Research Paper Volume 13, Issue 1 pp 1001—1016

Conditional deletion of Wntless in granulosa cells causes impaired corpora lutea formation and subfertility

WntlessFlox/Flox, Amhr2-Cre females displayed subfertility with normal oocyte and embryonic development. (A) Comparison of the accumulative number of pups per WntlessFlox/Flox, WntlessFlox/-, Ddx4-Cre, WntlessFlox/Flox, Gdf9-Cre, and WntlessFlox/Flox, Amhr2-Cre females (n = 6 for each group). (B) Histological ovarian images of WntlessFlox/Flox and WntlessFlox/Flox, Amhr2-Cre females. Scale bar = 100 μm. The ovaries were collected from 6-wk-old mice with a random cycle. (C) 345 and 400 GV oocytes from WntlessFlox/Flox and WntlessFlox/Flox, Amhr2-Cre females were matured in M16 medium, respectively. After maturing for 3 and 16 h, oocytes were counted, and the rates for GVBD and PB1 were calculated, respectively. Representative images of mouse oocytes at GVBD and PB1 stages are shown above the bar graph. (D) Zygotes collected from WntlessFlox/Flox (n=160) and WntlessFlox/Flox, Amhr2-Cre (n=193) females with an obvious vaginal plug after mating with wildtype male mice were cultured in KSOM medium for 1 and 3 days to calculate the rates of 2-cell and blastocyst, respectively. Representative images of mouse 2-cell and blastocyst are shown above the bar graph. Experiments were repeated a minimum of three times; the data are presented as mean ± SEM.

Figure 3. WntlessFlox/Flox, Amhr2-Cre females displayed subfertility with normal oocyte and embryonic development. (A) Comparison of the accumulative number of pups per WntlessFlox/Flox, WntlessFlox/-, Ddx4-Cre, WntlessFlox/Flox, Gdf9-Cre, and WntlessFlox/Flox, Amhr2-Cre females (n = 6 for each group). (B) Histological ovarian images of WntlessFlox/Flox and WntlessFlox/Flox, Amhr2-Cre females. Scale bar = 100 μm. The ovaries were collected from 6-wk-old mice with a random cycle. (C) 345 and 400 GV oocytes from WntlessFlox/Flox and WntlessFlox/Flox, Amhr2-Cre females were matured in M16 medium, respectively. After maturing for 3 and 16 h, oocytes were counted, and the rates for GVBD and PB1 were calculated, respectively. Representative images of mouse oocytes at GVBD and PB1 stages are shown above the bar graph. (D) Zygotes collected from WntlessFlox/Flox (n=160) and WntlessFlox/Flox, Amhr2-Cre (n=193) females with an obvious vaginal plug after mating with wildtype male mice were cultured in KSOM medium for 1 and 3 days to calculate the rates of 2-cell and blastocyst, respectively. Representative images of mouse 2-cell and blastocyst are shown above the bar graph. Experiments were repeated a minimum of three times; the data are presented as mean ± SEM.