Chicken Gonadotropin-releasing Hormone Research Articles

Sequence identification and expression characterization of leptin in the spotted scat, Scatophagus argus.

Scatophagus argus is an important marine culture fish in South and South-East Asia, including Southeast coastal areas of China. Artificial propagation technology for S. argus is not optimum; thus further studies on its reproduction biology are required. Although previous studies have shown that leptin (Lep) can regulate fish reproduction, the role of lep genes in S. argus is unknown. Herein, in silico analysis showed that S. argus has two lep genes (lepa and lepb). Protein 3D-structure prediction showed that Lepa has four α-helices (similar to mammals), while Lepb only has three. Tissue distribution analysis showed that lepa is highly expressed in the liver, whereas lepb was not detected in any tissue. Notably, lepr was expressed in all tissues. Lepa mRNA expression levels in the liver and serum Lep, estradiol (E2) and vitellogenin (Vtg) levels of female fish were significantly higher in ovaries at stage IV than in ovaries at stage II. Serum E2 levels were significantly positively correlated with Vtg levels in female fish at different development stages, while serum E2 was not correlated with Lep levels. Consistently, in vitro incubation of the liver with E2 significantly up-regulated vtga, while it did not affect lepa expression. Recombinant Lep (10nM) significantly up-regulated chicken gonadotropin-releasing hormone (cGnRH/GnRH-II) in the hypothalamus and GnRH receptor (GnRHR) and luteinizing hormone beta (Lhb) in the pituitary. These results suggest that lepa regulates female reproduction in S. argus.

Exogenous cortisol and red light irradiation affect reproductive parameters in the goldfish Carassius auratus.

Reproductive hormones play essential roles in the control of reproduction and gonadal maturation in fish. The purpose of this study was to determine the effects of cortisol administration (10 µg/g or 50 µg/g) or red light irradiation at two intensities (0.5 W/m2 or 1.0 W/m2) on the reproductive hormones in goldfish (Carassius auratus). The effects of different treatments were analyzed by determining the mRNA expression levels of gonadotropin-inhibitory hormone receptor (GnIH-R), chicken gonadotropin-releasing hormone (cGnRH-II), salmon GnRH (sGnRH), FSHβ, LHβ, and plasma testosterone and the level of 17β-estradiol for 48 h. Additionally, by double immunofluorescence staining, we detected the expression of both GnIH and GnRH in the diencephalons of goldfish brains. The mRNA expression of GnIH-R was significantly higher in the cortisol group and red light-irradiated group from 3 to 48 h than in the control group. Additionally, the mRNA levels of cGnRH-II, sGnRH, FSHβ, LHβ, testosterone, and 17β-estradiol were significantly lower in the cortisol group than in the other groups from 3 to 48 h. These results indicated that both cortisol and red light-emitting diode (LED) light increased GnIH expression and inhibited GnRH expression. In particular, red light irradiation suppressed reproductive responses as much as the cortisol treatment at 48 h. Thus, it could be an alternative method for suppressing reproductive responses in future aquacultures.

Is the use of recombinant cGnRH may be a future alternative to control the fish spawning? Let us go with the goldfish example.

The use of recombinant gonadotropin-releasing hormone (rGnRH) has very rarely been tested in fish to promote spawning. This study evaluated the impact of recombinant chicken gonadotropin-releasing hormone (rcGnRH) with metoclopramide on the release of sex steroids and final maturation induction in goldfish (Carassius auratus) broodstock. For this purpose, goldfish broodstock was divided into four groups and treated with 0.9% NaCl with 20mg/kg metoclopramide (Met) (C); 10μg/kg body weight (BW) rcGnRH with 20mg/kg metoclopramide (rcGn10); 15μg/kg BW rcGnRH with 20mg/kg metoclopramide (rcGn15); and 20μg/kg BW rcGnRH with 20mg/kg metoclopramide (rcGn20). The capability of the rcGnRH for eliciting biological response was tested in vivo by evaluating the changes of 17β estradiol (E2), testosterone (T), and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) and the induced spawning. Blood samples were obtained at 0h, 12h, and 24h after injection. The rcGn10, rcGn15, and rcGn20 treatments induced lower E2 concentration, especially 24h post-injection. T levels were significantly higher in rcGn10, rcGn15, and rcGn20 treatments 12h post-injection than at 0h and then decreased at 24h post-injection. Furthermore, the rcGnRH tested significantly enhanced DHP secretion in rcGn10, rcGn15, and rcGn20 treatments 12h post-injection before a decline at 24h post-injection. No significant difference between the sampling times was found in the C treatment for the 3 sex steroids tested. The results also displayed that rcGnRH at 10-20µg/kg of body weight can trigger spawning with the highest speed and efficiency of spawning at 20µg/kg. The obtained results represent a possible strategy for enhancing the artificial reproduction and ovulation of broodstock fish by rGnRH and further support the use of recombinant hormones to promote reproduction in aquaculture.

Nesfatin-1-like peptide suppresses hypothalamo-pituitary-gonadal mRNAs, gonadal steroidogenesis, and oocyte maturation in fish†.

Nucleobindin (Nucb)-1 and Nucb2 are DNA and Ca2+ binding proteins with multiple functions in vertebrates. Prohormone convertase-mediated processing of Nucb2 results in the production of biologically active nesfatin-1. Nesfatin-1 is involved in the regulation of reproduction in many vertebrates, including fish. Our lab originally reported a nesfatin-1-like peptide (Nlp) encoded in Nucb1 that exhibits nesfatin-1-like metabolic effects. We hypothesized that Nlp has a suppressive role in the reproductive physiology of fish. In this research, whether Nlp regulates reproductive hormones and oocyte maturation in fish were determined. Single intraperitoneal (IP) injection of goldfish Nlp (50ng/g body weight) suppressed salmon and chicken gonadotropin-releasing hormone (sgnrh and cgnrh2), gonadotropin-inhibiting hormone (gnih) and its receptor (gnihr), and kisspeptin and brain aromatase mRNA expression in the hypothalamus of both male and female goldfish. In the pituitary, Nlp decreased mRNAs encoding lhb, fshb and kisspeptin and its receptor, while a significant increase in gnih and gnihr was observed. In the gonads, lh (only in male fish) and fsh receptor mRNAs were also significantly downregulated in Nlp-injected fish. Sex-specific modulation of gnih, gnihr, and kisspeptin system in the gonads was also observed. Nlp decreased sex steroidogenic enzyme encoding mRNAs and circulating levels of testosterone and estradiol. In addition, incubation of zebrafish ovarian follicles with Nlp resulted in a reduction in oocyte maturation. These results provide evidence for a robust role for Nlp in regulating reproductive hormones in goldfish and oocyte maturation in zebrafish, and these effects resemble that of nesfatin-1.

Feeding of phytosterols reduced testosterone production by modulating GnRH and GnIH expression in the brain and testes of male Japanese quail (Coturnix coturnix japonica)

Phytosterols (PS), or plant sterols used as cholesterol-lowering agents, have been shown to act as endocrine-disrupting chemicals in some laboratory animals. Moreover, dietary PS efficiently pass through the blood-brain barrier and accumulate in brain cell membranes. We asked whether the accumulation of PS affects reproduction through the hypothalamic-pituitary-gonadal axis. Thirty male quail chicks were randomly divided into 3 groups (control, 80mg/kg BW, and 800mg/kg BW), and daily single doses of PS or vehicle were gavaged into the crop sac from 15 to 100 d of age. At the end of the entire period, half of each group was injected intramuscularly with either 10μg of chicken gonadotropin-releasing hormone 1 (cGnRH-1) or phosphate-buffered saline solution (PBS) as the vehicle. Blood was collected before and 30min after cGnRH-1 challenge by jugular venipuncture and decapitation, respectively. The results indicated that testosterone concentrations were low (P <0.05) before (800mg/kg BW) and after GnRH challenge in PS-treated quails compared with controls (P <0.001). However, luteinizing hormone (LH) levels were not different among the groups before cGnRH-1 challenge. In addition, PS-gavaged animals failed to manifest increased LH levels after cGnRH-1 injection (P <0.01). The same trends were observed in pituitary LH levels at 800mg/kg BW PS after cGnRH-1 injection (P <0.05). Real-time PCR results revealed that PS (800mg/kg BW) feeding reduced expression of GnRH-1 in the brain and testes compared to controls. However, gonadotropin-inhibitory hormone (GnIH) expression was significantly elevated before and after GnRH-1 challenges in the brain and testes. Collectively, these results suggest that brain-mediated effects of PS on gonadal function occurs via the induction of GnIH gene expression, and these indirect effects are less potent than direct effects.

GnRH mRNA levels in male three-spined sticklebacks, Gasterosteus aculeatus, under different reproductive conditions

In vertebrates, reproduction is regulated by the brain–pituitary–gonad (BPG) axis, where the gonadotropin-releasing hormone (GnRH) is one of the key components. However, very little is known about the possible role of GnRH in the environmental and feedback control of fish reproduction. To investigate this, full-length gnrh2 (chicken GnRH II) and gnrh3 (salmon GnRH) sequences of male three-spined sticklebacks (Gasterosteus aculeatus), which are clustered with the taxa of the same GnRH type as other Euteleostei, were cloned and annotated. gnrh1 is absent in this species. The mRNA levels of gnrh2 and gnrh3 in the sticklebacks' brain were measured under breeding and post-breeding conditions as well as in castrated and sham-operated breeding fish and castrated/sham-operated fish kept under long-day (LD 16:8) and short-day (LD 8:16) conditions. Fully breeding males had considerably higher mRNA levels of gnrh2 and gnrh3 in the thalamus (Th) and in the telencephalon and preoptic area (T+POA), respectively, than post-breeding males. Sham-operated breeding males have higher gnrh3 mRNA levels than the corresponding castrated males. Moreover, higher gnrh2 mRNA levels in the Th and higher gnrh3 mRNA levels in the T+POA and hypothalamus (HypTh) were also found in long-day sham-operated males than in sham-operated fish kept under an inhibitory short day photoperiod. Nevertheless, gnrh2 and gnrh3 mRNA levels were not up-regulated in castrated males kept under long-day photoperiod, which suggests that positive feedbacks on the brain-pituitary-gonad axis are necessary for this response.

Characterization of Schizothorax prenanti cgnrhII gene: fasting affects cgnrhII expression.

In this study, the role of chicken gonadotropin-releasing hormone II (cgnrhII) in feeding regulation was investigated in Schizothorax prenanti. First, the full-length S. prenanti cgnrhII cDNA consisted of 693 bp with an open reading frame of 261 bp encoding a protein of 86 amino acids. Next, cgnrhII was widely expressed in the central and peripheral tissues. Last, there were significant changes in cgnrhII mRNA expression in the fasted group compared to the fed group in the S. prenanti hypothalamus during 24 h fasting (P < 0.05). Furthermore, the cgnrhII gene expression presented a significant decrease in the fasted group compared with the fed group (P < 0.05) on days 3, 5 and 7, after re-feeding, there was no significant changes in cgnrhII mRNA expression level between refed and fed group on day 9 (P > 0.05). Thus, the results suggest that cGnRH II expression is influenced by fasting and the gene may be involved in feeding regulation in S. prenanti.

Effects of various photoperiods on Kisspeptin and reproductive hormones in the goldfish, Carassius auratus

The study aimed to test differences in the hormones of the hypothalamus–pituitary–gonad axis, Kisspeptin (Kiss) and sex steroids, and the sexual maturation of goldfish (Carassius auratus) according to various photoperiods: 10 h light (L):14 h dark (D) (short day), 12L:12D (control), and 14L:10D (long day). To investigate the sexual maturation under various photoperiods, quantitative real-time PCR (QPCR) assays for salmon gonadotropin-releasing hormone, chicken gonadotropin-releasing hormone, gonadotropin hormones (GTHs), Kisspeptin 1 (Kiss1), and its receptor, G protein-coupled receptor 54 (GPR54), were developed. mRNA expression was monitored in cultured pituitary cells (in vitro) of various photoperiod groups. Furthermore, we injected or treated the animals with Kiss and found that Kiss treatment increases the mRNA expression levels of GTHs. We measured the plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), and 17α-hydroxypregnenolone levels. The gonadotropin-releasing hormones (GnRHs), GTHs, Kiss1/GPR54 mRNA expression, plasma FSH, LH, and 17α-hydroxypregnenolone levels in the 14L:10D group were significantly higher than in the other groups. These results suggest that light length regulates sex maturation by GnRH and Kiss1 in the brain of the goldfish.

Gonadotropin-releasing hormone 2 suppresses food intake in the zebrafish, Danio rerio

Gonadotropin-releasing hormone (GnRH) is an evolutionarily conserved neuropeptide with 10 amino acid residues, of which several structural variants exist. A molecular form known as GnRH2 ([His5 Trp7 Tyr8]GnRH, also known as chicken GnRH II) is widely distributed in vertebrates except for rodents, and has recently been implicated in the regulation of feeding behavior in goldfish. However, the influence of GnRH2 on feeding behavior in other fish has not yet been studied. In the present study, therefore, we investigated the role of GnRH2 in the regulation of feeding behavior in a zebrafish model, and examined its involvement in food intake after intracerebroventricular (ICV) administration. ICV injection of GnRH2 at 0.1 and 1 pmol/g body weight (BW) induced a marked decrease of food consumption in a dose-dependent manner during 30 min after feeding. Cumulative food intake was significantly decreased by ICV injection of GnRH2 at 1 pmol/g BW during the 30-min post-treatment observation period. The anorexigenic action of GnRH2 was completely blocked by treatment with the GnRH type I receptor antagonist Antide at 25 pmol/g BW. We also examined the effect of feeding condition on the expression level of the GnRH2 transcript in the hypothalamus. Levels of GnRH2 mRNA obtained from fish that had been provided excess food for 7 days were higher than those in fish that had been fed normally. These results suggest that, in zebrafish, GnRH2 acts as an anorexigenic factor, as is the case in goldfish.

Development, validation, and utilization of a novel antibody specific to the type III chicken gonadotropin-releasing hormone receptor

Two gonadotropin-releasing hormone receptors (GnRH-Rs) have been characterized in chickens to date: cGnRH-R-I and cGnRH-R-III, with cGnRH-R-III being the predominant pituitary form. The purpose of the present study was to first validate a novel antibody for the specific detection of cGnRH-R-III and second, using this antibody, detect changes in cGnRH-R-III protein levels in the pituitary gland of male and female chickens during a reproductive cycle. The localization of cGnRH-R-III within the anterior pituitary gland was also determined. Western blotting of pituitary extracts and transiently transfected COS-7 cell lysates revealed that our antibody is highly specific to cGnRH-R-III protein. Similarly, when used in immunocytochemistry, this antibody specifically detects cells expressing cGnRH-R-III and not cGnRH-R-I. Western blot analyses of chicken pituitary gland homogenates show that cGnRH-R-III protein levels are significantly greater in sexually mature birds than in immature birds or birds at the end of a reproductive cycle ( P < 0.0001). A similar pattern was observed for both males and females. Additionally, the antibody was able to detect cGnRH-R-III in cells along the periphery of the cephalic and caudal lobes of the anterior pituitary where the cells containing the gonadotropins are located. In summary, we successfully validated a novel antibody to cGnRH-R-III and showed levels of cGnRH-R-III protein in the pituitary fluctuate with respect to the reproductive status in both male and female chickens.

Expression of GnRH genes is elevated in discrete brain loci of chum salmon before initiation of homing behavior and during spawning migration

Our previous studies suggested the importance of gonadotropin-releasing hormones (GnRHs) for initiation of spawning migration of chum salmon, although supporting evidence had been not available from oceanic fish. In farmed masu salmon, the amounts of salmon GnRH (sGnRH) mRNAs in the forebrain increased in the pre-pubertal stage from winter through spring, followed by a decrease toward summer. We thus hypothesized that gene expression for GnRHs in oceanic chum salmon changes similarly, and examined this hypothesis using brain samples from winter chum salmon in the Gulf of Alaska and summer fish in the Bering Sea. They were classified into sexually immature and maturing adults, which had maturing gonads and left the Bering Sea for the natal river by the end of summer. The absolute amounts of GnRH mRNAs were determined by real-time PCRs. The amounts of sGnRH mRNA in the maturing winter adults were significantly larger than those in the maturing summer adults. The amounts of sGnRH and chicken GnRH mRNAs then peaked during upstream migration from the coast to the natal hatchery. Such changes were observed in various brain loci including the olfactory bulb, terminal nerve, ventral telencephalon, nucleus preopticus parvocellularis anterioris, nucleus preopticus magnocellularis and midbrain tegmentum. These results suggest that sGnRH neurons change their activity for gonadal maturation prior to initiation of homing behavior from the Bering Sea. The present study provides the first evidence to support a possible involvement of neuropeptides in the onset of spawning migration.

Effect of chicken gonadotropin-releasing hormone (cGnRH-II) on plasma steroid hormone, maturation and ovulation in African catfish, Clarias gariepinus (Burchell)

Nowadays, many studies on the effects of native GnRHs and various agonists have been evaluated on the spawning of African catfish, Clarias gariepinus. For the current study, three experiments have been conducted to determine the effectiveness of chicken gonadotropin-releasing hormone II (cGnRH-II) in stimulation of maturation and ovulation of African catfish. The first experiment concerned the graded dosage treatment; the second experiment was designed specifically to determine the response of dopamine inhibitor to accelerate the maturation process. Finally the third experiment was carried out to compare the effectiveness of cGnRH-II against leutinising hormone releasing-hormone analogue (LHRHa) and salmon gonadotropin-releasing hormone analogue (sGnRHa) in inducing maturation and ovulation. Plasma steroid levels (17s-estradiol (E2) and testosterone) were determined and germinal vesicle (GV) was observed prior to hormone administration at (0), 6, 12 and 24 h post injection. From the first experiment, cGnRH-II 200
g/kg was found to be effective to induce ovulation in African catfish. 100% ovulation was also observed for the fish treated with cGnRH-II 200
g/kg with the combination of pimozide based on the second experiment. Finally in the third experiment, sGnRHa was proven to be more efficient compared to cGnRH-II, only 20
g/kg was administered intramuscularly in single injection. For the group that showed ovulation, that is cGnRH 200
g/kg alone and with the combination of pimozide and also sGnRHa, plasma level of E2 and T increased significantly with the association of germinal vesicle migration and ultimately ovulation. The concentration reached the peaked after 12 h, then decreased dramatically at 24 h post treatment. In contrast to the non-ovulated group that is cGnRH-II 2 and 20
g/kg, the plasma level showed small increased after the injection and throughout the experiment. The present study indicates that administration of cGnRH-II is effective for sexual maturation and ovulation in African catfish.

Major Genomic Events and Their Consequences for Vertebrate Evolution and Endocrinology

Comparative studies of proteins often face the problem of distinguishing a true orthologue (species homologue) from a paralogue (a gene duplicate). This identification task is particularly challenging for endocrine peptides and neuropeptides because they are short and usually have several invariant positions. For some peptide families, this has led to a terminology with peptide names relating to the first species where a specific peptide sequence was determined, such as chicken or salmon gonadotropin-releasing hormone, or names that highlight amino acid differences, e.g., Lys-vasopressin. With accumulating information from multiple species, such a terminology becomes almost impenetrable for nonexperts and difficult even for aficionados. The sequenced genomes offer a new way to distinguish orthologues and paralogues, namely by location of the genes relative to neighboring genes on the chromosomes. In addition, the genome databases can ideally provide a complete listing of the family members in each species. Many vertebrate gene families have expanded in the two basal tetraploidizations (2R) and the teleost fish third tetraploidization (3R), after which some vertebrate lineages have lost some of the duplicates. We review here some peptide families (neuropeptide Y, oxytocin-vasopressin, and somatostatin) where genomic information helps simplify nomenclature. This approach is useful also for other gene families, such as peptide receptors.

Induction of final oocyte maturation in Cyprinidae fish by hypothalamic factors: a review

Gonadotropin-releasing hormone in Cyprinidae as in other Vertebrates functions as a brain signal which stimulates the secretion of luteinizing hormone from the pituitary gland. Two forms of gonadotropin-releasing hormone have been identified in cyprinids, chicken gonadotropin-releasing hormone II and salmon gonadotropin-releasing hormone. Hypohysiotropic functions are fulfilled mainly by salmon gonadotropin-releasing hormone. The only known factor having an inhibitory effect on LH secretion in the family Cyprinidae is dopamine. Most cyprinids reared under controlled conditions exhibit signs of reproductive dysfunction, which is manifested in the failure to undergo final oocyte maturation and ovulation. In captivity a disruption of endogenous gonadotropin-releasing hormone stimulation occurs and sequentially that of luteinizing hormone, which is indispensible for the final phases of gametogenesis. In addition to methods based on the application of exogenous gonadotropins, the usage of a method functioning on the basis of hypothalamic control of final oocyte maturation and ovulation has become popular recently. The replacement of natural gonadotropin-releasing hormones with chemically synthesized gonadotropin-releasing hormone analogues characterized by amino acid substitutions at positions sensitive to enzymatic degradation has resulted in a centuple increase in the effectiveness of luteinizing hormone secretion induction. Combining gonadotropin-releasing hormone analogues with Dopamine inhibitory factors have made it possible to develop an extremely effective agent, which is necessary for the successful artificial reproduction of cyprinids.

Interactions between gonadotropin-releasing hormone (GnRH) and orexin in the regulation of feeding and reproduction in goldfish ( Carassius auratus)

Links between energy homeostasis and reproduction have been demonstrated in vertebrates. As a general rule, abundant food resources favor reproduction whereas low food availability induces an inhibition of reproductive processes. In both mammals and fish, gonadotropin-releasing hormone (GnRH) and orexin (OX) are hypothalamic neuropeptides that play critical roles in the regulation of sexual behavior and appetite, respectively. In order to assess possible interactions between orexin and GnRH in the control of feeding and reproduction in goldfish, we examined the effects of chicken GnRH (cGnRH-II) intracerebroventricular (ICV) injection on feeding behavior and OX brain mRNA expression as well as the effects of orexin ICV injections on spawning behavior and cGnRH-II brain mRNA expression. Treatment with cGnRH-II at doses that stimulate spawning (0.5 ng/g or 1 ng/g) resulted in a decrease in both food intake and hypothalamic orexin mRNA expression. Treatment with orexin A at doses that stimulate feeding (10 ng/g) induced an inhibition of spawning behavior and a decrease in cGnRH-II expression in the hypothalamus and optic tectum-thalamus. Our results suggest that the anorexigenic actions of cGnRH-II in goldfish might be in part mediated by OX and that orexin inhibits reproductive behavior in part via the inhibition of the GnRH system. Our data suggest the existence of a coordinated control of feeding and reproduction by the orexin and GnRH systems in goldfish.

Inhibitory effect of chicken gonadotropin-releasing hormone II on food intake in the goldfish, Carassius auratus

Gonadotropin-releasing hormone (GnRH) is an evolutionarily conserved neuropeptide with 10 amino acid residues, which possesses some structural variants. A molecular form known as chicken GnRH II ([His 5 Trp 7 Tyr 8] GnRH, cGnRH II) is widely distributed in vertebrates, and has recently been implicated in the regulation of sexual behavior and food intake in an insectivore, the musk shrew. However, the influence of cGnRH II on feeding behavior has not yet been studied in model animals such as rodents and teleost fish. In this study, therefore, we investigated the role of cGnRH II in the regulation of feeding behavior in the goldfish, and examined its involvement in food intake after intracerebroventricular (ICV) administration. ICV-injected cGnRH II at graded doses, from 0.1 to 10 pmol/g body weight (BW), induced a decrease of food consumption in a dose-dependent manner during 60 min after treatment. Cumulative food intake was significantly decreased by ICV injection of cGnRH II at doses of 1 and 10 pmol/g BW during the 60-min post-treatment observation period. ICV injection of salmon GnRH ([Trp 7 Leu 8] GnRH, sGnRH) at doses of 0.1–10 pmol/g BW did not affect food intake. The anorexigenic action of cGnRH II was completely blocked by treatment with the GnRH type I receptor antagonist, Antide. However, the anorexigenic action of cGnRH II was not inhibited by treatment with the corticotropin-releasing hormone (CRH) 1/2 receptor antagonist, α-helical CRH (9−41), and the melanocortin 4 receptor antagonist, HS024. These results suggest that, in the goldfish, cGnRH II, but not sGnRH, acts as an anorexigenic factor, as is the case in the musk shrew, and that the anorexigenic action of cGnRH II is independent of CRH- and melanocortin-signaling pathways.

Gonadotropin-Releasing Hormone in the Ovary

Gonadotropin-releasing hormone (GnRH) plays a pivotal role in the physiology of reproduction in mammals. GnRH acts by binding to the GnRH receptor (GnRHR). In humans, only 1 conventional GnRH receptor subtype (type I GnRH receptor) has been found. In the human genome, 2 forms of GnRH have been identified, GnRH-I (mammal GnRH) and GnRH-II (chicken GnRH II). Both forms and their common receptor are expressed, apart from the hypothalamus, in various compartments of the human ovary. Gonadal steroids, gonadotropins, and GnRH itself controls the regulation of the GnRH/GnRHR system gene expression in the human ovary. The 2 types of GnRH acting paracrinally/autocrinally influence ovarian steroidogenesis, decrease the proliferation, and induce apoptosis of ovarian cells. In this review, the biology of GnRH/GnRHR system in humans, the potential roles of GnRH, and the direct effects of GnRH analogues in ovarian cells are discussed.

Associations of Gonadotropin-Releasing Hormone Receptor (GnRHR) and Neuropeptide Y (NPY) Genes' Polymorphisms with Egg-Laying Traits in Wenchang Chicken

Single nucleotide polymorphisms (SNP) of chicken gonadotropin-releasing hormone receptor (GnRHR) and neuropeptide Y (NPY) were selected to identify the genotypes of Wenchang (Chinese indigenous breed) chicken with restricton fragment length polymorphisms. The associations of the SNPs with the total egg production (NE), average days of continual laying (ADCL), and number of double-yolked eggs (DYE) traits were analyzed. The frequency of restriction enzyme A/a alleles in the population was for GnRHR 0.69 ( Bpul102 I A) and 0.31 ( Bpul102 I a) and for NPY 0.46 ( Dra I B) and 0.54 ( Dra I b). Trait data from a total of 120 hens, which were purebred introduced from Hainan Province, China from one generation were recorded. Two significant effects of genes' marker were found: for GnRHR and number of eggs (dominant; t=2.67, df= 116) and NPY and number of eggs (additive; t= 1.97, df= 116). The current research supports the effects of GnRHR and NPY genes on egg-laying traits of chickens.

Identification of a Novel Pituitary-Specific Chicken Gonadotropin-Releasing Hormone Receptor and Its Splice Variants1

In all vertebrates, GnRH regulates gonadotropin secretion through binding to a specific receptor on the surface of pituitary gonadotropes. At least two forms of GnRH exist within a single species, and several corresponding GnRH receptors (GNRHRs) have been isolated with one form being pituitary specific. In chickens, only one type of widely expressed GNRHR has previously been identified. The objectives of this study were to isolate a chicken pituitary-specific GNRHR and to determine its expression pattern during a reproductive cycle. Using a combined strategy of PCR and rapid amplification of cDNA ends (RACE), a new GNRHR (chicken GNRHR2) and two splice variants were isolated in domestic fowl (Gallus gallus domesticus). Full-length GNRHR2 and one of its splice variant mRNAs were expressed exclusively in the pituitary, whereas mRNA of the other splice variant was expressed in most brain tissues examined. The deduced amino acid sequence of full-length chicken GNRHR2 reveals a seven transmembrane domain protein with 57%-65% homology to nonmammalian GNRHRs. Semiquantitative real-time PCR revealed that mRNA levels of full-length chicken GNRHR2 in the pituitary correlate with the reproductive status of birds, with maximum levels observed during the peak of lay and 4 wk postphotostimulation in females and males, respectively. Furthermore, GnRH stimulation of GH3 cells that were transiently transfected with cDNA that encodes chicken GNRHR2 resulted in a significant increase in inositol phosphate accumulation. In conclusion, we isolated a novel GNRHR and its splice variants in chickens, and spatial and temporal gene expression patterns suggest that this receptor plays an important role in the regulation of reproduction.

Rapid inhibition of female sexual behavior by gonadotropin-inhibitory hormone (GnIH)

Gonadotropin-releasing hormone (GnRH) is largely responsible for the initiation of sexual behaviors; one form of GnRH activates a physiological cascade causing gonadal growth and gonadal steroid feedback to the brain, and another form is thought to act as a neurotransmitter to enhance sexual receptivity. In contrast to GnRH, gonadotropin-inhibitory hormone (GnIH) inhibits gonadotropin release. The distribution of GnIH in the avian brain suggests that it has not only hypophysiotropic actions but also unknown behavioral actions. GnIH fibers are present in the median eminence (ME) and are in apparent contact with chicken GnRH (cGnRH)-I and -II neurons and fibers. In birds, cGnRH-I regulates pituitary gonadotropin release, whereas cGnRH-II enhances copulation solicitation in estradiol-primed females exposed to male song. In the present study, we determined the effects of GnIH administered centrally to female white-crowned sparrows. A physiological dose of GnIH reduced circulating LH and inhibited copulation solicitation, without affecting locomotor activity. Using rhodaminated GnIH, putative GnIH binding sites were seen in the ME close to GnRH-I fiber terminals and in the midbrain on or close to GnRH-II neurons. These data demonstrate direct effects of GnIH upon reproductive physiology and behavior, possibly via separate actions on two forms of GnRH.