Evidence that RF-amide related peptides are inhibitors of reproduction in mammals

Gonadotropin releasing hormone (GnRH) secretion represents the final common pathway in the control of the reproductive axis. Dogma has been that GnRH is solely responsible for the control of gonadotropin secretion, but emerging data presents a strong case for the existence of a gonadotropin inhibitory hormone in mammals. This evidence arose from initial work in avian species to isolate and identify a factor that inhibited gonadotropin release, which is known as gonadotropin inhibitory hormone (GnIH). The mammalian ortholog of avian GnIH is named RF-amide related peptide (RFRP). There are two forms of RFRP in mammals, RFRP-1 and RFRP-3 encoded by a single gene, but there has been skepticism and controversy as to whether these peptides play a significant role in the regulation of gonadotropin secretion. There is now a significant body of evidence that one or more RFRP exists in mammals and acts as an inhibitor of GnRH and/or gonadotropin secretion. Moreover, RFRP-producing neurons have been shown to transmit information to GnRH cells and/or gonadotropes in relation to seasonal status and to coordinate events around the preovulatory luteinizing hormone surge. This review will focus on the significant advances in RFRP research in mammalian species.     

The influence of chemosensory input and gonadotropin releasing hormone on mating behavior circuits in male hamsters

Chemosensory input is important for mating behavior in male hamsters. Chemosignals found in female hamster vaginal fluid activate regions of the brain that receive input from the vomeronasal/accessory olfactory system and are important for mating behavior. Mating or exposure to these chemosignals produces increased Fos protein expression in the amygdala, bed nucleus of the stria terminalis, and medial preoptic area (MPOA). These brain regions contain cell bodies and/or fibers of gonadotropin releasing hormone (GnRH) neurons, suggesting potential relationships between chemosensory systems and GnRH. GnRH is released naturally when male rodents (mice and hamsters) encounter female chemosignals, and intracerebrally injected GnRH restores mating behavior in sexually naive male hamsters after removal of the vomeronasal organs. We report here that the combination of pheromone exposure and intracerebrally-injected GnRH increases Fos expression in the MPOA above the increase seen in pheromone-exposed males, or in males given only the exogenous GnRH. In males with vomeronasal organs removed (VNX), there was an also an increment in Fos expression in the MPOA when these pheromone exposed males were injected with GnRH, provided they had previous sexual experience. Males with vomeronasal organs removed and without sexual experience showed increased Fos expression in the medial amygdala when pheromone exposure and GnRH injection were combined, but not in the medial preoptic area.     

Gonadotropin hormone releasing hormone agonists alter prefrontal function during verbal encoding in young women

Gonadotropin hormone releasing hormone agonists (GnRHa) are commonly used in clinical practice to suppress gonadal hormone production in the management of various gynaecological conditions and as a treatment for advanced breast and prostate cancer. Animal and human behavioural studies suggest that GnRHa may also have significant effects on memory. However, despite the widespread use of GnRHa, the underlying brain networks and/or stages of memory processing that might be modulated by GnRHa remain poorly understood. We used event-related functional magnetic resonance imaging to examine the effect of GnRHa on verbal encoding and retrieval. Neuroimaging outcomes from 15 premenopausal healthy women were assessed at baseline and 8 weeks after Gonadotrophin Releasing Hormone analogue (GnRHa) treatment. Fifteen matched wait-listed volunteers served as the control group and were assessed at similar intervals during the late follicular phase of the menstrual cycle. GnRHa was associated with changes in brain response during memory encoding but not retrieval. Specifically, GnRHa administration led to a change in the typical pattern of prefrontal activation during successful encoding, with decreased activation in left prefrontal cortex, anterior cingulate, and medial frontal gyrus. Our study suggests that the memory difficulties reported by some women following GnRHa, and possibly at other times of acute ovarian hormone withdrawal (e.g. following surgical menopause and postpartum), may have a clear neurobiological basis; one that manifest during encoding of words and that is evident in decreased activation in prefrontal regions known to sub-serve deep processing of to-be-learned words.     

Effect of manipulating central catecholamines on puberty and the surge of luteinizing hormone and gonadotropin releasing hormone induced by pregnant mare serum gonadotropin in female rats

We have investigated the effect of manipulating central catecholamines on the timing of puberty (as assessed by vaginal opening) in female rats and the surge of luteinizing hormone (LH) and gonadotropin releasing hormone (GnRH) induced by pregnant mare serum gonadotropin (PMSG) in immature female rats. Manipulation of the catecholamines was carried out with either 6-hydroxydopamine (6-OHDA) administered with or without either desipramine (DMI) or pargyline, orα-methyl-p-tyrosine (α-MPT).The neonatal administration of 6-OHDA delayed puberty, an effect which was potentiated by pretreatment with DMI and was associated with a reduction in the rate of body growth. Catecholamine fluorescence in animals aged 60–65 days that had been treated with DMI followed by 6-OHDA was diminished only in the caudatus-putamen; treatment with 6-OHDA alone resulted in diminished fluorescence in the hypothalamus and in the intermediate but not the external layer of the median eminence. The neonatal administration of α-MPT had no significant effect on either the growth rate or the timing of puberty. Regular oestrous cycles occured after puberty in animals treated with either 6-OHDA or α-MPT.The PMSG-induced LH surge was significantly enhanced by 6-OHDA (administered i.v.) plus DMI, and reduced by 6-OHDA injected into the lateral ventricle (v). The inhibitory effect of 6-OHDA (v) wasreduced by DMI, but in animals given 6-OHDA (i.v.) after pargyline there was a marked reduction in the height of the LH surge. There was a good correlation between the changes in the concentrations of LH in peripheral plasma and the concentrations of GnRH in pituitary stalk plasma in that the PMSG-induced surge of GnRH was significantly increased by 6-OHDA (i.v.) plus DMI and reduced by 6-OHDA (v). In animals treated with 6-OHDA (i.v.) plus DMI catecholamine fluorescence was reduced only in the external layer of the median eminence, while after 6-OHDA (v) plus DMI degeneration was seen in the medial forebrain bundle.These results demonstrate a marked difference between the long-term and acute effects of 6-OHDA on the gonadotropin control system. Neonatal treatment with 6-OHDA plus DMI significantly delays puberty and the rate of body growth, but does not affect cyclical gonadotropin release and has no persistent effect on the hypothalamic catecholaminergic systems. The acute administration of 6-OHDA, depending upon the route of administration and whether it is given after DMI, can either potentiate or inhibit the PMSG-induced surge of GnRH and consequently LH by mechanisms which involve destruction, respectively, of either dopaminergic terminals in the median eminence or catecholaminergic fibres in the dorsal hypothalamus.     

Control of GnRH secretion: one step back

The reproductive system is controlled by gonadotropin releasing hormone (GnRH) secretion from the brain, which is finely modulated by a number of factors including gonadal sex steroids. GnRH cells do not express estrogen receptor α, but feedback is transmitted by neurons that are at least ‘one step back’ from the GnRH cells. Modulation by season, stress and nutrition are effected by neuronal pathways that converge on the GnRH cells. Kisspeptin and gonadotropin inhibitory hormone (GnIH) neurons are regulators of GnRH secretion, the former being a major conduit for transmission of sex steroid feedback. GnIH cells project to GnRH cells and may play a role in the seasonal changes in reproductive activity in sheep. GnIH also modulates the action of GnRH at the level of the pituitary gonadotrope. This review focuses on the role that kisspeptin and GnIH neurons play, as modulators that are ‘one step back’ from GnRH neurons.     

男性肝豆状核变性下丘脑-垂体-睾丸轴功能研究

目的了解男性肝豆状核变性（ＷＤ）病人下丘脑－垂体－睾丸轴激素分泌功能。方法用放射免疫法测定２６例病人血清垂体、性激素水平。结果孕酮（Ｐ）显著低于正常对照组（Ｐ＜０．０５），生长激素（ＧＨ）、黄体生成素（ＬＨ）、雌二醇（Ｅ２）亦显著降低（Ｐ＜０．０１），性激素结合球蛋白（ＳＨＢＧ）则显著高于正常对照组（Ｐ＜０．００１），２０例患者进行促性腺激素释放激素（ＧｎＲＨ）试验，发现腺垂体对外源性ＧｎＲＨ的刺激反应迟钝。结论ＷＤ垂体、性腺细胞中部分酶的活性降低，使二者激素分泌功能降低；同时肝的铜沉积扰乱了蛋白代谢，使ＳＨＢＧ升高，进一步降低了性激素的生物效应     

Dual projections of gonadotropin releasing hormone containing neurons to the interpeduncular nucleus and to the vasculature in the female rat

Immunofluorescence for gonadotropin releasing hormone (GnRH) in combination with retrograde labeling from the interpeduncular nucleus, as well as from the vasculature confirms that, in the rat, certain GnRH neurons project from the septum-diagonal band to the interpeduncular nucleus. However, about one half of these GnRH neurons also project to fenestrated capillaries as evidenced by uptake and retrograde transport of both peripherally injected Fluoro-Gold and centrally injected rhodamine labeled microspheres. The results indicate that the endocrine effects of GnRH are exerted in part by neurons which simultaneously project to neurohemal contact zones and to areas in the brain which are involved in the regulation of certain behaviors. It is suggested that certain GnRH neurons can directly couple endocrine events with other intracerebral events, such as regulation of lordosis behavior.     

Effects of aging on N-methyl-d-aspartate (NMDA)-induced GnRH and LH release in female rats

In order to evaluate if the changes of the hypothalamic-pituitary-ovary axis that induce a decrease in fertility and modifications in the sexual cycles during senescence involve modifications in the regulatory action of excitatory amino acid neurotransmission on GnRH neurons, we measured the in vitro effects of NMDA on GnRH release by the anterior preoptic and medial basal hypothalamic areas (APOA-MBH) of castrated aging (18 months old) and young (90 days of age) rats. In a second series of experiments the in vivo LH release response to intrahypothalamic (push-pull) administration of NMDA to aged and young castrated female rats was also determined. A similar rate of basal GnRH release was observed in old and young rats during the incubation time. The addition of NMDA to the medium significantly increased GnRH release in both groups; nevertheless, the GnRH release response to NMDA was significantly lower in old ( P < 0.01) than in young rats (Young: Basal: 50 ± 10; NMDA 15′: 410 ± 63, 22,5′: 1,469 ± 300; Old; Basal: 47±10 NMDA 15′:210 ± 30; 22,5′: 350 ± 65 ng/GnRH/mg . protein). The LH levels measured throughout the in vivo experiments indicated that basal LH concentrations were significantly lower in the aged group. The mean LH concentrations (fractions 1 to 6) was significantly lower in the aged group (Young: 3.9 ± 0.07, Old: 2.4 ± 0.03 ng/ml, P < 0.01). The LH release response to NMDA measured 10 min after the intrahypothalamic administration of the glutamate agonist was significantly lower in aged rats (4 .2 ± 1.6 ng/ml) as compared to young animals (18.0 ± 6.1 ng/ml; P < 0.05). LH levels in young rats increased to 580% vs., and only 47% in aged rats as compared to previous basal values. In conclusion, present results demonstrate that the GnRH responses to NMDA neurotransmission, which has a predominantly excitatory effects on GnRH neurons, is significantly decreased in old rats, these data give further support to the hypothesis that a decrease in the excitatory inputs to GnRH neurons could be directly involved in the reduction of the hypothalamic-pituitary-ovary axis activity observed during aging.     

Guidelines for pubertal suspension and gender reassignment for transgender adolescents

Pubertal suppression at Tanner stage 2 should be considered in adolescents with persistent gender identity disorder (GID). Issues related to achievement of adult height, timing of initiating sex steroid treatment, future fertility options, preventing uterine bleeding, and required modifications of genital surgery remain concerns. Concerns have been raised about altering neuropsychological development during cessation of puberty and reinitiation of puberty by the sex steroid opposite those determined by genetic sex. Collaborative assessment and treatment of dysphoric adolescents with persistent GID resolves these concerns and deepens our understanding of gender development.     

The sex-specific patterns of changes in hypothalamic-pituitary-gonadal axis during experimental autoimmune encephalomyelitis

Multiple sclerosis develops during reproductive years in a sex-specific manner. Various neuroendocrine changes have been described in this inflammatory, demyelinating, and debilitating disease. We here aimed to determine the extent and sex specificity of alterations in the hypothalamic-pituitary-gonadal axis in the rat model of multiple sclerosis named experimental autoimmune encephalomyelitis. During the disease course, the hypothalamic tissue showed transient upregulation of inflammatory marker genes Gfap, Cd68, Ccl2, and Il1b in both sexes, but accompanied by sex-specific downregulation of Kiss1 (in females only) and Gnrh1 (in males only) expression. In females, the expression of gonadotrope-specific genes Lhb, Cga, and Gnrhr was also inhibited, accompanied by decreased basal but not stimulated serum luteinizing hormone levels and a transient arrest of the estrous cycle. In contrast, Fshb expression and serum progesterone levels were transiently elevated, findings consistent with the maintenance of the corpora lutea, and elevated immunohistochemical labeling of ovarian StAR, a rate limiting protein in steroidogenic pathway. In males, downregulation of Gnrhr expression and basal and stimulated serum luteinizing hormone and testosterone levels were accompanied by inhibited testicular StAR protein expression. We propose that inflammation of hypothalamic tissue downregulates Kiss1 and Gnrh1 expression in females and males, respectively, leading to sex-specific changes downstream the axis.     

Effect of gonadotropin releasing hormone on thermoregulatory vasomotor activity in ovariectomized female rats

To investigate the effect of gonadotropin releasing hormone (GnRH) on thermoregulatory skin vasomotion, we injected GnRH into various brain regions in both anesthetized and unanesthetized ovariectomized female rats. Local warming of preoptic area (PO) elicited skin vasodilation in anesthetized rats. Injection of 2 μg GnRH into the septal area lowered the threshold hypothalamic temperature for skin vasodilation at least for 2 h. Similar injections of 2 μg GnRH into the lateral ventricle (LV) and PO were ineffective. Although this vasodilative effect was also obtained after the injection of 20 ng GnRH into the septal area, injections of 2 ng GnRH were without effect. Not only injections of 20 ng Antide, a potent GnRH antagonist, but also injections of the mixed solution of 20 ng GnRH and 20 ng Antide were also without effect. In unanesthetized and unrestrained rats at an ambient temperature of 17°C, injections of 20 ng GnRH into the septal area elicited tail vasodilation lasting for 30 minutes, whereas vehicle injections were ineffective. Injections of 20 ng GnRH into LV and PO were also ineffective. These results indicate that GnRH can elicit thermoregulatory skin vasomotion by acting on GnRH receptors in the septal area. This thermoregulatory vasodilative effect of GnRH might be possibly related to the etiology of climacteric hot flush.     

Vomeronasal organ: electrical stimulation activates Fos in mating pathways and in GnRH neurons

Electrical stimulation of the vomeronasal organ in male hamsters activated Fos expression in neurons of the chemosensory pathways, as in experiments where animals were stimulated with female chemical stimuli. Fos was also activated in gonadotropin hormone releasing hormone (GnRH, or LHRH) neurons in the rostral medial preoptic region of the brain, a possible substrate for GnRH influence on chemosensory-dependent reproductive behavior.     

Growth Hormone (GH)-Releasing Peptide and GH Releasing Hormone Stimulate GH Release from Subpopulations of Somatotrophs in Rats

The synthetic hexapeptide GH-releasing peptide (His-D-Trp-Ala-Trp-D-Phe-Lys-NH2; GHRP-6) and GH releasing hormone (GHRH) are both potent stimulators of GH release in rats. Using reverse hemolytic plaque assay (RHPA), we have compared the effects of human GHRH and GHRP-6 on GH release from the dispersed individual cells of rat anterior pituitary. In a single RHPA, we quantified the percentage of plaque forming cells (% PFC) and their mean plaque area (MPA) after 30 min-incubation, and calculated a total secretion index (TSI) by multiplying % PFC and MPA. 10 nM GHRH and 100 nM GHRP-6 each caused a significant increase in % PFC (%) (GHRH 39.15, GHRP-6 29.4, vs vehicle 24.3, P<0.01), MPA (×10^?2 μm²) (GHRH 124.04, GHRP-6 94.80, vs vehicle 44.57, P<0.01) and TSI (×10^?2) (GHRH 54.46, GHRP-6 32.87, vs vehicle 10.84, P<0.01). Simultaneous addition of both secretagogues caused a further increase in GH release (%PFC 46.4, MPA 142.55, TSI 69.82, P<0.01 vs vehicle), although the effect was additive but not synergistic. Somatostatin analog, SMS201–995 (SMS) partially suppressed all parameters in GH secretion after stimulation by GHRH and/or GHRP-6. A double RHPA was then performed to test whether all somatotrophs respond equally to GHRH and GHRP-6 or some cells formed plaques only by either GHRH or GHRP-6. There were somatotrophs responsive to only GHRH (23.3%vs control 6.2%, P<0.01), those responsive to only GHRP-6 (11.9%vs control 6.1%, P<0.01), and those responsive to both GHRH and GHRP-6 (7.8%vs control 0.2%, P<0.01). These results confirmed the previous findings that GHRP-6 and GHRH directly but independently stimulate GH release from the pituitary cells, and further suggest the presence of at least three functionally distinct somatotroph subpopulations concerning the responsiveness to GHRP-6 and GHRH in rats.     

Distribution of luteinizing hormone-releasing hormones I and II (LHRH-I and -II) in the quail and chicken brain as demonstrated with antibodies directed against synthetic peptides

Polyclonal antibodies were raised in rabbits against polypeptides corresponding to the N-terminal part (heptapeptides) of the two avian gonadotropin-releasing hormones, chicken (c) LHRH-I and -II. These peptides, which were synthesized by the continuous-flow technique, were selected because they contained the smallest number of common amino acid residues. The pGlu-His-Trp-Ser sequence at the C-terminal was suppressed to avoid possible cross-reactions between the antisera. The antisera generated in this way were tested for specificity by solid and liquid phase absorption as well as by antigen spot tests. The antiserum raised against cLHRH-I recognized this peptide preferentially though not exclusively. Some cross-reaction with cLHRH-II was observed in the absorption test, although spotting tests suggested a total specificity. The anti cLHRH-II appeared to be completely specific in all tests. These two antibodies were then used to study the distribution of cLHRH-I and -II immunoreactive structures in the quail and chicken brain. cLHRH-I immunoreactive perikarya were observed in a fairly wide area covering the preoptic-anterior hypothalamic and septal region. By contrast, cLHRH-II cells were confined to a single group located in the dorsal aspects of the occulomotor nuclei, at the junction of the di- and mesencephalon. A sex difference in the number of cLHRH-I cells was detected in the anterior lateral preoptic region of the quail. Fibers immunoreactive for either cLHRH-I or cLHRH-II were widely distributed in the telencephalon, diencephalon, and mesencephalon but showed a specific pattern of anatomical localization. In particular, a high density of cLHRH-I fibers were seen in the external layer of the median eminence, while cLHRH-II fibers were less prominent at this level. Contrary to previous reports, a significant amount of cLHRH-II fibers were however seen throughout the median einence (mostly external layer). The extensive distribution of both cLHRH-I and -II fibers in the quail and chicken brain is consistent with the potential role played by these peptides in the gonadotropin secretion and in the control of reproductive behavior. The specific role of cLHRH-II remains however elusive at present. © 1993 Wiley-Liss, Inc.     

Maturation of Sleep–Wake Gonadotrophin‐Releasing Hormone Secretion Across Puberty in Girls: Potential Mechanisms and Relevance to the Pathogenesis of Polycystic Ovary Syndrome

Neuroendocrine mechanisms underlying the progression of sleep–wake gonadotrophin‐releasing hormone (GnRH) pulse secretion across puberty have remained enigmatic. Here, the changes of sleep–wake luteinising hormone (LH) (and, by inference, GnRH) pulse secretion across puberty in normal girls are reviewed, primarily focusing on available human data. It is suggested that the primary control of GnRH pulse frequency changes across puberty, with sex steroid feedback exerting minimal control during childhood, but primary control during adulthood. A working model is proposed regarding how such a transfer of GnRH pulse frequency control may partly account for the prominent day–night differences of GnRH pulse frequency characteristic of puberty. How this model may be relevant to the genesis of abnormal GnRH secretion in peripubertal girls with hyperandrogenaemia is then described.     

Effect of season on neuropeptide Y and galanin within the hypothalamus of the ewe in relation to plasma luteinizing hormone concentrations and the breeding season: an immunohistochemical analysis

Within the hypothalamus, neurones that express neuropeptide Y (NPY) and galanin have been implicated in the regulation of gonadotropin-releasing hormone (GnRH) and gonadotropin secretion. We aimed to determine the extent to which the expression of these two neuronal systems is linked to the seasonal reproductive cycle, and the effect of chronic oestrogen treatment. Immunohistochemical analysis was used to examine changes between the breeding season and anestrus in ovariectomized (OVX) ewes with or without oestrogen treatment (s.c. implants for 2 weeks). Serial blood sampling established plasma luteinizing hormone (LH) profiles, and the ewes were subsequently killed and the brains perfused for immunohistochemistry. In OVX ewes, the amplitude of LH pulses was greater in the nonbreeding season than in the breeding season. Oestrogen treatment caused a marked reduction in plasma LH concentrations during anestrus, but not in the breeding season. The number of cells in the arcuate nucleus/median eminence region (ARC-ME) that stained for NPY was lower in ewes killed in anestrus (September) than in ewes killed in the breeding season (May), but there was no seasonal change in the number of galanin-stained cells. Within season, oestrogen treatment did not affect NPY- or galanin-cell number. There was no effect of season or oestrogen on the area of varicose fibres/terminals for either peptide in the ARC-ME, but galanin immunostaining was more intense during the breeding season. We conclude that the amount of NPY in cell bodies of the ARC-ME is lower in ewes in the nonbreeding season; this could reflect a steroid-independent effect of photoperiod. We also conclude that the long-term negative-feedback effect of oestrogen on GnRH/LH secretion does not appear to be mediated by NPY- or galanin-containing neurones in the ewe.     

Stimulation of CRH-Mediated ACTH Secretion by Central Administration of Neurotensin: Evidence for the Participation of the Paraventricular Nucleus

Central administration of neurotensin (NT) stimulates hypothalamic-pituitary-adrenal (HPA) activity in freely-moving rats. Increases in adrenocorticotropin hormone (ACTH) and corticosterone (B) were observed 15 rnin following central NT administration and remained elevated for up to 4 h. Of the two NT fragments tested, NT_1–8 and NT_8–13 only NT_8–13 was found to significantly elevate ACTH and B levels. Moreover, NT_8–13 activated the HPA axis with a temporal profile similar to NT_1–13, suggesting an interaction with the pharmacologically and molecularly characterized NT receptor. Animals pre-treated intravenously with the corticotropin-releasing hormone (CRH) antagonist, α-helical CRH, showed attenuated plasma ACTH and B responses to central NT administration. This indicates that CRH receptor activation is necessary for the stimulatory effects of NT on HPA function. Bilateral lesions of the paraventricular nucleus (PVN) of the hypothalamus significantly reduced NT-induced stimulation of ACTH and B release suggesting that the PVN is essential for NT's stirnulatory action. Median eminence content studies indicated that acute central NT administration stimulates CRH, but not arginine vassopressin (AVP), release in animals examined 60 min following NT injection. Taken together, these findings suggest that the stimulatory effects of NT on HPA activity occur via specific NT receptors and that one site of action of NT is likely at the level of the PVN where NT elicits the release of CRH.     

卡马西平与TRH抗癫痫作用相互关系的研究

目的探讨卡马西平与促甲状腺素释放激素（ＴＲＨ）抗癫痫作用的相互关系。方法采用放射免疫分析法测定戊四氮（ＰＴＺ）急性致病大鼠海马ＴＲＨ的含量。结果急性致痫后,海马内ＴＲＨ表达水平在２小时未见明显变化,４８小时显著升高。卡马西平预处理组在痫性发作后４８小时海马内ＴＲＨ表达水平明显升高,与对照组及假模型组相比均有显著性差异。结论卡马西平能显著升高痫性发作对ＴＲＨ的诱导表达,ＴＲＨ与卡马西平的作用可能有重