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Expression of Fertilin and CD9 in Bovine Trophoblast and Endometrium During Implantation¹

^a Department of Plant and Animal Sciences, Nova Scotia Agricultural College, Truro, Nova Scotia, Canada B2N 5E3

The superficial placentation of cattle involves the development of fetal binucleate cells that arise from the chorion and migrate between adjacent cell tight junctions to fuse with maternal epithelium. Thus, the temporal and spatial patterns of expression of the cell migration, adhesion, and fusion molecules fertilin and CD9 were investigated in bovine trophoblast and endometrium. Bovine fertilin and fertilin ß messenger RNA sequences were amplified by reverse transcriptase-polymerase chain reaction in testis (positive control), peri-implantation (Days 18, 19, and 21), and postimplantation (Days 35–40) trophoblast RNA, but not in caruncular endometrium (Day 40). Northern blot analysis indicated that the transcript hybridizing to fertilin in trophoblast RNA was approximately 4.0 kilobases (kb), whereas in testis, 2 transcripts of approximately 3.3 and 3.8 kb were indicated. The transcript hybridizing to the fertilin ß probe was also larger in trophoblast than in testis (3.8 vs. 2.4 kb, respectively). In situ hybridization revealed that fertilin ß mRNA was expressed by trophoblast cells, including binucleate cells. Immunohistochemical study of CD9, a member of the transmembrane-4-superfamily which is thought to be involved in sperm-egg fusion, showed that CD9 was present on the apical surface of uterine epithelium and in a subpopulation of binucleate cells of the trophoblast. Immunoprecipitation followed by Western blot analysis showed association between CD9 and integrin ₃ in endometrium. The results support the hypothesis that fertilin and CD9 are involved in bovine binucleate cell migration and fusion.

First decision: 1 November 2001.

¹ Funded by grants from the Nova Scotia Agri-Focus 2000 Technology Development Program and the Natural Sciences and Engineering Research Council.

² Correspondence: Leslie MacLaren, Department of Plant and Animal Sciences, Nova Scotia Agricultural College, 58 River Rd., Box 550, Truro, NS, Canada B2N 5E3. FAX: 902 895 6734; lmaclaren{at}nsac.ns.ca

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