Colony-stimulating factor 1 (CSF-1) is a hematopoetic cytokine that also plays an important role in placental physiology. We report here the molecular cloning of two alternative splice variants of the bovine gene coding for a putative secreted and a membrane-bound form of the cytokine and the dynamic regulation of expression in the reproductive tract of cattle during the estrous cycle and pregnancy. Bovine CSF-1 was expressed mainly as the 3- and 4-kilobase (kb) transcripts, but 1.4- and 0.8-kb mRNAs were also detected in Day 50–70 pregnant uterine tissue. During the estrous cycle, both the 4- and 3-kb mRNAs were present, but the 3-kb putative membrane-bound form was more abundant than the 4-kb secreted form during diestrus. This pattern of expression was reversed in pregnancy, so that the exponential increase in CSF-1 expression seen during pregnancy was due predominantly to increased abundance of the 4-kb transcript. The change in the 4-kb:3-kb ratio was detected between Day 14 and Day 17, approximately the time of maternal recognition of pregnancy. Thus, CSF-1 was identified as one gene whose expression in the uterus might be altered early in response to the presence of the conceptus. CSF-1 was also expressed in the extraembryonic membranes of the conceptus and in the trophoblastic cells of the fetal cotyledons after the formation of the placentomes. The high level of CSF-1 expression during bovine pregnancy in uteroplacental tissues is consistent with its proposed role in placental physiology.