Estrogen has numerous effects on immunoreactive levels of oxytocin (OXT) centrally, particularly in the preoptic lateral subcommissural nucleus (LSN). In this study in situ hybridization of a 38-base oligodeoxyribonucleotide (38mer) complementary to OXT mRNA revealed that estrogen treatment altered the pattern of OXT production in the rostral LSN and the more caudal anterior commissural nucleus. Rats were injected with 20 pg estradiol benzoate or sesame oil vehicle im 4 and 5 days after ovariectomy. On the sixth day all animals were perfused with paraformaldehyde-glutaraldehyde and their brains sectioned to 10 pm thickness in a -10 "C cryostat. Coronal brain sections were taken from four parallel levels of the preoptic-anterior hypothalamic area. These sections were mounted and hybridized insifu to a [1'25]-labeled 38mer for 16 h at 37 "C. Washed and dried slides were processed for autoradiography and analyzed with a light microscope. The effect of estrogen on OXT production differed between the rostral and caudal sections in both the LSN and periventricular (PeV) areas. Estrogen significantly increased OXT mRNA levels in LSN cells while decreasing hybridization in the anterior commissural nucleus cells. Changes in frequency patterns in the PeV paralleled those in the LSN with a significant drop of hybridization in the caudal PeV. Neurons hybridizing 38mer probe were also found in several other areas including the ventral medial preoptic area, lateral hypothalamus, bed nucleus of the stria terminalis and the nucleus triangularis septi. OXT mRNA levels were affected by estrogen treatments and this effect differed between the preoptic area and anterior hypothalamus. The sensitivity of LSN oxytocinergic cells to estrogen has implications for estrogen-sensitive OXT-enhanced reproductive behaviorsThe ovarian steroid estrogen influences oxytocin (OXT) physiology in the CNS. Estrogen treatments increase OXT release from Ihe posterior pituitary (1) and increase firing of oxytocinergic neurons in the paraventricular nucleus (PVN) (2). Levels of immunoreactive OXT in the PVN vary with estrous state in ways {hat suggest estrogen's influence (3). OXT immunoreactive levels :tho increase in the PVN and supraoptic nucleus (SON) peripar-I um as estrogen levels increase, while OXT levels in more rostral synthesis sites are decreasing (4).The preoptic area (POA) sites of OXT synthesis are particularly sensitive to estrogen influence. Estrogen implants increased OXT immunoreactive content in the POA (5). Short-term treatment with silastic implants of estradiol increased the number of OXT i~nmunostained cells in the periventricular (PeV) and dorsolateral I' OA as well as in several other sites (6). The collection of oxytocinergic cells ventrolateral of the anterior commissure in the dorsolateral POA and overlapping the septohypothalamic nucleus and bed nucleus of the stria terminalis (BnST) has been called the lateral subcommissural nucleus (LSN) (6, 7). Oxytocinergic cells in the LSN showed increased immunostaining after s...