Selective concentration of [(1)H]progesterone or its metabolites is observed in nuclei of neurons in certain hypothalamic regions of the guinea pig 15 minutes after injection of [1,2,6,7-(3)H]progesterone, by use of dry-mount autoradiography. Highest concentrations of progestin target neurons exist in the nucleus arcuatus, the nucleus preopticus periventricularis, and the nucleus preopticus suprachiasmaticus. Previous administration of unlabeled progesterone inhibits the nuclear concentration of radioactivity, but cortisol has no effect. Estradiol priming enhances the nuclear uptake of radioactivity. The results demonstrate the existence of progestin target sites in the hypothalamus and suggest a direct action of progestin on certain hypothalamic structures.