Adverse effects of diethyl phthalate and butyl benzyl phthalate on circadian rhythms and sleep patterns in zebrafish larvae

Abstract

The zebrafish, a diurnal vertebrate, is commonly used in circadian rhythm studies due to its genetic and neurological similarities to humans. Circadian rhythms, which regulate sleep, neurotransmitter, behavior, and physiological responses to environmental changes, can be disrupted by various environmental factors. Phthalic acid esters (PAEs) are pervasive endocrine disruptors that individuals are frequently exposed to in daily life. However, the impact of PAEs on circadian rhythms during early development remains poorly understood. This study aimed to investigate the effects of exposure to diethyl phthalate (DEP) and butyl benzyl phthalate (BBzP) on the behavior and circadian rhythms of developing zebrafish larvae using a series of layered assays. Zebrafish larvae were exposed to the two PAEs from less than 2 hour post-fertilization (hpf) until 96 hpf. The results demonstrated a concentration-dependent reduction in tail coiling (TC), touch-evoked response (TER), and locomotor activity, alongside an increase in sleep time and alterations in sleep bouts and sleep latency during both 24-hour and Light1/Dark/Light2 (7/10/7-hour) periods. Additionally, exposure to BBzP led to increased acetylcholinesterase (AChE) and dopamine (DA) levels, and a decrease in 5-hydroxytryptamine (5-HT) levels. Gene expression analysis revealed that DEP and BBzP exposure increased the expression of circadian rhythm and light-response-related genes. In conclusion, exposure to these PAEs disrupts the circadian rhythm of zebrafish larvae, providing novel insights into the developmental impact of these common environmental contaminants. Further research is needed to understand the broader implications of these findings for human health and environmental safety.

Keywords

• Diethyl phthalate
• Butyl benzyl phthalate
• Circadian rhythm
• Sleep bout
• Sleep latency