Zebrafish is a useful model organism in neuroscience; however, its gene expression atlas in the adult brain is not well developed. In the present study, we examined the expression of 38 neuropeptides, and glutamatergic neuron marker gene mix (slc17a6a, slc17a6b, slc17a7a, and slc17a7b) in the adult zebrafish brain using in situ hybridization. The results are summarized as an expression atlas in 19 coronal planes of the forebrain. Furthermore, the scan data of all sections were made publicly available as a database. Based on these data, we performed detailed neuroanatomical analyses of the hypothalamus. By scrutinizing and comparing the expression patterns of neuropeptides, we found that several regions described as one nucleus in the reference zebrafish brain atlas contain two or more subregions with significantly different neuropeptide/neurotransmitter expression profiles, and we proposed them as novel subnuclei. Subsequently, the expression data obtained in this study were compared with those in mice, and a cluster analysis was performed to examine the similarities. As a result, several nuclei in zebrafish and mice were clustered in close vicinity: zebrafish ventral part of the anterior part of the parvocellular preoptic nucleus (PPav)/magnocellular preoptic nucleus (PM) and mouse paraventricular hypothalamic nucleus (Pa), zebrafish posterior part of the parvocellular preoptic nucleus (PPp) and mouse medial preoptic area (MPA), zebrafish dorsal part of the ventral zone of periventricular hypothalamus (Hvd)/anterior tuberal nucleus (ATN) and mouse ventromedial hypothalamic nucleus (VMN). The present expression atlas, database, and anatomical findings will contribute to future neuroscientific research using zebrafish.