NMDA Receptors in the Medial Zona Incerta Stimulate Luteinizing Hormone and Prolactin Release

1. The aim of the present work is to demonstrate the interaction between the glutamatergic/NMDA and dopaminergic systems in the medial zona incerta on the control of luteinizing hormone and prolactin secretion and the influence of reproductive hormones. 2. Proestrus and ovariectomized rats were primed with estrogen and progesterone to induce high or low levels of luteinizing hormone and prolactin. 2-Amino-7-phosphonoheptanoic acid, an NMDA receptor antagonist, and dopamine were injected in the medial zona incerta. Blood samples were withdrawn every hour between 1,600 and 2,000 hours or 2,200 hours via intracardiac catheter from conscious rats. Additional groups of animals injected with the NMDA receptor antagonist were killed 1 or 4 h after injection. Dopamine and its metabolite 3,4-dihydroxyphenylacetic acid were measured in different hypothalamic regions. 3. 2-Amino-7-phosphonoheptanoic acid blocked the ovulatory luteinizing hormone surge in proestrus rats. 2-Amino-7-phosphonoheptanoic acid also blocked the increase in luteinizing hormone induced by ovarian hormones in ovariectomized rats, an effect that was partially reversed by dopamine injection. Conversely, the increased release of luteinizing hormone and prolactin induced by dopamine was prevented by 2-amino-7-phosphonoheptanoic acid. We found that the NMDA antagonist injection decreased the dopaminergic activity--as evaluated by the 3,4-dihydroxyphenylacetic acid/dopamine ratio--in the medio basal hypothalamus and increased in the preoptic area. 4. Our results show an stimulatory role of NMDA receptors on the ovulatory luteinizing hormone release and on luteinizing hormone release induced by sexual hormones and demonstrate that the stimulatory effect of dopamine on luteinizing hormone and prolactin is mediated by the NMDA receptors. These results suggest a close interaction between the glutamatergic and dopaminergic incertohypothalamic systems on the control of luteinizing hormone and prolactin release.     

In vitro sensitivity of rat ovarian follicles to the direct action of various hormones applied in particular stages of sexual cycle

The direct action of various hormones on the ovary of adult rat ovaries has been investigated. The ovaries were surgically exposed at particular stages of sexual cycle i.e. in estrus, metestrus, diestrus and proestrus, and were injected with various doses of gonadotropic hormones. Injected ovaries were excised 56 hrs after injection and examined histologically. The reactivity of the ovaries to hormone applied was dependent on stage of the cycle. The considerable increase of the amount of t1 and t2 follicles was observed after injection of gonadotropins in estrus stage. In metestrus stage the influence of gonadotropins on the follicles more advanced in development was pronounced. Numerous follicles t3 appeared in diestrus under the influence of all used hormones.     

神经内分泌因子调控鱼类生殖和生长的相互作用

脊椎动物的生长与生殖活动有着密切的联系并相互作用。许多调节生长和代谢活动的内分泌因子对青春期或者性腺的发育产生影响。同样,调节生殖活动的许多激素亦同时对生长和代谢产生影响。近年来,我们和其他学者对鱼类生长和生殖的神经内分泌调节的相互作用进行了研究,主要的进展是：①在促进性腺的激素影响生长方面,发现促性腺激素释放激素（ＧｎＲＨ）和多巴胺都能和脑垂体生长激素细胞的特异性受体结合而刺激生长激素释放,并能     

Early manipulation of juvenile hormone has sexually dimorphic effects on mature adult behavior in Drosophila melanogaster

Hormones are critical for the development, maturation, and maintenance of physiological systems; therefore, understanding their involvement during maturation of the brain is important for the elucidation of mechanisms by which adults become behaviorally competent. Changes in exogenous and endogenous factors encountered during sexual maturation can have long lasting effects in mature adults. In this study, we investigated the role of the gonadotropic hormone, juvenile hormone (JH), in the modulation of adult behaviors in Drosophila. Here we utilized methoprene (a synthetic JH analog) and precocene (a JH synthesis inhibitor) to manipulate levels of JH in sexually immature male and female Drosophila with or without decreased synthesis of neuronal dopamine (DA). Locomotion and courtship behavior were assayed once the animals had grown to sexual maturity. The results demonstrate a sexually dimorphic role for JH in the modulation of these centrally controlled behaviors in mature animals that is dependent on the age of the animals assayed, and present DA as a candidate neuronal factor that differentially interacts with JH depending on the sex of the animal. The data also suggest that JH modulates these behaviors through an indirect mechanism. Since gonadotropic hormones and DA interact in mammals to affect brain development and later function, our results suggest that this mechanism for the development of adult behavioral competence may be evolutionarily conserved.     

Behavioural and morphological effects of testosterone and gonadotropins in the young male domestic duck (Anas platyrhynchos L.)

Young male domestic ducks 20–72 days old were successively injected with two hormonal preparations. The first hormone treatment included males injected with testosterone propionate (TP), human chorionic gonadotropin (HCG), pregnant mare serum (PMS), control oil injected males (C) and males injected with TP and submitted at the same time to a permanent intense light. During the second hormonal treatment all males except controls were injected with TP.Almost no behavioural effects were observed in any group of males following the first treatment. The second one, however, induced intense social display and sexual behaviour in the four TP-injected groups. Some qualitative and quantitative differences were found between groups according to the first hormone treatment to which they had been submitted. This suggests a possible role of gonadotropic hormones in the control of social behaviour in ducks. Experimental data supporting this hypothesis are briefly reviewed and discussed.     

Inhibition of epinephrine and gonadotropic hormone induced ornithine decarboxylase activity by phenoxybenzamine in the testis of immature rat

The effect of α and β adrenergic receptor blockers on epinephrine and gonadotropic hormone induced ornithine decarboxylase (ODC) activity in the testis of immature rats was studied. Intratesticular injection with phenoxybenzamine at 15 min before treatment with epinephrine or gonadotropic hormones blocked ODC activity. Similar injection with propranolol or practolol had no effect on ODC activity. These results show that α adrenergic receptors are involved in the action of epinephrine and gonadotropic hormones in the testis.     

Gonadotropin production and release in female goldfish (Carassius auratus) after administration of pimozide and an LHRH analogue as studied by electron microscopy and radioimmunoassay

Summary The effect of pimozide and an LHRH-analogue (LHRH-A) on gonadotropic cells of the goldfish pituitary gland were described qualitatively and quantitatively. A scale of four categories was devised to reflect various ultrastructural appearances of the cells. Experimental animals were divided into a control group, a group injected with LHRH-A alone, pimozide alone, and groups receiving these two substances in combination. Fish injected with the single substance were killed 12 h after injection while the groups receiving the combined treatments were killed at 4, 12 and 48 h. Serum levels of gonadotropin measured by radioimmunoassay were used to indicate whether an increase in hormone release had occurred. An immunocytochemical technique, the protein A-gold procedure, assured that the cells studied were gonadotropes. The control group showed variation in the profiles of gonadotropic cells. The single treatment groups showed some increase in secretory inclusions. At 4 h after injection the combined treatment caused a significant increase in hormone granules; at 12 and 48 h there was a gradual decrease in content of secretory products, and an increase in vacuolization. The results indicate that the combined pimozide and LHRH-A treatment stimulated gonadotropin production as well as release.     

Induced spawning in carp with fractionated fish pituitary extract

Three different fractions of fish pituitary extract obtained by molecular sieving of the whole pituitary extracts on Sephadex G-100 (Sinha, 1969 a ) have been tested for effectiveness in inducing spawning of members of the carp family. Fish differing in their spawning habits were chosen-bighead carp and silver carp do not spawn at all in captivity, whereas Puntius and common carp spawn freely. Gravid fish injected with the second fraction alone showed courtship behaviour and ovulated viable eggs. None of the fish injected with the first fraction or the third fraction showed any courtship behaviour nor did they ovulate. The second fraction from the pituitary extract of free spawner or non-spawner, immature or mature, male or female was found effective in inducing spawning in both the free spawner and non-spawner gravid females. Clearly this suggested that this fraction contains either an active gonadotropin or a gonadotropin releasing factor. However, unlike the free spawner, these non-spawners do not breed in captivity unless injected with an additional amount of the second fraction or the whole pituitary material. The cause of this is still difficult to explain. Synthetic mammalian hormones such as FSH, LH, mixed mammalian anterior pituitary and chorionic hormone, ACTH and posterior pituitary hormones have failed to induce courtship behaviour and ovulation. Oxytocic activity of the fractions showed that there was no correlation between spawning activity and the rat uterus activity. Also these fractions did not contain any appreciable amount of arginine vasotocin. Thus it is suggested that isotocin/arginine vasotocin is not the principle responsible for spawning in these fish.     

Control of gonadotropic hormone release in trout: Influence of synthetic LHRH and LHRH analogues invivo and invitro

Studies were conducted to determine the influence of some LHRH analogues on gonadotropic hormone (GtH) secretion in two species of trout. The observations indicated that synthetic LHRH and various stimulatory LHRH analogues are approximately equipotent in these fish. This is an unexpected result considering the superactive properties of these analogues demonstrated in mammals. An inhibitory LHRH analogue was also tested in the trout with the result that powerful inhibition of LHRH induced GtH release was obtained.     

Role of calcium in the regulation of adenohypophysial hormone release.

It is now accepted by most investigators that the initial action of most peptide hormones involves an interaction with a specific receptor on (or in) the plasma membrane of the target cell. A cascade of intracellular events results and culminates in the physiological response characteristic of the interaction of the particular hormone with its target cell. The regulation of hormone release from the adenohypophysis by the hypothalamic releasing hormones is presumed to occur via a similar process. The nature of the interaction at the cell surface as well as the details and sequence of the subsequent intracellular events are largely unknown. We do know, however, that two of the key factors regulating the intracellular secretory machinery in most cells are 1) the adenylate cyclase — cyclic AMP — protein kinase system and 2) the divalent cation, calcium. Since there have been several recent reviews (1–3) which have covered the role of the cyclic nucleotides in pituitary hormone secretion, this discussion will be restricted to a consideration of the regulatory role played by calcium.As was the case with tissues, the early work regarding calcium and the adenohypophysis followed the pattern of determining the ability of secretagogues to release pituitary hormones subsequent to various manipulations designed to remove what was often implicity considered to be extracellular calcium.     

Dynamics of juvenile hormone-mediated gonadotropism in the lepidoptera

Reproduction in moths and butterflies is a dynamic process that is influenced by various endogenous physiological processes and exogenous factors. The Lepidoptera may be divided into four distinct groups based on their gonadotropic hormones and other reproductive and biological characteristics, regardless of phylogenetic relationships. Some species have integrated their reproduction tightly with the endocrine events of metamorphosis, the waxing or waning ecdysteroid levels. Other species rely instead on juvenile hormone (JH), in part or solely. Species of Lepidoptera that rely on JH as the gonadotropic signal also exhibit polyandry. Several mechanisms have been suggested for the occurrence of polyandry, including availability of male-transferred nutrients (gonadotrophic effect), need for additional sperm, and increased genetic variability. We propose an additional reason for polyandry observed in some lepidopterans. If a female remains a virgin, her endogenous gonadotropic signal diminishes, and eggs that have been produced already may be resorbed to increase longevity. During copulation, the male may trigger a neural/humoral response in the female, thus stimulating release of her endogenous gonadotropic signal, JH, and/or inhibiting degradation of the same, whence she matures new eggs. The mating effect appears to act humorally on the cephalic structures in several species. Whether this change in JH titer is due to an effect on synthesis and release by corpora allata only or occurs in conjunction with inhibition of JH degradation is unknown. Arch. Insect Biochem. Physiol. 35:539–558, 1997. © 1997 Wiley-Liss, Inc.     

New approaches to the therapy of various tumors based on peptide analogues.

The discovery of hypothalamic hormones was briefly reviewed. The development of new hormonal methods for the therapy of various cancers based on analogues of hypothalmic hormones is then presented. My group isolated luteininzing hormone-releasing hormone (LH-RH), also known as Gn-RH, from pig hypothalmi, elucidated its amino acid sequence, and synthesized it in 1971. The interest in medical applications of LH-RH led to the synthesis of LH-RH analogues by various groups. LH-RH agonists substituted in positions 6 or 10 including Decapeptyl, Leuprolide and Zoladex are much more active than LH-RH and on continuous administration produce inhibition of pituitary and gonads. Chronic administration of LH-RH agonists is being utilized for the treatment of prostate and breast cancer. Octapeptide analogues of somatostatin have various applications in Oncology. In 1980 we developed a new endocrine therapy for advanced prostate cancer based on agonists of LH-RH, which is now preferred by 70-90% of prostate cancer patients for primary treatment. LH-RH antagonists such as Cetrorelix can be used for therapy of BPH. On the basis of the presence of specific receptors for hypothalamic peptides on human cancers, we developed targeted cytotoxic analogues of LH-RH, somatostatin, and bombesin/GRP linked to doxorubicin or 2-pyrrolinodoxorubicin. These analogues inhibit the growth of experimental human prostate, breast, ovarian and endometrial cancer, renal cell carcinoma, pancreatic, colorectal and gastric cancers, small cell lung carcinoma (SCLC) and non-SCLC, brain tumors, melanomas, and lymphomas. Cytotoxic LH-RH analogues are now in clinical trials. Recently we demonstrated that growth hormone-releasing hormone (GH-RH) also serves as an autocrine growth factor in many cancers. Antagonistic analogues of GH-RH synthesized in our laboratory inhibit the growth of diverse tumors. The discovery of LH-RH and somatostatin has led to clinical use of their analogues in the field of cancer treatment and GH-RH antagonists also show a great promise.     

Early pituitary follicle stimulating hormone response to gonadotrophin releasing hormone in ewes

The object of our experiments was to characterize the response of plasma follicle stimulating hormone (FSH) within minutes of an i.v. injection of high or low doses of gonadotrophin releasing hormone (GnRH), especially in relation to contemporary changes in luteinizing hormone (LH) concentrations. In the deep anoestrous period (June), three intact ewes and two ovariectomized ewes were injected with 1 mug synthetic GnRH followed 2 h later by a second identical injection. A week later, the same regimen was repeated with the same sheep but with 50 mug GnRH after an interval of 5 h 20 min. Blood samples were collected every 15 sec for 15 min after each injection (early release), then at longer intervals (main release) till the next treatment, followed by sampling for a further 6-h period after the second treatment. FSH was released as soon as the second minute after GnRH injection in all ewes. The mean pituitary FSH response, during this early release, in intact and ovariectomized ewes was similar after either 1 or 50 mug GnRH. However, the main release was less pronounced in the ovariectomized sheep and was not stimulated after the second treatment in all sheep. Three other ewes were injected with 40 mug GnRH and sampled every 15 sec for seven, 6-min periods during the period of release to compare FSH and LH secretion. The profiles reflected a similarity in sensitivity and responsiveness to GnRH, especially soon after GnRH injection. Increases in both hormones were formed by several grouped associated spikes. It is suggested that a readily releasable pool of FSH exists in the ewe. There are probably differences in the mechanisms of synthesis and/or release between pituitary FSH and LH.     

Studies on sex-organ development. The hormonal regulation of steroidogenesis and adenosine 3'':5''-cyclic monophosphate in embryonic-chick ovary.

1. We investigated the production of steroid hormones by the ovaries of the developing embryonic chick under conditions of organ culture. Radioimmunoassay techniques were used to measure the amount of steroid hormone released into the culture medium. Stimulation of the production of steroid hormones by choriogonadotropin from the urine of pregnant human was dose-dependent. Oestradio and testosterone production was optimal when 20 i.u. of gonadotropic hormone was present in the culture medium 2. During development, both left and right ovaries responded to gonadotropic hormone stimulation with a 2.5-3-fold increase in oestrogen production. However, the right ovary was twice as efficient in testosterone production as the left one. The presence of dibutyryl cyclic AMP in the culture medium of the embryonic ovaries mimicked the effect of the gonadotropic hormone. 3. The human choriogonadotropic hormone stimulated cyclic AMP production in the embryonic ovarian tissue. Thyrotropin, growth hormone and insulin had no stimulating effect. 3-Isobutyl-1-methylxanthine potentiated the gonadotropic hormone effect by increasing the concentration of cyclic AMP in the ovarian tissue. 4. The amount of cyclic AMP synthesized in the embryonic ovary was gradually increased (from 1.2 to 6.5 pmol/mg of tissue) when incubated with increasing doses of human choriogonadotropic hormone in vitro. The newly synthesized cyclic AMP reached the maximum concnentration after 30 min of incubation, then decreased at 2 h of incubation. A portion of the newly synthesized cyclic AMP was released into the culture medium. 5. At various developmental stages, both left and right embryonic-chick ovaries responded to stimulation by gonadotropic hormone with an increase in cyclic AMP production. The cyclic AMP concentration in the right ovary was 80% higher than that in the corresponding left ovary.     

A model for the control of testosterone secretion

We produce here a model to explain the control of testosterone secretion. In this model the hypothalamic secretion of the hormone LHRH (luteinizing hormone releasing hormone) is controlled by a combination of local testosterone concentration and of the local concentration of the pituitary hormone LH (luteinizing hormone). Since LHRH stimulates the release of LH, and LH in turn stimulates the release of testosterone, the three hormones constitute a three-component "feedback" network. We show how this model is able to account for the pulsatility of the release of these three hormones. Furthermore, the model is consistent with results obtained from a wide range of experimental manipulations of the system. For example, it accounts for the changes observed in hormone release patterns after castration. In particular, it follows that no "neural clock", or "neural pulse-generator", is required to force the system into pulsatile behaviour.     

New medical approaches in pituitary adenomas

Recently, the medical approach to patients with secreting and clinically non-functioning pituitary adenomas has received great impulse thanks to the availability of new, selective and long-lasting compounds with dopaminergic activity, such as cabergoline, and of somatostatin analogues provided in slow-release formulations, such as lanreotide and octreotide long acting release (LAR). In particular, the use of cabergoline has induced control of hyperprolactinaemia and tumour shrinkage in the great majority of patients with micro- and macroprolactinomas. Cabergoline treatment restores fertility both in women and men, and partially improves osteoporosis, one of the major complications of hyperprolactinaemia. In acromegaly, disease control (growth hormone [GH] <2.5-1.0 microg/l as a fasting or glucose-suppressed value, respectively, together with age-normalised insulin-like growth factor [IGF]-I) is achievable in more than half of patients receiving treatment with lanreotide or octreotide-LAR. Improvement in cardiomyopathy, sleep apnoea and arthropathy has been reported during GH/IGF-I suppression after pharmacotherapy. A synthetic GH analogue, B2036-PEG, that antagonises endogenous GH binding to its receptor-binding sites and a GH-releasing hormone antagonist that blocks the effect of this releasing factor on the hypothalamus and pituitary are presently under investigation in acromegaly. Preliminary studies have clearly demonstrated the effectiveness of the GH receptor antagonist in suppressing IGF-I levels in acromegalic patients previously unresponsive to somatostatin analogues. Beneficial effects of subcutaneous octreotide and lanreotide have also been reported in adenomas secreting thyroid-stimulating hormone, while the results of treatment with dopamine agonists or somatostatin analogues remain disappointing in patients with clinically non-functioning adenomas. In these patients the possibility of visualising in vivo the expression of D(2) receptors using specific radiotracers such as (123)I-methoxybenzamide has allowed selection of patients likely to respond to cabergoline. Scant effects of pharmacotherapy have also been reported in patients with adenomas secreting adrenocorticotropic hormone. However, some preliminary data suggest a potential use of cabergoline in combination with ketoconazole, or alone, in selected cases of Cushing's disease or Nelson's syndrome.     

Quantification of the Effects of Resperine on Gonadotroph Expression in the Pituitary of Goldfish (Carassius auratus)

In many teleosts, the control of gonadotropin II (or luteinizing hormone) secretion is under the dual control of stimulatory and inhibitory neuroendocrine factors. The principal stimulating factor is gonadotropin-releasing hormone and the main inhibitor is dopamine. Inhibiting the activities of dopamine by antidopaminergic drugs potentiates the actions of exogenous gonadotropin-releasing hormone analogs, resulting in a surge release of luteinizing hormone and ovulation and spawning in a number of different species. As the effects of blocking the inhibitory actions of dopamine on gonadotroph cytology have not been studied, goldfish were treated with 2, 4, 6 or 8 injections of reserpine (0.1 mg/kg body weight), at 48 h intervals, and the numbers of gonadotrophic cells studied at 48 h following last injection. After two injections, the number of gonadotrophic cells increased by 189% over controls; after four injections the increase was 234%; after six injections the increase was 259% and after eight injections, 288%. The results suggest that dopamine has an inhibitory influence on the numbers of gonadotrophs.     

Neuroendocrine control of gonadotropin secretion

Luteinizing hormone releasing hormone (LHRH), a hypothalmic peptide that is concentrated in granules of neurons, has the capacity to release gonadotropins (luteinizing hormone (LH) and follicle stimulating hormone) from the pituitary gland. LHRH has been found in hypophysial portal blood of rats, monkeys, and rabbits. Antibodies to LHRH depress plasma LH concentrations in castrated animals and evoke testicular atrophy, but passive immunization against LHRH does not block the LH surge induced by estrogen in monkeys. Estrogens, progestin, prolactin, and dopamine have marked effects on LH secretion, yet an association between these effects and altered hypophysial portal blood concentrations of LHRH is not established. In view of the paucity of evidence demonstrating such a cause and effect relationship, two alternative proposals have become tenable. One, hormones and neurotransmitters may not alter the levels of portal blood LHRH, but rather alter the frequency of pulsatile LHRH secretion. Two, hormones, such as estrogens, progesterone, and prolactin, may alter the responsiveness of the gonadotropin-secreting cells to LHRH by affecting the secretion of dopamine.     

Changes in pituitary hormone levels induced by met-enkephalin in man—The role of dopamine

The interaction of dopamine with the effects of the opiate agonist peptide D-Ala²-MePhe⁴-met-enkephalin-O-o1 (DAMME) on anterior pituitary hormone secretion was investigated in normal male subjects. DAMME produced clear elevations in prolactin, growth hormone and thyroid-stimulating hormone, while inhibiting the release of luteinising hormone and cortisol. There was no change in follicle stimulating hormone. The elevations in prolactin and TSH were enhanced by the dopamine antagonist, domperidone, and blocked by an infusion of dopamine. Neither dopamine nor domperidone modulated the changes in growth hormone, luteinising hormone or cortisol. The data are comptible with the association of the release of prolactin and TSH by opiate peptides with decreased hypothalamic dopaminergic activity; changes in the other anterior pituitary hormones seem to involve different mechanisms.     

Fine Structure of the Pars Distalis of the Pituitary Gland in the Female Mink,Mustela vison

Pituitary glands of intact and experimental adult females of mink, Mustela vison, were examined by electron microscopy. Conventional methods involving removal of endocrine glands (ovaries and adrenals), administration of radioactive isotope, ¹³¹I, blocking agents (thiouracil and metopirone) and hormones (thyroxine, hydrocortisone, thyrotropin and luteinizing hormone releasing hormones) were employed. Five categories of granular cells were distinguished both by their ultrastructural characteristics and qualitative changes throughout the year and following different treatments. The cell types are described and their functions discussed. From conventional electron microscopical studies it proved difficult to draw any satisfactory conclusions about the gonadotropic cells. Further investigation by means of immunocytochemistry and radioimmunoassay techniques is required to determine, whether the presumptive gonadotropic cell type produces both FSH and ICSH or only one of these hormones. Morphologically two types of agranular cells were identified. Their morphological inter-relationship and function are discussed briefly.