Tuberoinfundibular Dopaminergic Neurons Research Articles

Nicotinic control of tuberoinfundibular dopaminergic neuron activity and prolactin secretion: diurnal rhythm and involvement of endogenous opioidergic system

The possible involvement of cholinergic and opioidergic neurons in the control of diurnal changes of tuberoinfundibular dopaminergic (TIDA) neuronal activity was reported. Adult Sprague-Dawley rats ovariectomized and treated with estrogen were used. All drugs were administered centrally through preimplanted intracerebroventricular cannula, and both TIDA neuronal activity and serum prolactin level were determined. Nicotine (10 ng/3 μl/rat) given at 10:00 h significantly inhibited TIDA neuronal activity from 5 to 30 min and stimulated serum PRL levels at 5 and 15 min. Co-administration of either mecamylamine (1 μg) or naloxone (2.5 μg) prevented both nicotine's effects. A dose-related (0.1–100 ng) effect of nicotine on TIDA neuronal activity and serum PRL level was also observed in the morning when TIDA neuronal activity is high and serum PRL level is low, but not in the afternoon when the former activity is low and the latter is high. When atropine (20 μg), naloxone (25 μg) or Nor-BNI (20 μg) was given at 14:00 h all increased the lowered TIDA neuronal activity in the afternoon. When atropine was co-administered with either naloxone or Nor-BNI, however, no additive effect was observed. Submaximal doses of atropine (0.2 μg), mecamylamine (0.1 μg) or naloxone (0.25 μg) was also effective in stimulating the afternoon levels of TIDA neuronal activity and inhibiting serum PRL, and no additive effect was observed either. Moreover, simultaneous injection of morphine (15 μg) prevented atropine's effect in the afternoon. These results indicate that cholinergic neurons may act through activating the endogenous opioidergic neurons to exhibit an inhibitory effect on TIDA neuronal activity and a stimulatory one on prolactin secretion. A diurnal difference in its endogenous activity between morning and afternoon was also implicated.

Atrial natriuretic peptide negatively modulates the stimulatory effects of angiotensin II on tuberoinfundibular dopaminergic neuronal activity. Neurochemical and electrophysiological studies.

The effects of angiotensin II (AII) and natriuretic peptide (ANP), alone or in combination, on tuberoinfundibular dopaminergic (TIDA) neuronal activity and on serum PRL levels were examined in this study. The TIDA neuronal activity was determined by measuring 3,4-dihydroxyphenylacetic acid (DOPAC) concentration in the median eminence using high-performance liquid chromatography plus electrochemical detection, and serum PRL levels were determined by radioimmunoassay. Intracerebroventricular injection of AII induced both time (5-60 min)- and dose (0.01-1 microg)-dependent effects by stimulating TIDA neuronal activity and inhibiting serum PRL levels in ovariectomized, estrogen-treated rats. ANP in 0.01-10 microg doses, on the other hand, had no significant effect on TIDA neurons, nor on serum PRL at 30 min. When ANP (in 0.1-10 microg doses) was co-administered with AII (1 microg dose), it dose-dependently attenuated the effects of AII on TIDA neuronal activity and on serum PRL levels. In a separate study using single-unit recording of neurons of the dorsomedial arcuate nucleus (dmARC) in brain slices, where most TIDA neurons reside, AII stimulated 68.0% of 72 units recorded. Few (5.6%) units were inhibited and the remaining ones were not responsive. ANP alone was mostly ineffective on dmARC neurons (63.0% of 54 units), and it stimulated and inhibited 22.2 and 14.8% of them, respectively. When ANP was co-administered with AII to AII-responsive units, however, it significantly attenuated the effects of AII in 81.5% of 27 units. Other neurons were unaffected. Thus, results from the in vivo study indicate that ANP can negatively modulate the stimulatory effect of AII on hypothalamic TIDA neurons and the inhibitory effect on serum PRL levels; and the in vitro electrophysiological data substantiates this observation by showing that ANP exerts a similar modulation on dmARC neurons.

Inhibitory Effect of Dopamine on Dorsomedial Arcuate Neurons in Rat Brain Slices: Potentiation by Coadministration of Cocaine

Whether dopamine (DA) can have a direct effect on the tuberoinfundibular dopaminergic neurons has been a controversial issue. The present report used single-unit recording of neurons in dorsomedial region of the arcuate nucleus, where most tuberoinfundibular dopaminergic neurons are located, to study this question. By focusing our recording in this region, we found that DA in 25–250 nmol ranges inhibited a significant number of arcuate neurons tested (74.2% of 182 units). The inhibitory effect of DA was not only prominent in most cases, it also persisted in low Ca2+, high Mg2+ solution in several trials. Cocaine, a drug of abuse whose main effect is due to its inhibition of DA transporters and increasing the DA concentration in synaptic clefts, also inhibited a significant number of arcuate neurons by itself (51.5% of 97 units), although its effects were lesser than those of DA. Nevertheless, when coadministered with DA, cocaine significantly potentiated the inhibitory effect of DA in 82% of DA-responsive units (n = 39). These results clearly demonstrate that DA exhibits a predominantly inhibitory effect on presumed DA neurons in dorsomedial arcuate nucleus. The effects of cocaine further support this notion.

Chronic Hyperprolactinemia and Changes in Dopamine Neurons

The tuberoinfundibular dopaminergic (TIDA) system is known to inhibit prolactin (PRL) secretion. In young animals this system responds to acute elevations in serum PRL by increasing its activity. However, this responsiveness is lost in aging rats with chronically high serum PRL levels. The purpose of this study was to induce hyperprolactinemia in rats for extended periods of time and examine its effects on dopaminergic systems in the brain. Hyperprolactinemia was induced by treatment with haloperidol, a dopamine receptor antagonist, and Palkovits' microdissection technique in combination with high-performance liquid chromatography was used to measure neurotransmitter concentrations in several areas of the brain. After 6 months of hyperprolactinemia, dopamine (DA) concentrations in the median eminence (ME) increased by 84% over the control group. Nine months of hyperprolactinemia produced a 50% increase in DA concentrations in the ME over the control group. However, DA response was lost if a 9-month long haloperidol-induced hyperprolactinemia was followed by a 11 12 month-long extremely high increase in serum PRL levels produced by implantation of MMQ cells under the kidney capsule. There was no change in the levels of DA, norepinephrine (NE), serotonin (5-HT), or their metabolites in the arcuate nucleus (AN), medial preoptic area (MPA), caudate putamen (CP), substantia nigra (SN), and zona incerta (ZI), except for a decrease in 5-hydroxyindoleacetic acid (5-HIAA) in the AN after 6-months of hyperprolactinemia and an increase in DA concentrations in the AN after 9-months of hyperprolactinemia. These results demonstrate that hyperprolactinemia specifically affects TIDA neurons and these effects vary, depending on the duration and intensity of hyperprolactinemia. The age-related decrease in hypothalamic dopamine function may be associated with increases in PRL secretion.

Inhibition of tuberoinfundibular dopaminergic neural activity during suckling: involvement of mu and kappa opiate receptor subtypes.

Previous studies have shown that mu (mu) and kappa (kappa) opioid antagonists inhibit suckling-induced prolactin release. Prolactin responses elicited by pup suckling or opioid administration are mediated, at least in part, by suppression of dopamine (DA) release from tuberoinfundibular dopaminergic (TIDA) neurons in the hypothalamus. We examined the effects of the mu opiate receptor antagonist, beta-funaltrexamine (beta-FNA), and the kappa opiate receptor antagonist, nor-binaltorphimine (nor-BNI) on the activity of TIDA neurons in lactating rats. TIDA neuronal activity was determined by measuring DOPA accumulation in the caudate putamen (CP) and median eminence (ME). The effects of opioid antagonist treatment were determined in pup-deprived (low circulating prolactin levels) or pup-suckled rats (high circulating prolactin levels). The accumulation of 5-hydroxytryptophan (5-HTP) in the medial preoptic area (MPOA), the anterior hypothalamus (AH) and the median eminence (ME) was quantified as an index of serotonergic activity in the same animals for comparative purposes. In vehicle treated rats, suckling caused a significant and selective decrease in DOPA accumulation in the ME. beta-FNA (5 micrograms, i.c.v.) pretreatment significantly increased DOPA accumulation in the ME of pup-deprived and pup-suckled rats. beta-FNA pretreatment also prevented the suckling-induced suppression of DOPA accumulation in the ME. In contrast to the actions of beta-FNA, pretreatment with nor-BNI (8 micrograms, i.c.v.) did not significantly affect the activity of the TIDA neurons in pup-deprived or pup-suckled rats. Suckling alone did not alter 5-HTP accumulation in any of the brain regions examined, and neither opioid antagonist had appreciable effects on 5-HTP accumulation. These results demonstrate that the EOP tonically inhibit the TIDA neurons in both pup-deprived and pup-suckled, post-partum female rats by acting through the mu, but not the kappa, opiate receptor subtype. Furthermore, the suckling-induced inhibition of TIDA neurons is also mediated through the EOP acting at mu, but not kappa opioid receptors.

Stimulatory effect of central oxytocin on tuberoinfundibular dopaminergic neuron activity and inhibition on prolactin secretion: neurochemical and electrophysiological studies.

The posterior pituitary hormone, oxytocin (OT), has been shown to have either a stimulatory or an inhibitory effect on PRL secretion depending on the route of administration. Whether its central inhibitory effect involves the tuberoinfundibular dopaminergic (TIDA) neuron was the focus of this study. Adult female Sprague-Dawley rats ovariectomized for 1 week, implanted with sc estrogen-containing capsules and intracerebroventricular cannulas for 6 more days were used. TIDA neuron activity was determined by measuring the concentration of 3,4-dihydroxyphenylacetic acid or 3,4-dihydroxyphenylalanine in the median eminence by HPLC with electrochemical detection. Intracerebroventricular injection of OT induced both dose (0.01-1 microgram/rat)- and time (30-90 min)-dependent increases in 3,4-dihydroxyphenylalanine or 3,4-dihydroxyphenylacetic acid levels in the median eminence. Serum PRL levels were also decreased 30 min after the injection. The use of a specific OT antagonist, [d(CH2)5, Tyr(Me)2, Orn8]vasotocin, not only blocked the effect of OT on TIDA neuron activity, it further lowered it to below control levels, indicating the existence of an endogenous OT activity. When 1 microgram OT was administered at 1200 h, it also reversed the diurnal decrease in TIDA neuron activity at 1500 h. The effects of OT on the electrical activities of dorsomedial arcuate neurons were also tested using single unit recording in brain slices. In 33 neurons tested with OT, 66.7% were stimulated by OT in 0.5- to 50-nmol doses, and no inhibitory effect was observed. The rest were not responsive. In conclusion, both neurochemical and electrophysiological studies demonstrated that central OT may play a stimulatory role in regulating the TIDA neurons and, in turn, inhibition of PRL secretion.

Intraventricular administration of estradiol modulates rat prolactin secretion and synthesis.

The effect of estradiol (E2) on rat tuberoinfundibular dopaminergic (TIDA) neurons was examined in vivo, employing chronic intraventricular (i.c.v.) infusion technique using an osmotic mini-pump. The activity of TIDA neurons was assessed by the release and synthesis of prolactin (PRL) in the rat pituitary gland and by the changes in the 3, 4-dihydroxyphenylacetic acid (DOPAC) and dopamine (DA) levels and in the DOPAC/DA ratio in the rat hypothalamus. We also examined the [3H] E2 binding in the rat hypothalamus. Ovariectomized female Wistar rats with E2 replacement were treated with daily i.c.v. infusion of 1 microM of E2 or saline vehicle for 1, 3, and 7 days using the Alzet osmotic mini-pump and brain infusion kit. At 1 day of i.c.v. infusion of E2, the serum PRL level was significantly decreased compared with that in the vehicle group. Northern blot analysis of the total RNA isolated from the pituitary glands demonstrated a decrease in the PRL gene transcript level in the E2 group. At 3 days of E2 treatment, however, the serum PRL level was significantly increased compared with that of the vehicle-injected group and Northern blot analysis also demonstrated that the PRL gene transcript level was increased in the E2 group. At 7 days of E2 administration, there were no significant differences between the E2 and vehicle groups in either serum PRL or PRL gene transcript levels. There was a significant increase in the DOPAC/DA ratio after 1 day in the E2 group. However, no significant effects of E2 on this ratio were observed at 3 and 7 days of treatment. The DOPAC concentration in the E2 group was significantly increased at day 1 and significantly decreased at day 3, compared with that of the respective time in vehicle group. At day 7 there was no significant change in DOPAC concentration in either groups. The DA concentrations in the hypothalamus was not changed on any day in either group. Specific [3H] E2 binding was observed in the rat hypothalamus. These data suggest that E2 may have a biphasic effect on the accumulation of PRL gene transcripts and on the PRL secretion in the rat pituitary by first stimulating and then inhibiting the TIDA neuronal activity.

Stimulatory effect of central oxytocin on tuberoinfundibular dopaminergic neuron activity and inhibition on prolactin secretion: neurochemical and electrophysiological studies

Stimulatory effect of central oxytocin on tuberoinfundibular dopaminergic neuron activity and inhibition on prolactin secretion: neurochemical and electrophysiological studies

Evidence that D 2 receptor-mediated activation of hypothalamic tuberoinfundibular dopaminergic neurons in the male rat occurs via inhibition of tonically active afferent dynorphinergic neurons

The purpose of the present study was to determine if D 2 receptor-mediated activation of hypothalamic tuberoinfundibular dopaminergic (TIDA) neurons occurs via afferent neuronal inhibition of tonically active inhibitory dynorphinergic neurons in the male rat. To this end, the effects of either surgical deafferentation of the mediobasal hypothalamus or administration of a κ opioid receptor agonist (U-50,488) or antagonist (nor-binaltorphimine (NOR-BNI)) on D 2 receptor-mediated activation of TIDA neurons were assessed. For comparison, the activity of mesolimbic DA neurons was also determined in these studies. TIDA and mesolimbic DA neuronal activities were estimated by measuring dopamine synthesis (accumulation of 3,4-dihydroxyphenylalanine (DOPA) following decarboxylase inhibition) and metabolism (concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC)) in terminals of these neurons in the median eminence and nucleus accumbens, respectively. Intraperitoneal administration of the D 2 receptor agonist quinelorane caused a dose-dependent increase in DOPAC in the median eminence and a decrease in DOPAC in the nucleus accumbens; surgical deafferentation of the mediobasal hypothalamus prevented the effect of quinelorane in the median eminence, but not the nucleus accumbens. Activation of κ opioid receptors with U-50,488 had no effect per se, but blocked quinelorane-induced increases in median eminence DOPA. In contrast, U-50,488 had no effect on DOPA in the nucleus accumbens of either vehicle- or quinelorane-treated rats. Blockade of κ opioid receptors with NOR-BNI increased median eminence DOPA, and prevented the stimulatory effects of quinelorane on dopamine synthesis. Administration of prolactin also increased median eminence DOPA, but did not alter the ability of quinelorane to stimulate dopamine synthesis. Neither NOR-BNI nor prolactin had any effect on DOPA in the nucleus accumbens of vehicle- or quinelorane-treated rats. These results suggest that D 2 receptor-mediated activation of TIDA neurons occurs via an afferent neuronal mechanism involving, at least in part, inhibition of tonically active inhibitory dynorphinergic neurons in the male rat.

Prolonged pertussis toxin treatment affects morphine's action on tuberoinfundibular dopaminergic neuron activity and on prolactin secretion

The effects of pertussis toxin (PTX) pretreatment on basal and morphine-affected changes of tuberoinfundibular dopaminergic (TIDA) neuron activity and serum prolactin level were tested in this study. Adult female Sprague-Dawley rats, ovariectomized and treated with a long-acting estrogen (polyestradiol phosphate, 0.1 mg/rat, s.c.), were used. The activity of TIDA neurons was determined by measuring the turnover rate of dopamine (DA), and the concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) or 3,4-dihydroxyphenylalanine (DOPA) in the median eminence. Acute (30–90 min) treatment of PTX had no significant effect on any of the parameters measured. Prolonged (24 h) treatment of PTX significantly reduced morphine's inhibitory effect on TIDA neuron activity (using DOPA, but not DOPAC as the index), and stimulatory effect on PRL release. Basal TIDA neuron activity as determined by median eminence DOPAC concentration, DOPA accumulation, or DA rate constant was not significantly altered by PTX. Median eminence DA level, however, was significantly reduced. These results suggest that a pertussis toxin-sensitive GTP-binding protein may be responsible for the maintenance of TIDA neurons, and for mediation of the inhibitory effect of morphine on TIDA neuron activity, and in turn, the stimulation of prolactin secretion.

Prolonged pertussis toxin treatment affects morphine's action on tuberoinfundibular dopaminergic neuron activity and on prolactin secretion

The effects of pertussis toxin (PTX) pretreatment on basal and morphine-affected changes of tuberoinfundibular dopaminergic (TIDA) neuron activity and serum prolactin level were tested in this study. Adult female Sprague-Dawley rats, ovariectomized and treated with a long-acting estrogen (polyestradiol phosphate, 0.1 mg/rat, s.c.), were used. The activity of TIDA neurons was determined by measuring the turnover rate of dopamine (DA), and the concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) or 3,4-dihydroxyphenylalanine (DOPA) in the median eminence. Acute (30–90 min) treatment of PTX had no significant effect on any of the parameters measured. Prolonged (24 h) treatment of PTX significantly reduced morphine's inhibitory effect on TIDA neuron activity (using DOPA, but not DOPAC as the index), and stimulatory effect on PRL release. Basal TIDA neuron activity as determined by median eminence DOPAC concentration, DOPA accumulation, or DA rate constant was not significantly altered by PTX. Median eminence DA level, however, was significantly reduced. These results suggest that a pertussis toxin-sensitive GTP-binding protein may be responsible for the maintenance of TIDA neurons, and for mediation of the inhibitory effect of morphine on TIDA neuron activity, and in turn, the stimulation of prolactin secretion.

Central administration of 8-OH-DPAT and MCPP stimulates prolactin secretion in ovariectomized, estrogen-treated rats: lack of an effect on tuberoinfundibular dopaminergic neuron activity

The effects of central administration of two serotonin receptor agonists, 8-OH-DPAT and mCPP, on tuberoinfundibular dopaminergic (TIDA) neuron activity and serum prolactin (PRL) levels in ovariectomized, estrogen-treated rats were determined. 8-OH-DPAT dose-dependently (0.1–10 μg/rat, icv) stimulated serum PRL levels, and depressed serotonergic neuron activity in 30 min. However, the TIDA neuron activity was not affected at all. Similar treatment of mCPP was less effective than 8-OH-DPAT: only the highest dose of mCPP (10 μg) stimulated PRL secretion and inhibited serotonergic neuron activity. No change in TIDA neuron activity was observed either. We conclude that central serotonin acts on 5-HT 1 receptors to stimulate the PRL secretion, which may not involve the TIDA neurons.

Prolactin- and testosterone-induced inhibition of LH secretion after orchidectomy: role of catecholaminergic neurones terminating in the diagonal band of Broca, medial preoptic nucleus and median eminence.

Central catecholaminergic neurones projecting to specific hypothalamic structures are involved in stimulating and inhibiting the activity of the GnRH-containing neurosecretory neurones. Both testosterone and elevated circulating prolactin (PRL) levels inhibit postcastration LH release. Three groups of adult male rats were orchidectomized and adrenalectomized, received corticosterone replacement and were: (i) administered purified ovine PRL (oPRL; 2400 microgram/s.c. injection) or (ii) its diluent, polyvinylpyrrolidone (PVP), every 12 h, or (iii) received physiological testosterone replacement for 2 days. At 0, 2 and 6 days postcastration, norepinephrine (NE), epinephrine (E) and dopamine (DA) turnover were estimated by the alpha-methyl-p-tyrosine method in three micro-dissected hypothalamic structures: the diagonal band of Broca at the level of the organum vasculosum of the lamina terminalis (DBB(ovlt)), the medial preoptic nucleus (MPN) and the median eminence (ME). In control (PVP-treated) rats, serum LH concentrations increased eightfold at 2 and 6 days postcastration and this rise was prevented by testosterone. oPRL treatment transiently suppressed LH secretion at 2 but not 6 days postcastration. Castration significantly decreased basal rat PRL (rPRL) levels at 2 and 6 days and testosterone administration partially prevented this effect. NE turnover in the ME and E turnover in the MPN increased markedly at 2 and 6 days postcastration, and testosterone replacement for 2 days prevented these increases. Thus, noradrenergic neurones innervating the ME and adrenergic neurones innvervating the MPN may drive postcastration LH secretion by providing stimulatory afferent input to the GnRH neurones. It was striking to observe that oPRL blocked the increases in both ME NE and MPN E turnover at 2 but not 6 days postcastration. Hence, oPRL may transiently suppress LH release by an inhibitory action on these NE and E neurones. DA turnover in the DBB(ovlt) was significantly decreased by 6 days postcastration. Testosterone-treated (2 days postcastration) and oPRL-treated (2 and 6 days postcastration) rats exhibited turnover values indistinguishable from day 0 controls. Hence, the A14 dopaminergic neurones, which synapse on GnRH neurones in the rostral preoptic area and may exert an inhibitory effect on them, are positively regulated by PRL and perhaps by testosterone as well. Autoregulatory feedback suppression of endogenous rPRL secretion by oPRL was observed both 2 and 6 days postcastration. In contrast to the A14 dopaminergic neurones, turnover in the A12 tuberoinfundibular dopaminergic (TIDA) neurones innervating the ME increased significantly by 6 days postcastration in control rats while oPRL administration further increased ME DA turnover at both 2 and 6 days. Hence, autofeedback regulation of rPRL secretion persists through at least 6 days of oPRL exposure temporally associated with markedly increased turnover in the TIDA neurones. In summary, our results support the hypothesis that the inhibitory effect of PRL on postcastration LH release is mediated by suppression of the activity of NE neurones innervating the ME and E neurones terminating in the MPN which, with time, become refractory to continued PRL exposure.

The responsiveness of tuberoinfundibular dopaminergic neurons to prolactin feedback is diminished between early lactation and midlactation in the rat.

Lactation is a state of hyperprolactinemia resulting in part from suppressed tuberoinfundibular dopaminergic (TIDA) neuronal activity. The suckling stimulus contributes to this suppression despite the fact that the TIDA neurons are a potential site for PRL feedback to increase neuronal activity. This study examined the influence of PRL feedback and the suckling stimulus on tyrosine hydroxylase (TH), the rate-limiting enzyme in dopamine biosynthesis, during early and midlactation. On day 3 or 10 of lactation, rats were injected intracerebroventricularly with medium (control) or MMQ cells (200,000 cells), a PRL-secreting cell line. On day 6 of lactation, TH activity in the stalk-median eminence was increased 2- or 1.4-fold by MMQ cells or prior treatment with ovine PRL (oPRL; 4 mg/kg, sc), respectively. Removal of pups for 24 h increased TH activity 70% above levels in pup-exposed rats, and MMQ cells or oPRL caused an additional 60% increase. TH messenger RNA (mRNA) levels in the arcuate nucleus were increased 3-fold after removing the pups, but MMQ cells did not alter mRNA levels in either pup-exposed or pup-deprived dams. In contrast to early lactation, MMQ cells did not alter TH activity or mRNA levels in the pup-exposed dams on day 13 and only marginally increased enzyme activity in pup-derived dams. Circulating PRL levels were markedly reduced after removing pups. MMQ cells suppressed circulating PRL levels in both groups of dams on day 6 and in pup-deprived dams on day 13, but had no effect in pup-exposed dams at this time. In a second experiment, pup-exposed dams were injected with bromocriptine, a dopamine agonist, and killed after 4 or 12 h on day 5 or after 12 h on day 12. In some rats, PRL was replaced by injecting oPRL simultaneously with and 8 h after bromocriptine treatment. On day 5 of lactation, bromocriptine reduced and oPRL restored TH activity, whereas on day 12, oPRL was not able to reverse the effect of bromocriptine. These data indicate that the suckling stimulus suppresses TH activity and gene expression in the TIDA neurons in pup-exposed dams. The high endogenous PRL levels associated with the suckling stimulus may activate TH activity in TIDA neurons during early lactation. However, the responsiveness of the TIDA neurons to PRL feedback is attenuated by day 13 of lactation.

17β-estradiol inhibits the production of dopamine by the tuberoinfundibular dopaminergic neurons of the male rat

Previous work in our laboratory suggests that the indirect stimulation of prolactin gene transcription observed in male rats between 3 and 6 h following estrogen injection results from an inhibition of dopamine release from the tuberoinfundibular dopaminergic neurons. To evaluate further the role of dopamine in the regulation of prolactin gene transcription by estrogens, we have examined the acute effects of 17β-estradiol (E 2) on dihydroxyphenylalanine accumulation in the arcuate median eminence region and the amount of dopamine associated with the arcuate median eminence region. Dihydroxyphenylalanine accumulation in the arcuate median eminence region was unaffected when examined 2 h following a single injection of E 2 (10 μg), and was reduced by 60% when examined 4 h following E 2 treatment. When examined 4 h after E 2 injection, dopamine content in the arcuate median eminence region was significantly increased. These data suggest that estrogens exert acute inhibitory effects on the tuberoinfundibular dopaminergic neurons of the male rat, and further support the assertion that the indirect stimulatory actions of estrogens on prolactin gene transcription result from inhibition of hypothalamic dopamine production.

Antagonism by progesterone of diethylstilbestrol-induced pituitary tumorigenesis in Fischer 344 rats: effects on sex steroid receptors and tyrosine hydroxylase mRNA.

It is known that chronic exposure of F344 rats to diethylstilbestrol (DES) induces prolactin (PRL)-secreting pituitary tumors composed of proliferating mammotropic cells. In the present work, we studied the effects of progesterone (P4) on several parameters stimulated in the pituitary tumors (DES-T), such as nuclear estrogen receptors (NE2R), cytosolic progestin receptors (CP4R) and serum PRL. Additionally, we have measured in hypothalamus the mRNA levels for tyrosine hydroxylase (TH), the rate-limiting enzyme for synthesis of dopamine, the main PRL-inhibitory factor. We found that pellet implantation of P4 during 1 month significantly reduced weight, ligand binding to NE2R and CP4R and serum PRL in the tumorous glands. Reductions in sex steroid receptor binding were due to changes in Bmax without changes in Kd, as observed after Scatchard plot analysis. Receptor binding data, therefore, suggests a pituitary site of action of P4. TH mRNA expression was studied in tuberoinfundibular dopaminergic (TIDA) neurons by in situ hybridization techniques employing a 35S-labeled oligonucleotide probe. Mean number of grains/cell decreased significantly in DES-T, an effect partly reversed by P4 treatment. Frequency histograms were constructed by plotting the number of cells versus the number of grains/cell and examined by x2 test and analysis of residuals. We found that DES-T presented significantly more cells with less grains whereas in control glands, P4-treated rats and DES-T receiving P4, cells with a higher grain number prevailed. These results suggest that in addition to a direct pituitary effect, P4 may also antagonize DES-induced tumorigenesis acting on mRNA for TH and presumably on the activity of TIDA neurons of the hypothalamus. The use of DES-T as a model for hyperprolactinemia may allow further assessment of P4 effects in pituitary adenomas in humans.

Stimulatory effects of vasoactive intestinal peptide and pituitary adenylate cyclase-activating peptide on tuberoinfundibular dopaminergic neuron activity in estrogen-treated ovariectomized rats and their correlation with prolactin secretion.

The effects of central administration of vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP) on hypothalamic tuberoinfundibular dopaminergic neuron activity and serum prolactin (PRL) levels were reported. Adult female Sprague-Dawley rats overiectomized for 2 weeks, implanted with subcutaneous estrogen-containing capsules and intracerebroventricular (i.c.v.) cannulae for 6 days were used for experiments. I.c.v. injections of VIP or PACAP were performed in conscious rats in the morning, and the injected rats were decapitated at various times afterwards. Serum sample and the median eminence tissue were collected from each rat. The levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), and 3,4-dihydroxyphenylalanine (DOPA) in the median eminence were determined by high-performance liquid chromatography with electrochemical detection. Serum PRL levels were determined by radioimmunoassay. I.c.v. administration of VIP (1 microgram/3 microliter) significantly increased median eminence DOPAC/DA ratio at 30 min, and serum PRL level at 15 min. The same dose of VIP (1 microgram), but not higher (10 micrograms) or lower (0.1 microgram), was also effective in stimulating the median eminence DOPA accumulation 35 min after the injection. The effect of VIP (1 microgram) on median eminence DOPA could be blocked by coadministration of a VIP antagonist, VIP6-28 in 10- and 30-, but not in 1- or 0.1-microgram doses. On the other hand, i.c.v. administration of PACAP (10 micrograms) stimulated median eminence DOPAC and lowered serum PRL levels at 30 min. All doses of PACAP used (0.1, 1 and 10 micrograms/ rat, i.c.v.) significantly increased median eminence DOPA concentrations at 60 min. The stimulatory effect of PACAP (0.1 microgram) on median eminence DOPA could also be blocked by coadministration of a PACAP antagonist, PACAP6-38 (in 10 to 100 x higher doses). In summary, central administration of either VIP or PACAP exhibited a stimulating effect on TIDA neuron activity through specific receptors. Serum PRL levels, however, were stimulated and inhibited by VIP and PACAP, respectively.

The responsiveness of tuberoinfundibular dopaminergic neurons to prolactin feedback is diminished between early lactation and midlactation in the rat

The responsiveness of tuberoinfundibular dopaminergic neurons to prolactin feedback is diminished between early lactation and midlactation in the rat

Hypoprolactinemia decreases tyrosine hydroxylase activity in the tuberoinfundibular dopaminergic neurons acutely by protein dephosphorylation and chronically by changes in gene expression

This study evaluated the roles of protein dephosphorylation or suppressed gene expression in reducing tyrosine hydroxylase activity in tuberoinfundibular dopaminergic neurons after acute or chronic bromocriptine (BROMO) administration. Diestrous or ovariectomized rats were injected with BROMO (3 mg/kg, s.c.) at 1000 h and were sacrificed 4 h later or were injected with BROMO at 12 h intervals for 3 days.In vitro tyrosine hydroxylase activity was assessed by incubating hypothalamic explants with brocresine, an L-aromatic amino acid decarboxylase inhibitor, and measuring 3,4-dihydroxyphenylalanine (DOPA) accumulation in the stalk-median eminence (SME). The incubation medium also contained either 2 μM okadaic acid, a protein phosphatase 1 and 2A inhibitor, or its vehicle (0.25% dimethylsulfoxide). Acute (4 h) and chronic (3 days) BROMO treatment suppressed circulating PRL levels from 10-12 ng/ml to<1 ng/ml and reduced tyrosine hydroxylase activity in the SME by 60% or 40% in diestrous or ovariectomized rats, respectively. Okadaic acid increased tyrosine hydroxylase activity in the SME 2-fold in control diestrous or ovariectomized rats. The reduced tyrosine hydroxylase activity in the SME after acute BROMO treatment was increased by okadaic acid 5-or 3-fold in diestrous or ovariectomized rats respectively, to a value similar to the controls. In sharp contrast, after chronic BROMO treatment, the response to okadaic acid was blunted. As assessed byin situ hybridization, tyrosine hydroxylase mRNA signal levels in the arcuate nucleus of diestrous rats were not altered after acute BROMO treatment, but were reduced by 70% after chronic BROMO treatment. The acute BROMO-induced decrease in tyrosine hydroxylase activity was reversed by co-administration of oPRL or rPRL, indicating that the action of BROMO is via a reduction in PRL. The data suggest that protein dephosphorylation may be a primary mechanism for the rapid BROMO-dependent suppression of tyrosine hydroxylase activity, whereas suppression of tyrosine hydroxylase gene expression may contribute to the lower tyrosine hydroxylase activity after chronic BROMO treatment.

Bombesin acts in the suprachiasmatic nucleus to affect circadian changes in tuberoinfundibular dopaminergic neuron activity and prolactin secretion

Central administration of bombesin has been shown to block the afternoon PRL surge. A circadian change in the activity of tuberoinfundibular dopaminergic (TIDA) neurons that coincides with the onset of the PRL surge has also been found. Whether bombesin acts on the TIDA neuron to affect the PRL surge and where bombesin acts were the focuses of this study. Bombesin (0.75 microgram/3 microliters.rat) was first injected into the lateral cerebroventricle of estrogen-primed ovariectomized rats at 1200 h through preimplanted cannulas. Both the PRL surge and the circadian changes in TIDA neuron activity (using both 3,4-dihydroxyphenylacetic acid concentration and 3,4-dihydroxyphenylalanine accumulation in the median eminence as indexes) were blocked by the treatment. In suprachiasmatic nucleus-lesioned rats, however, both rhythms were absent, and the injection of bombesin exhibited no significant effect. Using microinjection of bombesin (50 ng/0.2 microliter) bilaterally into either the suprachiasmatic or arcuate nuclei at 1200 h, only the former injection effectively blocked both TIDA neuron activity and the PRL surge. We conclude that bombesin may act on the rhythm generation center, the suprachiasmatic nucleus, to exert its effect to disrupt TIDA neuron activity and the PRL surge.