Evaluation of fentanyl toxicity and metabolism using a zebrafish model

Abstract

The increased abuse of novel drugs has created a critical need for cheap and rapid in vivo models to understand whole organism drug‐induced toxicity and metabolic impacts. One such model is zebrafish, which share many similarities to human. Assays have been developed for behavioral, toxicity, and metabolism elucidation following chemical exposure. The zebrafish model provides the advantage of assessing these parameters within a single study. Previous zebrafish studies have evaluated the behavioral effects of fentanyl, but not developmental toxicity and its relation to metabolism. In this study, we evaluate the effects of fentanyl on the development of wild‐type (TL strain) zebrafish and its metabolism over 4 days. Fertilized eggs were exposed to six concentrations of fentanyl (0.01, 0.1, 1, 10, 50, and 100 μM) through embryo media incubated at 28–29°C. Observations included egg coagulation, somite formation, heartbeat, tail and yolk morphology, pericardial formation, and swim bladder inflation. The incubation media was analyzed for the presence of metabolites using a targeted metabolomics approach. Fentanyl concentration caused significant effects on survival and development, with notable defects to the tail, yolk, and pericardium at 50 and 100 μM. Despropionyl fentanyl (4‐ANPP), β‐hydroxy fentanyl, and norfentanyl were detected in zebrafish larvae. We present a single in vivo model to assess toxicity and metabolism of fentanyl exposure in a vertebrate model system. Our findings provide a foundation for further investigations into fentanyl's mechanism of action and translation to human drug exposure.

Zebrafish larvae: A new model to study behavioural effects and metabolism of fentanyl, in comparison to a traditional mice model

“…In particular, 100% of larvae exposed to fentanyl 50 and 100 μM showed an abnormal pericardial edema (PE), yolk sac edema (YSE), jaw defect (J) and spinal curvature (SC), as shown in Figure 1. These malformations have already been highlighted for fentanyl, 18,19 other fentanyl analogs, 16 cannabinoids 20 and environmental toxins. 21 Furthermore, at those concentrations, larvae seemed to be sedated and suffering, consequently it was decided to reduce the drug concentrations to 10 μM (which induces malformations only in 1/3 of cases) and 1 μM (that caused no malformations) for further experiments.…”

The metabolism of valerylfentanyl using human liver microsomes and zebrafish larvae

“…Studies have been performed to investigate metabolite markers for synthetic cannabinoids, 19 human performance‐enhancing drugs, 20,21 synthetic cathinones, 22 and opioids 23,24 using the zebrafish model. More recently, we utilized the zebrafish model as a single assay for the toxicity and metabolism of fentanyl and detected norfentanyl, β‐hydroxyfentanyl, and 4‐anilino‐ N ‐phenethylpiperidine (4‐ANPP) from 24 to 96 h postfertilization, 25 providing additional support for their ability to metabolize opioids.…”

“…More recently, we utilized the zebrafish model as a single assay for the toxicity and metabolism of fentanyl and detected norfentanyl, β-hydroxyfentanyl, and 4-anilino-N-phenethylpiperidine (4-ANPP) from 24 to 96 h postfertilization, 25 providing additional support for their ability to metabolize opioids.…”

Fentanyl Induces Novel Conditioned Place Preference in Adult Zebrafish, Disrupts Neurotransmitter Homeostasis, and Triggers Behavioral Changes

“…Compared to rodent models, zebrafish models require less space for behavioral testing, less expenditure on experimental individuals, and less requirements for animal welfare. It was found that the fentanyl metabolic pathway in zebrafish was similar to that of rats, implying that toxicity results from zebrafish could be verified by other models [ 18 ]. In addition, zebrafish larvae have been used to validate cardiotoxicity due to fentanyl exposure [ 19 ] and respiratory depression toxicity [ 20 ].…”