Spermatogenesis is a highly productive and intricate process occurring in testes that produces functional haploid sperm capable of fertilization therefore sustaining lifelong male fertility. A cornerstone of spermatogenesis is primitive spermatogonia, including spermatogonial stem cells (SSCs), that are able to self-renew and differentiate. The molecular mechanisms for spermatogonial self-renewal and differentiation in large domestic animals such as pigs, in comparison with their counterparts in mice, are poorly understood. In this study, we explored the expression pattern of ZBTB16 (a key transcription factor also known as PLZF) and its regulatory repertoire in porcine immature spermatogonia. We first co-stained ZBTB16 with spermatogonial/proliferative markers (DBA, SALL4, UCHL1 or Ki67) on testis sections from four ages of boars, demonstrating that ZBTB16