GPR54 System Research Articles

The Mechanism of Bushen Zhuyun Prescription in Regulating Luteal Phase Deficiency via Intervention of the Kiss1/GPR54 System in the Hypothalamus

Luteal phase deficiency (LPD) is a significant contributor to infertility and miscarriage. Bushen Zhuyun Prescription (BSZY) is a traditional Chinese medicine formula that has potential to treat LPD. To investigate the effect of BSZY in treating LPD, SD rats with complete estrous cycles were divided into blank, model, dydrogesterone (DYD), high-dose BSZY (BSZY-HD), and low-dose BSZY (BSZY-LD) groups. All the groups were received mifepristone gavage except for the blank group. The pathological changes were observed in the hypothalamus, and the analysis of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), progesterone (P), gonadotropin-releasing hormone (GnRH), and cyclic adenosine monophosphate (cAMP) levels in serum. Additionally, the mRNA and their protein expression levels of estrogen receptor α (ERα), ERβ, G protein-coupled receptor 30 (GPR30), kisspeptin 1 (Kiss1), and GPR54 were assessed; and the key molecules expression in cAMP/protein kinase A (PKA) pathway were analyzed. The results revealed that mifepristone treatment caused a decrease in FSH and cAMP levels and an increase in E2 and LH levels, which were normalised in the BSZY treatment groups. The expression of ERα and ERβ mRNA and protein in the hypothalamus was higher in the BSZY groups compared to the blank group. BSZY also resulted in increased expression of Kiss-1, GPR54, and PKA, as well as decreased expression of cAMP-response element binding protein (CREB), c-Fos, phospho-CREB (p-CREB), and p-c-Fos, compared to model group. No significant difference observed in GPR30 expression between the mifepristone and BSZY groups. The mRNA and protein of Kiss1 and GPR54 were decreased in model group and increased after BSZY treatment, while the mRNA and protein expression of PKA, CREB, c-Fos, p-CREB, and p-c-Fos were also decreased significantly. In conclusion, the findings demonstrate that BSZY could effectively regulate hypothalamic ER, interfere with the Kiss1/GPR54 signal transduction system, activate the cAMP/PKA pathway, and regulate key transduction molecules expression of PKA, CREB, c-Fos, there by effecting the level of reproductive hormones and has the potential to treat LPD-related infertility.

Effects of high-fat diet and treadmill running on the hypothalamic Kiss-1–GPR54 signaling pathway in male growing rats

BackgroundKiss-1 neuron, one of the metabolic sensors in the hypothalamus, is necessary for puberty initiation. It acts through G protein-coupled receptor, known as GPR54. In this study, the mechanism of the hypothalamic Kiss-1–GPR54 signaling pathway in a high-fat diet and exercise was investigated in growing male rats.MethodsA total of 135 3-week-old male weaned rats were kept on a high-fat diet (HFD) and exercise (60–70% dot{mathrm V}{mathrm O}_{2max}, 1 h/day, 5 days/week). They were randomly divided, as follows: control group (C); normal diet + exercise group (CE); HFD group (H); and HFD + exercise group (HE). Hypothalamus, testis, and serum samples of each group were collected on postnatal day (PND) 21 (early childhood), 43 (puberty), and 56 (maturity). Immunofluorescence, quantitative real-time PCR, hematoxylin and eosin staining, and chemiluminescent immunoassays were used in the study. ANOVA was used to analyze the effects of age (PNDs 21, 43, and 56), exercise (exercise and sedentariness), and diet (high-fat and normal) on the biological indices of rats.ResultsmRNA and protein expression of Kiss-1 and GPR54 in the hypothalamus gradually increased along with growth and peaked at PND 43, while those in serum testosterone increased and peaked at PND 56. The high-fat diet increased the expression of the Kiss-1–GPR54 system in the hypothalamus, whereas the serum testosterone decreased during different stages of growth. Exercise decreased the expression of Kiss-1 at PND 56 and increased it at PND 43. Meanwhile, it decreased testosterone and the deposition of lipid droplets in the testis at all ages of development.ConclusionsThe expression of Kiss-1–GPR54 in male rats showed fluctuating changes during growth and development. The high-fat diet was able to upregulate the expression of the Kiss-1–GPR54 system in the hypothalamus. The exercise was able to correct the adverse effect of the high-fat diet on the Kiss-1–GPR54 signaling pathway in the hypothalamus and the function of the hypothalamic-pituitary–gonadal (HPG) axis, but had age-specific effects on the male rats’ development.

The effect of high-fat diet and exercise on KISS-1/GPR54 expression in testis of growing rats

PurposeTo find the expression of KISS-1 and G protein-coupled receptor 54 in rats testis from PND 21st to 56th.Method128 three-week-old weaned rats underwent high-fat diet and exercise (60–70% VO2max, 1 h/day, 5 days/week) intervention and were randomly divided into group C, CE, HC, or HE. Sample time points were set on the PND 21st, 35th, 43rd, and 56th. The testicular testosterone and the mRNA content, and protein content of KISS-1 and GPR54 in testis tissue were detected by ELISA, RT-qPCR, and Western blotting.Result(1) The protein of KISS-1 and GPR54 increased gradually during the growing period. KISS-1 mRNA peaked at 35D and GPR54 peaked at 43D. (2) High-fat diet affected the expression of the KISS-1/GPR54 system in rat testis and reduced the expression level of KISS-1 protein. (3) 60–70% VO2max exercise decreased the KISS-1/GPR54 expression level. Exercise intervention improved testicular development in rats with a high-fat diet.ConclusionThe expression of KISS-1/GPR54 increased during the growing period. High-fat diet can downregulate the protein and gene expression of KISS-1/GPR54 and change the expression trend. 60–70% VO2max exercise decreased the expression of KISS-1/GPR54, which may be involved in the effects of exercise on high-fat dietary sex hormone disorders.

Effects Of Exercise On The Expression Of Kiss-1/GPR54 In Testis Of High-fat Diet Rats In Growth Phase

PURPOSE: To explore the effect of exercise on localization and expression of KISS-1 and GPR54 in the growth period (PND 21st to 56th day) of high-fat diet rat testes. METHODS: 21D old weanling rats were randomly divided into group HC (n=32) and group HE (n=32). HE group took 5-weeks trained (60-70% vVO2max, 1h/day, 5days/week). Groups HC and HE were fed with high-fat feed, which was purchased from Beijing Huafukang Biotechnology Co. Ltd. (LOT No: D12451).The rats of two groups were killed on the 21st D, 35th D, 43rd D, and 56th D old. The localization, mRNA expression and protein expression of KISS-1/GPR54 in the testis of each group were tested. RESULTS: The high-fat diet intervention resulted in a decrease in testicular index from pre-puberty(p<0.05); testicular tissue testosterone decreased significantly from prepuberty(p<0.01), and endocrine function was impaired; rat testicular tissue structure was loose, and the number of mature sperm decreased. The high-fat diet intervention had no significant effect on the localization of KISS-1/GPR54 system in rat testis, but affected the expression of KISS-1/GPR54 in rat testis and down-regulated the expression of KISS-1 protein(p<0.01). Exercise intervention can improve testicular development in rats with high-fat diet, especially to improve sex hormone disorders caused by high-fat diet. Under exercise intervention, the testicular tissue testosterone decline caused by high-fat diet was improved(p<0.05). Comparing with HC group, the expression of GPR54 protein increased significantly(p<0.01),but the KISS-1 protein expression had no obvious change(p>0.05). CONCLUSION: High-fat diet can inhibit the testicular development of male rats in the growth phase, and can also down-regulate the protein and gene expression levels of KISS-1/GPR54 system in testis tissue, and change the expression of KISS-1/GPR54 system, and its role needs further study. 60~70% VO2max moderate-intensity aerobic exercise change the inhibitory effect of high-fat diet on testicular development in male rats, and up-regulate KISS-1/GPR54 in prepubertal stage and whether KISS-1/GPR54 in testicular tissue participates in its regulation remains to be further study.

Potent Vasoconstrictor Kisspeptin-10 Induces Atherosclerotic Plaque Progression and Instability: Reversal by its Receptor GPR54 Antagonist.

BackgroundKisspeptin‐10 (KP‐10), a potent vasoconstrictor and inhibitor of angiogenesis, and its receptor, GPR54, have currently received much attention in relation to pre‐eclampsia. However, it still remains unknown whether KP‐10 could affect atherogenesis.Methods and ResultsWe evaluated the effects of KP‐10 on human umbilical vein endothelial cells, human monocyte‐derived macrophages, human aortic smooth muscle cells in vitro, and atherosclerotic lesions in apolipoprotein E–deficient (ApoE−/−) mice in vivo. KP‐10 significantly increased the adhesion of human monocytes to human umbilical vein endothelial cells, which was significantly inhibited by pretreatment with P234, a GPR54 antagonist. KP‐10 stimulated mRNA expression of tumor necrosis factor‐α, interleukin‐6, monocyte chemotactic protein‐1, intercellular adhesion molecule‐1, vascular adhesion molecule‐1, and E‐selectin in human umbilical vein endothelial cells. KP‐10 significantly enhanced oxidized low‐density lipoprotein–induced foam cell formation associated with upregulation of CD36 and acyl‐CoA:cholesterol acyltransferase‐1 in human monocyte‐derived macrophages. In human aortic smooth muscle cells, KP‐10 significantly suppressed angiotensin II–induced migration and proliferation, but enhanced apoptosis and activities of matrix metalloproteinase (MMP)‐2 and MMP‐9 by upregulation of extracellular signal‐regulated kinase 1 and 2, p38, Bcl‐2‐associated X protein, and caspase‐3. Four‐week‐infusion of KP‐10 into ApoE−/− mice significantly accelerated the development of aortic atherosclerotic lesions with increased monocyte/macrophage infiltration and vascular inflammation as well as decreased intraplaque vascular smooth muscle cells contents. Proatherosclerotic effects of endogenous and exogenous KP‐10 were completely canceled by P234 infusion in ApoE−/− mice.ConclusionsOur results suggest that KP‐10 may contribute to accelerate the progression and instability of atheromatous plaques, leading to plaque rupture. The GPR54 antagonist may be useful for prevention and treatment of atherosclerosis. Thus, the KP‐10/GPR54 system may serve as a novel therapeutic target for atherosclerotic diseases.

Relationships between leptin, the KiSS-1/GPR54 system and thyrotropic axis activity in ewe lambs predisposed to the delayed puberty

In the present study we compared body weight gains, some factors involved in the initiation of sexual maturation (pituitary expression of KiSS-1 and G protein-coupled receptors (GPR54), plasma concentrations of kisspeptin-10 (KiSS-10), leptin, thyroid-stimulating hormone (TSH), free thyroxin (fT4)) and the time of the first ovulation in ewe lambs predisposed to delayed puberty and control animals. The experiment was carried out on 114 ewes and 64 female lambs divided according to their birth type and body weight of their mothers. All ewe lambs were weighed at birth and every two weeks thereafter, until eight months of postnatal age. From four to eight months of postnatal age at monthly intervals, blood samples were collected from the jugular vein of all ewe lambs. Then, daily weight gains were recorded and plasma KiSS-10, leptin, TSH and fT4 concentrations were analysed by ELISA using species-specific antibodies. Starting from six months of age, the pituitary expression of KiSS-1/GPR54 mRNA was determined by Real Time-PCR. The activity of the ovaries was estimated using laparoscopy. The results obtained showed that the time of the onset of puberty in the ewe lambs depended on the mother’s body mass, the plasma leptin level, and the birth type (singleton/twin). It was observed that the elevation of leptin concentration up to 3.35±0.26–3.60±0.19ng/mL was associated with the initiation of puberty. Conversely, the hyperphysiological leptin levels found in ewe lambs, which were twin offspring of fatty sheep, were correlated with puberty delayed until the age of ten months. Moreover, it was found that a significant increase in pituitary KiSS-1 mRNA expression (1.40±0.12–1.63±0.22) (relative KiSS-1 mRNA expression level, ratio of KiSS-1 mRNA/GAPDH mRNA) and plasma KiSS-10 concentration (31.26±1.54ng/mL–32.24±2.25ng/mL) was connected with the occurrence of the first ovulation. On the other hand, GPR54 mRNA expression in the pituitary decreased around the time of the first ovulation. Also, the increase in thyroid gland activity was dependent on the mother’s body mass as well as birth type and occurred around the time of the first ovulation.

Development and Aging of the Kisspeptin–GPR54 System in the Mammalian Brain: What are the Impacts on Female Reproductive Function?

The prominent role of the G protein coupled receptor GPR54 and its peptide ligand kisspeptin in the progression of puberty has been extensively documented in many mammalian species including humans. Kisspeptins are very potent gonadotropin-releasing hormone secretagogues produced by two main populations of neurons located in two ventral forebrain regions, the preoptic area and the arcuate nucleus. Within the last 2 years a substantial amount of data has accumulated concerning the development of these neuronal populations and their timely regulation by central and peripheral factors during fetal, neonatal, and peripubertal stages of development. This review focuses on the development of the kisspeptin–GPR54 system in the brain of female mice, rats, sheep, monkeys, and humans. We will also discuss the notion that this system represents a major target through which signals from the environment early in life can reprogram reproductive function.

Evidence for a role of KISS-1/GPR54 system in decreased luteinizing hormone (LH) secretion in fasted prepubertal ewes

Evidence for a role of KISS-1/GPR54 system in decreased luteinizing hormone (LH) secretion in fasted prepubertal ewes

The role of kisspeptin in the control of gonadotrophin secretion

Kisspeptins, and their cognate receptor gpr-54, were first found to regulate the hypothalamic-pituitary-gonadal (HPG) axis in 2003, when two groups demonstrated that mutations in gpr-54 cause idiopathic hypogonadotropic hypogonadism characterized by delayed or absent puberty. This review aims to highlight discoveries in the KiSS-1/gpr-54 system, focusing on their regulation of the HPG axis in male and female reproductive systems of both mammalian and non-mammalian vertebrates. A search of PUBMED and the authors' files was done without limitations by language or species for citations relevant to kisspeptin, reproduction and signal transduction. Kisspeptins and gpr-54 are critical for puberty and the regulation of reproduction. Kisspeptins have been implicated in mediating many of the important signals relayed to the gonadotrophin-releasing hormone (GnRH) neuron such as positive and negative feedback, metabolic input and photoperiod. The ability of kisspeptin neurons to co-ordinate different signals impinging on the HPG axis makes it one of the most important regulators of GnRH and the reproductive axis. Kisspeptins are pivotal regulators of the HPG axis and reproduction, with the ability to integrate signals from both internal and external sources. Knowledge about the signalling mechanisms involved in kisspeptin stimulation of GnRH would help improve the understanding of the importance of this critical pathway in reproduction.

The KiSS‐1/GPR54 system: putative target for endocrine disruption of reproduction at hypothalamic–pituitary unit?

The hypothalamic-pituitary (HP) unit, key element in the control of development and function of the reproductive axis, is highly sensitive to the organizing and activational effects of endogenous and exogenous compounds with sex steroid activity. Thus, inappropriate sex steroid input during early critical periods of their maturation can induce immediate or delayed defects in the neuroendocrine systems governing reproduction, which might eventually lead to alterations in the timing of puberty onset and/or infertility. Similarly, later inadequate exposures can cause dysfunctions of the gonadotropic axis. In recent years, kisspeptins (the products of KiSS-1 gene that operate through the G protein-coupled receptor 54) have emerged as fundamental regulators of puberty onset and gonadotropin secretion, and neurons in the hypothalamus expressing KiSS-1 have been demonstrated as essential conduits for the dynamic control of the gonadotropic axis by a number of hormonal factors. In this context, the hypothalamic KiSS-1 system has been proven sensitive to the early organizing effects, as well as to the acute regulatory actions, of gonadal steroids: phenomena which are likely to play fundamental roles in the sexual differentiation of gonadotropin secretion, and its feedback regulation by androgen and oestrogen. These observations raise the question on whether the hypothalamic KiSS-1 system might constitute a potential target for endocrine disruption of puberty onset and reproductive function at the HP unit by compounds with oestrogenic, androgenic or anti-androgenic activity. We review herein the physiological basis and initial, albeit so far scarce, experimental evidence supporting such possibility.

Regulation of Hypothalamic Expression of KiSS-1 and GPR54 Genes by Metabolic Factors: Analyses Using Mouse Models and a Cell Line

It is well established that reproductive function is metabolically gated. However, the mechanisms whereby energy stores and metabolic cues influence fertility are yet to be completely deciphered. Recently, the hypothalamic KiSS-1/GPR54 system has emerged as a fundamental regulator of the gonadotropic axis, which conveys the modulatory actions of sex steroids to GnRH neurons. Evidence is also mounting that KiSS-1 neurons may also represent the link between systemic metabolic signals and central control of reproduction. To further explore this possibility, we examined the impact of changes in energy status and key metabolic regulators on the hypothalamic expression of KiSS-1 and GPR54 genes, using different mouse models and the hypothalamic cell line N6. Time-course analysis of the effects of short-term fasting revealed a rapid (12- and 24-h) decline in KiSS-1 and GPR54 mRNA levels, which preceded that of GnRH (48 h). In contrast, diet-induced obesity or obesity associated with leptin deficiency (ob/ob vs. wild-type mice) failed to induce overt changes in hypothalamic expression of KiSS-1 and GPR54 genes. However, leptin infusion of ob/ob mice evoked a significant increase in KiSS-1 and GPR54 mRNA levels compared with pair-fed controls. Moreover, leptin, but not insulin or IGF-I, stimulated KiSS-1 mRNA expression in the mouse hypothalamic cell line N6. In addition, neuropeptide Y (NPY) null mice showed decreased KiSS-1 mRNA levels at the hypothalamus, whereas exposure to NPY increased expression of KiSS-1 in hypothalamic N6 cells. In sum, our present data further characterize the functional relevance and putative key mediators (such as leptin and NPY) of the metabolic regulation of the hypothalamic KiSS-1 system in the mouse.

Neuroendocrine factors in the initiation of puberty: The emergent role of kisspeptin

Puberty is the end-point of a complex series of developmental events, defined by the dynamic interaction between genetic factors and environmental cues, ultimately leading to the attainment of reproductive capacity. The neuroendocrine basis of puberty has been the subject of extensive investigation in the last decades, and identification of the trigger(s) of puberty onset has drawn considerable attention. In this context, recognition of the fundamental role of kisspeptin (encoded by the KiSS-1 gene) and its receptor GPR54 as major gatekeepers of gonadotropic function in general, and puberty onset in particular, has been a major breakthrough in contemporary Neuroendocrinology. Indeed, during the last 3 years, the so-called KiSS-1/GPR54 system has been substantiated as pivotal regulator of puberty in mammals; the lack of GPR54 signaling being coupled to sexual immaturity (impuberism) in mice and humans. In this review, we will summarize the most salient experimental data (mostly obtained in laboratory animals) demonstrating the key roles of hypothalamic KiSS-1 neurons in the activation of the reproductive axis at puberty, and its regulation by metabolic and, eventually, environmental factors. Whether the KiSS-1 system is the trigger for puberty onset and/or it operates as integrator and effector of up-stream regulatory factors warrants further investigation.

Kisspeptin Mediates the Photoperiodic Control of Reproduction in Hamsters

The KiSS-1 gene encodes kisspeptin, the endogenous ligand of the G-protein-coupled receptor GPR54. Recent data indicate that the KiSS-1/GPR54 system is critical for the regulation of reproduction and is required for puberty onset. In seasonal breeders, reproduction is tightly controlled by photoperiod (i.e., day length). The Syrian hamster is a seasonal model in which reproductive activity is promoted by long summer days (LD) and inhibited by short winter days (SD). Using in situ hybridization and immunohistochemistry, we show that KiSS-1 is expressed in the arcuate nucleus of LD hamsters. Importantly, the KiSS-1 mRNA level was lower in SD animals but not in SD-refractory animals, which spontaneously reactivated their sexual activity after several months in SD. These changes of expression are not secondary to the photoperiodic variations of gonadal steroids. In contrast, melatonin appears to be necessary for these seasonal changes because pineal-gland ablation prevented the SD-induced downregulation of KiSS-1 expression. Remarkably, a chronic administration of kisspeptin-10 restored the testicular activity of SD hamsters despite persisting photoinhibitory conditions. Overall, these findings are consistent with a role of KiSS-1/GPR54 in the seasonal control of reproduction. We propose that photoperiod, via melatonin, modulates KiSS-1 signaling to drive the reproductive axis.

The roles of kisspeptins and G protein-coupled receptor-54 in pubertal development

This review summarizes the experimental data demonstrating the fundamental role of kisspeptins and their G protein-coupled receptor GPR54 in the control of reproduction, with special emphasis on their function at puberty. Kisspeptins, products of the KiSS-1 gene, were originally identified as metastasis suppressor peptides with the ability to bind G protein-coupled receptor GPR54. In late 2003, loss-of-function mutations of the GPR54 gene were found in patients suffering from hypogonadotropic hypogonadism. This finding kicked off the analysis of the role of the KiSS-1/GPR54 system in the control of reproduction. Kisspeptins are very potent elicitors of gonadotropin secretion, primarily through stimulation of gonadotropin-releasing hormone release. Enhanced expression of KiSS-1 and GPR54 genes, as well as increased GPR54 signaling, are detected at the hypothalamus during pubertal development, and activation of GPR54 by administration of kisspeptin is sufficient to induce precocious activation of the gonadotropic axis in immature rodents and monkeys. Hypothalamic KiSS-1 also functions as an essential integrator for peripheral inputs, including gonadal steroids and nutritional signals, controlling gonadotropin-releasing hormone and gonadotropin secretion. Kisspeptins and their putative receptor, GPR54, have recently emerged as indispensable factors for pubertal development, with a key role as gatekeepers of gonadotropin-releasing hormone release neurons and, hence, of reproductive function.

GPR54 and kisspeptin in reproduction

Kisspeptins, the peptide products of the KiSS-1 gene, were identified in 2001 as natural ligands of the previously orphan G protein-coupled receptor, GPR54. They include, among others, metastin and kisspeptin-10. The known biological functions of kisspeptins were initially restricted to their ability to suppress tumour metastasis, hence the name of metastin. However, in late 2003, two groups independently reported that loss-of-function mutations of the GPR54 gene are linked to absence of puberty onset and hypogonadotrophic hypogonadism in humans--a phenotype that was reproduced in GPR54-null mice. Those seminal observations revealed a totally unexpected, fundamental role of the KiSS-1/GPR54 system in control of puberty and reproductive function and boosted an extraordinary interest for the characterization of these novel facets of kisspeptin physiology. Indeed, in the last 2 years, metastin and kisspeptin-10 have been demonstrated as very potent stimulators of the gonadotrophic axis, in a number of species and through different routes of administration. In addition, the hypothalamic KiSS-1/GPR54 system has been proven as an essential gatekeeper of GnRH neurons, involved in their activation at puberty and their regulation by gonadal steroids and (probably) metabolic factors. This review comprehensively examines the experimental evidence obtained to date supporting a pivotal role of kisspeptins and GPR54 in the control of reproduction.

Effects of KiSS-1 Peptide, the Natural Ligand of GPR54, on Follicle-Stimulating Hormone Secretion in the Rat

KiSS-1 was originally identified as a metastasis suppressor gene encoding an array of structurally related peptides, namely kisspeptins, which acting through the G protein-coupled receptor GPR54 are able to inhibit tumor progression. Unexpectedly, a reproductive facet of this newly discovered system has recently arisen, and characterization of the role of the KiSS-1/GPR54 system in the neuroendocrine control of gonadotropin secretion has been initiated. However, such studies have been so far mostly restricted to LH, and very little is known about the actual contribution of this system in the regulation of FSH release. To address this issue, the effects of KiSS-1 peptide on FSH secretion were monitored in vivo and in vitro under different experimental conditions. Intracerebroventricular administration of KiSS-1 peptide significantly stimulated FSH secretion in prepubertal and adult rats. Yet, dose-response analyses in vivo demonstrated an ED(50) value for the FSH-releasing effects of KiSS-1 of 400 pmol, i.e. approximately 100-fold higher than that of LH. In addition, systemic (ip and iv) injection of KiSS-1 significantly stimulated FSH secretion in vivo. However, KiSS-1 failed to elicit basal FSH release directly at the pituitary level, although it moderately enhanced GnRH-stimulated FSH secretion in vitro. Finally, mechanistic studies revealed that the ability of KiSS-1 to elicit FSH secretion was abolished by the blockade of endogenous GnRH actions, but it was persistently observed in different models of leptin insufficiency and after blockade of endogenous excitatory amino acid and nitric oxide pathways, i.e. relevant signals in the neuroendocrine control of gonadotropin secretion. In summary, our results extend previous recent observations on the role of KiSS-1 in the control of LH secretion and provide solid evidence for a stimulatory effect of KiSS-1 on FSH release, acting at central level. Overall, it is proposed that the KiSS-1/GPR54 system is a novel, pivotal downstream element in the neuroendocrine network governing gonadotropin secretion.

Characterization of the potent luteinizing hormone-releasing activity of KiSS-1 peptide, the natural ligand of GPR54.

Loss-of-function mutations of the gene encoding GPR54, the putative receptor for the KiSS-1-derived peptide metastin, have been recently associated with hypogonadotropic hypogonadism, in both rodents and humans. Yet the actual role of the KiSS-1/GPR54 system in the neuroendocrine control of gonadotropin secretion remains largely unexplored. To initiate such analysis, the effects of KiSS-1 peptide on LH secretion were monitored using in vivo and in vitro settings under different experimental conditions. Central intracerebroventricular administration of KiSS-1 peptide potently elicited LH secretion in vivo over a range of doses from 10 pmol to 1 nmol. The effect of centrally injected KiSS-1 appeared to be mediated via the hypothalamic LHRH. However, no effect of central administration of KiSS-1 was detected on relative LHRH mRNA levels. Likewise, systemic (i.p. and i.v.) injection of KiSS-1 markedly stimulated LH secretion. This effect was similar in terms of maximum response to that of central administration of KiSS-1 and might be partially attributed to its ability to stimulate LH secretion directly at the pituitary. Finally, the LH-releasing activity of KiSS-1 was persistently observed after blockade of endogenous excitatory amino acid and nitric oxide pathways, i.e. relevant neurotransmitters in the neuroendocrine control of LH secretion. In summary, our results provide solid evidence for a potent stimulatory effect of KiSS-1 on LH release, acting at central levels (likely the hypothalamus) and eventually at the pituitary, and further document a novel role of the KiSS-1/GPR54 system as a relevant downstream element in the neuroendocrine network governing LH secretion.