Comparative transcriptome analysis identified genes involved in testicular development in Takifugu rubripes

To identify candidate genes and pathways involved in testicular development in Takifugu rubripes, a comparative transcription analysis was conducted across the various developmental stages of the testis (stages II to V). A total of 9520 differentially expressed genes (DEGs) were identified among the different stages, and they were significantly clustered into six clusters (P < 0.05). One thousand four hundred eleven DEGs such as gndf, wnt1, and cyp17b1 were found to be decreased from stage II to V. In contrast, 994 DEGs such as fn1, ift81, and cdc25a were found to be increased from stage II to V. Six thousand three hundred eighteen DEGs (e.g., dmrt1, sdk2, and chrna1) were identified as being expressed at similar levels at stages II and III. However, they were subsequently found to be decreased from stage III to IV. Four hundred one DEGs exhibited a significant upregulation trend from stage II to III. These genes were expressed at similar levels in stages III, IV, and V, including chrnd, wnt4a, and cyp7a1. The highest expression levels of 200 DEGs (e.g., ccnb2, cdk1, and sycp2) were observed in stage IV, while 196 DEGs (e.g., chmp1b, hsd17b3, and zp3) exhibited the highest expression level in stage III. Those DEGs were mainly enriched in the pathways (e.g., neuroactive ligand-receptor interaction, cell adhesion molecules, and calcium signaling pathways) associated with testicular development. Quantitative polymerase chain reaction of eight randomly selected genes validated the RNA sequencing results. This study may provide new insights into the molecular regulatory mechanisms governing testicular development and spermatogenesis in T. rubripes.