Acute embryonic exposure of zebrafish to permethrin induces behavioral changes related to anxiety and aggressiveness in adulthood

doi:10.1016/j.jpsychires.2019.11.006

Journal of Psychiatric Research

Volume 121, February 2020, Pages 91-100

https://doi.org/10.1016/j.jpsychires.2019.11.006 Get rights and content

Abstract

Permethrin (PM) is one of the most used synthetic pyrethroid worldwide. Exposure to this compound during pregnancy and early childhood has been indicated as a risk factor for neurodevelopmental disorders. We evaluated the long-term effects of embryonic PM exposure in different stages of zebrafish development. Briefly, embryos (3 hpf) were exposed to sub-lethal concentrations of PM (25 and 50 μg.L⁻¹) during 24 h and then behavioral parameters were evaluated during embryonic (28 hpf), eleutheroembryonic (3 dpf), larval (7 dpf), and adult stages (90 dpf). PM exposure decreased spontaneous movement at 28 hpf and decreased thigmotaxis in eleutheroembryos. The long-term effects of PM include changes in non-motor behaviors such as fear and anxiety in larva and adults. Adults embryonically exposed to PM also showed a significant increase in aggressiveness parameters. These results demonstrated that embryonic exposure to PM induces persistent neurotoxic effects in adulthood, which can impair the cognitive and behavioral fitness of non-target species contributing to a rise in neurodevelopmental disorders.

Introduction

Neurodevelopmental disorders have significantly increased over the last decades (Grandjean and Landrigan, 2014). Pieces of evidence indicate that environmental pollutants cause injury to the developing brain either through direct toxicity or interactions with genome (Landrigan et al., 2012). The pesticides are considered significant risk factors involved in the initiation and progression of neurodevelopmental and neurodegenerative diseases (Parrón et al., 2011).

Permethrin (PM; 3-phenoxybenzyl-(1R,S)-cis, trans-3-(2,2-dichlorovinyl)-2,2- dimethylcyclopropane carboxylate) is a synthetic type I pyrethroid frequently detected in domestic environments, being the second most frequently detected pyrethroid (Tang et al., 2018). Besides that, PM concentrations in water column have been found to range from 0.05 to 811 μg.L⁻¹ (Yang et al., 2014). The primary molecular target of pyrethroids is the voltage-gated sodium channels (VDSC) (Soderlund, 2013). The PM widely use is commonly associated with low mammalian toxicity; however, evidences suggest that PM might induce neurotoxic effects (Nasuti et al., 2008a, Nasuti et al., 2008b) related to motor and sensorimotor dysfunction, such as whole-body tremor and convulsions (Wolansky and Harrill, 2008). Nevertheless, the chronic effects of PM in non-motor behaviors, as well as the persistence of these effects during the development, still need to be clarified.

In adult rats, the exposure to pyrethroids was shown to cause hypersensitivity and aggression, followed by stimulus-induced bouts of general tremor, convulsive twitching, coma, and death (Verschoyle and Barnes, 1972). In humans, prenatal exposure to pyrethroids was associated with worsening of behavioral regulation, emotional control, shifting, internalizing behaviors, depression, somatization, emotional control, inhibition, externalizing problems, and conduct problems in childhood (Furlong et al., 2017). Epidemiological studies have shown that the detection of urinary 3-phenoxybenzoic acid, a biomarker of exposure to pyrethroids, indicates that children and pregnant women may bioaccumulate higher levels of pyrethroids than adults (Watkins et al., 2016). Thus, children exposed during early developmental stages to PM have raised concerns mainly due to the vulnerability of developing brain to the effects of neurotoxic pesticides (Grandjean and Landrigan, 2014). Additionally, studies in mammals have been related to the persistence effects of perinatal exposure to subtoxic doses of pesticides with changes in anxiety, aggressive, and abnormal social behaviors during adult stage (Bjørling-Poulsen et al., 2008). Therefore, a better understanding of the potential impairing outcomes of pyrethroid exposure during early life is needed.

The zebrafish (Danio rerio) has become an outstanding tool for biological psychiatry (Levin et al., 2015) and neurodevelopmental studies (Roper and Tanguay, 2018), due to its fast embryonic ex-utero development and highly conserved genetic programming during early stages of life (Kalueff et al., 2013). Thus, this study aimed to evaluate the long-term effects of embryonic exposure to low concentrations of PM through behavioral endpoints across different developmental stages.

Section snippets

Animals maintenance and reproduction

Wild-type zebrafish of both sexes were maintained in a recirculation aquatic system (Zebtec®) under appropriate conditions and fed thrice daily on commercial flocked fish food as established elsewhere (Costa-Silva et al., 2018a, Costa-Silva et al., 2018b).

Zebrafish (2/1 male/female; 6–12 months-old) were placed overnight. The reproduction was induced by the light irradiation on the next morning. Fertilized eggs were washed with Hank's solution (HS) for several times and incubated in system

PM impairs embryonic motor behavior

Embryos at 28hpf exposed to PM during the first hours of development showed a significant decrease of spontaneous movement episodes (Fig. 2).

PM decrease sensory-motor capacity in eleutheroembryos

When the exposed embryos reached the stage of eleutheroembryo, it was possible to observe a significant increase in the number of stimuli needed to first scape response in embryos exposed to 50 μg.L⁻¹ (Fig. 3A). The percent of eleutheroembryos able to escape to periphery of the apparatus was significantly decreased in both exposed groups (Fig. 3B).

PM exposure alters non-motor behaviors during larval stage

Embryos

Discussion

This study aimed to examine the adverse effects of embryonic exposure of zebrafish to sub-lethal concentrations of PM along different stages of development. We used behavioral assays to assess such effects throughout zebrafish's early life and adulthood. We hypothesized that submitting zebrafish embryos to environmental concentrations of this compound during the first hours of development would result in neurological impacts in adulthood, and what we found was that PM treatment resulted in

Declaration of competing interest

On behalf of all authors listed below I declare no conflict of interest.

Acknowledgments

This study was supported by CAPES PROEX (23038.005848/2018-31), CAPES, CNPq (456207/2014-7), FAPERGS (2380-2551/14-8), and FAPERGS/PRONEX (16/25510000499-4). JLF is a recipient of CNPq fellowship. MEMN, IKM, GEM, and NRR are recipients of CAPES doctoral scholarship.

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Acute embryonic exposure of zebrafish to permethrin induces behavioral changes related to anxiety and aggressiveness in adulthood

Abstract

Introduction

Section snippets

Animals maintenance and reproduction

PM impairs embryonic motor behavior

PM decrease sensory-motor capacity in eleutheroembryos

PM exposure alters non-motor behaviors during larval stage

Discussion

Declaration of competing interest

Acknowledgments

Aquat. Toxicol

J. Physiol. Paris

Brain Res.

Brain Res.

Behav. Process.

Neurotoxicol. Teratol.

Psychoneuroendocrinology

Neurotoxicology

Lancet Neurol.

Aquat. Toxicol

Behav. Brain Res.

Pharmacol. Biochem. Behav

Behav. Brain Res.

Dev. Brain Res.

Environ. Int.

Toxicology

Toxicology

Toxicol. Appl. Pharmacol.

Chemosphere

Brain Res.

Chemosphere

Ecotoxicol. Environ. Saf.

Chemosphere

Pestic. Biochem. Physiol.