Abstract: The experiment was conducted to reveal whether the superior spermatic nerve (SSN) and the inferior spermatic nerve (ISN) were involved in regulating testicular development and spermatogenesis. The present study was thus designed to transect the SSN and ISN of rats on PD10 and then analyzed changes on PD25 and PD50. The results demonstrated that testicular denervation have no obvious influence to testis mass on PD25（P<0.05）, but significantly reduced testis mass, cauda epididymal sperm counts on PD50（P<0.01）. Proliferating cell nuclear antigen (PCNA) immunohistochemistry staining proved that the denervation had no influence on the proliferarion of germ cells on PD25, but obviously inhibited the proliferation on PD50. In addition, RT-PCR results showed that testis denervation significantly decreased testis 3β-hydroxy sterol dehydrogenase 1 (3β-HSD1) on PD25（P<0.05）and significantly decreased testis 3β-HSD1 and androgen binding protein (ABP) mRNA levels（P<0.01）, luteinizing hormone receptor (LHR) and follicle stimulating hormone receptor (FSHR) mRNA levels on PD50（P<0.05）. These results suggest that the testicular nerve supply plays an important role in supporting seminiferous tubule development and spermatogenesis after testis descend to scrotum.