Prolactin (PRL) receptors have been identified in both classical and nonclassical target organs. Actions of PRL on the secretion of anterior pituitary hormones have been reported. In order to confirm the presence of PRL receptors on specific cell types of the anterior pituitary, immunocytology on ultrathin frozen sections was carried out using monoclonal antibodies specific to the rat PRL receptor. Anterior pituitary glands were removed and fixed in 4% paraformaldehyde or 2.5% glutaraldehyde and postfixed in 1% osmium tetroxide. Tissues were frozen and ultrathin sections were made by cryoultramicrotomy. In anterior pituitary cells, all endocrine cell populations showed PRL-R-like immunoreactivity. The relative labeling frequency based on immunolabeling studies was somatotrophs > lactotrophs > thyrotrophs approximately corticotrophs > gonadotrophs. Within all endocrine cell types, no apparent differences could be observed in the subcellular localization of PRL-R-like immunoreactivity. At the subcellular level, gold particles, either isolated or in groups of two, were seen at the level of the plasma membrane, in the cytoplasmic matrix, in the vicinity of the endoplasmic reticulum and in the nucleus. The identification of receptors specific to PRL within different cells of the anterior pituitary raises the interesting possibility that PRL may act via an autocrine or paracrine network in regulating anterior pituitary function.