Tuberoinfundibular Dopamine Neuronal Activity Research Articles

The Roles of Dopamine and the Neurointermediate Lobe of the Pituitary in the Regulation of Prolactin Secretion During Late Pregnancy in Rats

The major hypothalamic control over prolactin secretion from the anterior pituitary gland is inhibitory by means of dopamine released from tuberoinfundibular dopamine (TIDA) neurones. We have previously shown a dissociation between activity of TIDA neurones and prolactin secretion during late pregnancy, suggesting involvement of additional regulatory factors. The aim of the present study was to investigate the role of dopamine and the neurointermediate lobe (NIL) of the pituitary in the regulation of prolactin secretion during late pregnancy. To determine whether dopamine maintains inhibition of prolactin during late pregnancy, the D(2) receptor antagonist domperidone was administered at 12.00 h on days 18 and 21 of pregnancy. These times are characterized by high and low TIDA neuronal activity, respectively, and low prolactin secretion. Domperidone produced an immediate increase in plasma prolactin compared to vehicle-treated controls on both days 18 and 21. Thus, dopaminergic inhibition of prolactin secretion is maintained despite reduced TIDA neuronal activity at the end of pregnancy. The contribution of NIL-derived dopamine in regulating prolactin secretion was then examined by investigating the effect of surgical removal of the NIL. NIL removal produced significantly increased basal prolactin concentrations, indicating that dopamine from the NIL contributes to the suppression of prolactin before the antepartum prolactin surge. Furthermore, NIL removal also completely prevented the antepartum prolactin surge compared to sham-operated controls, which is consistent with the hypothesis that the NIL supplies a prolactin-releasing factor to the anterior pituitary to induce the antepartum prolactin surge.

Opioid receptor subtypes involved in the regulation of prolactin secretion during pregnancy and lactation.

Afferent endogenous opioid neuronal systems facilitate prolactin secretion in a number of physiological conditions including pregnancy and lactation, by decreasing tuberoinfundibular dopamine (TIDA) inhibitory tone. The aim of this study was to investigate the opioid receptor subtypes involved in regulating TIDA neuronal activity and therefore facilitating prolactin secretion during early pregnancy, late pregnancy and lactation in rats. Selective opioid receptor antagonists nor-binaltorphimine (kappa-receptor antagonist, 15 micro g/5 micro l), beta funaltrexamine (mu-receptor antagonist, 5 microg/5 microl) and naltrindole (delta-receptor antagonist, 5 microg/5 microl) or saline were administered intracerebroventricularly (i.c.v.) on day 8 of pregnancy during a nocturnal prolactin surge, on day 21 of pregnancy during the ante partum prolactin surge or on day 7 of lactation before the onset of a suckling stimulus. Serial blood samples were collected at regular time intervals, via chronic indwelling jugular cannulae, before and after drug administration and plasma prolactin was determined by radioimmunoassay. TIDA neuronal activity was measured using the 3,4-dihydroxyphenylacetic acid (DOPAC) : dopamine ratio in the median eminence 2 h 30 min after i.c.v. drug injection. In each experimental condition, plasma prolactin was significantly inhibited by both kappa- and mu-receptor antagonists, whereas the delta-receptor antagonist had no effect compared to saline-injected controls. Similarly, nor-binaltorphimine and beta funaltrexamine significantly increased the median eminence DOPAC : dopamine ratio during early and late pregnancy, and lactation whereas naltrindole had no effect compared to saline-injected controls. These data suggest that TIDA neuronal activity, and subsequent prolactin secretion, is regulated by endogenous opioid peptides acting at both kappa- and mu-opioid receptors during prolactin surges of early pregnancy, late pregnancy and lactation.

Opioid control of prolactin secretion in late pregnant rats is mediated by tuberoinfundibular dopamine neurons

Prolactin (PRL) secretion from the anterior pituitary is tonically inhibited by tuberoinfundibular dopamine (TIDA) neurons in the arcuate nucleus of the hypothalamus. During late pregnancy, TIDA neuronal activity is reduced allowing the expression of an antepartum PRL surge. We show here that continuous infusion of the opioid receptor antagonist naloxone (10 mg/h) during the night preceding parturition completely abolished the antepartum PRL surge and significantly increased TIDA neuronal activity. These data indicate that endogenous opioid neurons facilitate PRL secretion at the end of pregnancy by suppressing TIDA neuronal activity.

Feedback regulation of PRL secretion is mediated by the transcription factor, signal transducer, and activator of transcription 5b.

PRL secretion from the anterior pituitary gland is inhibited by dopamine produced in the tuberoinfundibular dopamine neurons of the hypothalamus. The activity of tuberoinfundibular dopamine neurons is stimulated by PRL; thus, PRL regulates its own secretion by a negative feedback mechanism. PRL receptors are expressed on tuberoinfundibular dopamine neurons, but the intracellular signaling pathway is not known. We have observed that mice with a disrupted signal transducer and activator of transcription 5b gene have grossly elevated serum PRL concentrations. Despite this hyperprolactinemia, mRNA levels and immunoreactivity of tyrosine hydroxylase, the key enzyme in dopamine synthesis, were significantly lower in the tuberoinfundibular dopamine neurons of these signal transducer and activator of transcription 5b-deficient mice. Concentrations of the dopamine metabolite dihydroxyphenylacetic acid in the median eminence were also significantly lower in signal transducer and activator of transcription 5b-deficient mice than in wild-type mice. No changes were observed in nonhypothalamic dopaminergic neuronal populations, indicating that the effects were selective to tuberoinfundibular dopamine neurons. These data indicate that in the absence of signal transducer and activator of transcription 5b, PRL signal transduction in tuberoinfundibular dopamine neurons is impaired, and they demonstrate that this transcription factor plays an obligatory and nonredundant role in mediating the negative feedback action of PRL on tuberoinfundibular dopamine neurons.

Dissociation of prolactin secretion from tuberoinfundibular dopamine activity in late pregnant rats.

This study investigated whether the PRL surge that precedes parturition is accompanied by a decrease in activity of hypothalamic tuberoinfundibular dopamine (TIDA) neurons, as occurs during the PRL surges of early pregnancy. Serial blood samples were collected at regular intervals during early and late pregnancy via chronic indwelling jugular cannulae, and concentrations of plasma PRL were determined by RIA. In addition, pregnant rats were killed at either 1200 and 0300 h on different days throughout pregnancy. Levels of TIDA neuronal activity were estimated using concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence as an index of dopamine metabolism. During early pregnancy, plasma PRL concentrations showed characteristic diurnal and nocturnal surges peaking at 1700 and 0300 h, respectively, whereas during late pregnancy, there was a broad nocturnal surge throughout the night preceding parturition. During early pregnancy, DOPAC was elevated at 1200 h, associated with suppressed plasma PRL, whereas at 0300 h, during the nocturnal PRL surge, DOPAC was significantly reduced (P < 0.05). On the last day of pregnancy DOPAC levels were significantly reduced at both 1200 and 0300 h compared with those at 1200 h in early pregnancy regardless of the PRL concentration. This experiment was repeated with additional groups to further characterize the timing of the fall in TIDA activity during late pregnancy. DOPAC concentrations were elevated throughout the second half of pregnancy, then fell significantly between 0300-1200 h on day 21, approximately 36 h before parturition. As in the previous experiment, the timing of changes in DOPAC concentrations in the median eminence was dissociated from the antepartum PRL surge. These data indicate that the regulation of PRL secretion during late pregnancy is different from that of early pregnancy. Despite the prolonged reduction in activity of TIDA neurons during late pregnancy, PRL secretion still occurs as a nocturnal surge, suggesting that dopamine is not the only regulator of PRL secretion at this time.

Effects of chronic bromocriptine treatment on tyrosine hydroxylase (TH) mRNA expression, TH activity and median eminence dopamine concentrations in ageing rats.

The purpose of this study was to investigate the age-related changes in the responsiveness of tuberoinfundibular dopamine (TIDA) neurones to chronic hypoprolactinemia induced by treatment with bromocriptine, a dopamine receptor agonist. In one experiment, TIDA neuronal activity after acute hypoprolactinemia or exogenous prolactin was monitored by measuring tyrosine hydroxylase (TH) activity in the stalk median eminence of middle-aged cycling female rats (10-12 months), old constant oestrous rats (18-20 months) and old pseudopregnant rats (22-24 months). In another experiment, middle-aged cycling (10-12 months) rats were treated with bromocriptine for 6 or 12 months. TH activity was measured in the stalk median eminence, TH mRNA levels were measured in the arcuate nucleus and dopamine concentrations were measured in the arcuate nucleus and median eminence. Responsiveness of TIDA neurones to exogenous prolactin and to the withdrawal of bromocriptine in these rats was also tested. While the TIDA neurones in all three age groups responded to acute hypoprolactinemia by showing a reduction in TH activity, older rats failed to respond to exogenous prolactin administration. In contrast, chronic hypoprolactinemia for 12 months enabled the rats to retain TIDA neuronal responsiveness to exogenous prolactin. It also decreased TIDA neuronal function as measured by dopamine concentrations in the median eminence, TH activity in the stalk median eminence and TH mRNA in the arcuate nucleus of ageing rats. The restoration of the responsiveness of these neurones to prolactin stimulation in older rats demonstrates for the first time that hypoprolactinemia produced by chronic bromocriptine treatment indeed provides a neuroprotective effect on TIDA neurones. These results indicate that maintaining a low level of neuronal activity by lowering prolactin levels may be a contributing factor in retaining the plasticity of TIDA neurones.

Hypothyroidism increases prolactin secretion and decreases the intromission threshold for induction of pseudopregnancy in adult female rats

In order to understand the mechanism by which thyroid hormones alter prolactin (PRL) secretion, we investigated the role of tuberoinfundibular dopamine (TIDA) neurons and pituitary and hypothalamus vasoactive intestinal peptide (VIP) in thiouracil- (0.03% in drinking water for 16 days) induced–hypothyroid adult female rats. The intromission threshold for induction of pseudopregnancy also was examined to evaluate the PRL response to coital stimulation in hypothyroid rats. Hypothyroidism in adult female rats did not affect TIDA neuronal activity as measured by tyrosine hydroxylase activity (DOPA accumulation 30 min after administration of m-hydroxybenzylhydrazine dihydrochloride, 100 mg/kg, i.p.) in the stalk-median eminence compared with that in euthyroid rats, whereas pituitary concentration of VIP was dramatically increased. Plasma concentration of PRL was higher at 1100 h of proestrus and estrus in hypothyroid rats as compared with that of euthyroid rats. The proportion of female rats exhibiting pseudopregnancy was higher in hypothyroid animals (100%) receiving seven intromissions than in euthyroid animals (43%). Administration of l-thyroxine in hypothyroid rats decreased the proportion of pseudopregnancy (40%) to the level of euthyroid animals. These results indicate that the increased level of pituitary VIP probably affects PRL secretion in a paracrine or autocrine manner and account for the hyperprolactinemia induced in hypothyroid female rats. No role for TIDA neurons in PRL elevation can be ascribed. A decrease in the intromission threshold for induction of pseudopregnancy might be due to increased levels of PRL in hypothyroid female rats.

Prolactin regulation of tuberoinfundibular dopaminergic neurons: immunoneutralization studies

The purpose of this study was to determine the effects of acute hypoprolactinemia on tuberoinfundibular dopamine (DA) neurons using a rabbit anti-rat prolactin antiserum (PRL-AB) to immunoneutralize circulating prolactin under basal conditions and at various times after haloperidol-induced hyperprolactinemia. The specificity of PRL-AB for prolactin was determined by examining the ability of unlabelled hormone to displace binding of 125 I -labelled prolactin to PRL-AB. Tuberoinfundibular DA neuronal activity was estimated by measuring the concentrations of the DA metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence which contains terminals of these neurons. Systemic (i.v.) administration of 200 μl of PRL-AB decreased plasma prolactin concentrations below detectable levels for at least 4 h, and this was accompanied by a pronounced decrease in DOPAC concentrations in the median eminence of females, but not males. Central (i.c.v.) administration of 2 μl PRL-AB diluted up to 1:100 mimicked the inhibitory effect of systemic administration of PRL-AB on median eminence DOPAC concentrations suggesting that the tonic stimulatory effect of prolactin on the basal activity of tuberoinfundibular DA neurons in females occurs via a central site of action. In male rats, blockade of anterior pituitary DA receptors with haloperidol (1 mg/kg; s.c.) caused an prompt (by 1 h) increase in plasma prolactin concentrations which was maintained for at least 12 h. Haloperidol-induced hyperprolactinemia also caused a delayed (at 6 and 12 h) increase in median eminence DOPAC concentrations in these animals which was blocked by PRL-AB. Exposure of rats to initial priming periods of endogenous hyperprolactinemia of up to 6 h duration (followed by 6 h or more of PRL-AB-induced hypoprolactinemia) failed to alter median eminence DOPAC concentrations unless prolactin exposure was reinstated by an i.c.v. injection of prolactin. These results confirm that prolactin mediates the stimulatory effects of haloperidol on tuberoinfundibular DA neurons, and reveal that delayed induced activation of these neurons by prolactin is dependent upon a priming period of sustained hyperprolactinemia longer than 3 h for initiation and maintenance of this response.

Pharmacological Evidence that Neurotensin Mediates Prolactin- Induced Activation of Tuberoinfundibular Dopamine Neurons

The purpose of this study was to investigate the role of neurotensin (NT) receptors in mediating the stimulatory effects of prolactin on the activity of tuberoinfundibular dopamine (TIDA) neurons in male and female rats. TIDA neuronal activity was estimated by measuring concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) in terminals of these neurons in the median eminence (ME). Haloperidol activates TIDA neurons indirectly by blocking D₂ receptors on pituitary lactotropes, thereby increasing secretion of prolactin. Twelve hours after administration of haloperidol (1 mg/kg, s.c.), DOPAC concentrations in the ME were increased. Blockade of NT receptors with the selective antagonist SR-48692 had no effect per se on basal DOPAC concentrations in the ME but produced a dose-related (10–1,000 µg/kg, i.p.; 1 h) reversal of haloperidol-induced increases in ME DOPAC concentrations. In contrast, SR-48692 had no effect on either basal or haloperidol-induced increases in plasma prolactin. SR-48692 also blocked the stimulatory effects of prolactin (10 µg/rat, i.c.v.; 12 h) on ME DOPAC concentrations. SR-48692 was equally effective in blocking the stimulatory effects of haloperidol and prolactin on TIDA neurons in male and female rats. These results suggest that NT mediates the induced stimulatory effect of hyperprolactinemia on the activity of TIDA neurons in both males and females, whereas the tonic regulation of these neurons by prolactin in females occurs via an NT-independent mechanism.

Opposing roles for dopamine D 1 and D 2 receptors in the regulation of hypothalamic tuberoinfundibular dopamine neurons

The purpose of the present study was to characterize pharmacologically dopamine D 1 receptor-mediated inhibition of tuberoinfundibular dopamine neurons in males rats, and to determine if inhibitory dopamine D 1 receptors oppose stimulatory dopamine D 2 receptors and account for the inability of mixed dopamine receptor agonists to alter the activity of these neurons. Tuberoinfundibular dopamine neuronal activity was estimated by measuring the concentrations of the dopamine metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence, the region of the hypothalamus containing terminals of these neurons. Administration of the dopamine D 1 receptor agonist (±)-1 phenyl-2,3,4,5-tetrahydro-(1 H)-3-benzazepine-7,8-diol (SKF38393) decreased median eminence DOPAC and increased plasma prolactin concentrations, whereas administration of the dopamine D 1 receptor antagonist ((−)- trans,6,7,7a,8,9,13b-hexahydro-3-chloro-2-hydroxy- N-methyl-5 H-benzo[ d]naphtho-[2,1 b]azepine (SCH39166) increased median eminence DOPAC concentrations but had not effect on plasma prolactin. The inhibitory effect of SKF38393 on median eminence DOPAC concentrations was blocked by SCH39166. These results demonstrate that acute activation of dopamine D 1 receptors inhibits the activity of tuberoinfundibular dopamine neurons and thereby increases prolactin secretion, and that under basal conditions dopamine D 1 receptor-mediated inhibition of tuberoinfundibular dopamine neurons is tonically active. Administration of the dopamine D 2 receptor agonist (5a R- trans)-5,5a,6,7,8,9,9a,10-octahydro-6-propyl-pyridol[2,3- g]quinazolin-2-amine (quinelorane) increased median eminence DOPAC concentrations, and SKF38393 caused a dose-dependent reversal of this effect. Administration of the mixed dopamine D 1/D 2 receptor agonist R(−)-10,11-dihydroxy-apomorphine (apomorphine) had no effect per se, but blocked quinelorane-induced increases in DOPAC concentrations in the median eminence. These results reveal that concurrent activation of dopamine D 1 and D 2 receptors nullifies the actions of each of these receptors on tuberoinfundibular dopamine neurons, which likely accounts for the lack of an acute effect of mixed dopamine D 1/D 2 receptor agonists on these hypothalamic dopamine neurons.

Effects of selective activation of dopamine D 2 and D 3 receptors on prolactin secretion and the activity of tuberoinfundibular dopamine neurons

Dopamine agonists with activity at both dopamine D 2 and D 3 receptor subtypes stimulate tuberoinfundibular dopamine neurons and inhibit prolactin secretion from the anterior pituitary. The purpose of the present study was to identify the dopamine receptor subtypes mediating these effects using recently developed selective agonists for dopamine D 2 (PNU-95,666) and D 3 (PD128907) receptors. The activity of tuberoinfundibular dopamine neurons was estimated by measuring either the synthesis (accumulation of 3,4-dihydroxyphenylalanine [DOPA] following inhibition of decarboxylase activity) or metabolism (3,4-dihydroxyphenylacetic acid [DOPAC] concentrations) of dopamine in the median eminence, the region of the hypothalamus containing axon terminals of these neurons. In one experiment, the activity of mesolimbic dopamine neurons was also determined by measuring DOPA accumulation in terminals of these neurons in the nucleus accumbens. Activation of dopamine D 2 receptors with PNU-95,666 caused dose- and time-related increases in DOPAC concentrations in median eminence which were temporally correlated with decreases in plasma prolactin concentrations. Activation of dopamine D 3 receptors with PD128907 decreased DOPA concentrations in the nucleus accumbens, but had no effect on concentrations of DOPAC or DOPA in the median eminence or prolactin in plasma. These results reveal that tuberoinfundibular dopamine neurons are regulated by dopamine D 2 rather than D 3 receptors, and suggest that the ability of mixed dopamine D 2/D 3 receptor agonists to increase the activity of these neurons is mediated by an action at dopamine D 2 receptors. Furthermore, these results confirm that tuberoinfundibular dopamine neurons are not regulated by inhibitory dopamine D 2 or D 3 autoreceptors.

Seasonal changes in median eminence dopamine in male syrian hamsters: Role of the gonads and duration of exposure to short days

Median eminence dopamine concentrations are reduced in male hamsters housed in a short photoperiod for 12 weeks, but tyrosine hydroxylase activity and dopamine metabolism are unaffected. The mechanism responsible for this decrease in dopamine is unclear. The purpose of the experiments presented hare was to examine the effects of gonadal steroids on this decline in median eminence dopamine concentrations, and to characterize the time course over which this change occurs. Experiment 1 examined the effects of short day exposure on median eminence dopamine concentrations and tuberoinfundibular dopamine neuronal activity (i.e., accumulation of 3,4-dihydroxyphenylanins) in castrated male hamsters. The results of that experiment suggest that the short-day induced decline in median eminence dopamine concentrations is maintained in the absence of gonadal steroids. In Experiments 2 and 3, tuberoinfundibular dopamine neuronal activity (accumulation of 3,4dihydroxyphenylalanine and 3,4-dihydroxyphenylacetic acid concentrations, respectively), median eminence dopamine concentrations and circula ting prolactin levels were measured in animals housed In short days for varying lengths of time. This was done to determine when the decline in median eminence dopamine concentrations occurs and if the decline is preceded by a change In tuberoinfundibular dopamine neuronal activity. The results of these two experiments suggest that median eminence dopamine concentrations and circulating prolactin levels decline after approximately 4 weeks of short-day exposure, and that time photoperiod induced changes are not associated with changes In tubsrcintundibular dopaminergic neuronal activity.

Evidence that amplification of norepinephrine-induced LH release by morphine is indirectly due to suppression of tuberoinfundibular dopamine secretion

We previously observed that morphine markedly amplifies LH secretion following intracerebroventricular (i.c.v.) norepinehrine (NE) infusions. Based on additional evidence, we hypothesized that perhaps these morphine effects were due to suppression of tuberoinfundibular dopamine (TIDA) secretion thus allowing NE to evoke a greater release of LHRH from axon terminals in the median eminence than would otherwise occur. In the present studies, we examined whether apomorphine (a DA receptor agonist) would suppress and haloperidol (a DA receptor antagonist) would mimic these enhancing effects of morphine on NE-induced LH secretion. Estrogen-treated ovariectomized rats were used in these studies. NE, when infused i.c.v. (45 μg) evoked a modest increase in plasma LH(1.1 ± 0.2to2.2 ± 0.2ng/ml) within 15 min. When morphine sulfate (10 mg/kc s.c.) was given 15 min prior to NE, LH peak values of11 ± 2ng/ml were obtained by 60 min. Treatment of rats with apomorphine (1.5 mg/kg s.c.) at −15 min, morphine at 0 min and i.c.v. NE at 15 min resulted in a significant blunting of morphine's effect on NE-induced LH release. Moreover, in all morphine-treated rats, plasma prolactin (PRL) increased significantly within 10 min, peaked at 30 min and declined towards basal values by 90 min. Apomorphine completely blocked this morphine effect of PRL release. Haloperidol (HAL; 2.5 mg/kg s.c.) treatment had no effect on basal LH release but resulted in a significant increase in PRL which remained elevated up to 180 min. When HAL was given at 0 min and NE was infused i.c.v. at 15 min, a rapid, progressive and highly significant temporal increase in LH secretion occurred with peak values of20.4 ± 4ng/ml being reached by 90 min. Since our other recent studies have shown that i.c.v. DA infusions neither amplify nor suppress NE-induced LH release nor do they affect basal LH levels when given alone, we could not attribute morphine's action on NE release to alterations in A15 or A14 DA neuronal activity nor to increases in hypothalamic incertohypothalamic (A13) DA secretion. Rather, the suppression of TIDA (A12) neuronal activity or blockade of DA receptors by morphine or HAL, respectively, suggest that amplification of NE-induced LH release may be due to the suppression of inhibitory DA influences on the release of LHRH from axon terminals within the median eminence. The implications of these opiate actions on preovulatory LH surges are discussed.

Effects of acute and chronic cocaine on the activity of tuberoinfundibular dopamine neurons in the rat

We investigated the effects of acute and chronic cocaine on dopamine (DA) synthesis in tuberoinfundibular dopamine (TIDA) neurons by measuring the accumulation of 3,4-dihydroxyphenylalaine (DOPA) in the median eminence (ME). Cocaine-induced prolactin (PRL). Chronic cocaine did not alter were evaluated for comparison. Acute cocaine inhibited DA synthesis in the ME and decreased circulating PRL. Chronic cocaine did not alter basal DOPA levels in TIDA nerve terminals or PRL levels in plasma. After repeated cocaine injections, cocaine's inhibitory effect on DA synthesis was abolished in the ME while the PRL response to drug was unchanged. These results suggest that TIDA neurons become tolerant to cocaine-induced suppression of DA synthesis after chronic cocaine exposure, but the altered sensitivity of TIDA neurons is not revealed by plasma PRL measures.

Serotonin-induced decrease in hypothalamic tyrosine hydroxylase activity and corresponding increase in prolactin release are abolished at midpregnancy and by transplants of rat choriocarcinoma cells.

We investigated the effect of central serotonin (5-hydroxytryptamine, 5-HT) administration on hypothalamic tuberoinfundibular dopamine neurons and related changes in neuronal activity to circulating PRL levels in two physiological models: 1) pregnant rats expressing (day 8) or not expressing (days 11 and 16) PRL surges, and 2) ovariectomized rats transplanted with rat choriocarcinoma cells, which secrete functional placental lactogen-I. Over a 4-min period between 0900 and 1400 h, rats were administered either vehicle or 5-HT (20 micrograms/6 microliters) through lateral ventricular cannulae. Plasma PRL levels were determined by RIA. NSD 1015 (25 mg/kg intraarterial), a dihydroxyphenylalanine (DOPA) decarboxylase inhibitor, was injected 20 min after initiation of ventricular infusion. Ten min later, the stalk-median eminence (SME) was dissected. The rate of DOPA accumulation, determined by measuring DOPA levels in the SME by HPLC, was used as an index of tyrosine hydroxylase catalytic activity, indicating tubero infundibular dopamine neuronal activity. In day-8 pregnant rats 5-HT reduced DOPA accumulation to 57% of vehicle-injected controls and increased circulating PRL levels 13-fold. In contrast, on days 11 and 16 of pregnancy 5-HT did not alter DOPA accumulation in the SME or plasma PRL levels. In nonpregnant rats ovariectomized for 24 h, 5-HT decreased DOPA accumulation in the SME to 43% of vehicle-infused controls and increased PRL levels approximately 26-fold. However, in nonpregnant rats with rat choriocarcinoma cells, 5-HT produced no changes in either DOPA accumulation in the SME or in circulating PRL levels. The inability of 5-HT to reduce tyrosine hydroxylase activity after mid-pregnancy may account for the lack of a PRL response. Placental lactogens secreted at midpregnancy, particularly placental lactogen-1, may induce this loss of 5-HT effect.

Evidence that prolactin mediates the stimulatory effects of estrogen on tuberoinfundibular dopamine neurons in female rats.

The effects of ovariectomy and estrogen on prolactin secretion and/or the activity of tuberoinfundibular dopamine (TIDA) neurons were examined by either concurrently measuring concentrations of prolactin in plasma and 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence of female rats or by determining the rate of DA synthesis (accumulation of 3,4-dihydroxyphenylalanine (DOPA) after the administration of a decarboxylase inhibitor) in the median eminence. For comparison, concentrations of alpha-melanocyte-stimulating hormone (alpha MSH) in plasma and DOPAC in the intermediate lobe of the pituitary (an index of the activity of tuberohypophysial DA neurons) were also determined. Ovariectomy produced a time-dependent decrease in the accumulation of DOPA and the concentrations of DOPAC in the median eminence and prolactin in plasma with maximal effects occurring by 7 days. Estrogen administration to ovariectomized rats increased plasma prolactin and median eminence DOPAC concentrations to levels comparable to those in diestrous controls. In contrast, neither ovariectomy nor estrogen replacement altered the concentrations of alpha MSH in plasma or DOPAC in the intermediate lobe. Administration of the DA agonist bromocriptine blocked the ability of estrogen to increase plasma prolactin and median eminence DOPAC concentrations. Also, administration of antiserum to rat prolactin blocked the stimulatory action of estrogen on median eminence DOPAC concentrations. Taken together, these results indicate that the stimulatory effect of estrogen on the activity of TIDA neurons is mediated by prolactin.

Endogenous Opioids Mediate the Nocturnal Prolactin Surge in the Pregnant Rat*

Endogenous opioids stimulate PRL release in the rat during proestrus, stress, and lactation. This study investigated the potential role of opioids in regulating the daily nocturnal PRL surges that occur during the first half of pregnancy. On day 8 of pregnancy, infusion of naloxone (NAL; 0.25, 1.0, or 2.0 mg/10 min), an opioid receptor antagonist, from 0100-0400 h blocked the nocturnal surge in a dose-dependent manner. PRL remained at basal levels throughout the infusion when the highest dose of NAL was used. A PRL surge was still present in this group, but its delayed onset occurred 1 h after termination of the NAL infusion. In another experiment NAL was used to determine whether opioids are needed to maintain the nocturnal surge or only to initiate the surge. On day 8 of pregnancy, infusion of NAL (2.0 mg/10 min) was started at 0300 h, after initiation of the surge had occurred, and was continued until 0600 h. After 1 h of infusion, PRL levels were less than 20 ng/ml, compared to more than 200 ng/ml in controls (P less than 0.002) and remained low. A PRL surge occurred approximately 2-3 h after the infusion was discontinued, at a time when the surge was completed in controls (P less than 0.01). Experimental animals did not differ in their surges relative to controls on the next day when no infusion was given. Another set of experiments was performed to determine whether blockade of endogenous opioids inhibited the nocturnal PRL surge by stimulating tuberoinfundibular dopamine (TIDA) neurons. Day 8 pregnant rats infused with NAL (2.0 mg/10 min) from 0100-0400 h demonstrated no nocturnal PRL surge. Subsequently, the medial basal hypothalamus in NAL-treated animals and saline controls was incubated in vitro with the DOPA decarboxylase inhibitor brocresine (100 microM). The greater amount of resultant DOPA accumulation in the stalk median eminence of NAL-treated animals reflected increased TIDA neuronal activity compared to that in SAL-infused controls. Similar results were obtained when rats were treated in vivo with the DOPA decarboxylase inhibitor NSD 1015 (25 mg/kg BW) just before termination of the NAL infusion. Based on these results, we conclude that endogenous opioid peptides are essential in initiating and maintaining nocturnal PRL surges in pregnant rats. Moreover, opioids appear to exert their effects by decreasing TIDA neuronal activity.

Role of Testosterone in the Regulation of Tuberoinfundibular Dopaminergic Neurons in the Male Rat

The effects of testosterone on the tuberoinfundibular dopamine (DA) neuronal activity was examined by determining the rate of DA synthesis-accumulation of 3,4-dihydroxyphenylalanine (DOPA) after administration of a decarboxylase inhibitor and the concentration of a DA metabolite,--3,4-dihydroxyphenylacetic acid (DOPAC)--in the median eminence in the male rat. Within 1 week after orchidectomy, there was an increase in the accumulation of DOPA and the concentration of DOPAC in the median eminence, but there was no change in the concentration of DA. Conversely, 1 day after testosterone administration to orchidectomized rats, the elevated DOPAC concentrations in the median eminence returned to levels comparable to those in gonadally intact rats. Neither orchidectomy nor testosterone replacement had any effect on plasma prolactin concentrations, but inhibition of prolactin secretion following administration of the DA agonist bromocriptine blocked the increase in DOPA accumulation in the median eminence of orchidectomized rats; this latter effect was reversed by intracerebroventricular administration of prolactin. On the other hand, intracerebroventricular injection of prolactin caused a similar increase in the accumulation of DOPA in the median eminence of gonadally intact, orchidectomized, and testosterone-treated orchidectomized rats. Immobilization stress decreased the accumulation of DOPA and the concentration of DOPAC in the median eminence of orchidectomized rats, but had no effect in intact or testosterone-treated orchidectomized rats. These results indicate that testosterone inhibits the basal activity of tuberoinfundibular DA neurons and blocks the inhibitory effects of physical restraint on these neurons, but does not alter the ability of these neurons to respond to delayed activation by prolactin.

Stress-Induced Suppression of the Prolactin Afternoon Surge in Ovariectomized, Estrogen-Treated Rats and the Nocturnal Surge in Pseudopregnant Rats Are Accompanied by an Increase in Median Eminence Dihydroxyphenylacetic Acid Concentrations

Experiments were performed to determine whether the suppression of prolactin (PRL) surges during restraint was accompanied by changes in the activity of tuberoinfundibular dopamine (TIDA) neurons in the median eminence. Animals were either ovariectomized and estrogen-treated (OVX-PEP) or cervically stimulated to induce pseudopregnancy (PSP). Restraint stress was administered by tying the hind legs together with plastic-coated bell wire. Animals were decapitated following 15 or 30 min of restraint stress or immediately after removal from the animal room (control) when PRL levels were basal (10.00 h), at the peak of the afternoon PRL surge in OVX-PEP animals (17.00 h) or the nocturnal PRL surge in PSP animals (05.00 h). Median eminence dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) were significantly decreased in control rats at 17.00 h when compared to control rats at 10.00 h (103.1 +/- 3.7 vs. 85.8 +/- 3.3 and 11.4 vs. 7.1 +/- 0.4 pg/micrograms protein, respectively) and plasma PRL was markedly elevated. Restraint stress at 10.00 h resulted in a significant increase in serum PRL, but this increase was not accompanied by a change in DA or DOPAC when compared to control animals (103.1 +/- 3.7 vs. 107.9 +/- 4.8 and 11.4 +/- 0.4 vs. 10.4 +/- 0.6 pg/micrograms protein, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Activation of tuberoinfundibular dopamine neurons following the acute administration of atypical antipsychotics

The activity of tuberoinfundibular dopamine neurons, as judged from dihydroxyphenylacetic acid (DOPAC) concentrations or the accumulation of dihydroxyphenylalanine (DOPA) in the median eminence after the inhibition of DOPA decarboxylase, was increased following the acute administration of the purported atypical antipsychotics clozapine, thioridazine, melperone, setoperone, and RMI 81582. In contrast, the activity of these hypothalamic dopamine neurons was not acutely altered by the typical antipsychotics haloperidol, chlorpromazine, fluphenazine, and cis-flupentixol or by SCH 23390. The acute stimulatory effect of the atypical antipsychotics on the activity of tuberoinfundibular dopamine neurons was effectively antagonized by the D1 agonist SKF 38393 but not by the D2 agonist quinpirole. The production of an acute activation of tuberoinfundibular dopamine neurons, which appears to be sensitive to D1 receptor activation, may be an effect that distinguishes typical and atypical antipyschotic agents.