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ARS Home » Plains Area » Clay Center, Nebraska » U.S. Meat Animal Research Center » Livestock Bio-Systems » Research » Publications at this Location » Publication #150616
Title: REGULATION OF BLASTOCYST MIGRATION, APPOSITION, AND INITIAL ADHESION BY A CHEMOKINE, INTERFERON GAMMA-INDUCIBLE PROTEIN 10 KDA (IP-10), DURING EARLY GESTATION

Author
item NAGAOKA, KENTARO - UNIV TOKYO, JAPAN
item NOJIMA, HISASHI - UNIV TOKYO, JAPAN
item WATANABE, FUMIKO - HIGETA SHOYU CO, JAPAN
item CHANG, KYU-TAE - KOREA RES INST BIOSCI BIO
item Christenson, Ronald
item SAKAI, SENKITI - UNIV TOKYO, JAPAN
item IMAKAWA, KAZUHIKO - UNIV TOKYO, JAPAN

Submitted to: Journal of Biological Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/19/2003
Publication Date: 8/1/2003
Citation: NAGAOKA, K., NOJIMA, H., WATANABE, F., CHANG, K., CHRISTENSON, R.K., SAKAI, S., IMAKAWA, K. REGULATION OF BLASTOCYST MIGRATION, APPOSITION, AND INITIAL ADHESION BY A CHEMOKINE, INTERFERON GAMMA-INDUCIBLE PROTEIN 10 KDA (IP-10), DURING EARLY GESTATION. JOURNAL OF BIOLOGICAL CHEMISTRY. 2003. v. 278(31). p. 29048-29056.

Interpretive Summary: Biochemical communications between the developing conceptus and the mammalian uterus are important for successful implantation and recognition of pregnancy. Interferon-tau (IFN-t), a protein produced by the conceptus prior to implantation in ruminants, is considered to possess immune regulatory effects that permit a foreign protein (conceptus/embryo) to be implanted successfully in the uterus. The objective of this study was to examine the effect of an endometrial chemokine, interferon-gamma-inducible protein 10 kDa (IP-10), on conceptus (trophoblast cells) migration to the endometrial epithelium in the Shiba goat. Immunofluorescent analysis revealed that IP-10 receptor, CXCR3, was localized in the trophoblast cells to which biotinylated-recombinant caprine IP-10 bound, suggesting that CXCR3 receptor expressed in the trophoblast cells is functioning. Chemotaxis assays indicated IP-10 stimulated the migration of trophoblast cells, the effects were neutralized by an anti-IP-10 antibody. These findings support the postulate that interaction of IFN-t, IP-10, and CXCR3 receptor is important in the establishment of immunological communication between the conceptus and maternal uterine tissue at the time of implantation. Improved implantation can increase embryo survival and improve reproduction rates for the ruminant livestock industries.

Technical Abstract: For a pregnancy to be established, initial apposition and adhesion of the blastocyst to maternal endometrium must occur in a coordinated manner, however, a key factor(s) that mediates the trophoblast cell migration and attachment to the apical surface of the endometrium has not been identified. In this study, we examined the effect of an endometrial chemokine, interferon-gamma-inducible protein 10 kDa (IP-10), on conceptus migration to the endometrial epithelium. We first studied endometrial IP-10 mRNA expression, which was localized in the subepithelial stromal region, and detected the protein in the uterine flushing media during early pregnancy. Expression of IP-10 mRNA by the endometrium of cyclic animals was stimulated by the addition of a conceptus factor interferon-tau (IFN-t). Immunofluorescent analysis revealed that IP- 10 receptor, CXCR3, was localized in the trophoblast cells, to which biotinylated-recombinant caprine IP-10 (rcIP-10) bound, suggesting that CXCR3 receptor expressed in the trophoblast cells is functioning. Chemotaxis assay indicated that rcIP-10 stimulated the migration of trophoblast cells, and the effects of rcIP-10 were neutralized by the pretreatment with an anti-IP-10 antibody. Adhesion assay revealed that adhesive activity of trophoblast cells to fibronectin was promoted by rcIP-10, and the effect was inhibited by the use of anti-IP-10 antibody. Further adhesion experiments demonstrated that binding of trophoblast cells to fibronectin was completely inhibited by a peptide of the Arg-Gly-Asp (RGD) sequence, which binds to integrins alpha-5-beta-1, alpha-V-beta-1, alpha-V-beta-3 and alpha-V-beta-5, whereas non-binding peptide containing Arg-Gly-Glu (RGE) had minimal effects. More importantly, rcIP-10 promoted the adhesion of trophoblast cells to primary cells isolated from endometrial epithelium. Furthermore, rcIP-10 stimulated the expression of integrin alpha-5, alpha-V and beta-3 subunit mRNA in trophoblast cells. These findings suggest that endometrial IP-10 regulates the establishment of apical interactions between trophoblast and epithelial cells during early gestation.