Rongyu Yao, Fadi Li, Xiuxiu Weng, Wanhong Li
α-linolenic acid (ALA) has been reported to benefit testicular function. However, dietary ALA is extensively metabolized in the rumen, whereas isolated cell models do not retain seminiferous tubule structure or interactions among testicular cells. This pilot study examined the effects of ALA on seminiferous tubule morphology, UCHL1-positive cells, and gene expression in cultured sheep testicular tissue fragments. Tissue from one 3-month-old Hu ram (n = 1) was cultured for 50 d in control medium (CK) or medium containing 5 μM ALA. Three independently cultured fragments per condition were analyzed. Histology, UCHL1 immunofluorescence, RNA sequencing, and qRT-PCR were performed. ALA-treated fragments showed greater seminiferous tubule diameter, a lower proportion of empty space, and more UCHL1-positive cells per seminiferous tubule. Gene sets related to ATP-dependent chromatin remodeling, DNA replication, and male meiotic nuclear division were enriched in ALA-treated fragments, whereas immune gene sets were enriched in CK fragments. The relative expression levels of SYCP3 and MKI67 were higher in ALA-treated fragments than in CK fragments. Overall, these preliminary findings suggest that ALA treatment was associated with morphological and transcriptional changes in cultured sheep testicular tissue. Further studies using tissues from multiple rams are needed to confirm these findings.