The aim of this study was to detect the seasonal expressions of oxytocin (OT), oxytocin receptor (OTR), extracellular signal-regulated kinase1 and 2 (ERK1/2) and phospho-ERK1/2 (pERK1/2) in the epididymis of the wild ground squirrels (Citellus Dauricus Brandt) during the breeding season and non-breeding season. Histological results showed that size, weight, cell number and lumen diameter of epididymis underwent acute seasonal changes, which were all peaked in the breeding season. Immunohistochemical results suggested that strong staining of OT, OTR, ERK1/2, and pERK1/2 were observed in the epithelial layer in the whole epididymis, along with intense OT and OTR signal in smooth muscle cell (Smc) in caudal epididymis in the breeding season. The protein expression levels of OTR, ERK1/2, and pERK1/2 in the epididymis were higher in the breeding season than those of the non-breeding season. Besides, hormone assay revealed that there was no significant serum concentration of OT in these two periods, while epididymal concentration showed higher value in the breeding season. In summary, the identified localization and local concentration of OT in the epididymis in the wild ground squirrel suggested that epididymis may assume as a source of OT, and OT could act via OTR to activate ERK1/2 signaling to regulate seasonal epididymal functions.