Tuberoinfundibular Dopaminergic System Research Articles

Immunocytochemical colocalization of hypothalamic progestin receptors and tyrosine hydroxylase in steroid-treated monkeys.

Progesterone (P)-induced PRL secretion in estradiol (E)-primed monkeys is not due to direct pituitary stimulation, because lactotropes do not express progestin receptors (PR). However, the hypothalamus, particularly the tuberoinfundibular dopaminergic system (TIDA), plays a major role in the regulation of PRL secretion. To determine whether hypothalamic dopamine neurons are progestin target cells, the colocalization of PR and tyrosine hydroxylase (TH), a phenotypic marker of dopaminergic neurons, was examined with double immunocytochemistry. Two methods for visualizing the antigens were applied; the first was a dual peroxidase method, and the second was a peroxidase-alkaline phosphatase method. In addition, the question of whether E induces PR in dopamine neurons was explored. Spayed female monkeys were treated with empty Silastic capsules, E-filled capsules for a period of 28 days, or E capsules supplemented with P capsules for the last 14 days of E treatment. Only the E- plus P-treated monkeys exhibited an increase in serum PRL during the P treatment period. Frontal sections at the level of the optic chiasm and arcuate nucleus were examined for the colocalization of TH and PR. After E treatment, hypothalamic PR-positive cells increased in both intensity and number. Neurons expressing both TH and PR were detected in the rostral hypothalamus, lateral to the third ventricle (A11-rostral) and in a discrete subventricular population (A11-subvent). The lateral population continued caudally (A11-caudal). The A11-subvent population exhibited little steroid regulation. Of the remaining A11 TH neurons, approximately 20% exhibited PR in the spayed and E-treated groups. Addition of P doubled the percentage of PR-containing TH neurons in this group. Although very few TH-positive neurons in the ventral arcuate nucleus contained PR (A12-ventral), many double labeled neurons were observed in the dorsal arcuate region (A12-dorsal). Ventral arcuate TIDA neurons were not regulated by steroids, but E plus P increased PR expression in A12-dorsal. Double labeled cells were rarely seen in the zona incerta (A13) or the emerging ventral tegmental area (A10). In summary, P probably does not act directly on ventral arcuate TIDA neurons to stimulate PRL secretion. However, the frequency of PR-positive dopamine neurons in the A11-rostral, A11-caudal, and A12-dorsal groups increased with E and P treatment. Therefore, the contribution of the PR-positive periventricular dopamine neurons to progestin-stimulated PRL secretion may be important.

Immunocytochemical colocalization of hypothalamic progestin receptors and tyrosine hydroxylase in steroid-treated monkeys

Progesterone (P)-induced PRL secretion in estradiol (E)-primed monkeys is not due to direct pituitary stimulation, because lactotropes do not express progestin receptors (PR). However, the hypothalamus, particularly the tuberoinfundibular dopaminergic system (TIDA), plays a major role in the regulation of PRL secretion. To determine whether hypothalamic dopamine neurons are progestin target cells, the colocalization of PR and tyrosine hydroxylase (TH), a phenotypic marker of dopaminergic neurons, was examined with double immunocytochemistry. Two methods for visualizing the antigens were applied; the first was a dual peroxidase method, and the second was a peroxidase-alkaline phosphatase method. In addition, the question of whether E induces PR in dopamine neurons was explored. Spayed female monkeys were treated with empty Silastic capsules, E-filled capsules for a period of 28 days, or E capsules supplemented with P capsules for the last 14 days of E treatment. Only the E- plus P-treated monkeys exhibited an increase in serum PRL during the P treatment period. Frontal sections at the level of the optic chiasm and arcuate nucleus were examined for the colocalization of TH and PR. After E treatment, hypothalamic PR-positive cells increased in both intensity and number. Neurons expressing both TH and PR were detected in the rostral hypothalamus, lateral to the third ventricle (A11-rostral) and in a discrete subventricular population (A11-subvent). The lateral population continued caudally (A11-caudal). The A11-subvent population exhibited little steroid regulation. Of the remaining A11 TH neurons, approximately 20% exhibited PR in the spayed and E-treated groups. Addition of P doubled the percentage of PR-containing TH neurons in this group. Although very few TH-positive neurons in the ventral arcuate nucleus contained PR (A12-ventral), many double labeled neurons were observed in the dorsal arcuate region (A12-dorsal). Ventral arcuate TIDA neurons were not regulated by steroids, but E plus P increased PR expression in A12-dorsal. Double labeled cells were rarely seen in the zona incerta (A13) or the emerging ventral tegmental area (A10). In summary, P probably does not act directly on ventral arcuate TIDA neurons to stimulate PRL secretion. However, the frequency of PR-positive dopamine neurons in the A11-rostral, A11-caudal, and A12-dorsal groups increased with E and P treatment. Therefore, the contribution of the PR-positive periventricular dopamine neurons to progestin-stimulated PRL secretion may be important.

Repeated ECT and prolactin release in depressed patients

Whereas sham electroconvulsive therapy (ECT) increases the blood concentration of prolactin (PRL) up to two-fold after 15 m~n, real ECT increases PRL concentration several fold (Deakin et al 1983). Sequential study of the effects of electrode placement has established that bilateral ECT releases 25% (Swartz and Abrams 1984)-50% (Papakostos et al 1986) more PRL than does unilateral ECT which may demonstrate a greater hypothalan,ic-stimulating effect of bilateral ECT (Swartz and Abrams 1984). Electroencephalographic (EEG) study of the relationship between the length of cerebral seizure activity and PRL release has been inconclusive (Abrams and Swartz 198~b; Robin et at .~985), as has been the study of the relationship betw~,~ :, PRL release and recovery from depression (Deakin et al 1983; Abrams and Swartz 1985a). It has been suggested that the study of pituitary hormone release over a course of ECT may provide important information about alterations in central monoaminergic transmission in depressed patients. (Whalley et al 1982; Aperia et al 1985). The major role of the hypothalamus in the regulation of PRL release is inhibitory through the tuberoinfundibular dopaminergic system. A PRL releasing factor, such as thyrotropin releasing hormor ~ or vasoactive intestinal l~eptide, may be involved in stimulating PRL release and this may involve serotonin (5HT) (Leong et al 1983). After ECT, PRL release has been reported to be significantly reduced later on in a course of treatment for depression as compared to the first treatment (Deakin et al 1983; Abrams and Swartz 1985a; Aperia et al 1985; Cooper et al 1989), suggesting an increase in the responsiveness of post-synapuc dopamine receptors (Abrams and Swartz 1985a; Aperia et al 1985). This finding has not always been confirmed (Whalley et al 1982; Linnoila et al 1984; Haskett et al 1985). The intake of neuroleptic drugs increases both basal and ECl'-induced PRL release (Swartz 1985a; Aperia et a! 1985), but only one report concerned depressed patients free of neuroleptic drugs in the 2 weeks before ECT (Abrams and Swartz 1985a). O~:r aim was to study PRL release over a course of ECT in depressed patients who had not takeL', neuroleptic drugs during their index illness. ECT was monitored by 6-channel EEG because the ~gpatial distribution of epileptic seizures in hum,ms influences PRL release in that generalized se~ :ures release more PRL than focal seizures (Spc.ding et al 1986).

Chronic estradiol administration did not cause loss of hypothalamic LHRH or TIDA neurons in young or middle-aged C57BL/6J mice

Age-related decline in estrous cycle frequency and impaired pre-ovulatory gonadotropin surges at mid-life are modelled in young C57BL/6J mice by chronic (3 months) oral administration of estradiol (E 2). However, the cellular events that induce damage to the neuroendocrine center that regulate gonadotropins with age or following E 2 treatment are unclear. To address this issue, possible neuron loss was examined in relation to the loss of estrous cyclicity in E 2-treated mice, in particular neurons of the hypothalamic luteinizing hormone releasing hormone (LHRH) and/or tuberoinfundibular dopaminergic (TIDA) systems. By immunocytochemical methods, there was no change in the number of LHRH or TIDA neurons in mice that have become acyclic due to age or E 2 treatment. We conclude that the onset of acyclicity at middle-age or following chronic E 2 treatment is not associated with loss of LHRH or TIDA neurons and that other neuroendocrine changes must be considered for the cause of acyclicity, particularly those involved in the synaptic regulation of LHRH secretion.

Haloperidol Inhibits Maternal Retrieval and Licking, but Enhances Nursing Behavior and Litter Weight Gains in Lactating Rats

Abstract To determine whether the motorically-active and -inactive components of maternal behavior in rats are differentially affected by dopamine, we administered haloperidol, a dopamine receptor antagonist, to lactating rats on day 7 (+/-1) post-partum, 3 h after dam-litter separation and 1 h before their reunion. Compared to treatment with saline or domperidone, a peripherally-active dopamine antagonist, haloperidol treatment inhibited retrieval and licking of pups in a dose-dependent manner and hastened the onset of nursing behavior, but impaired the high crouch posture at the highest doses (8 and 10 mg/kg). Also, litters of dams treated with 0.2, 0.4, or 1.0 mg/kg haloperidol gained almost twice as much weight as litters of controls or of dams treated with 5 to 10 mg/ kg haloperidol. To determine whether the milk ejection pattern accounts for differential litter weight gain after moderate and high doses of haloperidol, milk ejections were assessed by pup stretch and nipple-switching responses for 30 min after the onset of crouching following a 4 h dam-litter separation on day 12 or 13 post-partum. The litters of dams treated with 1 or 3 mg/kg haloperidol had substantially greater litter weight gains than litters of control dams or of dams treated with 5 mg/kg haloperidol. The smaller litter weight gain of 5 mg/kg haloperidol litters was due to a retardation of milk ejections in their dams, while the smaller gain of control litters was due, at least in part, to greater behavioral activation by their dams. Non-dopaminergic effects of large doses of haloperidol (>/=5 mg/kg) may have reduced the display of the high crouching posture during nursing and contributed to the retardation of milk ejections. Thus, dopamine is necessary for the motorically-active components of maternal behavior, but its inhibition may be necessary for the assumption of the immobile nursing postures and for maximal litter weight gains. We hypothesize that extra-hypothalamic as well as tuberoinfundibular dopaminergic systems are inactivated by the suckling stimulus.

Evidence of Altered Dopaminergic Modulation of Prolactin and Thyrotropin Secretion in Patients with Polycystic Ovary Syndrome

In order to evaluate the functional activity of the tubero-infundibular dopaminergic system in polycystic ovary syndrome (PCOS), we analysed the prolactin (PRL) and thyrotropin (TSH) responses to the dopamine antagonist sulpiride. We studied 8 euthyroid women affected by PCOS and 7 normal women. The mean baseline PRL values were normal in both groups. After sulpiride administration the incremental area under the PRL profile in PCOS was significantly lower than in normal subjects (p less than 0.01). The mean basal plasma TSH levels were significantly higher in the PCOS than in the control group (p less than 0.01). After sulpiride administration the incremental area under the TSH profile was significantly lower in PCOS patients than in normal women (p less than 0.01). The higher basal plasma levels of TSH, the blunted response of PRL and the lack of response of TSH to sulpiride in PCOS suggest a relative decrease of the dopaminergic activity in PCOS patients.

Estrogen effects on the tuberoinfundibular dopaminergic system in the female rat brain

Estrogen effects on tyrosine hydroxylase (TH), monoamine oxidase types A and B (MAO), and dopamine (DA) in microdissected regions of the hypothalamus, preoptic area and substantia nigra (SNR) of the female rat brain were investigated. Ovariectomized (OVX) young adult female rats were implanted with single silastic capsules containing 100% estradiol alerate (EV). Control rats received empty silastic capsules. Two weeks following capsule insertion, EV decreased TH activity and DA concentration in the arcuate nucleus (AN) while no significant changes in TH activity or DA concentration were observed in the SNR, ventromedial nucleus (VMN), suprachiasmatic nucleus, paraventricular nucleus, medial preoptic nucleus, or the periventricular preoptic nucleus. Although estrogen suppressed TH and DA in the AN, 2 weeks following removal of the estrogen containing capsules, TH activity and DA concentration were restored to control (OVX) levels. Suppression of MAO activity occurred in both the AN and the VMN of rats implanted with EV capsules and returned to OVX levels following the removal of the estradiol load. These results revealed that estrogen effects on TH and MAO activities and DA concentration in the midbrain are region specific and reversible; and that among the dopaminergic systems studied, estrogen effects on TH and DA are confined to the tuberoinfundibular dopaminergic system (TIDAS). Furthermore, these results support our hypothesis that estrogen is a key regulator of DA function in the TIDAS via effects on TH. The importance of these findings to the control of gonadotropin secretion and reproductive cyclicity is discussed.

A central action of α-melanocyte-stimulating hormone on serum levels of LH and prolactin in rats

We have investigated the effect of administration of alpha-MSH into the median eminence (ME) of rats on the release of LH and prolactin. Continuous infusion of alpha-MSH (0.5 micrograms/h) into the ME from the afternoon of the second day of dioestrus and over the 24 h of pro-oestrus inhibited the preovulatory LH and prolactin surge and the occurrence of ovulation. This inhibitory effect on LH and prolactin release was also observed in chronically ovariectomized rats given a single injection of alpha-MSH (1 micrograms/ml per rat) into the ME (blood samples were collected 0, 20, 60, 90, 105 and 120 min after injection). The intraperitoneal injection of the dopamine receptor blocker, haloperidol (2 mg/kg), 30 min before the injection of alpha-MSH into the ME prevented the inhibitory effect of alpha-MSH on the release of LH and prolactin. These results suggest that hypothalamic alpha-MSH might be involved in the regulation of LH and prolactin release via the tuberoinfundibular dopaminergic system and that this system also modifies the serum concentrations of alpha-MSH.

Effects of clonidine and naloxone on the dopamine levels in the rat mediobasal hypothalamus measured by in vivo voltammetry.

We examined the effect of alpha 2-adrenergic stimulation on dopamine (DA) levels in the mediobasal hypothalamus (MBH) by means of in vivo voltammetry in the rat. Intravenous (i.v.) injection of an alpha 2-agonist, clonidine (15 micrograms/100 g b.wt.) caused a decrease of MBH DA levels (76 +/- 3% of basal levels). Pretreatment with phenoxybenzamine (500 micrograms/100 g b.wt.i.v.), an alpha-antagonist, blunted the decrease of MBH DA levels induced by clonidine. Naloxone (125 micrograms/100 g b.wt.i.v.), an opioid antagonist, blunted the decrease of MBH DA levels induced by clonidine. These results suggest that alpha 2-adrenergic mechanisms may inhibit the tuberoinfundibular dopaminergic (TIDA) system through the activation of the endogenous opioid system.

2-hydroxyestradiol is not mediating the effects of estradiol on tuberoinfundibular dopaminergic neurons controlling prolactin secretion in female rats

Conversion of estrogens in their hydroxylated derivatives has been proposed as one of the mechanisms of action for these steroids in the brain. However, the actions of catechol estrogens on the hypothalamic control of the prolactin (PRL) release seem to be different to the effects caused by native estrogens. To clarify this question, we have examined the activity of tuberoinfundibular dopaminergic (TIDA) neurons in ovariectomized rats treated with estradiol (E 2) or 2-hydroxyestradiol (OHE 2), both administered alone or previously to progesterone (P). l-3,4 dihydroxyphenylacetic acid (DOPAQ/dopamine (DA) ratio in the medio basal hypothalamus (MBH), the anterior pituitary (AP) content of DA and the PRL release into the peripheral blood have been used as indexes of neural activity of TIDA system. Ovarieetomy caused a significant reduction in plasma PRL levels, with increases of the DOPAC/DA ratio in MBH and of the AP content of DA. These changes were reversed by administration of E 2, suggesting that this steroid exerted its stimulatory effect on PRL release by decreasing the activity of TIDA neurons. In contrast, OHE 2 was unable to alter plasma PRL levels, the DOPAC/DA ratio in the MBH and the AP content of DA. On the other hand, P was able to stimulate the PRL release, but it needed a previous priming action of E 2 since it was unable to modify plasma PRL levels when it was injected alone. This priming effect of E 2 for the P action was not exerted by OHE 2. Hence, we can conclude that the conversion of E 2 in its 2-hydroxylated derivative is not mediating the effects of this steroid, nor its priming action for the P effect, on the hypothalamic dopaminergic control of the PRL release.

Infertility in transgenic female mice with human growth hormone expression: Evidence for luteal failure

Introduction of the human growth hormone (hGH) gene fused with mouse metallothionein I promoter into domestic mice leads to ectopic synthesis of hGH, marked stimulation of somatic growth, and female sterility. Transgenic females (produced by mating transgenic males to normal females) mated but failed to become pregnant or pseudopregnant as evidenced by the recurrence of vaginal plugs every 5-7 days. Daily injections of 1 mg progesterone, starting on day 1 postcoitum (p.c.), maintained pregnancy, suggesting that the sterility of these animals is due to inadequate luteal function. In ovariectomized female transgenic mice, median eminence (ME) turnover of dopamine (DA) was increased, and plasma prolactin (PRL) levels were reduced, presumably because of the known lactogenic activity of hGH in rodents. From these observations we suspected that either 1) the corpora lutea of these animals are unresponsive to lactogenic hormones, or 2) hGH by stimulating tuberoinfundibular dopaminergic (TIDA) neurons interferes with the increase in PRL release that normally follows mating and this, in turn, leads to luteal failure. To distinguish between these possibilities, transgenic females were treated with PRL-secreting ectopic pituitary transplants from normal females of the same strain on day 1 p.c. Eight of ten treated females became pregnant and delivered litters. We conclude that infertility of transgenic female mice with hGH expression is due to activation of the TIDA system, suppression of endogenous PRL release, and luteal deficiency.

Histaminergic regulation of prolactin secretion: involvement of tuberoinfundibular dopaminergic neurons.

It has been shown that histamine (HA) stimulates prolactin (PRL) secretion via H2 receptors following intra-cerebroventricular infusion and via H1 receptors following systemic (intra-arterial) infusion. Since the effect of HA appears to be exerted at a suprapituitary level, we investigated the involvement of the tuberoinfundibular dopaminergic (TIDA) system in HA-induced PRL secretion in urethane-anesthetized male rats. HA infused intracerebroventricularly (30 micrograms) or intra-arterially (420 micrograms) decreased the dopamine (DA) concentration in pituitary portal blood by 30 and 23%, respectively. Blockade of DA receptors by pimozide did not prevent the stimulation of PRL secretion induced by intracerebroventricular infusion of HA or the H2 receptor agonist dimaprit. Furthermore, during DA receptor blockade intracerebroventricular infusion of the H1 receptor agonist 2-thiazolylethylamine inhibited PRL secretion. In contrast, pimozide prevented the stimulation of PRL secretion induced by intra-arterial infusion of HA and the H1 receptor agonist 2-thiazolylethylamine. In fact, under these conditions intra-arterial infusion of HA or the H2-receptor agonist dimaprit inhibited PRL secretion. During treatment with alpha-methyl-p-tyrosine, which reduced the hypothalamic DA content by 50%, HA infused intracerebroventricularly stimulated PRL secretion, while HA infused intra-arterially inhibited the secretion, which is in accordance with the results obtained during pimozide treatment. Cholinergic blockade by atropine did not prevent the HA-induced PRL release, excluding the possibility that the observed effect of pimozide is due to its anticholinergic property. We suggest that intracerebroventricular infusion of HA by activation of H2 receptors may stimulate PRL secretion partly via inhibition of the TIDA system and partly via other mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of tyrosine-hydroxylase (TH)-immunoreactive neurons in the diencephalon of the pigeon (Columba livia domestica).

The distribution of tyrosine-hydroxylase (TH)-immunoreactive cell bodies and fibers in the diencephalon has been investigated with immunohistological techniques in the pigeon. The results suggest that TH is present in a number of morphologically distinct neuronal systems. Preoptic and hypothalamic TH neurons were subdivided into a medial periventricular and a lateral group. The medial group starts with a rostral collection of small cells in the preoptic region. A significantly larger collection of TH neurons occupies the paraventricular nucleus (PVN) (stratum cellulare internum) and mainly consists of large multipolar cells. Further caudally, the main concentration of cells is in the hypothalamic posteromedial and the periventricular regions of the tuberoinfundibular (arcuate) nucleus. No TH neuron was found in the ventral and lateral parts of the tuberoinfundibular region, suggesting that the prominent tuberoinfundibular dopaminergic system described in mammals is absent in the pigeon. This further substantiated by the relative scarcity of TH immunoreactive fibers and varicosities in the neurohemal zone of the median eminence (ME). The caudalmost components of the medial group appear to be continuous with the large population of TH neurons distributed in the midline of the mesencephalon. Tyrosine-hydroxylase-immunopositive cells have not been found in the paraventricular organ. The lateral group consists of TH neurons loosely arranged in the lateral hypothalamus, including regions of the supraoptic nucleus and hypothalamic posterolateral nucleus. Tyrosine-hydroxylase containing neurons vary widely in size, shape, and dendritic arborization in each diencephalic region. However, it is possible to distinguish two main cell types. Small bipolar neurons with two simple arborizing dendrites were concentrated in the medial periventricular system. The second type of cell is large, multipolar with four to five branching dendrites. This latter cell type occurs mainly in the lateral system and in the PVN. Major fiber bundles containing TH immunoreactivity were identified in the lateral and periventricular hypothalamus. The paraventricular organ and the organum vasculosum laminae terminalis contained the densest arborization of fibers and varicosities. In the ME, dense innervation was found in the subependymal layer. Dense arborizations of TH positive fibers and varicosities were located in the septal nuclei and the paleostriatum augmentatum.

Testosterone inhibition of tyrosine hydroxylase expression in the hypothalamic arcuate nucleus

Immunohistochemistry was used to detect tyrosine hydroxylase (TH)-containing neurons in the hypothalamic arcuate nucleus in male rats. Two weeks following castration, the number of TH-positive cells was significantly greater than in intact controls. The castration-responsive TH-positive cells were uniformly distributed throughout the mediolateral extent of the arcuate nucleus. Treatment with testosterone significantly suppressed the castration response, whereas neither estradiol nor 5α-dihydrotestosterone were effective. The number of TH-positive arcuate neurons in the female was similar to that in the male. Ovariectomy did not affect the number of TH-positive neurons. These findings indicate that TH expression is tonically inhibited in the tuberoinfundibular dopaminergic system of the male rat by testosterone.

Electrophysiologic evidence for connections between the supraoptic and the arcuate/ventromedial hypothalamic nuclei in the rat

Extracellular action potentials were recorded from 48 single units located in the hypothalamic arcuate and ventromedial nuclei. Fifteen percent of the cells were identified as projecting to the median eminence and some of these cells may have belonged to the tuberoinfundibular dopaminergic systems. Responses of all cells to stimulation of the ipsilateral supraoptic nucleus were recorded; 17% of ventromedial nucleus neurons were antidromically identified as projecting to the supraoptic nucleus. None of the latter cells was also identified as projecting to the median eminence. Three of six identified tuberoinfundibular and eight unidentified ventromedial nucleus cells were found to be excited by stimulation of the supraoptic nucleus. One arcuate cell identified as projecting to the median eminence was nonresponsive to supraoptic stimulation. Orthodromic inhibitory responses were recorded from 17% of all cells recorded but no inhibitory responses were recorded from cells identified as projecting to the median eminence. We suggest that these results may provide some neurophysiologic explanations for the observed interrelationships between oxytocin and prolactin secretion, and between vasopressin and growth hormone secretion.

Effect of different organic and inorganic blockers of calcium entry on the release of endogenous dopamine from tuberoinfundibular neurones

In the present study the effect of different blockers of calcium entry belonging to different chemical classes on basal and K +-elicited release of endogenous dopamine (DA) from tuberoinfundibular dopaminergic neurones was studied in vitro. For this purpose fragments of hypothalamus containing arcuate-periventricular nuclei and median eminence were incubated in vitro and endogenous DA released into the medium was assayed by radioenzymatic assay. The organic blockers of calcium entry, nitrendipine, nimodipine, nifedipine, diltiazem and flunarizine did not modify basal or K +-evoked release of endogenous DA, unless very large concentrations (100μM) of nifedipine or diltiazem were used. The phenylalkylamine methoxyverapamil (D-600) consistently inhibited K +-stimulated release of endogenous DA in concentrations of 50 and 100 μM. Cobalt and lanthanum, two ions with an ionic radius similar to that of calcium and which are known to inhibit calcium fluxes through nerve membranes, significantly blocked release of endogenous DA elicited by 35 mM K +. In summary, the results of the present study showed that calcium channels in the tuberoinfundibular dopaminergic system displayed a different sensitivity to various classes of blockers of calcium entry. Inorganic blockers of calcium entry, like lanthanum and cobalt, appeared to be the most effective in blocking Ca 2+-dependent release of endogenous DA, whereas, among the organic calcium antagonists, phenylalkylamines seemed to possess a certain degree of effectiveness.

The fine structural organization of tyrosine hydroxylase immunoreactive neurons in rat arcuate nucleus

The fine structure of the tyrosine hydroxylase (TH) immunoreactive neurons of the hypothalamic arcuate nucleus was examined by means of immunocytochemistry [peroxidase-antiperoxidase (PAP) method], utilizing an antibody against TH. Immunolabeled axon terminals were observed infrequently and were located predominantly in the lateral region, whereas numerous labeled perikarya and dendrites were found throughout the nucleus. The labeled terminals, containing primarily clear and occasionally dense core vesicles, were never observed in synaptic contact. On the other hand, unlabeled axon terminals were frequently seen synapsing on labeled dendrites. In addition, the labeled dendrites were often seen in direct apposition to other neuronal elements such as both labeled and unlabeled perikarya. In contrast, unlabeled dendrites were never seen apposed to labeled perikarya. Labeled dendrites also occurred in direct contact with one another and with unlabeled dendrites. Moreover, numerous labeled dendrites were encountered along tanycytic processes. Dendrites engaged in tanycytic appositions were occasionally partially encompassed by thin sheaths emanating from the tanycytic process. The extensive contact made by the labeled dendritic profiles on both labeled perikarya and dendrites suggests that tubero-infundibular dopaminergic (TIDA) cells may communicate with each other by means of dendritic release of dopamine. The presence of appositions between labeled dendrites and both unlabeled perikarya and dendrites suggests that the TIDA system also influences other neuronal populations through its dendrites. Finally, the dendrotanycytic relationship suggests that the TIDA system may play some role in the regulation of tanycytic function.

Tuberoinfundibular dopaminergic system and anterior pituitary dopamine receptor in Shy-Drager syndrome

Tuberoinfundibular dopaminergic system (TIDA) together with anterior pituitary dopamine receptor was evaluated in 9 patients with Shy-Drager syndrome (SDS) through the responses of serum growth hormone (GH) and prolactin (PRL) to dopaminergic stimulation or dopamine receptor inhibition. A variety of abnormal responses of serum GH and/or PRL following the administration of these drugs suggests the TIDA involvement as well as the nigrostriatal system and moreover the different degeneration processes in each dopaminergic system in SDS.

Hypothalamic neurotransmitter function in experimentally induced hyperprolactinemia

It is known that animals or patients bearing a prolactin (PRL)-secreting tumor (PST) do not suppress PRL levels after administration of indirectly acting dopamine agonists, namely nomifensine (Nom), and are not responsive to the PRL releasing effect of antidopaminergic drugs and opioid peptides. Since the action of these drugs is mediated through the tuberoinfundibular dopaminergic (TIDA) system, these findings have been taken to indicate that animals and humans bearing prolactinomas have a defective TIDA function. Alternatively, PRL unresponsiveness to these drugs could be due to hyperfunction of TIDA system for the feedback action of high PRL levels. To clarify whether hypo- or hyperfunction of the TIDA system was responsible for such behaviour, we tested the effect of a synthetic opioid peptide (FK 33–824), a DA receptor antagonist, domperidone (Dom), and of Nom on PRL secretion in two experimental models of non-tumoral hyperprolactinemia, i.e. rats bearing ectopic pituitaries since 3 days (TP rats), or treated with ovine PRL (oPRL 250 μg, twice daily for 3 days), in which existence of an increased TIDA function has been demonstrated. FK 33–824 (0.5 mg/kg i.p.) increased significantly plasma PRL levels in control rats but failed to do so in TP rats and it elicited a significantly lower PRL response than in controls in rats treated with oPRL. In both experimental models, a PRL secretagogue, e.g. 5-hydroxytryptophan (50 mg/kg i.p.), elicited the same response as in controls, indicating that the pituitayr PRL pool was preserved. Kinetic characteristics of opioid binding sites in the hypothalamus of hyperprolactinemic rats did not differ from those of controls, and also hypothalamic Met-enkephalin concentrations were similar to those of controls. These findings would exclude a defect of endogenous opioid neurotransmission as responsible for the impaired PRL stimulation by the enkephalin analog. Supporting this were the findings that oPRL-treated rats exhibited a growth hormone response to FK 33–824 and a rise in plasma luteinizing hormone after naloxone (5 mg/kg s.c.) similar to those present in controls. These data indicate that the defective PRL responsiveness to opioids has to be attributed to the impairment of a neurotransmitter e.g. DA, functionally located at a site ‘downstream’ from opioid receptors. As already shown for oPRL-treated rats, TP rats exhibited a faster turnover rate of DA in the median eminence of the hypothalamus. In keeping with the idea that the hypothalamic dopaminergic system was involoved in the defective PRL responsiveness to opioids, Dom (50 μg/kg i.p.) induced aignificantly lower increase of PRL levels in TP and oPRL-injected rats than in controls. All these data demonstrate that opioid peptides and DA receptor antagonists are unable to discriminate the functional state of TIDA system. In fact, they fail to elicit a rise in plasma PRL in hyperprolactinemic states related to either TIDA hypo- (PST rats and humans) or hyperfunction (TP and oPRL-treated rats). In contrast to FK 33–824 and Dom, Nom (10 mg/kg i.p.), a drug which fails to lower PRL secretion in animals and patients with prolactinoma, did suppress plasma PRL levels of oPRL-treated rats as those of controls. The latter data indicate that indirectly acting DA agonists can discriminate the functional state of TIDA system; they fail to lower PRL secretion only when TIDA function is defective.

Regulation of prolactin secretion during suckling: involvement of the hypothalamo-pituitary GABAergic system.

Previous results demonstrated that GABA exerts a dual control on PRL secretion, one excitatory mediated in part by the impairment of the tubero-infundibular dopaminergic (TIDA) system function, the other inhibitory occurring at the level of the anterior pituitary (AP), where 3H-GABA and 3H-Muscimol (3H-M) recognition sites have been described. This report provides evidence for a physiological role of the tubero-infundibular GABAergic system (TI-GA-BA) on PRL secretion in the rat. In lactating rats separated for 4 h from their pups reinstitution of suckling for different periods resulted in an increase either in glutamic acid decarboxylase (GAD) activity in the mediobasal hypothalamus (MBH) or in AP-GABA content. Dynamic changes of the GABAergic function in the MBH-AP system seemed to have a certain degree of specificity because suckling did not affect GAD activity in the caudate nucleus. In lactating rats 2, 4, 8 and 24 h after removal of the offsprings AP-GABA concentrations and plasma PRL titers significantly decreased with respect to values present in rats never separated from their pups. Since it has been demonstrated that the PRL lowering effect of GABA is a receptor-mediated event, we have investigated the plasticity of AP-GABA receptors during suckling. The inhibitory action of GABA seems to be mediated mainly by the activation of the high affinity binding sites. This proposition is supported by the fact that in lactating rats, where only the high affinity receptor population is present, M was still able to decrease significantly plasma PRL concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)