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BIOLOGY OF REPRODUCTION 68, 766–771 (2003)

© 2003 by the Society for the Study of Reproduction, Inc.

Embryo

A Role for {alpha}vß3 Integrin During Implantation in the Rabbit Model1

Maria J. Illeraa, P.L. Lorenzoa, Yao-ting Guib, Stanley A. Beylerc, K.B.C. Apparaoc, and Bruce A. Lessey2,c

a Departamento de Fisiologia, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Spain b Department of Physiology, University of Manitoba, Winnipeg, Manitoba, Canada R2E 3J7 c Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599

The study of implantation has been facilitated by the identification of specific biomarkers that are associated with uterine receptivity. The {alpha}vß3 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}vß3 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}vß3 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}vß3 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 ß3 subunit at the implantation site, with reduced expression in nonimplantation sites and in nonpregnant adult and immature endometrium. Immunohistochemistry demonstrated little, if any, ß3 immunoreactivity on the endometrial epithelium. In contrast, in situ hybridization showed expression of the ß3 integrin subunit mRNA in the uterine myometrium and on the trophoblast. To further determine the functional significance of {alpha}vß3 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}vß3 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}vß3 antibody (P < 0.001) and the RGD peptides (P = 0.03). In the rabbit, the {alpha}vß3 integrin is present on the embryo and trophoblast and appears to be involved in early embryo-maternal interaction.

1 This research was supported by NICHD/NIH through cooperative agreement U54 HD-35041 (B.L.) as part of the Specialized Cooperative Centers Program in Reproduction Research, the National Cooperative Program on Markers of Uterine Receptivity for Blastocyst Implantation (HD 34824; B.L.), and the Fogerty International Fellowship award (Y.G. and K.B.C.A.).

2 Correspondence: Bruce A. Lessey, Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, CB #7570, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599. FAX: 919 966 5214; e-mail: lessey{at}med.unc.edu




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Copyright © 2003 by the Society for the Study of Reproduction.