Numerous cells containing P-450(F-1) were detected in the magnocellular and parvocellular neurons of the paraventricular nucleus of the hypothalamus. Electron microscopic analysis of immunoreactive neurons has shown that P-450(F-1) immunoreactivity is present on the Golgi apparatus and rough endoplasmic reticulum. In the paraventricular nucleus, the P-450(F-1)-positive magnocellular neurons frequently contained oxytocin and some of them also contained CRF. Vasopressin was colocalized with P-450(F-1), but these neurons did not express CRF. In the supraoptic nucleus, P-450(F-1) was colocalized with oxytocin or CRF in single neurons, but not with vasopressin. No cells exhibiting the colocalization of both P-450(F-1) and somatostatin were observed in these nuclei. The results of the present study concerning colocalization of P-450 and peptides suggest that P-450(F-1) is involved in the hypothalamo-hypophyseal neuroendocrine function in the female rat.