; Neuron primary culture; Neurotoxicology; Teleost