Rotenone Neurotoxicity Causes Dopamine Neuron Loss in Zebrafish
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Abstract
ABSTRACT:
Objectives: We sought to determine whether rotenone, a commonly used pesticide, exhibits neurotoxicity in zebrafish by causing dopamine neuron loss through rotenone-induced oxidative damage. Methods: We exposed transgenic zebrafish embryos expressing green fluorescent protein under the control of the cis-regulatory elements of dopamine transporter (dat) to rotenone to determine the neurotoxic effects of rotenone on dopamine neuron abundance and pattern distribution, as well as the presence of apoptotic markers. The oxidative stress potential of rotenone on embryos was assessed using a live MitoSOX Red assay, and behavioural testing on adult zebrafish was assessed using video recordings of midline crossing events. Results: Zebrafish embryos treated with rotenone displayed a 50% reduction in dopamine neurons in the ventral diencephalon when exposed to 30µM rotenone (n=6, p<0.001), and rotenone-exposed zebrafish raised to adulthood demonstrate an anxiety-like behaviour (n=5, p<0.01). Furthermore embryos exposed to rotenone also demonstrated a logarithmic increase in markers of oxidative damage (n=3, p<0.001) and apoptotic activity in their diencephalic neurons. Conclusions: These results show that rotenone can induce dopamine neuron loss in zebrafish, providing a useful model for studying the environmental causes of Parkinson’s disease.
RÉSUMÉ:
Objectif: Nous cherchons à déterminer si la roténone, un élément commun dans les pesticides, démontre de la neurotoxicité dans les poissons-zèbres en causant une perte de dopamine dans leurs neurones à travers le dommage oxydatif induit par la roténone. Méthode: Nous avons exposé des embryons de poissons-zèbres transgéniques qui expriment la protéine fluorescente verte sous le contrôle d’éléments cis-régulateurs des transporteurs sélectifs de dopamine (dat) à la roténone pour déterminer les effets neurotoxiques de ce dernier sur les niveaux dopaminergiques dans leurs neurones. De plus, nous avons évalué la présence de marqueurs apoptotiques. Le stress oxydatif potentiel de la roténone sur les embryons a été analysé par le « live MitoSOX Red assay » et les tests comportementaux sur les poissons-zèbres adultes furent analysés en utilisant des enregistrements vidéo. Résultats: Les embryons de poissons-zèbres qui ont été traités avec la roténone ont démontré une réduction de dopamine de 50% dans les neurones localisés dans le diencéphale ventral, quand exposés à 30µM de roténone (n=6, p<0.001). Ils ont également illustré une augmentation logarithmique dans les marqueurs de dommage oxydatif (n=3, p<0.001) et une activité apoptotique dans les neurones du diencéphale. Les poissons-zèbres exposés à de la roténone qui ont atteint l’âge adulte ont démontré des comportements d’anxiété (n=5, p<0.01). Conclusion: Les résultats démontrent que la roténone peut induire une perte dopaminergique dans les neurones des poissons-zèbres. Ces résultats s’avèrent utiles pour étudier davantage les causes environnementales reliées à la maladie de Parkinson.
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