The investigation of hyaluronic acid resertors in rat implantation model by immunohistochemical and alactron microscopy

Hyaluronic acid (HA) was known to play a role in fertilization, implantation, embryonic cavitation, embryo growing and tissue morphogenesis. HA suggests increasing embryo implantation rate in IVF applications. HA receptors (CD44, CD44s, RHAMM and versican) [1,2, 3] and HA synthase 2 (HAS2) are mediated HA functions, but the roles of HA arent known exactly on implantation. In this study, we aim to evaluate the role of HA on the implantation process.Thirty-six adult female rats were divided in six groups: as control (proosterus, estrous and diostreus) and experimental groups (after fertilization 4.day, 5. day, 6. day). Menstruation periods were detected with vaginal smear both control group (proestrous, estrous and diestrous) and experimental group. Experimental group animals were mated in the estrous period. Implantation regions of experimental group were determinated with Chicago blue dye and uterine samples were obtained. Tissue samples of both groups were stained with CD44, CD44s, RHAMM, versican and HAS2 polyclonal antibodies using immunohistochemical technique. Immunoreactivity was evaluated as weak, moderate or strong and results were evaluated comparatively using Mann Whitney statistical test. İn this stages, ultrastructure of pinopodes and uterin epithelial evaluate using transmission electron microscope.In evaluation as immunohistochemical, strong HAS2 immunoreactivity of epithelial and subepithelial was observed in proestrous and desidua on 5th, 6th days but moderate in diestrous cycle. Strong CD44 immunoreactivity of endometrial epithelial cells was seen in proestrous and estrous cycle but moderate in diestrous cycle and on 5th day. Strong CD44s immunoreactivity of deep stroma was seen in diestrous and desidua on 5th, 6th days but moderate in proestrous and estrous cycle. Strong RHAMM immunoreactivity of epithelial, subepithelial and deep stroma was seen in proestrous and desidua on 5th, 6th days but moderate in diestrous and estrous cycle. Strong Versican immunoreactivity of subepithelial and deep stroma was seen in diestrous and desidua on 6th day but moderate in proestrous and estrous cycle.In evaluation of thin sections, in proestrous and estrous was increased microvilli but decreased in the diestrous. In experimental group was observed bleb on 5th day.In tissue samples of experimental group, increase as statistical of HAS2, CD44s, RHAMM immunoreactivity in 5th and 6th days; CD44 immunoreactivity in 5th day and Versican immunoreactivity in 6th day, pinopodes in electron microscopy was observed in 5th day and decreased of silial, It was concluded that te HA play a important role during implantation.