SDF-1/CXCR4 axis maintains porcine prospermatogonia undifferentiated state through regulation of transcription suppressor PLZF

Prospermatogonia (ProSGs), the progenitors of spermatogonial stem cells in neonatal testes, undergo critical migration to the testicular microenvironment—a fundamental process for testicular development and subsequent spermatogenic capacity. The SDF-1/CXCR4 chemokine axis serves as an essential molecular guidance mechanism, directing ProSGs toward the basal membrane of seminiferous tubules. Nevertheless, the precise molecular mechanisms governing this axis remain incompletely understood. Utilizing a porcine in vitro model system, this investigation elucidated the molecular mechanisms underlying the SDF-1/CXCR4 axis in ProSGs fate determination. Through integrated molecular and transcriptomic analyses, we investigated the consequences of CXCR4 inhibition on ProSG cellular dynamics. Our findings demonstrated that the SDF-1/CXCR4 axis exerts regulatory control over ProSGs differentiation via the PI3K-AKT-AP-1 signaling cascade. This regulation significantly influences the transcriptional landscape of ProSGs, particularly modulating the expression of PLZF, a crucial suppressor of spermatogonial differentiation, and DMRT1, an essential mediator of germ cell differentiation. These findings elucidate the molecular mechanisms orchestrating ProSGs homing and emphasize the significance of maintaining male reproductive competence. Furthermore, this research could enhance our understanding of ProSGs biology and its relationship to boar fertility, while potentially facilitating the development of innovative reproductive technologies and sustainable livestock management strategies.