Appropriate integrin expression appears to be necessary for successful implantation of human embryos and varies considerably among species. The present study was undertaken to determine the distributions of integrin subunits α1, α3, and α6 as well as the extracellular matrix (ECM) components collagen IV and laminin in implanting bovine trophoblast and endometrium. Immunohistochemical staining of cryostat sections prepared from nonpregnant endometrium, of preattachment through to early villus development pregnant endometrium (Days 18, 21, 24, and 30), and of isolated trophoblast binucleate cells was performed. Trophoblast down-regulated the integrin α1 subunit as attachment proceeded, whereas reactivity scores for α6 antibody tended to increase from Day 18 through 24 and remained high. A subpopulation of trophoblast binucleate cells expressed the α3 integrin subunit. Uterine epithelium constitutively expressed α3 and α6 integrin subunits, but the α1 subunit was down-regulated as the luminal epithelium was modified. Collagen IV and laminin reactivity increased in the basal lamina and underlying subepithelial stroma as pregnancy proceeded. The results suggest that binucleate cell fusion with the maternal epithelium initiates integrin and ECM changes in the subepithelial stroma.