Alterations in the distribution of actin and its binding proteins in the porcine endometrium during early pregnancy: Possible role in epithelial remodeling and embryo adhesion

During early pregnancy, uterine epithelial cells undergo major transformations in their cytoskeleton that make the endometrium receptive for conceptus attachment. Actin binding proteins (ABPs) such as cofilin, gelsolin, and vinculin are involved in regulating actin polymerization, severing or crosslinking actin to integrins. However, whether ABPs are involved in epithelial remodeling or embryo adhesion in pigs is unknown. Therefore, the expression and distribution of these proteins were investigated in porcine endometrium on Days 10 and 13 (pre-implantation period), and 16 (attachment phase) of the estrous cycle or pregnancy. While day and pregnancy status had no effect on ABP gene expression, the protein abundance of vinculin was significantly higher on Day 13 than on Day 10 (p < 0.05) of the estrous cycle, and its abundance was highest on Day 16 in the pregnant endometrium. Immunofluorescent staining showed alterations in the distribution of these proteins depending on the day of the estrous cycle or early pregnancy examined. Double immunofluorescent staining for the ABPs and actin revealed that while cofilin co-localized with actin in the apical epithelium on Days 13 and 16 of the estrous cycle, in pregnant animals, it was strongly associated with actin in the sub-epithelial stroma of the endometrium. Gelsolin was also co-localized with actin in the apical epithelium on Days 13 and 16 of the estrous cycle, but this association was absent in the pregnant endometrium. Vinculin co-localized with actin in the sub-epithelial stroma on Days 13 and 16 irrespective of the reproductive status, but was additionally associated with actin in the apical epithelium on Day 16 of pregnancy. Vinculin interacted with phosphorylated focal adhesion kinase in the endometrial epithelium, and the interaction was dependent on estradiol-17β, a conceptus-secreted pregnancy-recognition factor in pigs. Furthermore, silencing vinculin in the endometrial epithelial cells negatively affected trophoblast adhesion to them. In conclusion, the influence of stage and reproductive status on the specific localization of actin and its binding proteins in the porcine endometrium suggests that they play a role in regulating the endometrial cytoskeleton. Moreover, vinculin may facilitate conceptus attachment to the epithelium by interacting with focal adhesion kinase.