Full-Length Transcriptome Analysis of Male and Female Gonads in Japanese Eel (Anguilla japonica)

Background: The Japanese eel (Anguilla japonica) holds significant economic value in East Asia, but limitations in understanding its reproductive biology have hindered advancements in artificial breeding techniques. Previous research has primarily focused on conserved sex differentiation genes, offering limited insights into the broader molecular mechanisms driving gonadal development and sexual dimorphism. To address these limitations, this study aims to investigate key genes and pathways involved in gonadal development through a comprehensive transcriptomic analysis of male and female eel gonads. Results: PacBio Iso-Seq and Illumina RNA-Seq technologies were combined to conduct a full-length transcriptome analysis of male and female Japanese eel gonads at a post-differentiation, pre-maturation stage. A total of 24661 unigenes were identified in ovaries and 15023 in testes, along with genomic regulatory elements such as transcription factors, simple sequence repeats, and long non-coding RNAs. Additionally, 1,210 differentially expressed genes were detected. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed significant pathways involved in cell cycle regulation, metabolic processes, apoptosis, and hormone activity. Notably, several reproductive-related genes, including bambi, ccnb1, cdc20, gdf9, prlh, ccdc39, chrebp, tspo, syce3, and ngb, demonstrated significant dimorphic expression in eel gonads. Conclusions: This study provides valuable insights into the molecular mechanisms of gonadal differentiation and sexual dimorphism in Japanese eels. The findings expand the genetic resources available for the eel breeding industry and could facilitate the development of improved artificial breeding techniques focused on reproductive development. In this experiment, we investigated sexual dimorphism and gonadal development in aquaculture-reared Japanese eels (Anguilla japonica) at the silver eel stage, where gonads had differentiated but remained immature without hormone induction. Ovarian tissues from three females and testicular tissues from three males were collected, forming two groups with three biological replicates each. Six RNA libraries were constructed and sequenced using Illumina RNA-seq for gene expression analysis, with PacBio Iso-seq data serving as a reference for transcript validation.