Neuropeptide Y (NPY) stimulates luteinizing hormone (LH) release in the rat by a dual action, one in the hypothalamus to excite LHreleasing hormone (LHRH) release and the other at the level of pituitary gonadotrophs to activate and/or potentiate LH release induced by LHRH. Because NPY produces effects similar to norepinephrine with which it may comprise an excitatory hypothalamic circuit, it was hypothesized that NPY concentrations in the hypothalamus would change in a time-and site-specific manner in association with the preovulatory LH surge on proestrus. Concentrations of NPY in individual nuclei of the preoptic-tuberal pathway and serum LH levels were estimated by radioimmunoassays in rats during diestrus 2 and proestrus. On proestrus, serum LH levels were basal between 1000 and 1400 h, rose significantly at 1500 h and plateaued between 1600 and 1800 h. Of the five neural sites examined, only NPY in the median eminence displayed marked fluctuations in close association with the LH surge. NPY concentrations were low between 1000 and 1300 h, and rose abruptly at 1400 h (P< 0.05) preceding the onset of LH rise at 1500 h. These elevated levels were maintained until 1600 h, during which time serum LH rose to a plateau and then fell at 1800 h to the low range seen between 1000 and 1300 h. In contrast, the pattern of changes in NPY levels in the arcuate nucleus, suprachiasmatic nucleus and medial preoptic area, three additional sites in the preoptic-tuberal pathway known to participate in the preovulatory LH surge, were markedly different from that seen in the median eminence. In each of these three sites, NPY levels rose significantly at 1800 h from the values at 1000 to 1200 h with a slightly different time-course of increment. None of these areas exhibited changes in NPY concentrations on diestrus 2; NPY concentrations also were unaltered on diestrus 2 or proestrus in the ventromedial nucleus. The present observations of site-specific, dynamic changes in NPY levels on proestrus, in a manner previously documented for LHRH, support the hypothesis that a subset Of NPY neurons terminating in the median eminence may be a component of excitatory neural circuitry that either independently or in co-action with the adrenergic system is responsible for the induction of preovulatory LH release.The preovulatory release of luteinizing hormone (LH) during the critical period on the afternoon of proestrus occurs as a result of increased episodic discharge of LH-releasing hormone (LHRH) from the hypothalamus (1-7). The timing and duration of LHRH-induced pituitary activation on proestrus has been fully characterized (2,3,8). In addition, it is generally believed that neurogenic stimuli entrained to the daily photoperiod clock abruptly activate LHRH secretion into the hypophysial portal veins during the critical period (2, 9). The origin and route of transmission of these neural signals to LHRH neurons localized in the preoptic-tuberal pathway (2, 3), have not yet been delineated.A number of hypothalamic neural systems h...