Gonadotropin-releasing Hormone System Research Articles

Lesions of the suprachiasmatic nucleus indicate the presence of a direct vasoactive intestinal polypeptide-containing projection to gonadotrophin-releasing hormone neurons in the female rat.

In non-seasonal breeders like the rat, the influence of the suprachiasmatic nucleus (SCN) on reproduction is most clearly expressed in the female. Complete lesions of the SCN induce persistent oestrus (anovulation) in intact female rats, whereas oestrogen implantation in ovariectomized rats results in daily luteinizing hormone surges. Vasoactive intestinal polypeptide (VIP), a peptide synthesized in cell bodies of the SCN, inhibits the increase in pulsatile luteinizing hormone release observed in ovariectomized female rats. In search of the anatomical basis for these observations, the present study employs an immunocytochemical double staining for VIP and gonadotrophin-releasing hormone (GnRH) at the light microscopical level. It was demonstrated that approximately 45% of the GnRH positive neurons in the diagonal band of Broca, the preoptic and anterior hypothalamic area of female rats are innervated by VIP-containing processes. To investigate whether these VIP-containing fibres represent a direct projection of the SCN to the GnRH system, unilateral thermic SCN lesions were made. Lesions that unilaterally destroyed the majority of the VIP synthesizing cells in the SCN resulted in at least a 50% decrease of the VIP innervation of GnRH cell bodies at the lesioned side compared to the intact side. Lesions not affecting the VIP synthesizing cell population in the SCN did not change the percentage of GnRH neurons innervated by VIP-containing fibres, while partial lesions resulted in intermediate effects. These results indicate that the majority of the light microscopical VIP-containing input on GnRH neurons in the hypothalamus is derived from the SCN.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypothalamo-anterior pituitary gonadotropin-releasing hormone and substance-P systems during the 17 beta-estradiol-induced plasma luteinizing hormone surge in the ovariectomized female monkey.

The present study examined the effects of 17 beta-estradiol benzoate (E2B) on the hypothalamic (HT) and anterior pituitary (AP) content of GnRH precursor (pro-GnRH-GAP), GnRH, GnRH-associated peptide (GAP), and substance-P (SP) during the various phases of the E2B-induced LH surge in cynomolgus monkeys. Changes in GnRH and GnRH-associated peptide (GAP) at both hypothalamic and AP levels were closely related at all times after E2B treatment. However, the pattern of change in the AP was very different from that in the HT. In the HT, pro-GnRH-GAP levels did not change significantly throughout the experimental period. In the AP, the pro-GnRH-GAP increased 48 h post-E2B treatment, the time of initiation of the LH surge. An 8-fold increase in AP GnRH occurred 30 h post-E2B treatment. There were no significant changes in the HT content of SP at any time after E2B treatment. However, there was a depletion of AP content by 48 h post-E2B, the time of the LH surge. These results demonstrate that E2 activates and deactivates in a coordinated manner the GnRH and SP systems of the HT-AP complex during initiation of the E2-induced LH surge. The observation that more significant changes occur in the AP than in the HT suggests that an important component of the E2 effect in inducing the LH surge may be directly at the AP level. This action involves changes in the contents of GnRH and SP in the AP.

Hypothalamo-anterior pituitary gonadotropin-releasing hormone and substance-P systems during the 17 beta-estradiol-induced plasma luteinizing hormone surge in the ovariectomized female monkey

Hypothalamo-anterior pituitary gonadotropin-releasing hormone and substance-P systems during the 17 beta-estradiol-induced plasma luteinizing hormone surge in the ovariectomized female monkey

Immunocytochemical and histochemical analyses of gonadotrophin releasing hormone, tyrosine hydroxylase, and cytochrome oxidase reactivity within the hypothalamus of chicks showing early sexual maturation

In order to determine the changes in gonadotrophin releasing hormone (GnRH) and dopaminergic activity within the brain during the onset of sexual precocity, a Halasz-like knife was developed to produce discrete parasagittal cuts in 2-week-old male broiler chicks. At 5 weeks of age, sexually precocious respondents were selected on the basis of advanced secondary sex characteristics and randomly paired with sham-operated controls. Each pair of birds was perfused with heparinized saline followed by 4% paraformaldehyde. Sections 40 microns thick, obtained throughout the hypothalamus, were immunostained with either anti-GnRH or anti-tyrosine hydroxylase (TH) to ascertain dopaminergic activity. Alternate sections from each pair of brains were also treated with cytochrome oxidase to determine metabolic activity levels or with Nissl stain to localize the knife cuts. Analysis revealed an increase in GnRH immunoreactivity within the bed nucleus of the pallial commissure (nCPa) and paraventricular nucleus (PVN), as well as the median eminence (ME). An increase in TH immunoreactivity was observed in the nucleus intramedialis (nI). Also an increase in metabolic activity was seen in the PVN as revealed by cytochrome oxidase reactivity. It is hypothesized that during the onset of puberty there is an increase in immunoreactive GnRH cell numbers as a result of a decrease in the inhibition of the GnRH system, possibly involving the nI and PVN. The source of the dopamine reported in the ME could be from the nI and other nearby nuclei. Dopamine from the tubero-infundibular area may be one of the putative neurotransmitters responsible for the increased activity of GnRH within the ME of chicks showing precocious puberty.

C.E.B.A.S.-AQUARACK: Second generation hardware and latest scientific results

A second version of the C.E.B.A.S.-AQUARACK laboratory prototype was developed which includes a new water recycling system and a “Botanical Component” with higher water plants for nitrate elimination. It was a modified by a compact commercial filter system which involves a semi-biological coarse filter, a bacteria filter, gas and heat exchangers and a UV sterilization device. A highly sophisticated process control system offers all possiblities to implement robotics and telescience. With the installation of the Botanical component into the system the first step of the realisation of the conception of a “Closed Equilibrated Biological Aquatic System” is performed. In the scientific frame program the reproductive biology of the main experimental animal, Xiphophorus helleri, plays an important role. After the establishment of a morphological reference system of the stages of sexual maturity in both sexes this is correlated with relevant physiological data as concentrations of sexual steroids in gonadal tissue and plasma. On the brain-pituitary level the ontogeny of the gonadotropin releasing hormone (GnRH) and the gonadotropin (GtH) systems were investigated by immunohistochemistry. Surprisingly gonadotropin was not only localized in the pituitary gland but also in brain neurons and fibers which undergo a distinct ontogenetic development and a maior part of which are in close structural relation to the GnRH system. A special chapter pays attention to the role of the C.E.B.A.S.-system in C.E.L.S.S. research.

Immunohistochemical Localization of Multiple Forms of Gonadotropin-Releasing Hormone in the Brain of the Adult Frog

Abstract Immunohistochemical mapping with antibodies against four different types of gonadotropin-releasing hormone (GnRH)-like neuro-peptides has been studied in the brain of adult Rana esculenta. This study confirms the earlier described distribution pattern of the immunoreactive mammaiian GnRH system in the frog brain, as well as revealing that this system of neuronal cell bodies and fibres is immunopositive to antisera for mammalian, chicken-I, chicken-II and salmon GnRH-like molecules. The results also indicate coexistence of the four GnRH variants in the same anatomical areas. The presence of immunoreactive fibre endings in the cerebellum is also described, perhaps for the first time in the vertebrate brain. In addition, it was found that many immunoreactive GnRH fibres arising in the anterior preoptic area and thalamus-periventricular area project posteriorly to reach the interpeduncular nucleus-tegmentum area, thus connecting the diencephalon with the rhombencephalon. These data provide further information on the complex GnRH system in the frog brain. What role(s) in vivo the non-mammalian forms of GnRH-like peptides may play in amphibian reproduction is briefly discussed, and in the light of paucity of data it is here stressed that more amphibian species should be studied.

Ontogeny of immunoreactive gonadotropin-releasing hormone neuronal systems in amphibians

The ontogeny of gonadotropin-releasing hormone (GnRH) systems was investigated in 3 anuran amphibians (genus Rana) by means of immunocytochemical (ICC) techniques and antibodies generated against 3 different forms of GnRH. Antisera that recognize primarily chicken II and mammalian GnRHs revealed two anatomically and developmentally distinct GnRH systems. One system, referred to here as the forebrain-spinal cord system, contained GnRH immunoreactive (ir) fibers extending from the rostral diencephalon through the ventromedial brainstem to the spinal cord. Intensity of labeling was robust in the youngest, premetamorphic tadpoles, but decreased with age. GnRH immunolabeling in the hypothalamic-pituitary tract was not detected until late prometamorphosis and increased with age. Development of GnRHir in the hypothalamic-pituitary tract coincided with first appearance of GnRHir in the terminal nerve in R. catesbeiana, but not in R. cascadae or R. aurora, suggesting species differences. Comparisons of results obtained with antisera to different forms of GnRH support the interpretation that the forebrain-spinal cord system, hitherto undescribed in amphibians, develops first and synthesizes a non-mammalian, chicken II-like GnRH, and that the hypothalamic-pituitary system develops later and synthesizes primarily mammalian GnRH. We speculate that the forebrain-spinal cord system may represent a GnRH innervation of frog sympathetic ganglia, and that the two GnRH systems are chemically and embryonically distinct.

Location of the Neuroendocrine Gonadotropin‐Releasing Hormone Neurons in the Monkey Hypothalamus by Retrograde Tracing and Immunostaining*,**

Abstract In order to localize neuroendocrine gonadotropin-releasing hormone (GnRH) neurons in the monkey hypothalamus, four juvenile cynomolgus macaques (one female, three males) were each given two or three microinjections (0.2 to 0.3 mul per site) of the retrograde tracer wheat germ agglutinin-apoHorseradish peroxidase-10 nm colloidal gold into the superficial, median eminence region of the infundibular stalk. Five to 15 days following surgery, the brains were fixed by perfusion and vibratomed at 40 mum in the frontal plane. Every 12th section was immunostained with rabbit anti-GnRH using the peroxidase anti-peroxidase technique with diaminobenzidine as the chromogen. Neuroendocrine GnRH neurons were easily identified in tissue sections as brown, immunostained cell bodies containing more than three distinct, dark blue, tracer-filled lysosomes. Neuronal counts from each complete series of sections were compiled by anatomical region, and the percentages of GnRH and neuroendocrine GnRH neurons determined. The highest proportion of neuroendocrine GnRH neurons (with projections to the median eminence) occurred in the ventral hypothalamic tract, especially in its medial third (71%), and in the supraoptic decussation just anterior to it. Proportions decreased moving laterally into the middle third (58%) and lateral third (25%) of the ventral hypothalamic tract. Further anterior and lateral, progressively smaller but significant neuroendocrine GnRH contributions were found in the supraoptic nucleus (57%) and lateral hypothalamus (33%), and in the medial preoptic area (26%). Although the medial preoptic area contained a greater percentage of the total GnRH-immunoreactive cell bodies (36%) than the ventral hypothalamic tract (27%), as a whole, the ventral hypothalamic tract contained 60% of the neuroendocrine GnRH neurons compared to only 25% from the medial preoptic area. Large numbers of GnRH cell bodies found in the diagonal band of Broca near the organum vasculosum of the lamina terminalis were not retrogradely labeled. GnRH neurons were not observed in the arcuate nucleus, the few in the paraventricular nucleus were not neuroendocrine, and the contribution from the periventricular zone was negligible. Our results here are the first to identify the neurons giving rise to the neuroendocrine GnRH system in juvenile monkeys. The data indicate that more GnRH neurons close to the infundibulum serve a neuroendocrine (perhaps hypophysiotropic) role than do those in more anterior areas. Furthermore, they suggest that the ventral hypothalamic tract is the most important, and perhaps most influential, neuroendocrine GnRH cell group in primates. The data substantiate the observed autonomy of the medial basal hypothalamus in controlling gonadotropin secretion and menstrual cyclicity in these animals. However, they also infer that perhaps 60% of the GnRH neurons do not project to the primate median eminence, and thus may serve other non-neuroendocrine functions.

Testing of Arg-8-gonadotropin-releasing hormone-directed antisera by immunological and immunocytochemical methods for use in comparative studies

Three polyclonal antisera raised in rabbits against the mammalian molecular form of gonadotropin-releasing hormone (GnRH) were tested in enzyme-linked immunosorbent assays for crossreactivity with naturally occurring GnRHs and with GnRH analogues. Antisera were then tested immunocytochemically in order (i) to identify amino acids essential for the binding of each antiserum, and (ii) to evaluate the specificity of the immunocytochemical reaction in brain sections from various species of cyclostomes, amphibians, reptiles, and birds. Antiserum GnRH 80/1, recognizing mainly a discontinuous determinant including the NH2- and COOH-termini, crossreacts with GnRHs the molecular bending of which enables the spatial approach of both terminal amino acid residues. Antiserum GnRH 80/2, by requiring the COOH-terminus for binding and not tolerating substitutions by aromatic amino acids in the middle region of the molecule, recognizes chicken I GnRH, however, not the salmon form. The use of this antiserum is appropriate in species synthesizing the mammalian and/or the chicken I form of GnRH. GnRH antiserum 81/1 is specific mostly for mammalian GnRH. The results obtained by ELISAs are confirmed by immunocytochemical studies. A comparison between the results obtained in ELISA and in immunocytochemistry involving mammalian-, chicken I-, chicken II-, salmon-, and lamprey-directed GnRH antisera resulted in the following conclusions: (1) An antiserum recognizing the discontinuous antigen determinant including both NH2- and COOH-termini may be reactive in most vertebrate brain sections thus being appropriate for phylogenetically directed immunocytochemical studies. (2) Moreover, this discontinuous determinant seems to be immunocytochemically reactive in all parts of the neurons in the GnRH system, whereas, in some species, determinants located in the middle region of the molecule(s) tend to become reactive only during the axonal transport. (3) A crossreaction between tissue-bound antigen and antibodies recognizing the above cited discontinuous determinant indicates an appropriate bending of the molecule even in case of severe molecular differences, e.g., in lamprey form of GnRH. (4) It follows that in phylogenetic studies, an immunologically well characterized antiserum can be substituted for a species-directed antiserum.

Pheromone detection and olfactory pathways in the brain of the female African catfish, Clarias gariepinus

The olfactory tract of the African catfish, Clarias gariepinus, consists of two tracts, the medial and lateral olfactory tract. Ovulated female catfish are attracted by male steroidal pheromones. Attraction tests with catfish in which the medial and lateral olfactory tract have been selectively lesioned show that the effects of these pheromones are mediated by the medial olfactory tract. The central connections of the medial and lateral olfactory tract have been studied by retro- and anterograde transport techniques using horseradish peroxidase as a tracer. Upon entering the forebrain, the medial olfactory tract innervates the posterior pars ventralis and pars supracommissuralis of the area ventralis telencephali and the nucleus preopticus periventricularis, the nucleus preopticus and the nucleus recessus posterioris. Application of horseradish peroxidase to the olfactory epithelium shows that part of the innervation of the area ventralis telencephali and the nucleus preopticus periventricularis can be attributed to the nervus terminalis, which appears to be embedded in the medial olfactory tract. The lateral olfactory tract sends projections to the same brain areas but also innervates the nucleus habenularis and a large terminal field in the area dorsalis telencephali pars lateralis ventralis. Furthermore, the medial olfactory tract carries numerous axons from groups of perikarya localized in the area dorsalis telencephali. Contralateral connections have been observed in the olfactory bulb, telencephalon, diencephalon and mesencephalon. It is suggested that processes of the medial olfactory tract innervating the preoptic region may influence the gonadotropin-releasing hormone system and in doing so may lead to behavioral and physiological changes related to spawning.

The gonadotropin-releasing hormone containing ventral hypothalamic tract in the fetal rhesus monkey (Macaca mulatta).

A well-defined, gonadotropin-releasing hormone (GnRH)-containing fiber pathway, the ventral hypothalamic tract (VHT), is described by immunostaining in fetal rhesus macaques (109-156 days gestation). The VHT arises above the lateral aspects of the optic chiasm near the supraoptic nucleus, and courses ventromedially close to the ventral hypothalamic surface to terminate in the infundibulum and zona externa of the median eminence. It is formed by the confluence of GnRH-immunopositive (GnRH+) axons from local neurons, from a few GnRH+ cells in the inferior thalamic peduncle, and probably from more anterior neurons in the septum and preoptic area. Bipolar GnRH+ neurons contributing directly to the VHT are grouped at its origin dorsolateral to the optic chiasm, dorsal and medial to the optic tracts, at the infundibular lip, and within the pathway between. At the infundibular lip, GnRH+ perikarya are generally lateral or ventral to the infundibular (arcuate) nucleus, and are rarely within the nucleus itself. Cell bodies here are sometimes tripolar, but GnRH+ intercellular contacts are seldom seen. A few VHT fibers extend to the ventral surface of the brain just beneath the pia mater. Abundant capillaries in the subarachnoid space suggest a possible route for delivery of GnRH to the adenohypophysis in early gestation, before maturation of the hypophysial portal system occurs. Posterior to the infundibulum, a few VHT fibers are joined by descending periventricular fibers forming a dense fiber band beneath the premammillary recess of the third ventricle. Totals of GnRH+ cell bodies in the prosencephalon of the fetal rhesus macaque are estimated to be 5,600 in females (n = 2) and 2,600 in males (n = 3). More than 60% of VHT neurons are located in the medial basal hypothalamus, and the majority of basal hypothalamic GnRH+ neurons (86%) are associated with the VHT. Furthermore, reports of the autonomy of the medial basal hypothalamic-hypophysial unit in control of gonadotropin secretion suggest that the VHT may be the most important GnRH system involved in primate reproduction. It is clear that fetal material may offer the best model to study the GnRH neuronal system in primates.

Sites of origin of gonadotropin releasing hormone containing projections to the amygdala and the interpenduncular nucleus

The sites of origin of gonadotropin releasing hormone (GnRH) containing projections to the amygdala and the interpenduccular nucleus were studied with immunofluorescence for GnRH in combination with retrograde transport of True blue. After small injections of True blue into the amygdala, retrogradely labeled GnRH producing neurons were identified in the rostral medial septum, the caudal roots of the nervus terminalus, diagonal band, nucleus triangularis septi, nucleus interstitialis striae terminalis, and in the ventrolateral hypothalamus. After injection of True blue into the interpendicular nucleus, a small GnRH producing cell group in the ventro-medial diagonal band, and certain GnRH-positive neurons in the ventral hypothalamus had retrogradely transported the dye. The results suggest a link between the amygdala and the nervus terminalis which is provided in part by GnRH-containing projections. The heterogeneity of the sites of origin of inputs to the amygdala and the interpenduncular nucleus indicates that the ‘behavioral’ component of the GnRH system is not restricted to a single brain nucleus, but it may be part of a diffuse network which has close anatomical ties with the ‘endocrine’ GnRH system.

The olfactory gonadotropin-releasing hormone immunoreactive system in mouse

The olfactory gonadotropin-releasing hormone (GnRH) system in mice was studied with immunofluorescence in combination with lesions of the olfactory bulb and retrograde transport of horseradish peroxidase (HRP) which was administered intravascularly, intranasally or into the subarachnoid space. GnRH-positive neurons were located in the two major branches forming the septal roots of the nervus terminalis, in the ganglion terminale, within the fascicles of the nervus terminalis throughout its extent, in a conspicuous band which connects the ventral neck of the caudal olfactory bulb with the accesory olfactory bulb in the nasal mucosa. GnRH-positive fibers were seen in all areas in which neurons were found, i.e. in the rostral septum, the ganglion and nervus terminalis and in the nasal subepithelium. In addition, a broad bundle of fibers was observed to surround the entire caudal olfactory bulb, connecting the rostral sulcus rhinalis with the ventrocaudal olfactory bulb. Fibers were seen in close association with the main and accessory olfactory bulb, with the fila olfactoria and with the nasal mucosa. Throughout the olfactory bulb and the nasal epithelium, an association of GnRH fibers with blood vessels was apparent. Intravascular and intranasal injection of HRP resulted in labeling of certain GnRH neurons in the septal roots of the nervus terminalis, the ganglion terminale, the nervus terminalis, the caudal ventrodorsal connection and in the accessory olfactory bulb. After placement of HRP into the subarachnoid space dorsal to the accessory olfactory bulb, about 50% of the GnRH neurons in the accessory olfactory bulb and in the ventrodorsal connection were labeled with HRP. Also, a few GnRH neurons in the rostral septum, the ganglion terminale and in the fascicles of the nervus terminalis had taken up the enzyme. Lesions of the nervus terminalis caudal to the ganglion terminale resulted in sprouting of GnRH fibers at both sites of the knife cut. Lesions rostral to the ganglion terminale induced sprouting mostly at the distal site of the knife cut while most but not all GnRH fibers proximal to the lesion had disappeared. The results of the present study indicate that the olfactory GnRH system is mostly associated with the nervus terminalis. This cranial nerve apparently projects to the central nervous system as well as the periphery. The results of the HRP uptake studies suggest that the GnRH neurons in the nervus terminalis have access to fenestrated capillaries in the subepithelial connective tissue of the nasal mucosa, to the nasal epithelium proper, and to the subarachnoid space. It is concluded that GnRH in the olfactory system of the mouse can influence different target sites either via the blood stream after release into the submucosal capillaries, or via the external cerebrospinal fluid or via synaptic/asynaptic contacts with, for example, the receptor cells in the nasal epithelium.

A reinvestigation of the Gn-RH (gonadotrophin-releasing hormone) systems in the goldfish brain using antibodies to salmon Gn-RH.

The organization of Gn-RH systems in the brain of teleosts has been investigated previously by immunohistochemistry using antibodies against the mammalian decapeptide which differs from the teleostean factor. Here, we report the distribution of immunoreactive Gn-RH in the brain of goldfish using antibodies against synthetic teleost peptide. Immunoreactive structures are found along a column extending from the rostral olfactory bulbs to the pituitary stalk. Cell bodies are observed within the olfactory nerves and bulbs, along the ventromedial telencephalon, the ventrolateral preoptic area and the latero-basal hypothalamus. Large perikarya are detected in the dorsal midbrain tegmentum, immediately caudal to the posterior commissure. A prominent pathway was traced from the cells located in the olfactory nerves through the medial olfactory tract and along all the perikarya described above to the pituitary stalk. In the pituitary, projections are restricted to the proximal pars distalis. A second immunoreactive pathway ascends more dorsally in the telencephalon and arches to the periventricular regions of the diencephalon. Part of this pathway forms a periventricular network in the dorsal and posterior hypothalamus, whereas other projections continue caudally to the medulla oblongata and the spinal cord. Lesions of the ventral preoptic area demonstrate that most of the fibers detected in the pituitary originate from the preoptic region.

Immunoreactive gonadotropin-releasing hormone-like material in the brain and the pituitary gland during the periovulatory period in the brown trout ( Salmo trutta L.): Relationships with the plasma and pituitary gonadotropin

In fish there are few data on the gonadotropin-releasing hormone (Gn-RH) neurosecretory activity, which could explain long- and short-term variations of the gonadotropin secretion. There is no biological species specificity between mammal and fish Gn-RH; although there is a structural difference, they are, on the contrary, characterized by a high immunological specificity which does not allow measurement of fish Gn-RH using radioimmunoassay for LH-RH. We have synthesized salmon Gn-RH according to the formula recently proposed by Sherwood ( N. Sherwood, L. Eiden, M. Brownstein, J. Spies, J. Rivier, and W. Vale, 1983. Proc. Natl. Acad. Sci. USA 80, 2794–2798). Its activity has been tested by its ability to stimulate the gonadotropin hormone (GtH) secretion in vivo in testosterone-implanted juvenile rainbow trout, and for the recognition of synthesized Gn-RH (s-Gn-RH) perykaria by a specific antibody raised against the s-Gn-RH in regions of the brain described as containing LH-RH immunoreactive-like material. A radioimmunoassay has been developed for the salmon Gn-RH, and its specificity to measure trout Gn-RH has been tested. Using this assay, the brain and pituitary Gn-RH contents have been measured throughout the final phases of maturation and ovulation. Brain Gn-RH increases from the end of vitellogenesis (8.9 ± 0.76 ng/brain) to ovulation (more than 15 ng/brain). Pituitary Gn-RH is lower (1.58 ± 0.69 ng/pituitary) at the end of vitellogenesis and follows a similar profile as in the brain, except for a significant decrease just prior the beginning of oocyte maturation. The correlations between Gn-RH levels and GtH pituitary and plasma levels show that total brain Gn-RH is never correlated to the GtH, suggesting that the increase in the brain Gn-RH content is related to a Gn-RH system closely related to maturation and ovulation, which remains to be investigated. On the contrary, pituitary Gn-RH levels are well correlated with pituitary and plasma GtH levels, indicating that pituitary Gn-RH levels might represent a good index of the Gn-RH neurosecretory activity in the fish hypothalamohypophysial complex, given the absence of a portal system in teleost.

Immunocytochemical localization of GnRH (gonadotropin releasing hormone) systems in the brain of a marine teleost fish, the sole

The GnRH system was studied in the brain of the sole by immunocytochemistry (peroxidase-antiperoxidase method) (PAP) using antibodies to synthetic salmon GnRH (s-GnRH). Two centers containing immunoreactive cell bodies were observed in the forebrain, one located at the junction between the olfactory bulbs and the telencephalon and the other in the preoptic area. Numerous immunoreactive fibers were found, especially in the telencephalon, hypothalamus, pituitary, optic tectum and retina.

Modulation of lordosis behaviour in the female rat by corticotropin releasing factor, β-endorphin and gonadotropin releasing hormone in the mesencephalic central gray

A possible functional relationship between corticotropin-releasing factor (CRF) and opiate peptide neuronal systems (β-endorphin, dynorphin 1–17 and Met-enkephalin) and their interactions with gonadotropin releasing hormone (GnRH) in the mesencephalic central gray (MCG) for the regulation of lordosis behaviour was assessed in ovariectomized, oestrogen-treated and oestrogen-progesterone-treated female rats. Lordosis behaviour triggered by male mounting was inhibited in a dose-dependent fashion by CRF microinfused into the MCG in both oestrogen-treated and oestrogen-progesterone-treated female rats. This CRF-induced inhibition of lordosis could be overcome by a pre-infusion of naloxone or anti-β-endorphin-globulin (anti-β-end-G) directly into the MCG but not by anti-Met-enkephalin globulin (anti-enk-G) or anti-dynorphin 1–17 globulin (anti-dynor-G). Supporting data indicate that the facilitation of lordosis behaviour induced by treatment with naloxone or anti-β-end-G alone but not with anti-enk-G or anti-dynor-G may be due to enhanced GnRH release. This results from the action of these substances in overcoming the inhibition of GnRH secretion mediated specifically by β-endorphin but not by Met-enkephalin or dynorphin 1–17 in the MCG. These studies together with previous data showing that GnRH can overcome the abolition of lordosis by β-endorphin in the MCG, indicate a close relationship between β-endorphin (but not Met-enkephalin or dynorphin) and GnRH systems in the MCG in the control of lordosis behaviour. Thus, the inhibition of lordosis by CRF and the complete reversal of this blockade by naloxone or anti-β-end-G may suggest that CRF could enhance the release of β-endorphin from fibres in the MCG; β-endorphin then inhibits lordosis by inhibiting the release of GnRH. However, a direct inhibitory effect of CRF on GnRH release is also likely since anti-CRF-γ-globulin (anti-CRF-G) infused into the MCG produced a long-lasting facilitation of lordosis which can be blocked by an antagonist analogue of GnRH; in addition, previous studies have shown that GnRH infused into the MCG completely overcame the CRF-induced abolition of lordosis and potentiated lordosis to high levels. These results suggest that there may be functional neuroanatomical relationships between CRF, β-endorphin and GnRH neuronal systems in the MCG in the control of female sexual behaviour. Neither Met-enkephalin nor dynorphin 1–17 appear to participate in such mechanisms.

Neuroendocrinology of opioid peptides and their role in the control of gonadotropin and prolactin secretion

Substantial evidence now exists to indicate that the endogenous hypothalamic opioidergic mechanism(s) represents one of the important controlling systems for release of gonadotropin-releasing hormone. Modulations of frequency and amplitude of the secretory activity of gonadotropin-releasing hormone appears to be mediated through an inhibitory action of endogenous opioids, and the functional coupling of the opioidergic and gonadotropin-releasing hormone systems is an ovarian steroid-dependent event. There is also evidence to implicate suprahypothalamic mechanism(s) that enhance endogenous opioid inhibition of secretion of gonadotropin-releasing hormone. Although exogenous opioid peptides and their synthetic analogs consistently induce the secretion of prolactin, blockade of opioid receptors in humans by naloxone failed to elicit a decrement in the levels of prolactin under a variety of conditions. On the contrary, naloxone induced a remarkable increment in the secretion of prolactin via an increased frequency of pulsatile release which is synchronized with pulses of luteinizing hormone. These observations suggest that a common neuroendocrine mechanism is involved in the opioidergic control of the secretion of both luteinizing hormone and prolactin in women.

Modulation of lordosis behavior of female rats by naloxone, beta-endorphin and its antiserum in the mesencephalic central gray: possible mediation via GnRH.

The role of endogenous opiate peptides in the mesencephalic central gray (MCG) and their possible interactions with gonadotropin-releasing hormone (GnRH) in the regulation of lordosis behavior was assessed in ovariectomized, estrogen-treated and estrogen-progesterone-treated female rats. Lordosis behavior triggered by male mounting was enhanced by microinfusion of naloxone and anti-beta-endorphin-globulin (anti-beta-end-G) but not by anti-met-enkephalin-globulin or anti-dynorphin-globulin into the MCG in both estrogen-treated and estrogen- (low dose) progesterone-treated females. The potentiating effects of naloxone and anti-beta-end-G could be blocked by a preinfusion of either anti-GnRH-globulin or an antagonist analog of GnRH directly into the MCG. However, two potent antagonist analogs of GnRH were not effective in blocking lordosis indicating a dissociation between their neural actions and their known inhibitory effects on luteinizing hormone release. Conversely, beta-endorphin but not met-enkephalin or dynorphin infused into the MCG inhibited lordosis behavior in both estrogen-treated and estrogen-progesterone-treated rats. This beta-endorphin-induced inhibition of lordosis in the estrogen-treated rats could be overcome by GnRH microinfused directly into the MCG which potentiated lordosis to high levels. These observations provide evidence that beta-endorphin may be the sole opiate peptide in the MCG involved in the control of lordosis behavior and also suggests a functional relationship with GnRH systems in the MCG in such a regulatory mechanism.

Anatomical relationships of dopaminergic and GABAergic systems with the GnRH-systems in the septo-hypothalamic area. Immunohistochemical studies.

Immunohistochemical double staining for gonadotropin releasing hormone (GnRH) and tyrosine hydroxylase (TH) or glutamic acid decarboxylase (GAD) reveals in the septo-preoptic-diagonal band complex of the rat brain close spatial associations between GnRH-immunoreactive perikarya and TH and GAD immunoreactive fibers. In the organum vasculosum laminae terminalis, no close spatial relationships could be observed between TH- or GAD-positive fibers and the GnRH-containing system. In contrast, in the median eminence substantial overlap exists in the distribution of GnRH with TH and GAD containing nerve fibers. This overlap is most intense for TH throughout the lateral palisade zone, while for GAD it is more restricted to the outermost portion of the external palisade zone. The results suggest that dopamine and GABA influence GnRH secretion via axosomatic contacts in the septo-preoptic-diagonal band complex, as well as via axo-axonic interactions in the median eminence, while no such interactions seem to exist in the organum vasculosum laminae terminalis. Since dopaminergic cell bodies in the ventral hypothalamus are closely apposed by GnRH and GAD containing fibers, the existence of feedback circuits among GnRH, dopamine and GABA systems is proposed.