Gonadotropin-releasing Hormone Gene Research Articles

Embryonic exposure to benzo(a)pyrene inhibits reproductive capability in adult female zebrafish and correlation with DNA methylation

This study was conducted to investigate the effects of embryonic short-term exposure to benzo(a)pyrene (BaP), a model polycyclic aromatic hydrocarbon, on ovarian development and reproductive capability in adult female zebrafish. In 1-year-old fish after embryonic exposure to BaP for 96 h, the gonadosomatic indices and the percentage of mature oocytes were significantly decreased in the 0.5, 5 and 50 nmol/L treatments. The spawned egg number, the fertilization rate and the hatching success were significantly reduced, while the malformation rate of the F1 unexposed larvae were increased. The mRNA levels of follicle-stimulating hormone, luteinizing hormone, ovarian cytochrome P450 aromatase cyp19a1a and cyp19b, estrogen receptor esr1 and esr2, and hepatic vitellogenin vtg1 and vtg2 genes, were down-regulated in adult female zebrafish that were exposed to BaP during embryonic stage. Both 17β-estradiol and testosterone levels were reduced in the ovary of adult females. The methylation levels of the gonadotropin releasing hormone (GnRH) gene gnrh3 were significantly increased in the adult zebrafish brain, and those of the GnRH receptor gene gnrhr3 were elevated both in the larvae exposed to BaP and in the adult brain, which might cause the down-regulation of the mRNA levels of gnrh3 and gnrhr3. This epigenetic change caused by embryonic exposure to BaP might be a reason for physiological changes along the brain–pituitary–gonad axis. These results suggest that short-term exposure in early life should be included and evaluated in any risk assessment of pollutant exposure to the reproductive health of fish.

Phoenixin: uncovering its receptor, signaling and functions.

Phoenixin (PNX) is a newly discovered peptide that has been linked to reproductive function, both in the hypothalamus and pituitary. This review will focus on the most recent discoveries related to this novel neuropeptide. Initially, it was found that PNX increased gonadotropin releasing hormone (GnRH)-stimulated luteinizing hormone (LH) release from pituitary cells. Importantly, knockdown of PNX in female rats extended the estrous cycle by 2.3 days. Using novel hypothalamic cell lines, we found that PNX has a stimulatory role on kisspeptin (Kiss) and GnRH gene expression and secretion. The PNX receptor was uncovered using siRNA knockdown of GPR173, an orphan receptor postulated to bind PNX. We have found that the PNX-R signaling through protein kinase A (PKA) in hypothalamic neurons. Althuogh a number of studies demonstrate that PNX plays an important role in reproductive function, there is also evidence that it may have other functions, regulating the heart, feeding, memory, and anxiety, both in the brain and the periphery.

Transcriptional interaction between cFOS and the homeodomain-binding transcription factor VAX1 on the GnRH promoter controls Gnrh1 expression levels in a GnRH neuron maturation specific manner

Gonadotropin-releasing hormone (GnRH) is required for pubertal onset and reproduction, thus the control of GnRH transcription is tightly regulated during development and adulthood. GnRH neuron development depends on transcription factors of the homeodomain family. For example, Ventral anterior homeobox 1 (Vax1) is necessary to maintain GnRH expression after embryonic day 13 in the mouse. To further our understanding of the mechanisms by which VAX1 regulates GnRH gene expression, we asked whether VAX1 interacts with other transcription factors to modify GnRH expression levels. Using the GnRH cell lines, GN11 and GT1-7, we found that activation of PKC enhances expression of the immediate early gene cFos in both GN11, and GT1-7, and represses expression of Vax1 in GT1-7. Further, VAX1 interacts with cFOS while bound to the GnRH promoter. In immature GN11 cells, VAX1 and cFOS enhance GnRH expression, whereas VAX1 and cFOS have a repressive role in the mature GT1-7 cells.

Pyrethroid Insecticide Cypermethrin Accelerates Pubertal Onset in Male Mice via Disrupting Hypothalamic-Pituitary-Gonadal Axis.

Pyrethroids, a class of insecticides that are widely used worldwide, have been identified as endocrine-disrupting chemicals (EDCs). Our recent epidemiological study reported on an association of increased pyrethroids exposure with elevated gonadotropins levels and earlier pubertal development in Chinese boys. In this study, we further investigated the effects of cypermethrin (CP), one of the most ubiquitous pyrethroid insecticides, on hypothalamic-pituitary-gonadal (HPG) axis and pubertal onset in male animal models. Early postnatal exposure to CP at environmentally relevant doses (0.5, 5, and 50 μg/kg CP) significantly accelerated the age of puberty onset in male mice. Administration of CP induced a dose-dependent increase in serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone in male mice. CP did not affect gonadotropin-releasing hormone (GnRH) gene expression in the hypothalamus, but CP at higher concentrations stimulated GnRH pulse frequency. CP could induce the secretion of LH and FSH, as well as the expression of gonadotropin subunit genes [chorionic gonadotropin α (CGα), LHβ, and FSHβ] in pituitary gonadotropes. CP stimulated testosterone production and the expression of steroidogenesis-related genes [steroidogenic acute regulatory (StAR) and Cytochrome p 450, family 11, subfamily A, polypeptide 1 (CYP11A1)] in testicular Leydig cells. The interference with hypothalamic sodium channels as well as calcium channels in pituitary gonadotropes and testicular Leydig cells was responsible for CP-induced HPG axis maturation. Our findings established in animal models provide further evidence for the biological plausibility of pyrethroid exposure as a potentially environmental contributor to earlier puberty in males.

Iron overload induces hypogonadism in male mice via extrahypothalamic mechanisms

IntroductionIron overload leads to multiple organ damage including endocrine organ dysfunctions. Hypogonadism is the most common non-diabetic endocrinopathy in primary and secondary iron overload syndromes. AimTo explore the molecular determinants of iron overload-induced hypogonadism with specific focus on hypothalamic derangements. A dysmetabolic male murine model fed iron-enriched diet (IED) and cell-based models of gonadotropin-releasing hormone (GnRH) neurons were used. ResultsMice fed IED showed severe hypogonadism with a significant reduction of serum levels of testosterone (−83%) and of luteinizing hormone (−86%), as well as reduced body weight gain, body fat and plasma leptin. IED mice had a significant increment in iron concentration in testes and in the pituitary. Even if iron challenge of in vitro neuronal models (GN-11 and GT1-7 GnRH cells) resulted in 10- and 5-fold iron content increments, respectively, no iron content changes were found in vivo in hypothalamus of IED mice. Conversely, mice placed on IED showed a significant increment in hypothalamic GnRH gene expression (+34%) and in the intensity of GnRH-neuron innervation of the median eminence (+1.5-fold); similar changes were found in the murine model HFE−/−, resembling human hemochromatosis. ConclusionsIED-fed adult male mice show severe impairment of hypothalamus-pituitary-gonadal axis without a relevant contribution of the hypothalamic compartment, which thus appears sufficiently protected from systemic iron overload.

Reproductive axis gene regulation during photostimulation and photorefractoriness in Yangzhou goose ganders

BackgroundThe Yangzhou goose is a long-day breeding bird that has been increasingly produced in China. Artificial lighting programs are used for controlling its reproductive activities. This study investigated the regulations of photostimulation and photorefractoriness that govern the onset and cessation of the breeding period.ResultsIncreasing the daily photoperiod from 8 to 12 h rapidly stimulated testis development and increased plasma testosterone concentrations, with peak levels being reached 2 months after the photoperiod increase. Subsequently, testicular activities, testicular weight, spermatogenesis, and plasma testosterone concentrations declined steadily and reached to the nadir at 5 months after the 12-hour photoperiod. Throughout the experiment, plasma concentrations of triiodothyronine and thyroxine changed in reciprocal fashions to that of testosterone. The stimulation of reproductive activities caused by the increasing photoperiod was associated with increases in gonadotropin-releasing hormone (GnRH), but decreases in gonadotropin-inhibitory hormone (GnIH) and vasoactive intestinal peptide (VIP) gene messenger RNA (mRNA) levels in the hypothalamus. In the pituitary gland, the levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) mRNA abruptly increased during the longer 12-hour photoperiod. The occurrence of photorefractoriness was associated with increased GnIH gene transcription by over 250-fold, together with increased VIP mRNA levels in the hypothalamus, and then prolactin and thyroid-stimulating hormone in the pituitary gland. FSH receptor, LH receptor, and StAR mRNA levels in the testis changed in ways paralleling those of testicular weight and testosterone concentrations.ConclusionsThe seasonal reproductive activities in Yangzhou geese were directly stimulated by a long photoperiod via upregulation of GnRH gene transcription, downregulation of GnIH, VIP gene transcription, and stimulation of gonadotrophin. Development of photorefractoriness was characterized by hyper-regulation of GnIH gene transcription in the hypothalamus, in addition of upregulation of VIP and TRH gene transcription, and that of their receptors, in the pituitary gland.

Gonadotropin-releasing hormone and adipokinetic hormone/corazonin-related peptide in the female prawn

Crustacean neuropeptides (NPs) play important roles in the regulation of most physiological activities, including growth, molting and reproduction. In this study, we have performed an in silico analysis of female prawn (Macrobrachium rosenbergii) neural transcriptomes to identify NPs not previously identified. We predict that approximately 1309 proteins are destined for the secretory pathway, many of which are likely post-translationally processed to generate active peptides. Within this neural secretome, we identified a gene transcript that encoded a precursor protein with striking similarity to a gonadotropin-releasing hormone (GnRH). We additionally identified another GnRH NP superfamily member, the adipokinetic hormone/corazonin-related peptide (ACP). M. rosenbergii GnRH and ACP were widespread throughout the nervous tissues, implicating them as potential neuromodulators. Furthermore, GnRH was found in non-neural tissues, including the stomach, gut, heart, testis and ovary, in the latter most prominently within secondary oocytes. The GnRH/corazonin receptor-like gene is specific to the ovary, whereas the receptor-like gene expression is more widespread. Administration of GnRH had no effect on ovarian development and maturation, nor any effect on total hemolymph lipid levels, while ACP administration decreased oocyte proliferation (at high dose) and stimulated a significant increase in total hemolymph lipids. In conclusion, our targeted analysis of the M. rosenbergii neural secretome has revealed the decapod GnRH and ACP genes. We propose that ACP in crustaceans plays a role in the lipid metabolism and the inhibition of oocyte proliferation, while the role of the GnRH remains to be clearly defined, possibly through experiments involving gene silencing.

Single Nucleotide Polymorphisms of GnRHR Gene and Its Relationship with Reproductive Performance in Egyptian Buffaloes

THIS WORK aimed to search for single nucleotide polymorphisms of gonadotropin releasing hormone (GnRHR) gene in Egyptian buffalo which involved in endocrine control of fertility. An ultrasonographic examinations of anestrum and repeat breeders’ buffaloes wereperformed prior to blood sampling through a transrectal ultrasonography. DNA was extracted from blood and the PCR and single-strand conformation polymorphism (SSCP) techniqueswereadopted to study of GnRHR gene polymorphism. The PCR amplified a fragment with 240-bp in size and the SSCP results showed that there isagenetic polymorphism with three different patterns (AA, BB and CC) in Egyptian buffalo. The CC genotype was associated with smooth inactive ovary,while BB genotype was associated with fertility in buffalo. Multiple sequence alignment of the three patterns sequences revealed that GnRHR had 5 single nucleotide polymorphisms including one nucleotide insertion, one nucleotide deletion and 3 nucleotide substitutions. Insertion was at position 4 in pattern III. The deletion was at position 189 in pattern III, while the 3 nucleotide substitutions were at positions 204 (T/G), 206 (T/A) and 207 (A/T). In conclusion, GnRHR gene could be used as a candidate marker for fertility in Egyptian buffaloes with its mutation is relatedto ovarian inactivity.

절식이 나일 틸라피아 Oreochromis niloticus의 Kiss2, GnRH I mRNA 발현 및 성 스테로이드 호르몬 농도에 미치는 영향

In many fish species, including Nile tilapia (Oreochromis niloticus), gonadal development occurs at the expense of stored energy and nutrients. Therefore, reproductive systems are inhibited by limited food supply. It has been well established that reproductive function is highly sensitive to both metabolic status and energy balance. Nothing is known about the possible mediated connection between energy balance and reproduction. Kisspeptin, a neuropeptide product of the Kiss gene has emerged as an essential gatekeeper of reproduction and may be possibly be linked to energy balance and reproduction in non-mammalians. Thus, in this study, the effect of fasting (10 days) on the expression of kisspeptin and the gonadotropin-releasing hormone (GnRH) gene were assessed in Nile tilapia (male and female) using qRTPCR. In addition, plasma levels of estradiol-17β (E₂) and 11-ketotestosterone (11-KT) in adult tilapia were measured by ELISA. In male tilapia, fasting reduced Kiss2 and GnRH I mRNA expression in the brain and 11-KT level in comparison with the fed tilapia (p 0.05). These data indicate the impact of nutritional states on kisspeptin as a potential regulatory mechanism for the control of reproduction in male Nile tilapia.

Morphological analysis of the early development of telencephalic and diencephalic gonadotropin-releasing hormone neuronal systems in enhanced green fluorescent protein-expressing transgenic medaka lines.

Teleosts possess two or three paralogs of gonadotropin-releasing hormone (GnRH) genes: gnrh1, gnrh2, and gnrh3. Some species have lost the gnrh1 and/or gnrh3 genes, whereas gnrh2 has been completely conserved in the teleost species analyzed to date. In most teleosts that possess gnrh1, GnRH1 peptide is the authentic GnRH that stimulates gonadotropin release, whereas GnRH2 and GnRH3, if present, are neuromodulatory. Progenitors of GnRH1 and GnRH3 neurons originate from olfactory placodes and migrate to their destination during early development. However, because of the relatively low affinity/specificity of generally available antibodies that recognize GnRH1 or GnRH3, labeling of these neurons has only been possible using genetic manipulation. We used a model teleost, medaka, which possesses all three paralogous gnrh genes, to analyze development of forebrain GnRH neurons composed of GnRH1 and GnRH3 neurons. Here, we newly generated transgenic medaka lines that express enhanced green fluorescent protein under the control of promoters for gnrh1 or gnrh3, to detect GnRH neurons and facilitate immunohistochemical analysis of the neuronal morphology. We used a combination of immunohistochemistry and three-dimensional confocal microscopy image reconstructions to improve identification of neurites from GnRH1 or GnRH3 neuronal populations with greater precision. This led us to clearly identify the hypophysiotropic innervation of GnRH1 neurons residing in the ventral preoptic area (vPOA) from as early as 10 days post hatching. Furthermore, these analyses also revealed retinopetal projections of nonhypophysiotropic GnRH1 neurons in vPOA, prominent during early developmental stages, and multiple populations of GnRH3 neurons with different origins and migratory pathways.

Cloning, expression, and polymorphism at the 5ʹ-flanking region of the GnRH gene and their association with laying traits in Muscovy duck (Cairina moschata)

Gonadotropin-releasing hormone (GnRH), a neuropeptide, plays a vital role in the hypothalamus–pituitary–gonadal (HPG) axis. In vertebrates, GnRH is crucial for the onset of sexual development and the entire reproductive process. The purpose of this study was to identify genetic factors associated with egg-laying traits of Muscovy ducks.The full-length cDNA (474 bp) of Muscovy duck GnRH was obtained and characterised. It encodes 92 amino acids containing a 1-amino acid signal peptide cleavage site. Phylogenetic analysis revealed that Muscovy duck GnRH has a close relationship with Anas platyrhynchos GnRH.GnRH showed significantly different expression profiles between 4 developmental periods in the hypothalamus, pituitary, and ovary. The expression of GnRH in the laying period (36 weeks) was higher than at other periods in the three tissues. GnRH was widely expressed in 12 examined tissues of nesting and laying Muscovy ducks. In the hypothalamus, pituitary and gonads, the expression of GnRH was higher than in other tissues.In laying Muscovy ducks, the expression of GnRH in the hypothalamus, pituitary, ovary, muscular stomach, pancreas, heart, duodenum and spleen was significantly higher than in nesting dusks. Differences were detected in the liver and glandular stomach between laying ducks and nesting ducks. Differences between the kidney and lung were not significant.In the pituitary, the GnRH and GnRH receptor (GnRHR) genes shared the same expression profiles during 4 time points. Both genes had the highest expression at 36 weeks of age.A mutation (g.206G > A) in the 5ʹ-flanking region was associated with egg-laying performance. Individuals with genotype GG had better egg-laying performance than the individuals with genotype AA. GnRH may be used as a marker gene for laying performance in the Muscovy duck.

Characterization of an abalone gonadotropin-releasing hormone and its effect on ovarian cell proliferation

In various animal groups, both vertebrates and invertebrates, it has been reported that gonadotropin-releasing hormone (GnRH) is an important regulator of reproductive function. Knowledge of the molecular identity of a GnRH and its function in the abalone is unknown. In this study, we have characterized the abalone GnRH genes and their putative precursor sequences from the neural ganglia of sexually mature female Haliotis asinina and Haliotis laevigata. The abalone GnRH genes demonstrate high similarity with other gastropods within the proposed bioactive GnRH peptide regions (H. asinina: QNYHFSNGWYPG; H. laevigata: QNYHFSNGWHA) but less in the GnRH-associated peptide region. An antibody generated against HasGnRH showed specific expression of the peptide within the cerebral and pleuropedal ganglia. HasGnRH was localized in type-1 neurosecretory cells that are widely distributed within the ventral and dorsal parts of the cerebral ganglion and also observed in nerve fiber bundles extending into the dorsal part of the ganglion. In the pleuropedal ganglion, HasGnRH immunoreactive cells were distributed in both ventral and dorsal horns. Nerve fibers containing HasGnRH immunoreactivity were also detected primarily within the ventral region of ventral horns of the ganglion. Synthetic HasGnRH and buserelin stimulated oogonia and oocyte proliferation in one-year-old female H. asinina as determined by in vivo and in vitro BrdU ovarian cell proliferation. These findings provide important knowledge into the molecular identity of the abalone GnRH and its function in stimulating oocyte proliferation that helps us to further understand the role of this peptide in molluscs which may be applied to aquaculture of this species.

GnRH level regulation in the hypothalamus of female rats of different age

In recent years the study of age-related derangements of the gonadotropin releasing hormone (GnRH) synthesis and secretion resulting from both the GnRH gene expression changes and interaction of glia with GnRH-ergic neurons of the hypothalamus is a focus of attention. It was demonstrated earlier that this could result from the decreased activity of monoaminergic and peptidergic systems that control the GnRH preovulatory secretion surge initiation, specifically from the loss of the signal coming from the suprachiasmatic nuclei (SCN) of the hypothalamus. This signal is critical for the emerging of the GnRH regular cyclic secretion, which is mediated prominently by vasoactive intestinal peptide (VIP). We have studied age-related changes in the biogenic amines and VIP content in the hypothalamic structures responsible for the GnRH synthesis and secretion. It has been shown by us that the GnRH level in the median eminence with the arcuate nuclei (ME-Arc) of the hypothalamus of 22-month-old rats is half as high compared to that of 7-8-month-old animals. Beside that, the VIP level in the SCN tended to decrease, with the norepinephrine, dopamine, and 5-hydroxyindoleacetic acid levels decreased significantly in the median preoptic area of the hypothalamus responsible for the GnRH synthesis and in the ME-Arc exercising its secretion into the portal vein of the pituitary. It has been shown that the initial phase of the reproductive failure with 13-14-month-old animals having irregular estrous cycles is characterized by gradual disappearance of the normal biogenic amine diurnal dynamics in the studied hypothalamic structures, which could be due to the loss of regulatory signals coming from the SCN.

Autoshortloop feedback regulation of pulsatile gonadotropin-releasing hormone (GnRH) secretion by its metabolite, GnRH-(1-5).

Given the central role of the decapeptide gonadotropin-releasing hormone (GnRH) in reproductive function, our long-term objective is to delineate the underlying mechanism regulating these reproductive processes. The outcome of GnRH secretion is in part dependent on the proteolytic metabolism of this decapeptide. In contrast to the belief that the metabolism of GnRH serves only as a degradative process that removes excess GnRH, we have shown that a metabolite of the decapeptide, GnRH-(1-5), can directly regulate GnRH gene expression and reproductive behavior. To further characterize the effect of GnRH-(1-5) on GnRH neuronal function, we determined whether GnRH-(1-5) can directly regulate GnRH secretion and pulsatility using an in vitro perifusion system. We compared the effect of GnRH-(1-5) on GnRH secretion in the immortalized GnRH neuron (GT1-7 cell line), whole rat hypothalamic explant, and enzymatically dispersed rat hypothalamic cells. Tissue preparations were perifused continuously for 9 h during which a 3-h challenge with GnRH-(1-5) was administered (4-6 h). The results show that treatment with GnRH-(1-5) increased (p < 0.05) the mean GnRH secretion and the amplitude of the pulses but not the pulse frequency. The present study supports the notion that GnRH-(1-5) is functionally capable of regulating the reproductive neuroendocrine system.

Brain aromatase (cyp19a1b) and gonadotropin releasing hormone (gnrh2 and gnrh3) expression during reproductive development and sex change in black sea bass (Centropristis striata)

Teleost fish exhibit diverse reproductive strategies, and some species are capable of changing sex. The influence of many endocrine factors, such as gonadal steroids and neuropeptides, has been studied in relation to sex change, but comparatively less research has focused on gene expression changes within the brain in temperate grouper species with non-haremic social structures. The purpose of the present study was to investigate gonadotropin releasing hormone (GnRH) and brain aromatase (cyp19a1b) gene expression patterns during reproductive development and sex change in protogynous (female to male) black sea bass (Centropristis striata). Partial cDNA fragments for cyp19a1b and eef1a (a reference gene) were identified, and included with known gnrh2 and gnrh3 sequences in real time quantitative PCR. Elevated cyp19a1b expression was evident in the olfactory bulbs, telencephalon, optic tectum, and hypothalamus/midbrain region during vitellogenic growth, which may indicate changes in the brain related to neurogenesis or sexual behavior. In contrast, gnrh2 and gnrh3 expression levels were largely similar among gonadal states, and all three genes exhibited stable expression during sex change. Although sex change in black sea bass is not associated with dramatic changes in GnRH or cyp19a1b gene expression among brain regions, these genes may mediate processes at other levels, such as within individual hypothalamic nuclei, or through changes in neuron size, that warrant further research.

Expression changes of mRNAs encoding kisspeptins and their receptors and gonadotropin-releasing hormones during early development and gonadal sex differentiation periods in the brain of chub mackerel (Scomber japonicus)

In recent years, brain kisspeptin system has been shown to be involved in diverse reproductive function, including sexual differentiation in vertebrates. Our previous reports demonstrated that the chub mackerel (Scomber japonicus) brain expresses two kisspeptin (kiss1, kiss2), two kisspeptin receptor (kissr1, kissr2) and three gonadotropin-releasing hormone (gnrh1, gnrh2, gnrh3) genes. In the present study, using quantitative real-time PCR (qRT-PCR) assays, we analysed expression changes of these genes during early development (0–30dphs) and gonadal sex differentiation periods (37–60dphs). Absolute expression level of kiss–kissr–gnrh in the whole head was higher between 0 and 15dphs, in comparison to later developmental periods. Histological analyses revealed presence of sexually differentiated males and females with testicular and ovarian features at 37, 45, and 60dphs. In both males and females, kiss2, kissr1, and kissr2 levels were higher at 37dph, in comparison to 45 and 60dphs, with kiss1 showing no significant differences. Levels of all three gnrh mRNAs were higher at 45dph, in comparison to 60dph. Changes in the expression level of kiss–kissr–gnrh mRNAs in different brain regions of sexually differentiated males and females indicated differences in their regional distribution. These results suggest possible involvement of Kiss–KissR–GnRH systems during early development and gonadal sex differentiation in the chub mackerel.

Selective targeting of GnRH-II neurons to block ovulation

BackgroundIn rhesus macaques, luteinizing hormone (LH) secretion appears to be regulated by two distinct gonadotropin-releasing hormone (GnRH) neuronal populations, which can be distinguished by their unique anatomical locations and because they express different molecular forms of GnRH (GnRH-I and GnRH-II). Study DesignThe effect of estradiol on GnRH gene expression was examined. ResultsEstradiol inhibited GnRH-I neurons but stimulated GnRH-II neurons, suggesting that GnRH-II neurons play the dominant role in mediating estradiol-positive feedback and triggering the midcycle preovulatory LH surge. ConclusionsSelective silencing of GnRH-II neurons in women could serve as a novel contraceptive, by blocking ovulation while leaving the rest of the reproductive axis relatively unperturbed.

Perfluorooctane sulfonate effects on the reproductive axis in adult male rats

Perfluorooctane sulfonate (PFOS) is a neurotoxic agent and it can disrupt the endocrine system activity. This work was undertaken to evaluate the possible effects of PFOS exposure on the hypothalamic–pituitary–testicular axis (HPT) in adult male rats, and to evaluate the possible morphological alterations induced by PFOS in the endocrine tissues of this axis. Adult male rats were orally treated with 0.5; 1.0; 3.0 and 6.0mg of PFOS/kg/day for 28 days. After PFOS exposure, hypothalamic noradrenaline concentration increased in the anterior hypothalamus and in the median eminence, not changing in the mediobasal hypothalamus. PFOS treated rats presented a decrease of the gonadotropin releasing hormone (GnRH) gene expression, increasing the mRNA levels of the luteinizing hormone (LH) in rats treated with all doses administered except with the dose of 6mg/kg/day. PFOS also induced a raise of the follicle stimulating hormone (FSH) gene expression in the animals exposed to 0.5 and 1.0mg of PFOS/kg/day. After PFOS exposure, hypothalamic GnRH concentration was modified, LH and testosterone release was inhibited and FSH secretion was stimulated. Moreover, PFOS induced several histopathological alterations in the hypothalamus, pituitary gland and testis. The results obtained in the present study suggest in general terms that PFOS can inhibit the physiological activity of the reproductive axis in adult male rats, which could be explained, at least in part, by the structural alterations showed in the animals exposed to this chemical: very dense chromatin, condensed ribosomes and a loss of the morphology in the hypothalamus; a degeneration of the gonadotrophic cells, as well as a loss and degeneration of the spermatozoids and a very marked edema in the testis.

Actions and Interactions of Alcohol and Transforming Growth Factor β1 on Prepubertal Hypothalamic Gonadotropin‐Releasing Hormone

Alcohol (ALC) diminishes gonadotropin-releasing hormone (GnRH) secretion and delays puberty. Glial transforming growth factor β1 (TGFβ1) plays a role in glial-neuronal communications facilitating prepubertal GnRH secretion. We assessed the effects of acute ALC administration on TGFβ1-induced GnRH gene expression in the brain preoptic area (POA) and release of the peptide from the medial basal hypothalamus (MBH). Furthermore, we assessed actions and interactions of TGFβ1 and ALC on an adhesion/signaling gene family involved in glial-neuronal communications. Prepubertal female rats were administered ALC or water via gastric gavage at 7:30 am. At 9:00 am, saline or TGFβ1 (100 ng/3 μl) was administered into the third ventricle. At 3:00 pm, the POA was removed and frozen for gene expression analysis and repeated for protein assessments. In another experiment, the MBH was removed from ALC-free rats. After equilibration, tissues were incubated in Locke's medium only or medium containing TGFβ1 with or without 50 mM ALC for measurement of GnRH peptide released in vitro. TGFβ1 induced GnRH gene expression in the POA, and this effect was blocked by ALC. We also described the presence and responsiveness of the TGFβ1 receptor in the POA and showed that acute ALC exposure not only altered the TGFβ1-induced increase in TGFβ-R1 protein expression but also the activation of receptor-associated proteins, Smad2 and Smad3, key downstream components of the TGFβ1 signaling pathway. Assessment of an adhesion/signaling family consisting of glial receptor protein tyrosine phosphatase beta and neuronal contactin-associated protein-1 (Caspr1) and contactin showed that the neuronal components were induced by TGFβ1 and that ALC blocked these effects. Finally, TGFβ1 was shown to induce release of the GnRH peptide in vitro, an action that was blocked by ALC. We have demonstrated glial-derived TGFβ1 induces GnRH gene expression in the POA and stimulates release of the peptide from the MBH, actions necessary for driving the pubertal process. Importantly, ALC acted at both brain regions to block stimulatory effects of TGFβ1. Furthermore, ALC altered neuronal components of an adhesion/signaling family previously shown to be expressed on GnRH neurons and implicated in glial-GnRH neuronal communications. These results further demonstrate detrimental effects of ALC at puberty.

Characterization of GnRH, ILGFRI and AR Genes in Sturgeon’s Genomics

Abstract- In this study, gonadotropin-releasing hormone (GnRH), insulin-like growth factor receptor I (ILGFRI), and androgen receptor (AR) genes coding, were examined by polymerase chain reaction (PCR) analysis and DNA sequencing in three Eurasian sturgeon species: Huso huso, Acipenser stellatus and A. gueldenstaedtii. Amplicons of 190 bp for GnRH, 180 bp for ILGFRI, and 200 bp for AR genes were amplified with specific primer pairs in all species genome. All amplification products were sequenced and compared to reference H. huso genome retrieved from NCBI database by BLASTN analysis. The ILGFRI gene had the most similar sequences (75.96-96.15%) in all species. Gene sequence similarity calculated for AR gene ranged from 48.5- 95.71%. The GnRH gene had more distant sequences (23.48-83.91%) between species. There was moderate similarity between H. huso and the reference H. huso samples. An unauthorized (a sample seized from poachers) sample was more closely related to A. stellatus. Partial coding region of AR gene was registered in NCBI database under Accession Number: KC172108. Five nucleotide changes were detected in this sequence, two of which resulted in changes in amino acid sequence. Also, a partial sequence consisted of two microsatellite regions of two and four nucleotide repeats. Three constructed dendrograms displayed the relationship among species according to certain genomic regions. These findings indicate a high level of diversity among and even within the same species. They also show that these three genes could be useful in species’ identification and characterization