Embryonic Zebrafish Irritant-evoked Hyperlocomotion (EZIH) as a high-throughput behavioral model for nociception

Abstract

Behavioral models have served a key role in understanding nociception, the sensory system by which animals detect noxious stimuli in their environment. Developing zebrafish (Danio rerio) are a powerful study organism for examining nociceptive pathways, given the vast array of genetic, developmental, and neuroscience tools available for these animals. However, at present there are few widely-adopted behavioral models for nociception in developing zebrafish. This study examines the locomotor response of hatching-stage zebrafish embryos to dilute solutions of the noxious chemical and TRPA1 agonist allyl isothiocyanate (AITC). At this developmental stage, AITC exposure induces a robust uniphasic hyperlocomotion response. This behavior was thoroughly characterized by determining the effects of pre-treatment with an array of pharmacological agents, including anesthetics, TRPA1 agonists/antagonists, opioids, NSAIDs, benzodiazepines, SSRIs, and SNRIs. Anesthetics suppressed the response to AITC, pre-treatment with TRPA1 agonists induced hyperlocomotion and blunted the response to subsequent AITC exposures, and TRPA1 antagonists and the opioid buprenorphine tended to reduce the response to AITC. The behavioral responses of zebrafish embryos to a noxious chemical were minimally affected by the other pharmacological agents examined. The feasibility of using this behavioral model as a screening platform for drug discovery efforts was then evaluated by assaying a library of natural product mixtures from microbial extracts and fractions. Overall, our results indicate that irritant-evoked locomotion in embryonic zebrafish is a robust behavioral model for nociception with substantial potential for examining the molecular and cellular pathways associated with nociception and for drug discovery efforts.