Abstract: Many physiological processes in fish are influenced by nicotinic acetylcholine receptors, such as neuromuscular signaling and parasympathetic neurotransmission. In the marine environment, predation on Amphiprion ocellaris by Conus is closely linked to the presence of nicotinic acetylcholine receptors. In this study, genes encoding different subunits of the nicotinic acetylcholine receptor were cloned using cDNAs from muscle, brain, and intestine tissues of Amphiprion ocellaris as templates. Subsequently, a bioinforma-tics analysis was conducted based on the genes and a phylogenetic tree of the nicotinic acetylcholine receptor of Amphiprion ocellaris was constructed. The results revealed the successful cloning of the gene sequences of the cholinergic receptor nicotinic alpha1 subunit（chrna1） and the cholinergic receptor nicotinic epsilon subunit（chrne） from muscle tissues of Amphiprion ocellaris. Additionally, the gene sequence of the cholinergic receptor nicotinic alpha1 subunit（chrna10） was obtained from brain tissues. The three-dimensional structure of the encoded protein was predicted using Colabfold software, and the results were highly similar to the structure of the nicotinic acetylcholine receptor obtained from previous studies. Fluorescence quantitative PCR was employed to examine the expression levels of the chrna, chrne and chrna10 genes in muscle, brain, and intestine tissues of Amphiprion ocellaris. The results indicated that the expression levels of the chrna and chrne genes were higher in muscle tissues, whereas chrna10 gene expression was lower in brain tissues and absent in muscle or intestinal tissues. Furthermore, evolutionary tree analysis demonstrated a high conservation of the acetylcholine receptor sequence across species, with significant homology to the model organism zebrafish（Danio rerio）.