Nitric oxide synthase (NOS) enzyme activity is present in rat decidua and metrial gland, and the activity decreases near the end of pregnancy. In this study, inducible and endothelial NOS isoforms were immunolocalized to rat granulated metrial gland (GMG) cells using anti-NOS antibodies proven to be isoform specific. These NOS-positive GMG cells are of the natural killer cell lineage as they stained positively for NKR-P1 cell surface receptor, and for perforin. The number of NOS-positive GMG cells corresponded with the level of decidual and metrial gland NOS enzyme activity. NOS activity declined when GMG cells containing NOS decreased in number. Uterine arteriolar vascular smooth muscle also stained positively for inducible NOS and the staining did not change with advancing gestation. Only a minority of uterine myocytes stained positively for inducible NOS and these were subjacent to the placental attachment site. Neuronal NOS immunostaining was not present in the decidua and the metrial gland.