FIGURE 3.

Sperm energy restriction and recovery (SER) treatment impacts on embryonic parameters and implantation potential after non-surgical embryo transfer. A fraction of the blastocysts derived from CAP or SER treatments as described in Figure 2 were used to evaluate the impact of these treatments in blastocyst potential. (A) Blastocysts derived from CAP- or SER-treated sperm were transferred to pseudo-pregnant recipient female mice. The total number of pups obtained from either CAP or SER treatment (n = 15) is indicated. Image shown is representative of a litter obtained after transfer of SER-derived embryos. (B) Percentage of pups born per number of embryos transferred is indicated. (C) Blastocysts derived from CAP- or SER-treated sperm were stained with Hoechst as indicated in the section “Materials and Methods.” The number of stained nuclei that represents the number of cells in each blastocyst was counted (n = 3, 35 blastocysts/treatment counted). Representative images of stained embryos are shown. Scale bar = 25 μm. (D) Blastocyst derived from CAP or SER-treated sperm were assayed for outgrowth in vitro as explained in the section “Materials and Methods” (n = 10, 205 and 210 blastocysts counted for CAP and SER-derived blastocysts, respectively). In each panel, significant differences are indicated as: ^∗p < 0.05; ^∗∗p < 0.01, and ^∗∗∗p < 0.001. Experiments represented in this figure were done using blastocysts obtained by heterologous fertilization.