Gonadotropin-inhibitory Hormone Research Articles

Immunoreactive distribution of gonadotropin-inhibitory hormone precursor, RFRP, in a temperate bat species (Eptesicus fuscus).

Gonadotropin-inhibitory hormone (GnIH, also known RFRP-3 in mammals) is an important regulator of the hypothalamic-pituitary-gonadal axis and downstream reproductive physiology. Substantial species differences exist in the localization of cell bodies producing RFRP-3 and patterns of fiber immunoreactivity in the brain, raising the question of functional differences. Many temperate bat species exhibit unusual annual reproductive patterns. Male bats upregulate spermatogenesis in late spring which is asynchronous with periods of mating in the fall, while females have the physiological capacity to delay their reproductive investment over winter via sperm storage or delayed ovulation/fertilization. Neuroendocrine mechanisms regulating reproductive timing in male and female bats are not well-studied. We provide the first description of RFRP-precursor peptide of GnIH -expression and localization in the brain of any bat using a widespread temperate species (Eptesicus fuscus, big brown bat) as a model. RFRP mRNA expression was detected in the hypothalamus, testes, and ovaries of big brown bats. Cellular RFRP-immunoreactivity was observed within the periventricular nuclei, dorsomedial nucleus of the hypothalamus, arcuate nucleus (Arc), and median eminence (ME). As in other vertebrates, RFRP fiber immunoreactivity was widespread, with the greatest density observed in the hypothalamus, preoptic area, Arc, ME, midbrain, and thalamic nuclei. Putative interactions between RFRP-ir fibers and gonadotropin-releasing hormone (GnRH) cell bodies were observed in 16% of GnRH-immunoreactive cells, suggesting direct regulation of GnRH via RFRP signaling. This characterization of RFRP distribution contributes to a deeper understanding of bat neuroendocrinology, which serves as foundation for manipulative approaches examining changes in reproductive neuropeptide signaling in response to environmental and physiological challenges within, and among, bat species.

Gonadotropin Inhibitory Hormone and Its Receptor: Potential Key to the Integration and Coordination of Metabolic Status and Reproduction.

Since its discovery as a novel gonadotropin inhibitory peptide in 2000, the central and peripheral roles played by gonadotropin-inhibiting hormone (GnIH) have been significantly expanded. This is highlighted by the wide distribution of its receptor (GnIH-R) within the brain and throughout multiple peripheral organs and tissues. Furthermore, as GnIH is part of the wider RF-amide peptides family, many orthologues have been characterized across vertebrate species, and due to the promiscuity between ligands and receptors within this family, confusion over the nomenclature and function has arisen. In this review, we intend to first clarify the nomenclature, prevalence, and distribution of the GnIH-Rs, and by reviewing specific localization and ligand availability, we propose an integrative role for GnIH in the coordination of reproductive and metabolic processes. Specifically, we propose that GnIH participates in the central regulation of feed intake while modulating the impact of thyroid hormones and the stress axis to allow active reproduction to proceed depending on the availability of resources. Furthermore, beyond the central nervous system, we also propose a peripheral role for GnIH in the control of glucose and lipid metabolism at the level of the liver, pancreas, and adipose tissue. Taken together, evidence from the literature strongly suggests that, in fact, the inhibitory effect of GnIH on the reproductive axis is based on the integration of environmental cues and internal metabolic status.

Neuroendocrine mechanisms of reproductive dysfunctions in undernourished condition.

It is well known that undernourished conditions disturb female reproductive functions in many species, including humans. These alterations are mainly caused by a reduction in gonadotrophin-releasing hormone (GnRH) secretion from the hypothalamus. Evidence from the literature suggests that some hypothalamic factors play pivotal roles in the coordination of reproductive functions and energy homeostasis in response to environmental cues and internal nutritional status. Generally, anorexigenic/satiety-related factors, such as leptin, alpha-melanocyte-stimulating hormone, and proopiomelanocortin, promote GnRH secretion, whereas orexigenic factors, such as neuropeptide Y, agouti-related protein, orexin, and ghrelin, attenuate GnRH secretion. Conversely, gonadotrophin-inhibitory hormone, which exerts anti-GnRH and gonadotrophic effects, promotes feeding behavior in many species. In addition, the activity of kisspeptin, which is a potent stimulator of GnRH, is reduced by undernourished conditions. Under normal nutritional conditions, these factors are coordinated to maintain both feeding behavior and reproductive functions. However, in undernourished conditions their activity levels are markedly altered to promote feeding behavior and temporarily suppress reproductive functions, in order to prioritize the survival of the individual over that of the species.

Seasonal variations in gonad morphology and hypothalamic GnRH-I and GnIH in Eurasian Tree Sparrow, a multi-brooded passerine

The hypothalamic-pituitary-gonadal (HPG) axis ubiquitously regulates seasonal reproduction, following the rhythmicity of a suite of environmental cues. Birds display prominent seasonal variations in gonad size regulated by two hypothalamic peptides, gonadotropin-releasing hormone-1 (GnRH-I) and gonadotropin-inhibiting hormone (GnIH). How multi-brooded avian species adjust GnRH-I and GnIH to regulate gonadal morphology seasonally remains largely unknown. Here, we studied the variations in the hypothalamic immunoreactivity (ir) of GnRH-I and GnIH, gonadal proliferation, and apoptosis in a typical multi-brooded species, the Eurasian Tree Sparrow (Passer montanus), across the pre-breeding (PB), first breeding (FB), second breeding (SB), pre-basic molt (PM), and wintering stages (WS). Our results showed that both sexes had higher preoptic area (POA)-GnRH-I-ir but lower paraventricular nucleus (PVN)-GnIH-ir neurons during the breeding stages (FB and SB) relative to other life-history stages, with no significant differences between two broods. The testes and ovaries were significantly larger during the breeding stages. Testicular volume increased during the two broods due to anincreased diameter of seminiferous tubules. Furthermore, there were more testicular apoptotic cells in PB and WS stages than in breeding stages, and in PB stage than in PM stage. Males had higher POA-GnRH-I expression than females during the breeding stages, but both sexes had comparable PVN-GnIH expression throughout the annual cycle. Both sexes of the sparrows may undergo a similar pattern of life-history stage-dependent variation in the hypothalamic GnRH-I, GnIH, and gonadal morphology, except that during breeding stages, males may display higher expression of POA-GnRH-I relative to females. The higher expression of POA-GnRH-I-ir in breeding male sparrows may be critical for male-dependent parental care.

Effects of undernutrition and low energy availability on reproductive functions and their underlying neuroendocrine mechanisms.

It has been well established that undernutrition and low energy availability disturb female reproductive functions in humans and many animal species. These reproductive dysfunctions are mainly caused by alterations of some hypothalamic factors, and consequent reduction of gonadotrophin-releasing hormone (GnRH) secretion. Evidence from literature suggests that increased activity of orexigenic factors and decreased activity of anorexigenic/satiety-related factors in undernourished conditions attenuate GnRH secretion in an integrated manner. Likewise, the activity of kisspeptin neurons, which is a potent stimulator of GnRH, is also reduced in undernourished conditions. In addition, it has been suggested that gonadotrophin-inhibitory hormone, which has anti-GnRH and gonadotrophic effects, may be involved in reproductive dysfunctions under several kinds of stress conditions. It should be remembered that these alterations, i.e., promotion of feeding behavior and temporary suppression of reproductive functions, are induced to prioritize the survival of individual over that of species, and that improvements in metabolic and nutritional conditions should be considered with the highest priority.

WITHDRAWN: Distribution of the Novel RFRP-3/receptors system in the epididymis of the seasonal desert rodent, Gerbillus tarabuli, during sexual activity

RFamide-related peptide (RFRP-3), the Mammalian ortholog of the Avian gonadotropin-inhibitory hormone (GnIH), is a novel neuropeptide known for its inhibitory regulatory effect on reproduction in various mammalian species. However, a stimulatory action has been reported. This paper aims to: i) study the histology of the epididymis (caput) of Gerbillus tarabuli during the breeding period; and ii) to determine the distribution of the “RFRP-3/receptors system” in the epididymis (caput) of this desert rodent during the active season, and thus, to inspect its potential local interfering in sperm maturation. For that, immunohistochemistry was performed to detect the epididymal immunolocalizations of the three molecules, RFRP-3, GPR147, and GPR74. This is the first report of the epididymis histology in Gerbillus tarabuli, as it is the first evidence of the existence of the RFRP-3/Receptor system in the same organ of the same species. During the breeding season, moderate immunostaining of the RFRP-3/receptors system was present in the caput epididymis’ epithelial parts (basal and principal cells) and spermatozoa. In contrast, these three molecules were absent in the peritubular and muscle coat's myoid cells and of the interstitial part of the caput epididymis. The results suggest that the epididymis is a potential source of RFRP-3 in the desert Rodent, Gerbillus tarabuli, which may function as a paracrine and/or autocrine factor affecting the main epididymis’ function: sperm maturation.

Gonadotropin inhibitory hormone receptors (GnIHRs): Molecular characterization and synergistic effect of different drugs in Indian major carp, Labeo catla

After the discovery of Gonadotropin-inhibitory hormone (GnIH) in birds in 2000, it showed different roles in different vertebrate classes and even in different species of same classes. In birds and mammals, GnIH inhibits the expression of gonadotropins during reproduction, while in fishes it exerts both inhibitory and stimulatory effect on reproduction. The current study evaluates the role of GnIH during reproduction in Labeo catla. The partial cDNA sequence of GnIHR1 and GnIHR3 receptor genes was identified by degenerate PCR. The mRNA expression analysis of GnIHRs during different reproductive phases showed that the expression of all three GnIH receptor genes is highest during spawning phase. The expression of GnIH receptors is detected in both brain and gonads except for GnIHR3 which only expressed in gonads. The in vivo experiments with GnIH antagonist, RF313 drastically reduced the expression level of reproduction related genes like LH, FSH, and GnRH at 1 h post-injection. In another experiment the surge induced by cGnIH-III peptide on gonadotropins gene expression is further increased when co-injected with LHRHa. However, co-injection of melatonin along with cGnIH-III peptide had opposite effects. These results showed that the GnIH/GnIHRs system has positive effect on reproduction in L. catla.

The Role of GnIH in Biological Rhythms and Social Behaviors.

Gonadotropin-inhibitory hormone (GnIH) was first discovered in the Japanese quail, and peptides with a C-terminal LPXRFamide sequence, the signature protein structure defining GnIH orthologs, are well conserved across vertebrate species, including fish, reptiles, amphibians, avians, and mammals. In the mammalian brain, three RFamide-related proteins (RFRP-1, RFRP-2, RFRP-3 = GnIH) have been identified as orthologs to the avian GnIH. GnIH is found primarily in the hypothalamus of all vertebrate species, while its receptors are distributed throughout the brain including the hypothalamus and the pituitary. The primary role of GnIH as an inhibitor of gonadotropin-releasing hormone (GnRH) and pituitary gonadotropin release is well conserved in mammalian and non-mammalian species. Circadian rhythmicity of GnIH, regulated by light and seasons, can influence reproductive activity, mating behavior, aggressive behavior, and feeding behavior. There is a potential link between circadian rhythms of GnIH, anxiety-like behavior, sleep, stress, and infertility. Therefore, in this review, we highlight the functions of GnIH in biological rhythms, social behaviors, and reproductive and non-reproductive activities across a variety of mammalian and non-mammalian vertebrate species.

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.

Widespread alterations to hypothalamic-pituitary-gonadal (HPG) axis signaling underlie high temperature reproductive inhibition in the eurythermal sheepshead minnow (Cyprinodon variegatus)

Fish experiencing abnormally high or prolonged elevations in temperature can exhibit impaired reproduction, even for species adapted to warm water environments. Such high temperature inhibition of reproduction has been linked to diminished gonadal steroidogenesis, but the mechanisms whereby hypothalamic-pituitary-gonadal (HPG) axis signaling is impacted by high temperature are not fully understood. Here, we characterized differences in HPG status in adult sheepshead minnow (Cyprinodon variegatus), a eurythermal salt marsh and estuarine species of eastern North America, exposed for 14 d to temperatures of 27 °C or 37 °C. Males and females at 37 °C had lower gonadosomatic index (GSI) values compared to fish at 27 °C, and females at 37 °C had fewer spawning capable eggs and lower circulating 17β-estradiol (E2). Gene transcripts encoding gonadotropin-inhibitory hormone (gnih) and gonadotropin-releasing hormone-3 (gnrh3) were higher in relative abundance in the hypothalamus of both sexes at 37 °C. While pituitary mRNAs for the β-subunits of follicle-stimulating hormone (fshβ) and luteinizing hormone (lhβ) were lowered only in males at 37 °C, Fsh and Lh receptor mRNA levels in the gonads were at lower relative levels in both the ovary and testis of fish at 37 °C. Females at 37 °C also showed reduced ovarian mRNA levels for steroid acute regulatory protein (star), P450 side-chain cleavage enzyme (cyp11a1), 3β-hydroxysteroid dehydrogenase (3βhsd), 17β-hydroxysteroid dehydrogenase (hsd17β3), and ovarian aromatase (cyp19a1a). Females at the higher 37 °C temperature also had a lower liver expression of mRNAs encoding estrogen receptor α (esr1) and several vitellogenin and choriogenin genes, but elevated mRNA levels for hepatic sex hormone-binding globulin (shbg). Our results substantiate prior findings that exposure of fish to high temperature can inhibit gonadal steroidogenesis and oogenesis, and point to declines in reproductive performance emerging from alterations at several levels of HPG axis signaling including increased hypothalamic Gnih expression, depressed gonadal steroidogenesis, and reduced egg yolk and egg envelope protein production in the liver.

LPXRFa down-regulates brain reproductive genes in yellowtail kingfish (Seriola lalandi)

Gonadotropin-inhibitory hormone (GnIH) is a hypothalamic peptide that inhibits the reproductive axis of mammals and birds. Little information is available in teleosts regarding its actions on the brain neuroendocrine systems controlling reproduction. Here, we analyzed the effects of intraperitoneal injection of the piscine GnIH ortholog LPXRFa peptides (LPXRFa-1, LPXRFa-2, and LPXRFa-3) on the expression of reproduction-related genes in the brain of yellowtail kingfish. Our results revealed an inhibitory effect of the three LPXRFa peptides on gnrh1, lpxrfa, kiss1r, kiss2r, and lpxrfa-r mRNA levels. Both LPXRFa-1 and LPXRFa-2 were found to suppress kiss2 expression, and LPXRFa-1 reduced gnrh2 expression. Moreover, LPXRFa-3 induced a decrease in kiss1 and gnrh2 expression. Overall, these data suggest the inhibitory actions of each LPXRFa peptide encoded by the same precursor on the brain reproductive genes in yellowtail kingfish.

LPXRFa and its receptor in yellowtail kingfish (Seriola lalandi): Molecular cloning, ontogenetic expression profiles, and stimulatory effects on growth hormone and gonadotropin gene expression

Despite its functional significance in mammals and birds, the biological role of gonadotropin-inhibitory hormone (GnIH) in reproduction is still far from being fully understood in teleosts. In the current study, we have identified LPXRFa, the piscine ortholog of GnIH, and its cognate receptor (LPXRFa-R) in yellowtail kingfish (YTK), which is considered as a promising species for aquaculture industry worldwide. The YTK cDNA sequence of lpxrfa was 534 base pair (bp) in length and encoded a 178-amino acids (aa) preprohormone. The LPXRFa precursor comprised three putative peptide sequences that included –MPMRF, –MPQRF, or -LPERL motifs at the C-termini, respectively. The YTK lpxrfa-r cDNA sequence was composed of 1265 bp that gave rise to a LPXRFa-R of 420 aa, encompassing the characteristic seven hydrophobic transmembrane domains. In males, both lpxrfa and lpxrfa-r transcripts could be detected at high levels in the brain and testis. In females, a noteworthy expression of lpxrfa was observed in the brain and ovary, while the expression of lpxrfa-r was especially evident only in the brain. To study the ontogeny of LPXRFa system, transcript levels were also investigated during early life stages. Variable expression of the LPXRFa system was observed during all stages of YTK embryogenesis. The highest expression of lpxrfa and lpxrfa-r were noticed at 7 dph and 15 dph, respectively. Furthermore, LPXRFa peptides stimulated growth hormone (gh), luteinizing hormone (lhβ) and follicle-stimulating hormone (fshβ) gene expression from the pituitary. Taken together, our results provide initial evidence for the existence of the LPXRFa system in yellowtail kingfish and suggest its possible involvement at early development and reproductive functions.

Leptin Modulates the mRNA Expression of Follicle Development Markers in Post-hatch Chicks in an Age-Dependent Manner.

Leptin is involved in regulating reproductive function in chickens, and the development of the leptin system is initiated during the early embryonic stage; however, whether leptin has a specific role in regulating the ovarian development in early post-hatch days is still not fully understood. This study investigated the expression of ovarian functional markers in growing juvenile chickens, along with the effects of leptin on gene expression in the hypothalamus–pituitary–gonadal (HPG) axis on specific ovarian-remodeling days. Leptin receptor (LEPR), follicle-stimulating hormone receptor (FSHR), and the mRNA expression of aromatase (CYP19A1) tended to increase with age in the ovaries of growing chicks. In the ovaries of 7-day-old chicks, intraperitoneally injected leptin significantly increased the mRNA expressions of LEPR, FSHR, and CYP19A1, and this resulted in the increased serum estradiol levels. However, leptin had no effect on hypothalamic LEPR, gonadotropin-releasing hormone 1 (GnRH1), or gonadotropin-inhibitory hormone (GnIH) mRNAs; however, in the pituitary gland, leptin significantly increased the mRNA expression of luteinizing hormone beta subunit (LHB) but had no effect on the mRNA expression of follicle-stimulating hormone beta subunit (FSHB). In 28-day-old chicks, hypothalamic and pituitary mRNAs were unaffected by leptin administration, except hypothalamic LEPR mRNA that was upregulated by a high dose of leptin. In the ovary, leptin dose-dependently decreased the mRNA expression of LEPR; low doses of leptin significantly increased the mRNA expression of FSHR, whereas high doses significantly decreased this expression; leptin did not affect the mRNA expression of CYP19A1; and high leptin doses significantly reduced the serum estradiol levels. Collectively, the results of this study show that leptin modulates ovarian development and folliculogenesis marker genes by primarily acting on ovaries on the specific ovarian-remodeling days in post-hatch chicks, which may alter folliculogenesis and ovarian development toward puberty in chicken.

Effects of LPXRFamide peptides on chub mackerel gonadotropin secretion†.

Gonadotropin-inhibitory hormone (GnIH), a neuropeptide, suppresses gonadotropin (GTH) secretion in birds and mammals. In fish, the GnIH homolog LPXRFamide (LPXRFa) produces mature peptides with species-dependent effects on sexual reproduction. Here, we investigated the effects of LPXRFa on GTH secretion in the chub mackerel (cm; Scomber japonicus). We cloned cmlpxrfa (603bp) and cmlpxrfa-r (1416bp). Additionally, we isolated lpxrfa from the bluefin tuna (Thunnus orientalis) to confirm the conservation of the LPXRFa mature sequence. Phylogenetic analysis showed that the LPXRFa precursor protein produces three mature peptides, LPXRFa-1, -2, and -3, in both species. Reverse transcription-quantitative polymerase chain reaction revealed that cmlpxrfa is expressed in the hypothalamus and thalamus and midbrain (T.MB), and sexual differences were observed. Receptor expression was observed in the preoptic area, hypothalamus, T.MB, and pituitary. Female hypothalamic lpxrfa expression did not change during puberty. Reporter gene assay showed that LPXRFa induced receptor activation via the CRE and SRE signaling pathways. However, in the presence of forskolin, an intracellular cyclic AMP enhancer, none of the LPXRFa could suppress receptor activity. The in vitro bioassay results showed that gonadotropin-releasing hormone-1 (GnRH1) had no effect on follicle-stimulating hormone (FSH) secretion, whereas the three LPXRFa significantly increased FSH secretion in pituitary cells from male chub mackerel. Contrarily, GnRH1 and three LPXRFa significantly increased luteinizing hormone secretion. The in vivo administration of LPXRFa had no effect on fshb and lhb expression in prepubertal and mature male chub mackerel. Overall, cmLPXRFa lacks the ability to suppress GTH secretion but can promote GTH secretion.

Interaction between thyroid hormones and gonadotropin inhibitory hormone in ex vivo culture of zebrafish testis: An approach to study multifactorial control of spermatogenesis

Reproduction is under multifactorial control of neurohormones, pituitary gonadotropins, as well as of local gonadal signaling systems including sex steroids, growth factors and non-coding RNAs. Among the factors, gonadotropin-inhibitory hormone (Gnih) is a novel RFamide neuropeptide which directly modulates gonadotropin synthesis and release from pituitary, and in the gonads, Gnih mediated inhibitory actions on gonadotropin response of zebrafish spermatogenesis. Thyroid hormones are peripheral hormones which are also known to interact with reproductive axis, in particular, regulating testicular development and function. This study investigated the interaction between Gnih and thyroid hormones in zebrafish spermatogenesis using in vivo and ex vivo approaches. Three experimental groups were established: “control” (non-treated fish), “methimazole” and “methimazole + T4”. Fish were exposed to goitrogen methimazole for 3 weeks; T4 (100 μg/L) was added in the water from the second week only in the “reversal treatment” group. After exposure, testes were dissected out and immediately incubated in Leibovitz's L-15 culture medium containing hCG, Gnih or hCG + Gnih for 7 days. Germ cell cysts and haploid cell population were evaluated by histomorphometry and flow cytometry, respectively. Our results showed that hypothyroidism affected germ cell development in basal and gonadotropin-induced spermatogenesis, in particular, meiosis and spermiogenesis. Hypothyroid testes showed lower amount of spermatozoa, and decreased potency of hCG. We also showed that goitrogen treatment nullified the inhibitory actions of Gnih on the gonadotropin-induced spermatogenesis. This study provided evidences that thyroid hormones are important regulatory factors for hCG- and Gnih-mediated functions in zebrafish spermatogenesis.

Changes in the Control of the Hypothalamic-Pituitary Gonadal Axis Across Three Differentially Selected Strains of Laying Hens (Gallus gallus domesticus).

Genetic selection for earlier sexual maturation and extended production cycles in laying hens has significantly improved reproductive efficiency. While limited emphasis has been placed on the underlying physiological changes, we hypothesize that modifications in the control of the hypothalamic-pituitary gonadal (HPG) axis have occurred. Thus, three strains of White leghorn derivatives were followed from hatch to 100 weeks of age (woa), including Lohmann LSL-lite (n = 120) as current commercial hens, heritage Shaver White leghorns (n = 100) as 2000s commercial equivalents, and Smoky Joe hens (n = 68) as 1960s commercial equivalents. Body weight (BW) and egg production were monitored, and blood samples were collected throughout to monitor estradiol (E2) concentrations. Tissue samples were collected at 12, 17, 20, 25, 45, 60, 75, and 100 woa to capture changes in mRNA levels of key genes involved in the HPG axis and monitor ovarian follicular pools. All hens, regardless of strain, age or photoperiod laid their first egg within a 64-gram BW window and, as E2 levels increased prior to photostimulation (PS) in Lohmann and Shaver hens, a metabolic trigger likely induced sexual maturation. However, increased levels of Opsin 5 (OPN5) were observed during the maturation period. Although an elevation in gonadotrophin-releasing hormone I (GnRH-I) mRNA levels was associated with early maturation, no changes in gonadotrophin-inhibitory hormone (GnIH) mRNA levels were observed. Nonetheless, a significant shift in pituitary sensitivity to GnRH was associated with maturation. Throughout the trial, Lohmann, Shaver, and Smoky Joe hens laid 515, 417, and 257 eggs, respectively (p < 0.0001). Results show that the extended laying persistency in Lohmann hens was supported by sustained pituitary sensitivity to GnRH-I, recurrent elevations in follicle-stimulating hormone (FSH) mRNA levels, and five cyclical elevations in E2 levels. This was also associated with a consistently higher pool of small white ovarian follicles. In summary, our results demonstrate first that, regardless of photoperiodic cues, meeting a specific narrow body weight threshold is sufficient to initiate sexual maturation in Leghorn chicken derivatives. Furthermore, recurrent increases in E2 and FSH may be the key to sustain extended laying period, allowing modern layers to double their reproductive capacity compared to their 1960s-counterparts.

Urban environment alter the timing of progression of testicular recrudescence in tree sparrow (Passer montanus).

Urbanization is a rapidly growing phenomenon that affects wildlife. Laboratory studies show the effects of night light on the physiology of the organisms. Limited studies have been conducted on birds in their natural habitat. Here, we studied the effects of the urban environment on reproduction-linked phenomenon and molecules involved in the regulation of seasonal breeding. Birds (N=5/time/site) were procured from urban and rural sites at specific times, i.e., in March (stimulatory phase), June (reproductive phase), September (refractory phase), and December (sensitive phase) of 2018. Immediately after procurement, birds were brought to the laboratory. Bodyweight, bill color, molt in body feathers, and testes size were recorded. The next day, all the birds were sacrificed in the middle of the day. Blood was collected and serum was used for ELISA of corticosterone, triiodothyronine (T3), and thyroxine (T4). mRNA levels of thyroid-stimulating hormone-β (Tshβ), type 2 deiodinase (Dio2), type 3 deiodinase (Dio3), gonadotropin-releasing hormone (GnRh), and gonadotropin inhibitory hormone (GnIh) were measured in hypothalamic tissue. Urban birds showed higher levels of corticosterone during the stimulatory phase. There was a delay in the initiation of testicular growth in urban birds and it was supported by reduced levels of T3 in blood plasma and relatively lower transcription of Dio2 and GnRH mRNA in urban birds. Our findings suggest that the urban environment delays the timing of reproduction in birds and could be the consequence of local environmental conditions.

An Inhibitory Circuit From Brainstem to GnRH Neurons in Male Mice: A New Role for the RFRP Receptor

RFamide-related peptides (RFRPs, mammalian orthologs of gonadotropin-inhibitory hormone) convey circadian, seasonal and social cues to the reproductive system. They regulate gonadotropin secretion by modulating gonadotropin-releasing hormone (GnRH) neurons via the RFRP receptor. Mice lacking this receptor are fertile but exhibit abnormal gonadotropin responses during metabolic challenges such as acute fasting, when the normal drop in gonadotropin levels is delayed. Although it is known that these food intake signals to the reproductive circuit originate in the nucleus tractus solitarius (NTS) in the brainstem, the phenotype of the neurons conveying the signal remains unknown. Given that neuropeptide FF (NPFF), another RFamide peptide, resides in the NTS and can bind to the RFRP receptor, we hypothesized that NPFF may regulate GnRH neurons. To address this question, we used a combination of techniques: cell-attached electrophysiology on GnRH-driven green fluorescent protein-tagged neurons in acute brain slices; calcium imaging on cultured GnRH neurons; and immunostaining on adult brain tissue. We found 1) NPFF inhibits GnRH neuron excitability via the RFRP receptor and its canonical signaling pathway (Gi/o protein and G protein-coupled inwardly-rectifying potassium channels), 2) NPFF-like fibers in the vicinity of GnRH neurons coexpress neuropeptide Y, 3) the majority of NPFF-like cell bodies in the NTS also coexpress neuropeptide Y, and 4) acute fasting increased NPFF-like immunoreactivity in the NTS. Together these data indicate that NPFF neurons within the NTS inhibit GnRH neurons, and thus reproduction, during fasting but prior to the energy deficit.

Acute social defeat stress upregulates gonadotrophin inhibitory hormone and its receptor but not corticotropin-releasing hormone and ACTH in the Male Nile Tilapia (Oreochromis niloticus)

Stress impairs the hypothalamic-pituitary-gonadal (HPG) axis, probably through its influence on the hypothalamic-pituitary-adrenal (= interrenals in the teleost, HPI) axis leading to reproductive failures. In this study, we investigated the response of hypothalamic neuropeptides, gonadotropin-inhibitory hormone (GnIH), a component of the HPG axis, and corticotropin-releasing hormone (CRH) a component of the HPI axis, to acute social defeat stress in the socially hierarchical male Nile tilapia (Oreochromis niloticus). Localization of GnIH cell bodies, GnIH neuronal processes, and numbers of GnIH cells in the brain during acute social defeat stress was studied using immunohistochemistry. Furthermore, mRNA levels of GnIH and CRH in the brain together with GnIH receptor, gpr147, and adrenocorticotropic hormone (ACTH) in the pituitary were quantified in control and socially defeated fish. Our results show, the number of GnIH-immunoreactive cell bodies and GnIH mRNA levels in the brain and the levels of gpr147 mRNA in the pituitary significantly increased in socially defeated fish. However, CRH and ACTH mRNA levels did not change during social defeat stress. Further, we found glucocorticoid type 2b receptor mRNA in laser captured immunostained GnIH cells. These results show that acute social defeat stress activates GnIH biosynthesis through glucocorticoid receptors type 2b signalling but does not change the CRH and ACTH mRNA expression in the tilapia, which could lead to temporary reproductive dysfunction.

RF-amide related peptide-3 (RFRP-3): a novel neuroendocrine regulator of energy homeostasis, metabolism, and reproduction.

A hypothalamic neuropeptide, RF-amide related peptide-3 (RFRP-3), the mammalian ortholog of the avian gonadotropin-inhibitory hormone (GnIH) has inhibitory signals for reproductive axis via G-protein coupled receptor 147 in mammals. Moreover, RFRP-3 has orexigenic action but the mechanism involved in energy homeostasis and glucose metabolism is not yet known. Though, the RFRP-3 modulates orexigenic action in co-operation with other neuropeptides, which regulates metabolic cues in the hypothalamus. Administration of GnIH/RFRP-3 suppresses plasma luteinizing hormone, at the same time stimulates feeding behavior in birds and mammals. Likewise, in the metabolically deficient conditions, its expression is up-regulated suggests that RFRP-3 contributes to the integration of energy balance and reproduction. However, in many other metabolic conditions like induced diabetes and high-fat diet obesity, etc. its role is still not clear while, RFRP-3 induces the glucose homeostasis by adipocytes is reported. The physiological role of RFRP-3 in metabolic homeostasis and the metabolic effects of RFRP-3 signaling in pharmacological studies need a detailed discussion. Further studies are required to find out whether RFRP-3 is associated with restricted neuroendocrine function observed in type II diabetes mellitus, aging, or sub-fertility. In this context, the current review is focused on the role of RFRP-3 in the above-mentioned mechanisms. Studies from search engines including PubMed, Google Scholar, and science.gov are included after following set inclusion/exclusion criteria. As a developing field few mechanisms are still inconclusive, however, based on the available information RFRP-3 seems to be a putative tool in future treatment strategies towards metabolic disease.