Journal of Jilin University(Medicine Edition) ›› 2022, Vol. 48 ›› Issue (2): 277-283.doi: 10.13481/j.1671-587X.20220202

• Research in basic medicine • Previous Articles     Next Articles

Expression levels of Wnt5a protein in ovary tissue of mice at different development stages and its effect on oocyte autophagy

Yabo MA1,2,Xiaotan YUAN1,2,Xianguo XIE1,2,Xinfeng LIU1,2,Jinrui XU1,2,Yi YANG1,2()   

  1. 1.Key Laboratory of Conservation and Utilization of Western Characteristic Biological Resources,Ministry of Education,Ningxia University,Yinchuan 750021,China
    2.Department of Microbiology and Molecular Biology,College of Life Sciences,Ningxia University,Yinchuan 750021,China
  • Received:2021-06-28 Online:2022-03-28 Published:2022-05-10
  • Contact: Yi YANG E-mail:yangyi@nxu.edu.cn

Abstract: Objective

To investigate the expression levels of Wnt5a protein in ovary tissue of the mice at different development stages, and to clarify its effect on oocyte autophagy.

Methods

The ovary tissue of mice at different developmental stages were collected, and Western blotting method was used to detect the expression levels of Wnt5a protein in perinatal ovary tissue of the mice. The ovarian tissue of the 1 d after partum (1 dpp) mice were selected for immunofluorescence staining. The oocyte cytoplasmic marker DDX4 and granulosa cell marker FOXL2 (red) were co-stained with Wnt5a (green) and nuclear marker Hoechst (blue), respectively,and the morphology and the number of oocytes were observed. The ovary tissue of the mice at 17.5 d of embryonic period(17.5 dpc)were divided into control group, 1 mol·L-1 overexpressing Wnt5a group (rWnt5a group), 1 mol·L-1 inhibitor IWP2 group and 1 mol·L-1 inhibitor BOX5 group. The expression levels of Wnt5a, LC3 and P62 proteins in ovary tissue of the mice in various groups were detected by Western blotting method, and the number of ovarian oocytes was observed by immunofluorescence method.

Results

Wnt5a was expressed in ovary tissue of the mice at different developmental stages. Compared with control group, the expression levels of Wnt5a protein in 17.5 dpc and 1 dpp groups were significantly increased (P<0.05). The fluorescence localization revealed that the target protein Wnt5a overlapped with the markers DDX4 and FOXL2. The Western blotting results showed that compared with control group, the expression level of Wnt5a protein in rWnt5a group was significantly increased (P<0.01), the expression levels of Wnt5a and LC3 proteins in inhibitor IWP2 group were significantly decreased (P<0.01), and the expression level of Wnt5a protein in inhibitor BOX5 group was significantly increased (P<0.01). Compared with rWnt5a group, the expression level of LC3 protein in inhibitor IWP2 group was significantly decreased(P<0.01),and the expression level of P62 protein was significantly increased (P<0.01). The immunofluorescence staining results showed that the number of oocytes in IWP2 inhibitor group was reduced compared with control group.

Conclusion

Wnt5a may regulate the oocyte survival by affecting the level of autophagy in ovary of the mice.

Key words: Wnt5a, Ovary tissue, Oocyte, Granulosa cell, Autophagy

CLC Number: 

  • Q955