Abstract In fish species, there is limited analysis of signature transcriptome profiles at the single‐cell level in gonadal cells. Here, the molecular signatures of distinct ovarian cell categories in adult Nile tilapia ( Oreochromis niloticus ) were analysed using single‐nucleus RNA sequencing (snRNA‐seq). We identified four cell types (oogonia, oocytes, granulosa cell, and thecal cell) based on their specifically expressed genes and biological functions. Similarly, we found some key pathways involved in ovarian development that may affect germline–somatic interactions. A cell‐to‐cell communication network between the distinct cell types was constructed. We found that the bidirectional communication is mandatory for the development of germ cells and somatic cells in fish ovaries, and the granulosa cells and thecal cells play a central regulating role in the cell network in fish ovary. Additionally, we identified some novel candidate marker genes for various types of ovarian cells and also validated them using in situ hybridization. Our work reveals an ovarian atlas at the cellular and molecular levels and contributes to providing insights into oogenesis and gonad development in fish.