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Regulated Expression of Osteopontin in the Peri-Implantation Rabbit Uterus¹

Department of Obstetrics and Gynecology,³ Division of Reproductive Endocrinology and Fertility, the University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7570 Department of Physiology,⁴ School of Veterinary Medicine, Universidad Complutense, 28040 Madrid, Spain Department of Cell Biology,⁵ Vanderbilt University School of Medicine, Nashville, Tennessee 37232 Department of Obstetrics and Gynecology,⁶ Women's Reproductive Health Research Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232 DuPont Pharmaceuticals Research Laboratories,⁷ Experimental Station, Wilmington, Delaware 19880-0400 Department of Obstetrics and Gynecology,⁸ Division of Maternal and Fetal Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7570

Blastocyst attachment to the lining of the mammalian uterus during early implantation involves the initial apposition of the trophoblast to the uterine epithelial surface. Osteopontin (OPN) is a glycoprotein component of the extracellular matrix that is secreted by the glandular epithelium of mammalian uteri at the time of implantation. This protein is recognized by several members of the integrin family and promotes cell-cell attachment and adhesion. In the present study, rabbit uteri were examined using Northern and in situ hybridization to evaluate the temporal and spatial distribution of OPN mRNA during early pregnancy. Northern blot analysis demonstrated a dramatic increase in OPN expression on Days 4–7 of pregnancy, corresponding to the rise in circulating progesterone and the time of initial embryo attachment in this species. In situ hybridization analysis revealed OPN mRNA expression on Day 6.75 of pregnancy, which was most prominent on endometrial epithelium. Using immunofluorescence, OPN protein was present on the glandular epithelium on Day 6.75 of pregnancy, but was absent on blastocysts. Further, no expression of OPN mRNA or protein was found in the nonpregnant endometrium. Induction of endometrial OPN expression was observed in unmated rabbits treated with progesterone alone and was prevented by cotreatment with the antiprogestin ZK137.316. Estradiol-17ß had no effect on OPN expression by itself, and estrogen priming was not necessary to demonstrate the stimulatory effect of progesterone. In The rabbit uterus, as in other mammalian species studied, OPN is expressed in a stage-specific manner by the endometrial glands during the peri-implantation period and is regulated by progesterone.

¹ Part of this work was presented at the 47th Annual Meeting of the Society for Gynecologic Investigation in Chicago, Illinois, March 22–25, 2000. This research was supported by NICHD/NIH through cooperative agreement U54 HD-35041 (B.A.L.) and HD-37321 (K.G.O., G.E.O.) 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.A.L.), and the Fogerty International Fellowship award (K.B.C.A.).

² Correspondence: Bruce A. Lessey, Department of OB/GYN, Division of Reproductive Endocrinology and Fertility, CB #7570, Old Clinic Building, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7570. FAX 919 966 5214; lessey{at}med.unc.edu

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