The effect of 17 beta-estradiol (17 beta E2) on anterior pituitary dopaminergic receptor (D2) content was studied in vitro in relation to PRL secretion. Anterior pituitaries from ovariectomized rats were incubated for short periods in medium 199, with or without the steroid. Dopamine (DA) receptors in partially purified pituitary membranes were quantified by equilibrium binding using [3H]spiperone; the PRL released into the incubation medium was analyzed by RIA. Addition of 10(-10) to 10(-6) M 17 beta E2 to the incubation medium of anterior pituitaries rapidly and reversibly decreased the number of DA receptors (P less than 0.01 to 0.001), while increasing PRL release, in a dose-related fashion. The maximal effect on both receptor numbers and PRL secretion was obtained with 10(-8) M 17 beta E2. This effect involved no change in receptor affinity (Kd = 0.11 +/- 0.01 nM in presence or in absence of 17 beta E2). This estrogen-induced decrease in DA-binding capacity was apparently not the result of the occupation of spiperone binding sites by the steroid, since after a 30-min incubation with 10(-8) M [3H]17 beta E2, no radioactivity was detectable on the partially purified membranes. Moreover, the presence of 17 beta E2 at the same time as the labeled D2 ligand did not modify the kinetics of association or dissociation of spiperone with pituitary membranes. This decrease in anterior pituitary DA receptor content and the increase in PRL release were already significant after a 7-min incubation in the presence of 10(-8) M 17 beta E2. Finally, these effects of 17 beta E2 were not mimicked by its 17 alpha-stereoisomer, nor by progesterone, or testosterone. These results suggest that the stimulatory effect of 17 beta E2 on PRL secretion may be due, at least in part, to the desensitization of anterior pituitary cells to DA. The steroid may produce this desensitization directly by decreasing the number of D2. The short latency of this effect likely discards the possibility of a genomic action of 17 beta E2.

An attempt was made to correlate the physiological or the dimethylbenz(a)anthracene (DMBA)-enhanced serum prolactin (PRL) surge, which occurs in the afternoon of proestrus in female Sprague-Dawley (SD) rats, with physiological or pathological changes in two biochemical estimates of the tuberoinfundibular dopaminergic (TIDA) neuron activity. Dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) concentrations as well as tyrosine hydroxylase (TH) activity were measured in the median eminence (ME) of control or DMBA-pretreated SD rats throughout the estrous cycle in relation to PRL secretion. In both groups of females, while the DA content was fairly constant, the DOPAC content and TH activity in the ME fluctuated markedly throughout the estrous cycle. Thus, in control animals, the DOPAC content, DOPAC/DA ratio and TH activity which were stable on the days of diestrus and morning of proestrus were markedly decreased at noon and early afternoon when serum PRL levels began to rise. Later in the afternoon of proestrus, when serum PRL levels were maximal, there was a marked but transient increase in the DOPAC content and DOPAC/DA ratio as well as a brief surge in TH activity. In the evening of the same day, when serum PRL returned to basal levels, the DOPAC content, DOPAC/DA ratio and TH activity were low. Finally on estrus morning, the DOPAC content, DOPAC/ DA ratio and TH activity increased again to reach the diestrus levels. In DMBA-pretreated females, similar fluctuations in TIDA neuronal activity occurred during the estrous cycle, but the dynamics of these changes was altered: the DOPAC/DA ratio and TH activity first showed a marked increase in the morning of proestrus day, before decreasing dramatically. No surge in TH activity was detectable in the afternoon of proestrus and the increase in the DOPAC/DA ratio was of lesser amplitude than that in the controls. In those females whose PRL levels were still higher than those of controls on the morning of estrus, the second increase in the DOPAC/DA ratio was not observed. These results suggest that the lowering in TIDA neuronal activity on the day of proestrus might explain the outset, but not the termination of the preovulatory PRL surge. The regulation involved in the latter process seems to be particularly impaired by DMBA pretreatment.

Using a high-performance liquid chromatography (HPLC) system coupled with an electrochemical detector, the concentrations of dopamine (DA) and 5-hydroxytryptamine (5-HT) and their major specific metabolites 3,4-dihydroxy-phenylacetic acid (DOPAC) and 5-hydroxyindole-3-acetic acid (5-HIAA), respectively, were measured in the median eminence (ME) throughout the rat estrous cycle. The ME DA content remained fairly constant throughout the estrous cycle except on estrus when Π.OOh values were significantly lower than 10.00 h values (40% decrease, p < 0.05). The ME 5-HT content determined at 10.00 h was higher on proestrus than on any other day of the cycle. The ME DOPAC concentrations did not differ between 10.00 and 17.00 h on diestrus I, diestrus II or estrus. On the contrary, there was an almost linear decline between 10.00 and 17.00 h on proestrus (36% decrease, p < 0.05). The ME 5-HIAA content did not differ between 10.00 and 17.00 h on any day of the estrous cycle. Significant changes were recorded for the DOPAC/DA and 5-HIAA/ 5-HT ratio in the ME on proestrus. There was a progressive decrease, starting from 10.00 in the DOPAC/DA ratio with minimal values (42% decrease, p < 0.05) at 16.00 h followed by an increase from 16.00 to 19.00 h. On the other hand, the 5-HIAA/5-HT ratio increased between 10.00 and 17.00 h (97% increase, p < 0.05) and subsequently declined until 19.00 h (67% decrease vs. 17.00 h, p < 0.05). These data show that a concomitant decrease in dopaminergic activity and increase in serotoninergic activity in the ME occur in association with the preovulatory prolactin and luteinizing hormone preovulatory surges.