Saxitoxins induce cytotoxicity, genotoxicity and oxidative stress in teleost neurons in vitro

Saxitoxins induce cytotoxicity, genotoxicity and oxidative stress in teleost neurons in vitro

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 concentrat...

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Bibliographic Details
Published in:Toxicon (Oxford) Vol. 86; pp. 8 - 15
Main Authors: Silva, Cesar Aparecido da, Morais, Elizabeth Cunha Penna de, Costa, Michele Dietrich Moura, Ribas, João Luiz Coelho, Guiloski, Izonete Cristina, Ramsdorf, Wanessa A., Zanata, Silvio Marques, Cestari, Marta M., Oliveira Ribeiro, Ciro Alberto, Magalhães, Valéria F., Trudeau, Vance L., Silva de Assis, Helena C.
Format: Journal Article
Language:English
Published: England Elsevier Ltd 01-08-2014
Subjects:
Animals
Assessments
Brain - cytology
Brain - drug effects
Cell Culture Techniques
Cellular
Culture
Environmental neuroscience
Flatfishes
Freshwater
Genotoxicity
Hoplias malabaricus
Neuron primary culture
Neurons
Neurons - cytology
Neurons - drug effects
Neurotoxicology
Oxidative Stress
Saxitoxin - toxicity
Sodium
Stress concentration
Stresses
Teleost
Teleostei
Teleost
Environmental neuroscience
Neuron primary culture
Neurotoxicology
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Summary: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. •A protocol for primary culture of neurons from freshwater fish was established.•The basic growth fibroblast factor showed better results to the cell culture.•Saxitoxin caused cytotoxicity, genotoxicity and oxidative stress.•New applications in fish ecotoxicology in the assessment of neurotoxic substances.
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ISSN:0041-0101
1879-3150
DOI:10.1016/j.toxicon.2014.04.016