Semen Quality, Testicular Cell Apoptosis, and Transcriptome Analysis Following Mild Scrotal Heat Stress in Wugu-Hu Crossbred and Hu Rams.

Hu and Wugu × Hu rams underwent scrotal insulation to simulate mild heat stress, resulting in a 3.0 ± 0.1 °C increase in scrotal surface temperature. Semen samples were collected every five days from day 11 to 56, and testis samples immediately after insulation. Both breeds experienced similar semen quality reductions and recovery trends, including reduced motility, concentration and the percentage of morphologically normal, but on days 41 and 46, Wugu-Hu rams exhibited significantly lower sperm motility than Hu rams ( < 0.05). Wugu-Hu rams demonstrate more transcriptomic changes. Further GO analysis revealed enrichment in spermatogenesis-related processes, while KEGG analysis identified Oocyte meiosis and cell cycle pathways, with a downregulation of key genes (CDK1, CDK2, CDC20, and PLK1) indicating impaired meiosis in Wugu-Hu rams. In contrast, Hu rams showed minimal transcriptional changes, contrary to the transcriptomic results. The significantly increased apoptosis rate of Wugu-Hu sheep testicular cells ( < 0.05) suggests compensatory or post-transcriptional mechanisms mitigating functional impacts caused by transcriptomic changes. The conclusion is that mild scrotal heat stress affects sperm quality and testicular gene expression. Wugu-Hu rams demonstrate greater transcriptomic sensitivity, but this does not show significant differences in semen quality recovery due to the compensatory mechanism of cell apoptosis.