Gonadotropin-releasing Hormone Content Research Articles

Habitat-induced changes in forage quality and implications for fitness in Plateau Pika (Ochotona curzoniae)

Abstract Plateau pikas (Ochotona curzoniae) play a keystone role in the alpine meadow ecosystem on the Qinghai–Tibetan Plateau (QTP). For decades, QTP grasslands have become degraded to some degree, largely as a result of heavy livestock grazing. Concomitantly, the abundance of plateau pikas has increased dramatically as grassland degradation has altered the vegetation community structure and dominant species, shifting the plant nutrient contents toward higher protein conditions that favor them. Considerable research supports the hypothesis that the quantity and quality of food limit herbivore populations. Here, we examined the relationship between the availability of essential amino acids in the diets of plateau pikas and the degree of meadow degradation associated with livestock grazing intensity through a field survey, as well as the fitness of individuals by laboratory feeding experiments with 2 pelleted chows containing 0.26% and 0.45% methionine. Sulfur-containing methionine and cystine were the most limiting amino acids in the diets of pikas. During the pika breeding season, the concentrations of most essential acids, particularly methionine and cystine, were higher in heavily degraded meadows than in lightly and moderately degraded meadows. Individuals fed 0.45% methionine exhibited enhanced cell-mediated immunity, reduced intensity of coccidian infection, and increased concentrations of gonadotropin-releasing hormone, luteinizing hormone, estradiol, progesterone, and testosterone compared to those fed 0.26% methionine. These results showed that heavily degraded meadows provided relatively high-quality food that improved individual fitness, suggesting that the high-methionine food in the heavily degraded meadows may be a key factor in the generation and maintenance of high-density populations of plateau pikas.

Effects of Pasturella Multocida B:2 and its immunogens (LPS and OMP) on reproductive hormones in Nili-Ravi Buffaloes.

Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P<0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.

Effects of sub-chronic exposure to microcystin-LR on the endocrine system of male rats

Microcystins (MCs) can cause reproductive and developmental toxicity and disrupt endocrine homeostasis in mammals. In the present study, male, Sprague-Dawley (SD) rats were administrated 3 or 30 μg MC-LR/kg, body mass (bm) per day via intraperitoneal (i.p.) injections for 6 weeks. Effects of MC-LR on histology, hormone concentrations, gene transcriptional profiles and protein expressions along the hypothalamic-pituitary-adrenal (HPA), ‐gonad (HPG) and -thyroid (HPT) axes were assessed. Sub-chronic administration with MC-LR caused histological damage to hypothalamus, pituitary, adrenal, testes and thyroid and affected relative masses of pituitary, adrenal and testes. The HPA axis was activated and serum concentrations of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were significantly augmented. Along the HPG axis, serum concentrations of gonadotropin-releasing hormone (GnRH) and dihydrotestosterone (DHT) were diminished, while concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T) and estradiol (E2) were augmented. For the HPT axis, only concentrations of free tetra-iodothyronine (fT4) were significantly diminished, while concentrations of thyrotropin-releasing hormone (TRH), thyroid-stimulating hormone (TSH) or free tri-iodothyronine (fT3) were not significantly changed. Also, several genes and proteins related to synthesis of steroid hormones were significantly altered. Findings of the present study illustrate that MC-LR can cause endocrine-disrupting effects through the disruption of synthesis and secretion of hormones along the HPA, HPG and HPT axes and negative feedback regulation. Also, there could be crosstalk among HPA, HPG and HPT axes. These findings elucidate mechanisms of endocrine-disrupting effects of MCs.

Gonadal Feedback Alters the Relationship between Action Potentials and Hormone Release in Gonadotropin-Releasing Hormone Neurons in Male Mice.

In vertebrates, the pulsatile release of gonadotropin-releasing hormone (GnRH) from neurons in the hypothalamus triggers secretion of anterior pituitary gonadotropins, which activate steroidogenesis, and steroids in turn exert typically homeostatic negative feedback on GnRH release. Although long-term episodic firing patterns of GnRH neurons in brain slices resemble the pulsatile release of GnRH and LH in vivo, neither the relationship between GnRH neuron firing and release nor whether this relationship is influenced by gonadal feedback are known. We combined fast-scan cyclic voltammetry and patch-clamp to perform simultaneous measurements of neuropeptide release with either spontaneous action potential firing or in response to neuromodulator or action-potential-spike templates in brain slice preparations from male mice. GnRH release increased with higher frequency spontaneous firing to a point; release reached a plateau after which further increases in firing rate did not elicit further increased release. Kisspeptin, a potent GnRH neuron activator via a Gq-coupled signaling pathway, triggered GnRH release before increasing firing rate, whether globally perfused or locally applied. Increasing the number of spikes in an applied burst template increased release; orchidectomized mice had higher sensitivity to the increased action potential number than sham-operated mice. Similarly, Ca2+ currents triggered by these burst templates were increased in GnRH neurons of orchidectomized mice. These results suggest removal of gonadal feedback increases the efficacy of the stimulus-secretion coupling mechanisms, a phenomenon that may extend to other steroid-sensitive regions of the brain.SIGNIFICANCE STATEMENT Pulsatile secretion of GnRH plays a critical role in fertility. The temporal relationship between GnRH neuron action potential firing and GnRH release remains unknown as does whether this relationship is influenced by gonadal feedback. By combining techniques of fast-scan cyclic voltammetry and patch-clamp we, for the first time, monitored GnRH concentration changes during spontaneous and neuromodulator-induced GnRH neuron firing. We also made the novel observation that gonadal factors exert negative feedback on excitation-secretion coupling to reduce release in response to the same stimulus. This has implications for the control of normal fertility, central causes of infertility, and more broadly for the effects of sex steroids in the brain.

&lt;i&gt;Corrigendum to&lt;/i&gt;: Effects of supplementation with different concentrations of L-citrulline on the plasma amino acid concentration, reproductive hormone concentrations, antioxidant capacity, and reproductive performance of Hu ewes

&lt;sec&gt; Context &lt;sc&gt;L&lt;/sc&gt;-citrulline (&lt;sc&gt;L&lt;/sc&gt;-Cit) does not degrade in the rumen and has the ability to form peptide bonds in the body; however, it does not participate in protein synthesis. &lt;/sec&gt; &lt;sec&gt; Aims This study aimed to evaluate the effects of &lt;sc&gt;L&lt;/sc&gt;-Cit on the reproductive performance of Hu ewes. &lt;/sec&gt; &lt;sec&gt; Methods In total, 30 ewes were randomly categorised into five groups. The control group was fed with a basic diet, whereas the Experimental Groups I, II, III, and IV were provided feed supplemented with 5, 10, 15, and 20g/day of &lt;sc&gt;L&lt;/sc&gt;-Cit respectively. Blood samples of ewes were collected 4h after feeding on Day 21 of the experiment and before feeding on Day 30. The optimal supplementary feeding dose was selected on the basis of blood biochemical indexes. Overall, ninety 2-year-old ewes were classified into two groups. The control group was fed with a basic diet and the experimental group was fed with a diet supplemented with 10g/day of &lt;sc&gt;L&lt;/sc&gt;-Cit. After 30days of supplementary feeding, reproductive performance of ewes was determined. &lt;/sec&gt; &lt;sec&gt; Key results The plasma concentrations of Cit, ornithine, and arginine in ewes increased linearly with an increase in the level of &lt;sc&gt;L&lt;/sc&gt;-Cit supplementation. The plasma concentrations of gonadotropin-releasing hormone, luteinising hormone, and follicle-stimulating hormone in the experimental group increased significantly compared with those in the control group. The plasma total antioxidant capacity and catalase, superoxide dismutase, and glutathione peroxidase in the experimental group were significantly higher than those in the control group, whereas the concentrations of malondialdehyde in all experimental groups were significantly lower than those in the control group. The conception, lambing, and double lambing rates of the experimental group were increased by 28.76%, 15.90%, and 40.21% respectively. &lt;/sec&gt; &lt;sec&gt; Conclusions Supplementation with different doses of &lt;sc&gt;L&lt;/sc&gt;-Cit can improve the concentrations of some plasma amino acids and reproductive hormones as well as antioxidant capacity of ewes. Supplementary feeding with 10g/day of &lt;sc&gt;L&lt;/sc&gt;-Cit could increase the lambing and double lambing rates of ewes. &lt;/sec&gt; &lt;sec&gt; Implication &lt;sc&gt;L&lt;/sc&gt;-Cit can improve the reproductive performance of ewes. &lt;/sec&gt;

Effects of supplementation with different concentrations of L-citrulline on the plasma amino acid concentration, reproductive hormone concentrations, antioxidant capacity, and reproductive performance of Hu ewes

Context L-citrulline (L-Cit) does not degrade in the rumen and has the ability to form peptide bonds in the body; however, it does not participate in protein synthesis. Aims This study aimed to evaluate the effects of L-Cit on the reproductive performance of Hu ewes. Methods In total, 30 ewes were randomly categorised into five groups. The control group was fed with a basic diet, whereas the Experimental Groups I, II, III, and IV were provided feed supplemented with 5, 10, 15, and 20 g/day of L-Cit respectively. Blood samples of ewes were collected 4 h after feeding on Day 21 of the experiment and before feeding on Day 30. The optimal supplementary feeding dose was selected on the basis of blood biochemical indexes. Overall, ninety 2-year-old ewes were classified into two groups. The control group was fed with a basic diet and the experimental group was fed with a diet supplemented with 10 g/day of L-Cit. After 30 days of supplementary feeding, reproductive performance of ewes was determined. Key results The plasma concentrations of Cit, ornithine, and arginine in ewes increased linearly with an increase in the level of L-Cit supplementation. The plasma concentrations of gonadotropin-releasing hormone, luteinising hormone, and follicle-stimulating hormone in the experimental group increased significantly compared with those in the control group. The plasma total antioxidant capacity and catalase, superoxide dismutase, and glutathione peroxidase in the experimental group were significantly higher than those in the control group, whereas the concentrations of malondialdehyde in all experimental groups were significantly lower than those in the control group. The conception, lambing, and double lambing rates of the experimental group were increased by 28.76%, 15.90%, and 40.21% respectively. Conclusions Supplementation with different doses of L-Cit can improve the concentrations of some plasma amino acids and reproductive hormones as well as antioxidant capacity of ewes. Supplementary feeding with 10 g/day of L-Cit could increase the lambing and double lambing rates of ewes. Implication L-Cit can improve the reproductive performance of ewes.

Acute exposure to microcystins affects hypothalamic-pituitary axes of male rats

Microcystins (MCs) produced by some cyanobacteria can cause toxicity in animals and humans. In recent years, growing evidence suggests that MCs can act as endocrine disruptors. This research systematically investigated effects of microcystin-LR (MC-LR) on endocrine organs, biosynthesis of hormones and positive/negative feedback of the endocrine system in rats. Male, Sprague-Dawley rats were acutely administrated MC-LR by a single intraperitoneal injection at doses of 45, 67.5 or 90 μg MC-LR/kg body mass (bm), and then euthanized 24 h after exposure. In exposed rats, histological damage of hypothalamus, pituitary, adrenal, testis and thyroid were observed. Serum concentrations of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and corticosterone (CORT), expressions of genes and proteins for biosynthesis of hormones were lesser, which indicated an overall suppression of the hypothalamus-pituitary-adrenal (HPA) axis. Along the hypothalamus-pituitary-gonadal (HPG) axis, lesser concentrations of gonadotropin-releasing hormone (GnRH) and testosterone (T), but greater concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and estradiol (E2) were observed. Except for greater transcription of cyp19a1 in testes, transcriptions of genes and proteins for T and E2 biosynthesis along the HPG axis were lesser. As for the hypothalamus-pituitary-thyroid (HPT) axis, after MCs treatment, greater concentrations of thyroid-stimulating hormone (TSH), but lesser concentrations of free tri-iodothyronine (fT3) were observed in serum. Concentrations of free tetra-iodothyronine (fT4) were greater in rats dosed with 45 μg MCs/kg, bm, but lesser in rats dosed with 67.5 or 90 μg MCs/kg, bm. Transcripts of genes for biosynthesis of hormones and receptors along the HPT axis and expressions of proteins for biosynthesis of tetra-iodothyronine (T4) and tri-iodothyronine (T3) in thyroid were significantly altered. Cross-talk among the HPA, HPG and HPT axes probably occurred. It was concluded that MCs caused an imbalance of positive and negative feedback of hormonal regulatory axes, blocked biosynthesis of key hormones and exhibited endocrine-disrupting effects.

Association of long-term effects of low-level sulfamethoxazole with ovarian lipid and amino acid metabolism, sex hormone levels, and oocyte maturity in zebrafish

Sulfamethoxazole (SMZ) is an important antibiotic used to prevent and treat infections in both clinical settings and animal husbandry. High levels of SMZ may exhibit endocrine toxicity. Environmental SMZ enters the human body via food and water; however, the toxicity of environmental doses of SMZ and its effects on reproductive health are unknown. In the present study, zebrafish were exposed to low concentrations of SMZ (1000 and 5000 ng/L) from 2 h post-fertilization to 120 d post-fertilization. Consequently, the proportion of mature oocytes in adult female zebrafish ovarian tissue increased by 98.2 %, indicating that SMZ promotes ovarian maturation. Metabolomics analysis revealed significant changes in ovarian lipid and amino acid levels after SMZ treatment. An enzyme-linked immunoassay used to detect sex hormones in the ovaries showed that SMZ exposure significantly increased the levels of estradiol, a follicle-stimulating hormone, and of luteinizing hormone. Furthermore, an association analysis showed that most of the differentially expressed metabolites in the ovary were strongly correlated with the levels of sex hormones secreted by the pituitary gland. Therefore, significantly increased transcript levels of gonadotropin-releasing hormone (GnRH) and follicle-stimulating hormone detected in brain tissue suggested that SMZ may exhibit ovarian toxicity via the hypothalamus. In vitro experiments were performed to demonstrate that SMZ targets neurons in the hypothalamus. Exposure to SMZ significantly increased the GnRH content in GnRH neurons. Finally, molecular docking simulations indicated the potential interaction of SMZ with G protein-coupled receptor 54; this molecular binding can activate, synthesize, and release GnRH in neurons. In conclusion, long-term environmental exposure to SMZ may induce ovarian toxicity by affecting the hypothalamus–pituitary–gonad axis.

Gynecological uses of GnRH antagonists: review article

Aim: to review and analyze the use of gonadotropin-releasing hormone (GnRH) antagonists in endometriosis, adenomyosis, non-menstrual pelvic pain, uterine fibroids, prostate cancer, female infertility, and precocious puberty. Methods: the information used to write this manuscript was obtained during a three-month period, between October and May 2022, from specialized literature, written in English and Spanish, related to the use and potential applications of GnRH antagonists in medicine, mainly published during the last five years, using journals found in the most relevant medical digital archives, including PubMed, SciELO, Google Scholar, Cochrane, and Elsevier. Among the keywords used for obtaining this updated information were gonadotropin-releasing hormone (GnRH) antagonists; GnRH receptors (GnRHR); elagolix; relugolix; cetrorelix; linzagolix; hypothalamic-pituitary-gonadal (HPG) axis; endometriosis; adenomyosis; pelvic pain; uterine fibroids; leiomyomas; infertility; precocious puberty; PP; and central PP. Results: GnRH antagonists induce a rapid onset of clinical effects, without the flare-up effect that is seen with agonists, and have immediate therapeutic effects; once treatment concludes, hormonal suppression rapidly ceases, with normalization of gonadal function within a few days, guaranteeing an increase of GnRH concentration, controlling non-menstrual pelvic pain and heavy menstrual bleeding, and serving as part of the treatment of infertility and precocious puberty. Conclusion: even though the aforementioned gynecological pathologies can be treated with the use of multiple drugs, GnRH antagonists have shown to be potential first lines of treatment, as long as their administration protocols are followed correctly.

Expression and functional analysis of GnRH at the onset of puberty in sheep

Gonadotropin-releasing hormone (GnRH) is a key factor at the onset of puberty. This decapeptide has been found in mammalian ovaries, but its regulatory mechanism in the ovary of sheep at the onset of puberty is not clear. This study investigated the coding sequence (CDS) of the GnRH gene in the ovary of Duolang sheep and the expression of GnRH mRNA in different tissues at the onset of puberty, and analyzed the effect of GnRH on ovarian granulosa cells (GCs) of Duolang sheep. The results showed that the GnRH CDS of sheep was cloned, the full length of the GnRH CDS in sheep ovary was 279 bp, and the nucleotide sequence was completely homologous to that in the hypothalamus. The expression of GnRH mRNA was highest in the hypothalamus and ovary. The expression of related hormones and receptors in GCs of Duolang sheep treated with different concentrations of GnRH for 24 h was affected. GnRH significantly inhibited LH synthesis and LHR expression in GCs. Low concentration (100 ng mL) had the most obvious therapeutic effect on follicle-stimulating hormone (FSH) and FSHR. Higher concentration (250 ng mL) significantly promoted estradiol and ER mRNA. These findings provide strong evidence that ovarian GnRH is an important regulatory factor at the onset of puberty in sheep.

Direct effect of RFRP‑3 microinjection into the lateral ventricle on the hypothalamic kisspeptin neurons in ovariectomized estrogen‑primed rats

RFamide-related peptide-3 (RFRP-3) may be involved in the inhibition of kisspeptin, but there is no direct evidence that RFRP-3 can directly act on kisspeptin neurons. The present study aimed to investigate the role and mechanism of RFRP-3 and kisspeptin in the hypothalamic-pituitary reproductive axis. In order to detect the expression and localization of RFRP-3 and kisspeptin in dorsomedial hypothalamic nucleus, double immunofluorescence method combined with confocal microscopy were performed. RFRP-3 was injected into the lateral ventricle of ovariectomized estrogen primed rats. Blood and brain tissues were collected at 60-, 120-, 240- and 360-min. Serum levels of gonadotropin-releasing hormone, luteinizing hormone and follicle-stimulating hormone were detected by ELISA. Kisspeptin expression in hypothalamus was detected by western blotting. Finally, surface plasmon resonance was used to verify whether RFRP-3 can directly interact with kisspeptin. Confocal images indicated that RFRP-3 and kisspeptin were co-expressed in the same neurons in the hypothalamus of ovariectomized estrogen-primed rats. Serum concentrations of gonadotropin-releasing hormone, luteinizing hormone and follicle-stimulating hormone were demonstrated to be significantly reduced following microinjection of RFRP-3 into the lateral ventricle for 60, 120, 240 and 360 min compared with the corresponding saline groups. The expression levels of kisspeptin in hypothalamus were gradually decreased following microinjection of RFRP-3 into the lateral ventricle. In addition, the affinity constant (KD) of RFRP-3 binding to kisspeptin was 6.005x10-5 M, indicating that RFRP-3 bound directly to kisspeptin in the range of protein-protein binding strength (KD, 10-3-10-6 M). In conclusion, RFRP-3 may regulate the hypothalamic-pituitary reproductive axis by inhibiting the expression of hypothalamic kisspeptin and direct binding.

Effects of ammonia on hypothalamic-pituitary-ovarian axis in female rabbits

BackgroundAs one of the most harmful gases in the livestock house, ammonia is recognized as an environmental stressor by Environmental Protection Agency (United States). The study aimed to explore the effect of ammonia on hypothalamic-pituitary-ovarian (HPO) axis of rabbits. A total of ninety two-month-old female IRA rabbits were randomly divided into three groups, and were kept in animal environment control rooms for four weeks at college of animal science and technology, Hebei Agricultural University (Baoding, China). The rabbits in the control group were kept under ammonia concentration of < 3 ppm. The two treatment groups were kept under ammonia concentration of 30 ppm and 50 ppm. Hypothalamus, pituitary, and ovary were collected for hematoxylin and eosin (HE) staining, immunohistochemistry, terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Serum was collected for enzyme-linked immunosorbent assay (ELISA). ResultsHistopathological examination revealed that exposed to excess ammonia damaged the morphology and structure of hypothalamus, pituitary, and ovary. TUNEL assay revealed that apoptosis rate increased in hypothalamus, pituitary, and ovary. The protein expression levels of Bcl-2associated X protein (Bax) and Caspase-9 increased, while B-cell lymphoma-2 (Bcl-2) decreased, resulting in apoptosis. Moreover, the concentration of gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and progesterone (PROG) reduced in plasma. The mRNA expression of FSH and LH in pituitary and follicle-stimulating hormone receptor (FSHR), E2, PROG in ovary as well as decreased, indicated hormone secretion disorder. ConclusionsThe results indicated that ammonia exposure damaged hypothalamus, pituitary, and ovary, caused hormone secretion disorder and apoptosis.

Hindlimb unloading-induced reproductive suppression via Downregulation of hypothalamic Kiss-1 expression in adult male rats

BackgroundSpaceflights-induced microgravity can alter various physiological processes in human’s body including the functional status of the reproductive system. Rodent model of tail-suspension hindlimb unloading is extensively used to stimulate the organs responses to the microgravity condition. This study explores the potential effects of hindlimb unloading on testicular functions and spermatogenesis in adult male rats and the underlying mechanism/s.MethodsTwenty Sprague-Dawley rats were allotted into two groups: normally loaded group (control; all arms were in touch with the grid floor) and hindlimb unloaded group (HU; only the forearms were in contact with the grid floor).ResultsFollowing 30 days of exposure, the HU group saw a decline in body weight, testicular and epidydimal weights, and all semen parameters. The circulating concentrations of gonadotropin-releasing hormone (GnRH), follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone significantly decreased, while levels of kisspeptin, corticosterone, inhibin, prolactin and estradiol (E2) increased in the HU group. Intratesticular levels of 5α-reductase enzyme and dihydrotestosterone (DHT) were suppressed, while the levels of aromatase and kisspeptin were significantly elevated in the HU group. Hypothalamic kisspeptin (Kiss1) mRNA expression levels were downregulated while its receptors (Kiss1R) were upregulated in the HU group. On the contrary, the mRNA expression levels of testicular Kiss1 were upregulated while Kiss1R were downregulated. The pituitary mRNA expression levels of FSHβ and LHβ decreased in the HU group. The levels of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and nitric oxide (NO) concentrations, and total antioxidant capacity (TAC) were elevated while malondialdehyde (MDA) concentrations declined in the testes of HU group. The testes of the HU rats showed positive immunostaining of caspase-3, heat shock protein 70 (HSP70) and Bcl2.ConclusionsAltogether, these results revealed an inhibitory effect of hindlimb unloading on kisspeptin signaling in the hypothalamic-pituitary-testicular axis with impaired spermatogenesis and steroidogenesis.

Effects of acute exposure to microcystins on hypothalamic-pituitary-adrenal (HPA), -gonad (HPG) and -thyroid (HPT) axes of female rats.

Microcystins (MCs) are common, well-known cyanobacterial toxins that can affect health of humans. Recently, it has been reported that MCs affect endocrine functions. In the present study, for the first time, histopathology, concentrations of hormones and transcription of genes along the hypothalamic-pituitary-adrenal (HPA), hypothalamic-pituitary-gonad (HPG) and hypothalamic-pituitary-thyroid (HPT) axes were examined in rats exposed to microcystin-LR (MC-LR). Female, Sprague-Dawley (SD) rats were exposed acutely to MC-LR by a single intraperitoneal (i.p.) injection at doses of 0.5, 0.75, or 1 median lethal dose (LD50), i.e. 36.5, 54.75, or 73μg MC-LR/kg body mass (bm) then euthanized 24 hours after exposure. Acute exposure to MC-LR significantly increased relative mass of adrenal in a dose-dependent manner, but relative mass of hypothalamus, pituitary, ovary and thyroid were not significantly different from respective mass in controls. However, damage to all these tissues was observed by histology. Along the HPA axis, lesser concentrations of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were observed in blood serum of exposed individuals, relative to controls. For the HPG axis, concentrations of gonadotropin-releasing hormone (GnRH) and estradiol (E2) were significantly less in rats treated with MC-LR, but greater concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone (T) were observed. Along the HPT axis, MC-LR caused greater concentrations of thyroid-stimulating hormone (TSH), but lesser concentrations of thyrotropin-releasing hormone (TRH), free tetra-iodothyronine (fT4) and tri-iodothyronine (fT3). Significant positive/negative correlations of concentrations of hormones were observed among the HPA, HPG and HPT axes. In addition, profiles of transcription of genes for synthesis of hormones along the endocrine axes and nuclear hormone receptors in adrenal, ovary and thyroid were significantly altered. Therefore, these results suggested that MC-LR affected HPA, HPG and HPT axes and exerted endocrine-disrupting effects. Effects of MC-LR on crosstalk among these three axes need further studies.

Protective effect of naringenin against cadmium-induced testicular toxicity in male SD rats

This study aimed to investigate the effect of naringenin (Nar) on cadmium (Cd)-induced testicular toxicity. Twenty-four male Sprague–Dawley (SD) rats aged 5 weeks were used. Rats were administered with 0.9% NaCl (control group), CdCl2 (2 mg/kg b.w. intraperitoneally), Nar (50 mg/kg b.w, orally), and CdCl2 + Nar (2 mg/kg b.w intraperitoneally and 50 mg/kg b.w. orally, respectively) for 4 weeks. Results showed that body weight, relative testis weights, and sperm quality decreased in the Cd-treated group, and Cd accumulated in serum and testes. Pathological examination showed that Cd can cause testicular damage. Cd decreased the serum concentrations of gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone. It also decreased the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Moreover, exposure to Cd resulted in decreased content of reduced glutathione (GSH) and total antioxidant capacity (T-AOC) concentrations, as well as increased malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents. Cd also provoked testis autophagy by upregulating the expression of the autophagy-related proteins P62 and LC3 II. However, the combined administration of Nar and Cd significantly attenuated the Cd-induced negative effects by increasing the body weight, relative testis weights, and sperm quality and by decreasing testicular damage. Simultaneous supplementation of Nar and Cd markedly restored the decreased levels of GnRH, FSH, LH, testosterone, GSH, and T-AOC and the activities of SOD, CAT, and GPx caused by Cd treatment. Nar further suppressed MDA and H2O2 production and protected the testes from Cd-induced autophagy by downregulating P62 and LC3 II expression. Therefore, Nar protected the testes from Cd-induced toxicity.

Dietary supplementation with Forsythia suspensa extract during late gestation improves reproductive performance, colostrum composition, antioxidant status, immunoglobulin, and inflammatory cytokines in sows and newborn piglets

This study aimed to evaluate the effects of dietary Forsythia suspensa extract (FSE) supplementation during late gestation on reproductive performance, colostrum composition, nutrient digestibility, antioxidant status, immune status, and inflammatory cytokines in sows and newborn piglets. Forty healthy Yorkshire × Landrace gestating sows (average parity of 3.28 ± 0.61; average body weight of 240.16 ± 6.81 kg) were randomly assigned to 2 dietary treatments with 20 sows per treatment based on body weight, back fat thickness and parity. From d 85 of gestation to farrowing, sows were fed one of two diets including a control diet (CON) and a FSE diet (CON + 100 mg/kg FSE). The results showed that sows fed FSE had increased (P < 0.05) litter birth weight, enhanced (P < 0.05) fat and protein contents in colostrum, and decreased (P < 0.05) rate of stillbirth compared with CON. Dietary FSE supplementation increased (P < 0.05) the apparent total tract digestibility of dry matter, crude protein, ether extracts, gross energy, and organic matter in gestating sows compared with CON. Dietary FSE supplementation in sows increased (P < 0.05) the contents of total antioxidant capacity and superoxide dismutase, decreased (P < 0.05) the content of malondialdehyde in colostrum, and increased (P < 0.05) the contents of catalase and glutathione peroxidase in serum of their offsprings compared with CON. Additionally, sows fed FSE showed higher immunoglobulin A (P < 0.05) in serum of their offsprings and a trend for higher immunoglobulin M (P = 0.08) in colostrum compared with CON. Sows fed FSE showed increased (P < 0.05) content of glucose in colostrum and decreased (P < 0.05) content of urea in serum of their offsprings compared with CON. Sows fed FSE had decreased (P < 0.05) content of interleukin (IL)-6 and increased (P < 0.05) content of IL-10 in serum and colostrum compared with CON. Sows supplemented with FSE had decreased (P < 0.05) contents IL-6 and IL-8 in serum of their offsprings compared with CON. Sows supplemented with FSE also had increased (P < 0.05) content of gonadotropin-releasing hormone in serum of their offsprings compared with CON. In conclusion, dietary supplementation with FSE in sows during late gestation could improve litter birth weight, colostrum fat and protein contents, nutrient digestibility, antioxidant status, immunoglobulin, inflammatory cytokines in sows and newborn piglets. Moreover, FSE could also potentially modulate reproductive hormones in sows.

Neuroendocrine disruption is associated to infertility in chronically stressed female rats

Infertility is a growing worldwide public health problem, and stress is a main factor exerting detrimental effects on female reproduction. However, knowledge regarding the neuroendocrine changes caused by chronic stress in females is limited. Therefore, this study assessed the effects of stress on hormones that control female reproduction during the proestrus and diestrus stages of the estrous cycle, as well as its effects on fertility. Adult females were assigned to either a control or a stress group. Stress consisted of exposure, for 15 min, to cold-water immersion daily for 30 days. Estrous cyclicity, female sexual behavior, as well as hypothalamic kisspeptin, gonadotropin releasing hormone (GnRH) content, serum luteinizing hormone (LH), estradiol (E2), progesterone (P4), corticosterone (CORT) and fertility were assessed after chronic stress. The results show that chronically stressed females exhibited disrupted estrous cyclicity, decreased receptivity, low pregnancy rates and lower numbers of fetuses. The content of Kisspeptin and GnRH in the Anteroventral Periventricular/medial Preoptic Area decreased during proestrus, while Kisspeptin increased in the Arcuate nucleus in proestrus and diestrus. Serum LH decreased only during proestrus, whereas E2 and P4 concentrations decreased during proestrus and diestrus, with a concomitant increase in CORT levels in both stages. As a whole, these results indicate that chronic stress decreases Kisspeptin content in AVPV nucleus and GnRH in POA in females, and might induce disruption of the LH surge, consequently disrupting estrous cyclicity and fertility, leading to lower rates of pregnancy and number of fetuses.

The content of gonadotropin-releasing hormone (GnRH), kisspeptin, and estrogen receptors (ERα/ERβ) in the anteromedial hypothalamus displays daily variations throughout the rat estrous cycle.

The content of gonadotropin-releasing hormone (GnRH), its mRNA, and estrogen receptor alpha (ERα) and beta (ERβ) in the hypothalamus varies throughout the estrous cycle. Furthermore, the abundance of these molecules displays asymmetry between the right and left side. In the present study, we investigated the changes in the content of ERα, ERβ, kisspeptin, and GnRH by western blot in the left and right anteromedial hypothalamus, at four different times during each stage of the rat estrous cycle. The serum levels of the follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were also measured. ERα and ERβ levels changed depending on the stage of the estrous cycle, meanwhile that of kisspeptin was modified according to both the hour of the day and the stage of the cycle. Except in estrus day, ERβ was higher in the right hypothalamus, while ERα was similar in both sides. During both proestrus and estrus, the content of kisspeptin and GnRH was higher in the right hypothalamus. The highest levels of FSH and LH occurred at 17:00h of proestrus. But at estrus, the highest FSH levels were observed at 08:00h and the lowest at 17:00h. Thus, the current results show that the content of ERα, ERβ, kisspeptin, and GnRH in the anteromedial hypothalamus are regulated as a function of the stage of the estrous cycle and the hour of the day. Furthermore, the content of these proteins is regularly higher in the right anteromedial hypothalamus, regardless of the stage of the cycle or time of the day.

Neoadjuvant chemotherapy modifies serum pyrrolidone carboxypeptidase specific activity in women with breast cancer and influences circulating levels of GnRH and gonadotropins.

Functional studies have demonstrated that gonadotropin-releasing hormone (GnRH) regulates cell proliferation, apoptosis, and tissue remodeling. GnRH is metabolized by the proteolytic regulatory enzyme pyrrolidone carboxypeptidase (Pcp) (E.C. 3.4.19.3), which is an omega peptidase widely distributed in fluids and tissues. We previously reported a decrease in both rat and human Pcp activity in breast cancer, suggesting that GnRH may be an important local hormonal factor in the pathogenesis of breast cancer. Recently, we have described that postmenopausal women with breast cancer show lower levels of serum Pcp activity than control postmenopausal women. To determine the effect of neoadjuvant chemotherapy (NACT) on serum Pcp specific activity and circulating levels of GnRH, luteinizing hormone (LH), follicle-stimulating hormone (FSH) and steroid hormones 17-ß-estradiol and progesterone in pre- and postmenopausal women diagnosed with infiltrating ductal carcinoma. Serum Pcp activity was measured fluorometrically using pyroglutamyl-ß-naphthylamide. Circulating GnRH levels were dosed using a commercial RIA kit. Circulating LH and FSH levels were measured by enzyme immunoassays. Levels of steroid hormones were measured in serum samples by dissociation-enhanced lanthanide fluorescence immunoassay. Our results show the effect of NACT on the hypothalamic-pituitary axis, with the consequent alteration of circulating gonadotropins in premenopausal women with breast cancer. However, the results obtained in postmenopausal women with breast cancer treated with NACT, that is, the significant decrease in the concentration of GnRH and FSH compared to control postmenopausal women, differ from those obtained for premenopausal women. The only difference between pre- and postmenopausal women is their hormonal profile at the beginning of the study, that is, the presence of menopause and the consequent alteration of the hypothalamic-pituitary-gonadal axis.

Hypothalamic concentrations of kisspeptin and gonadotropin‐releasing hormone during the breeding season and non‐breeding season in ewes

Current methods to quantify kisspeptin (KP) are limited. To this end, a radioimmunoassay (RIA) specific for KP was developed and validated. We hypothesized that use of a RIA would reveal multiple hypothalamic regions as targets of negative seasonal feedback of estradiol on KP production in sheep. Ovariectomized (OVX) ewes bearing a subcutaneous implant of estradiol were euthanized during the breeding season (BS) (n=4) and non-breeding season (NBS) (n=3). Coronal sections of preoptic area (POA), anterior hypothalamic area (AHA), and mediobasal hypothalamus (MBH) were collected, as well as the median eminence (ME), cortex, brain stem, and cerebellum. Amounts of KP and gonadotropin-releasing hormone (GnRH) in individual hypothalamic nuclei were quantified by radioimmunoassay. Concentration and content of KP were lower during the NBS than the BS in the MBH (P<0.01) and POA (P<0.01). Levels of KP in tissue adjacent to the POA and MBH were much lower, and neither concentration nor content of KP differed between the BS and NBS. Kisspeptin was also detected in the cortex, brain stem, and cerebellum, but concentrations were not affected by season. In addition, concentration and content of GnRH in the POA, AHA, MBH, and ME were similar between seasons. Our RIA results indicate that in addition to the MBH, the POA and AHA appear to be involved in the seasonal negative feedback of estradiol on KP expression.