Abstract: In order to explore the effect of fixed-time artificial insemination procedures on the expression of genes related to gonadotropin-releasing hormone（GnRH） and WNT signaling pathways in corpus luteum of replacement gilts, 12 replacement gilts with good body condition were selected and divided into four groups: D16-APG group、D16-C group and D25-APG group, D25-C group（gilts in D16-APG and D25-APG groups were treated according to the procedure of fixed-time artificial insemination）. The animals were slaughtered on the 16 th and 25 th day of pregnancy, and the corpus luteum tissues were collected for the detection of the relative expression levels of genes related to the GnRH and WNT signaling pathways by fluorescence quantitative PCR. The results showed that in the GnRH signaling pathway, compared with the D16-C group, the mRNA relative expression levels of NRAS, MMP2, GNA11, and PLCβ4 in D16-APG group were significantly upregulated on the 16 th days of pregnancy（P＜0.05）; compared with D25-C group, the mRNA relative expression levels of MMP2, NRAS, and GNA11 in D25-APG group were significant increase on the 25 th days（P＜0.05）; in the WNT signaling pathway, compared with the D16-C group, the mRNA relative expression levels of WNT10 B,PRKCB,CTNNB1 and ESR1 in D16-APG group were significantly increased on the 16 th days of pregnancy（P＜0.05）;compared with the D25-C group, the mRNA relative expression level of PRKCB in D16-APG group was significantly increased on the 25 th days of pregnancy（P＜0.05）. The results indicated that fixed-time artificial insemination could regulate the expression of related genes through GnRH and WNT signaling pathways.