Abstract
The study of implantation has been facilitated by the identification of specific biomarkers that are associated with uterine receptivity. The alpha(v)beta(3) integrin is a cell surface adhesion receptor, whose expression has been shown to be elevated in the endometrium at the time of implantation in both humans and other mammalian species; however, the distribution of alpha(v)beta(3) in the rabbit model is unknown. The rabbit has been shown to be an excellent model for the study of implantation. As an obligate ovulator, the timing of pregnancy can be precisely established, and embryonic attachment occurs through specialized trophoblast-endometrial structures known as trophoblastic knobs. In the present study, the expression of alpha(v)beta(3) integrin subunit in the rabbit uterus was examined by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and in situ hybridization. Expression of the alpha(v)beta(3) integrin was examined in Day 6.5 embryos, flushed from pregnant does. Immunofluorescence demonstrated strong immunostaining on the rabbit blastocyst (Day 6.5). RT-PCR analyses showed higher levels of mRNA for beta(3) subunit at the implantation site, with reduced expression in nonimplantation sites and in nonpregnant adult and immature endometrium. Immunohistochemistry demonstrated little, if any, beta(3) immunoreactivity on the endometrial epithelium. In contrast, in situ hybridization showed expression of the beta(3) integrin subunit mRNA in the uterine myometrium and on the trophoblast. To further determine the functional significance of alpha(v)beta(3) integrin expression during implantation, pregnant female rabbits that underwent ventral laparotomy on the morning of Day 6 received intrauterine injection of the following into the right uterine horn: 1) the monoclonal alpha(v)beta(3) neutralizing antibody (LM609), 2) arg-gly-asp (RGD) hexapeptides (GRGDSP), 3) non-RGD hexapeptides (GRGESP), and 4) IgG isotype matched control antibody. The left horn served as a control and received only saline injections. A significant reduction in the number of implantation sites was observed in the horns receiving anti-alpha(v)beta(3) antibody (P < 0.001) and the RGD peptides (P = 0.03). In the rabbit, the alpha(v)beta(3) integrin is present on the embryo and trophoblast and appears to be involved in early embryo-maternal interaction.
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