A role for connective tissue growth factor (CTGF) in reproductive function has been suggested from recent studies in the pig. To extend these findings, we have analyzed the immunohistochemical localization of CTGF during the estrous cycle and early pregnancy in mice. During the diestrous and early proestrous stages, CTGF was localized at high levels to both luminal and glandular uterine epithelial cells and at much lower levels in the stroma or myometrium. Epithelial expression of CTGF was considerably reduced at estrus. On Days 1.5-3.5 of pregnancy, CTGF was localized mainly to the uterine epithelial cells, which showed a substantially reduced level of CTGF on Day 4.5. On Days 5.5 and 6.5, CTGF was present at high levels in uterine decidual cells. CTGF was detected in the trophectoderm and inner cell mass of the preimplantation embryo on Day 4.5 and became preferentially localized to embryonic endoderm and mesoderm on Days 5.5-6.5. Multiple mass forms of CTGF (Mr 14 000-38 000) were present in endometrial extracts and uterine luminal flushings. Collectively, these data support a role for CTGF in uterine cell growth, migration, adhesion, and extracellular matrix production during the estrous cycle and early pregnancy, as well as in early development of the embryo.