Plasma Luteinizing Hormone Levels Research Articles

L-arginine supplementation attenuates ovarian oxidative stress in female rats subjected to chronic intermittent hypoxia.

Systemic and organ-specific oxidative stress triggered by hypoxia is suggested to play a key role in germ cell apoptosis and DNA damage. This study was designed to investigate the impact of chronic intermittent hypoxia (CIH) on female fertility and evaluate the potential antioxidant effect of L-arginine (L-Arg) supplementation. Adult female rats were allocated into three groups: controls (normoxic), hypoxic and hypoxic supplemented with L-Arg. After 12 weeks; hematocrit value was determined, body weight (BW) and ovarian weight were measured for the calculation of the gonado-somatic index. Plasma levels of luteinizing hormone (LH) and progesterone were estimated. Ovarian tissue malondialdehyde (MDA) and catalase were assessed, and caspase-3 enzyme expression was detected by immunohistochemistry. Compared to controls, CIH resulted in increased oxidative stress in the ovarian tissue, decreased ovarian weight, and increased frequency of irregular cycles and higher plasma level of LH in rats with either regular or irregular ovarian cycles. Histological examination of ovarian sections revealed areas of degeneration, atretic follicles, interstitial edema, congested vessels and inflammatory cell infiltration. Immunohistochemistry confirmed the presence of apoptosis by increased caspase-3 expression. Hypoxic rats pre-treated with L-Arg showed increased BW and ovarian weight, decreased ovarian tissue MDA and plasma LH accompanied by a lower incidence of irregular cycles and mortality. The histological picture was improved and caspase-3 expression was reduced. Oxidative stress caused by CIH is detrimental to the structure and function of the corpus luteum with an increased risk of reduced fertility. L-Arg supplementation increases antioxidant capacity and improves hypoxia-induced fertility disorders.

Central injection of neuropeptide B induces luteinizing hormone release in male and female rats

Neuropeptide B (NPB) has been identified as an endogenous peptide ligand for the orphan receptor NPBWR1. However, the effect of NPB on the central regulatory mechanisms of reproductive functions remains unclear. Our findings indicated the presence of Npb, Npw (which is another ligand for NPBWR1), and Npbwr1 mRNA in the hypothalamus of male and female rats at each stage of the estrous cycle. Npb mRNA expression was found to be significantly higher in diestrus compared to estrus. The expression of Npw mRNA was one order of magnitude lower than that of Npb mRNA, and Npw mRNA expression in diestrus was significantly higher than that in the other stages of the estrous cycle. Furthermore, Npbwr1 mRNA expression was found to be significantly higher in diestrus compared to the other stages of the estrous cycle and intact males. Notably, estrogen did not alter the expression of Npb, Npw, and Npbwr1 mRNAs in the hypothalamus of females. Central injection of NPB increased plasma luteinizing hormone (LH) levels in both intact males and estrogen-primed ovariectomized females but not in ovariectomized females. These results suggest that NPB-NPBWR1 signaling would be a facilitatory regulatory mechanism in the reproductive function of male and female rats. To the best of our knowledge, this study is the first report to describe the central role of NPB-NPBWR1 signaling in LH regulation in mammals.

The GLP-1 agonist, Exendin-4, stimulates LH secretion in female sheep.

Our previous studies showed that microinjection into the median eminence of the sheep of glucagon-like peptide- 1 (GLP-1) or its receptor agonist Exendin-4 stimulates luteinizing hormone (LH) secretion but it is unknown whether the same effect may be obtained by systemic administration of the same. The present study measured the response in terms of plasma LH concentrations, to intravenous (iv) infusion of Exendin-4. A preliminary study showed that infusion of 2mg Exendin-4 into ewes produced a greater LH response in the follicular phase of the oestrous cycle than the luteal phase. Accordingly, the main study monitored plasma LH levels in response to either 0.5mg or 2mg Exendin-4 or vehicle (normal saline) delivered by jugular infusion for 1h in the follicular phase of the oestrous cycle. Blood samples were collected at 10 min intervals before, during and after infusion. Both doses of Exendin-4 increased mean plasma LH concentrations and increased LH peripheral pulse amplitude. There was no effect on inter-pulse interval or timing of the preovulatory LH surge. These doses of Exendin-4 did not alter plasma insulin or glucose concentrations. Quantitative PCR of the gastrointestinal tract samples from a population of ewes confirmed expression of the preproglucagon gene (GCG). Expression increased aborally and was greatest in the rectum. It is concluded that endogenous GLP-1, most likely derived from the hind-gut may act systemically to stimulate LH secretion. The present data suggest that this effect may be obtained with levels of agonist that are lower than those functioning as an incretin.

Rare association of Kallman Morsier syndrome with a non-functioning pituitary microadenoma

Kallmann's syndrome (KS) is a form of hypogonadotropic hypogonadism associated with a defect in the sense of smell, due to abnormal migration of gonadotropin-releasing hormone-producing neurons. We report the case of a patient aged 16 years and 6 months who presented a micropenis with poor development of secondary sexual characteristics and anosmia. Plasma levels of luteinizing hormone, follicle stimulating hormone and testosterone were very low, while chromosomal analysis showed a 46XY karyotype. Hypothalamic-pituitary MRI showed bilateral agenesis of the olfactory bulbs associated with a 4 mm pituitary microadenoma. The patient was put on hormone replacement therapy with a good response.

Gestational exposure to continuous light impairs the development of the female reproductive system in adult Wistar rat offspring.

It has been suggested that maternal exposure to constant light during the gestational period could be considered as a chronic stressor, impairing offspring development by interfering in neuroendocrine and behavior responses. This study aimed to evaluate whether maternal exposure to continuous light during pregnancy affects the adult reproductive system in the female offspring. Pregnant Wistar rats were allocated into light-dark (LD) group, exposed to light and dark photoperiod during gestation, and the light-light (LL) group, exposed to a photoperiod of constant light during gestation. After birth, pups were maintained under normal light-dark photoperiod until adulthood. At postnatal day 90, blood was collected from the female offspring, to analyze plasma luteinizing hormone (LH) and progesterone levels, and the uterus and ovaries were harvested for morphometric, histological, and oxidative stress evaluations. Female exposure to continuous light during the intrauterine period resulted in the adult reduction of LH and increased progesterone plasma levels, and uterine injuries a higher number of endometrial glands and reduced levels of antioxidant enzymes, such as glutathione reductase and glutathione S-transferase. In these experimental conditions, gestational continuous light exposure disturbs sex hormone balance and reduces the antioxidant enzymatic activity in the uterus of female offspring in adult life.

Mutation pattern of cancer suppressor p53 gene in factory workers exposed to polypropylene and impact on their reproductive hormones.

Polypropylene is a thermoplastic polymer playing the role of an endocrine disruptor that interferes with the union, emission, transport or elimination of normal hormones. Epidemiological information indicated the relation of endocrine-disturbing chemicals with prostate cancer, testis tumor and diminished fertility. p53 is a key tumor silencer gene. The present study aimed to evaluate luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels and the risk of p53 mutations as a result of exposure to polypropylene in non-tumorous adult male factory workers. In total, 150 (controls = 35, workers = 115) subjects were recruited. Groups were maintained according to the tenure of exposure G1 (1-5 years), G2 (6-10years), G3 (11-15 years) and G4 (16-20 years). Concentrations of LH and FSH were determined through an enzyme-linked immunosorbent assay. Genotyping analysis was performed by polymerase chain reaction based gel electrophoresis followed by DNA sequencing. The structural and functional impact of the mutation on the p53 structure was evaluated using 50-ns molecular dynamics (MD) simulations and protein-DNA docking. Mean plasma LH levels were significantly decreased in G1 (p > 0.05) as well as the G2, G3 and G4 (p > 0.001) groups. Similarly, FSH levels were significant decrease in G1 (p > 0.05), G2 (p > 0.01), G3 (p > 0.001) and G4 (p > 0.001) compared to the control group. Sequencing results found three variants i.e. g.13450 T>G, g.13430C>T and g.13737G>A. One of them was predicted to be disease-causing others are polymorphisms. MD simulation of missense mutation R273H showed no structural impact on the protein structure in MD simulation, but it resulted in weaker binding of p53 with the DNA that might lower the gene expression of cell cycle regulatory proteins. These findings predict decreased fertility and risk of malignancies in the future. The spectrum of p53 mutations as a result of polypropylene exposure in the Pakistani population has not been investigated before. Further studies and meta-analyses are required to elucidate the role of different plasticizers in reproduction and cancer-causing risk factors in a larger population.

Enkephalin-δ opioid receptor signaling partly mediates suppression of LH release during early lactation in rats.

Gonadal function is often suppressed during lactation in mammals including rodents, ruminants, and primates. This suppression is thought to be mostly due to the inhibition of the tonic (pulsatile) release of gonadotropin-releasing hormone (GnRH) and consequent gonadotropin. Accumulating evidence suggests that kisspeptin neurons in the arcuate nucleus (ARC) play a critical role in the regulation of pulsatile GnRH/gonadotropin release, and kisspeptin mRNA (Kiss1) and/or kisspeptin expression in the ARC are strongly suppressed by the suckling stimuli in lactating rats. This study aimed to examine whether the central enkephalin-δ-opioid receptor (DOR) signaling mediates the suckling-induced suppression of luteinizing hormone (LH) release in lactating rats. Central administration of a selective DOR antagonist increased the mean plasma LH levels and baseline of LH pulses in ovariectomized lactating mother rats compared to vehicle-injected control dams on day 8 of lactation without affecting the number of Kiss1-expressing cells and the intensity of Kiss1 mRNA signals in the ARC. Furthermore, the suckling stimuli significantly increased the number of enkephalin mRNA (Penk)-expressing cells and the intensity of Penk mRNA signals in the ARC compared to non-lactating control rats. Collectively, these results suggest that central DOR signaling, at least in part, mediates the suppression of LH release induced by suckling stimuli in lactating rats via indirect and/or direct inhibition of ARC kisspeptin neurons.

Short-term depletion of plasma estrogen affects hypothalamic kisspeptin-neurokinin B-dynorphin A neurons, gonadotrophs, and pulsatile luteinizing hormone secretion in female rats

Kisspeptin-neurokinin B-dynorphin A (KNDy) neurons in the arcuate nucleus (ARC) regulate pulsatile luteinizing hormone (LH) secretion. These neurons express estrogen receptors and are negatively regulated by estrogen. This study aimed to determine whether estrogen supplementation after short-term ovariectomy-induced estrogen depletion has different effects on KNDy neurons depending on the timing of the supplementation. To decrease endogenous estradiol (E2) for a short time, adult female rats received a tube filled with E2 one week after ovariectomy and utilized it one week later (O1w + E). From the results of immunohistochemistry, the response to E2 was attenuated in KNDy neurons of O1w + E rats. Enlarged LH-secreting cells in the anterior pituitary were found in O1w + E rats; however, such enlarged LH cells were not found in ones without previous short-term E2 depletion. From the analysis of LH pulses, plasma LH levels were increased in O1w + E rats relative to ones without previous short-term E2 depletion. These results suggested that once endogenous sex steroids were depleted, the response to E2 in hypothalamic KNDy neurons did not fully recover in one week. Thus, short-term sex steroid depletion due to gonadectomy could alter the response to the sex steroids in KNDy neurons even though the period without sex steroids is only one week, and the alteration is likely to affect plasma hormone levels.

An electron microscopic and biochemical study of the potential protective effect of ginger against Cadmium-induced testicular pathology in rats.

Background: Cadmium (Cd) is a toxic heavy metal used in many industries. Since the second half of the 20th century, legislation on Cd use was put to limit the exponential rise in its environmental levels. This study aimed to investigate Cd’s functional and ultrastructural changes on rats’ reproductive systems and the role of Zingiber officinale (Ginger) in protecting against Cd-induced toxicity. Methods: Thirty adult male albino rats were randomly assigned into three equal groups (n = 10); control, Cd-exposed/untreated, and Cd-exposed/Gin-treated. Rat testes were weighed, and testicular tissue sections were examined under the electron microscope. Semen analysis, morphological examination of spermatozoa, and serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone were measured. In addition, testicular tissue homogenates were analyzed for malondialdehyde (MDA), nitric oxide (NO), and reduced glutathione (GSH) levels. Results: Cd-induced significant reduction in the mean testicular weight and GSH levels and plasma testosterone, LH and FSH levels with a concomitant increase in testicular MDA and NO levels. There was also a deterioration in semen analysis parameters and spermatozoa morphology, with testicular structural damage in the form of architecture distortion and necrosis of seminiferous tubules and testicular interstitial cells. Daily administration of ginger for 4 weeks protected against CD-induced toxicity, preserving tissue architecture, improved plasma levels of testosterone, LH and FSH and testicular levels of GSH, and reduced testicular levels of MDA, NO. Conclusion: Ginger has a protective effect on Cd-induced deterioration of testicular tissue’s structural and functional integrity by improving testicular tissue antioxidant capacity and steroid production, which ameliorates sex hormone levels in the blood.

Ameliorative Effect of Ethanolic Fruit Extract of Perssea americana and Prunus dulcis on Caffein-Testicular Damage in Male Wistar Albino Rats

This study investigated the effect if ethanolic fruit extracts of Perssea americana and Prunus dulcis on caffeine-testicular damage in male albino rats. Sixty rats weighing 120 and 170 g were used for this study. The rats were separated into 12 groups of five rats per group. Group 1 served as normal control, group 2 received 200mg/kg of caffeine, serving as negative control while group 3 served as positive control. Rats in group 4-6 were orally administered 200mg/kg caffeine + ethanolic 100, 200, and 400mg/kg of Prunus dulcis, group 7-12 received 200mg/kg caffeine + ethanolic 100, 200, and 400mg/kg, and Caffeine + ethanolic 100mg/kg Persea americana +100mg/kg Prunus dulcis fruit extract for 7, 14, and 21 days. Blood sample was collected from the tail of each rats at 7 days interval after treatment, for biochemical assays while one rats in each group was sacrificed and the testis was harvested for histological analysis. The plasma luteinizing hormone level of the negative control for 7, 14, and 21 days were 1.62 ± 0.01IU/ML, 1.71 ± 0.02IU/ML, and 1.05 ± 0.09IU/ML respectively. The plasma luteinizing hormone level of group 6 treated with 400mg/kg of Prunus dulcis fruit extract for 7, 14, and 21 days were 3.06. ± 0.05IU/ML, 3.41 ± 0.05IU/ML, and 3.63 ± 0.01IU/ML respectively, were significantly different from group 2 while those of group 9 treated with 400mg/kg of Persea americana fruit extract for 7, 14, and 21 days were 3.51 ± 0.05IU/ML, 3.82 ± 0.05IU/ML, and 3.90 ± 0.02IU/ML. Regeneration of damage testicular tissues occurred after treatment. The significant increases observed on the hormonal profile and regeneration of damaged testicular tissues of the extracts treated rats indicated testiculo-curative effect of P. dulcis and P. americana fruit on caffeine-induced testicular damage in rats.

Chemogenetic Depletion of Hypophysiotropic GnRH Neurons Does Not Affect Fertility in Mature Female Zebrafish.

The hypophysiotropic gonadotropin-releasing hormone (GnRH) and its neurons are crucial for vertebrate reproduction, primarily in regulating luteinizing hormone (LH) secretion and ovulation. However, in zebrafish, which lack GnRH1, and instead possess GnRH3 as the hypophysiotropic form, GnRH3 gene knockout did not affect reproduction. However, early-stage ablation of all GnRH3 neurons causes infertility in females, implicating GnRH3 neurons, rather than GnRH3 peptides in female reproduction. To determine the role of GnRH3 neurons in the reproduction of adult females, a Tg(gnrh3:Gal4ff; UAS:nfsb-mCherry) line was generated to facilitate a chemogenetic conditional ablation of GnRH3 neurons. Following ablation, there was a reduction of preoptic area GnRH3 neurons by an average of 85.3%, which was associated with reduced pituitary projections and gnrh3 mRNA levels. However, plasma LH levels were unaffected, and the ablated females displayed normal reproductive capacity. There was no correlation between the number of remaining GnRH3 neurons and reproductive performance. Though it is possible that the few remaining GnRH3 neurons can still induce an LH surge, our findings are consistent with the idea that GnRH and its neurons are likely dispensable for LH surge in zebrafish. Altogether, our results resurrected questions regarding the functional homology of the hypophysiotropic GnRH1 and GnRH3 in controlling ovulation.

Effect of ZhuLian's type Ⅱ inhibition needling technique on endocrine metabolism in rats with polycystic ovary syndrome and insulin resistance

To observe the effect of ZhuLian's type Ⅱ inhibition needling technique on sex hormones, insulin resistance and ovarian morphology in rats with polycystic ovary syndrome (PCOS), so as to explore its mechanism on regulating endocrine and metabolic dysfunction of PCOS. Twenty-four female SD rats were randomly divided into control, model and ZhuLian acupuncture groups (n=8 in each group). The PCOS model was established by continuous gavage of letrozole combined with high-fat diet. Rats of the ZhuLian acupuncture group were stimulated at "Guanyuan"(CV4) and bilateral "Guilai"(ST29), "Zusanli"(ST36), "Sanyinjiao"(SP6), and "Fenglong"(ST40) by ZhuLian's type Ⅱ inhibition needling technique for 30 min, continuously intervented for 28 days. The body weight was measured before and after modeling and after intervention. The ovarian volume was calculated. HE staining was used to observe the changes of ovarian histology. Plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) and fasting insulin (FINS) were measured by ELISA. LH/FSH ratio and insulin resistance index (HOMA-IR) were calculated. After modeling, the rats' body weight and the ovarian volume were increased (P<0.05), and the ovaries showed polycystic changes in the model group compared with the control group. And the FINS level, HOMA-IR, LH content and LH/FSH ratio were significantly increased (P<0.05), while the FSH level was decreased (P<0.05). After the intervention, the rats' body weight and ovarian volume were decreased (P<0.05), and a small number of preantral follicles and more antral follicles were observed under microscope, and the levels of LH, LH/FSH, FINS and HOMA-IR were significantly decreased (P<0.05), while FSH content was increased (P<0.05) in the ZhuLian acupuncture group than in the model group. ZhuLian's type Ⅱ inhibition needling technique can improve the endocrine and metabolic disorders in PCOS rats, which may be related to the regulation of endocrine system.

Vasoactive Intestinal Peptide Indirectly Elicits Pituitary LH Secretion Independent of GnRH in Female Zebrafish

Vasoactive intestinal peptide (Vip) regulates luteinizing hormone (LH) release through the direct regulation of gonadotropin-releasing hormone (GnRH) neurons at the level of the brain in female rodents. However, little is known regarding the roles of Vip in teleost reproduction. Although GnRH is critical for fertility through the regulation of LH secretion in vertebrates, the exact role of the hypophysiotropic GnRH (GnRH3) in zebrafish is unclear since GnRH3 null fish are reproductively fertile. This phenomenon raises the possibility of a redundant regulatory pathway(s) for LH secretion in zebrafish. Here, we demonstrate that VipA (homologues of mammalian Vip) both inhibits and induces LH secretion in zebrafish. Despite the observation that VipA axons may reach the pituitary proximal pars distalis including LH cells, pituitary incubation with VipA in vitro, and intraperitoneal injection of VipA, did not induce LH secretion and lhβ mRNA expression in sexually mature females, respectively. On the other hand, intracerebroventricular administration of VipA augmented plasma LH levels in both wild-type and gnrh3-/- females at 1 hour posttreatment, with no observed changes in pituitary GnRH2 and GnRH3 contents and gnrh3 mRNA levels in the brains. While VipA's manner of inhibition of LH secretion has yet to be explored, the stimulation seems to occur via a different pathway than GnRH3, dopamine, and 17β-estradiol in regulating LH secretion. The results indicate that VipA induces LH release possibly by acting with or through a non-GnRH factor(s), providing proof for the existence of functional redundancy of LH release in sexually mature female zebrafish.

Effects of aqueous and hydro-alcoholic extracts of Seidlitzia rosmarinus on male mouse reproductive system: An experimental study.

This study investigated the effects of aqueous and hydro-alcoholic extracts of Seidlitzia rosmarinus on reproductive hormones, sperm variables, and antioxidant enzymes level in the mice testis. In this experimental study, 24 three-month-old male NMRI mice weighing (25-30g) were divided into three groups: control, aqueous and hydro-alcoholic extracts of Seidlitzia rosmarinus 100mg/kg. Dissolved extracts were gavaged orally for 35 days. One day after receiving the last dose of the extract, the blood sample, testis, and the epididymis tail were taken for plasma hormonal, testicular antioxidants level, sperm count, and vitality assessments. Testicular level of malondialdehyde increased in aqueous and hydro-alcoholic extracts groups (p=0.04); total antioxidant capacity decreased in aqueous and hydro-alcoholic extracts groups (p=0.008); and the consumption of aqueous (p<0.001) and hydro-alcoholic (p=0.03) extracts decreased catalase in comparison with the control group. The plasma level of luteinizing hormone decreased in the aqueous extracts administrated group (p=0.009); the follicle-stimulating hormone increased in aqueous (p=0.03), and hydro-alcoholic extracts administered mice; and the testosterone level decreased in aqueous extract-treated animals versus the control group (p<0.001). The sperm count was increased in aqueous (p=0.04) and hydro-alcoholic (p=0.009) extracts groups, but its vitality was decreased (p=0.008) in comparison with the control group. In conclusion, Seidlitzia rosmarinus has an adverse effect on male reproductive hormones and sperm viability via increased lipid peroxidation and reduced antioxidant defense system performance.

Plasma levels of luteinizing hormone and prolactin in relation to double brooding in Great Tit (Parus major)

The reproductive behaviors of birds are mainly controlled by the hypothalamus-pituitary-gonad axis. Many studies have shown that reproductive hormones are tightly linked to the breeding sub-stages. However, only a few studies have examined the temporal trend of hormone levels among different reproductive stages in multiple brooded species. We investigated the changes in plasma luteinizing hormone (LH) and prolactin (PRL) concentrations during different reproductive stages of the facultative double-brooded Great Tit (Parus major). We found that the concentrations of LH and PRL in females were significantly higher than those in males. Females had significantly higher LH and lower PRL concentrations in the pre-breeding period than in the first/second brooding periods, and there were no significant changes between the first and second brooding periods. The concentrations of LH and PRL in males had no significant difference between the pre-breeding period and the first brooding periods, while LH and PRL concentrations in the second brooding period were significantly higher than those in the first brooding period. We conclude that there are sex-based differences between LH and PRL at different stages of reproduction. The changes in LH and PRL in both males and females should be related to their physiological functions. Especially for males, individuals with higher levels of LH and PRL are more likely to maintain second clutches.

The GnRH Antagonist Degarelix Suppresses Gonadotropin Secretion and Pituitary Sensitivity in Midgestation Sheep Fetuses.

The specific role of gonadotropin-releasing hormone (GnRH) on brain sexual differentiation remains unclear. To investigate whether gonadotropin and, in turn, testosterone (T) secretion is regulated by GnRH during the critical period for brain differentiation in sheep fetuses, we attempted to selectively suppress pituitary-testicular activation during midgestation with the long-acting GnRH antagonist degarelix. Fetuses received subcutaneous injections of the antagonist or vehicle on day 62 of gestation. After 2 to 3 weeks we examined consequences of the intervention on baseline and GnRH-stimulated plasma luteinizing hormone (LH) and T levels. In addition, we measured the effect of degarelix-treatment on messenger RNA (mRNA) expression for the pituitary gonadotropins and key gonadal steroidogenic enzymes. Baseline and GnRH-stimulated plasma LH levels were significantly suppressed in degarelix-treated male and female fetuses compared to control values. Similarly, T concentrations were suppressed in degarelix-treated males. The percentage of LHβ-immunoreactive cells colocalizing c-fos was significantly reduced by degarelix treatment indicating that pituitary sensitivity was inhibited. Degarelix treatment also led to the significant suppression of mRNA expression coding for the pituitary gonadotropin subunits and for the gonadal enzymes involved in androgen synthesis. These findings demonstrate that pharmacologic inhibition of GnRH early in gestation results in suppression of LH secretion and deficits in the plasma T levels of male lamb fetuses. We conclude that GnRH signaling plays a pivotal role for regulating T exposure during the critical period of sheep gestation when the brain is masculinized. Thus, disturbance to gonadotropin secretion during this phase of gestation could have long-term consequence on adult sexual behaviors and fertility.

Pituitary and ovarian hormones: is their plasma concentration affected by litter size in primiparous lactating rabbit does?

Genetic selection in commercial rabbit lines based on litter size has positively improved the number of kits suckling, presumably to weaning. Although it has been proven that the energetic balance of primiparous does is due to the need to satisfy pregnancy, lactation and growth requirements, litter size adjustment from 7 to 12 kits is applied as a routine in commercial rabbit farms. The suckling stimulus provokes a prolactin (PRL) secretion, which in turn can modulate the preovulatory release of luteinising hormone (LH) and, consequently, the ovulatory and productive responses of the does. This study aimed to determine if litter size of prolific primiparous rabbit does during lactation [Group HL, with high litter density (10-12 kits; n=21) and Group LL, with low litter density (7-9 kits; n=29)] influences plasma concentration of PRL. Blood samples from lactating does were taken weekly throughout lactation starting on day 4 post-partum, until day 32 post-partum, before and immediately after suckling. In addition, the does were re-inseminated after weaning (day 32 post-partum), and sampled at 0 and 60 min after induction of ovulation to determine whether litter size affected the peak of LH, progesterone (P4) concentrations and the main productive parameters of their second pregnancy. All hormones were determined by enzyme immunoassay. Statistical analysis of the results revealed that the PRL concentrations of hyperprolific rabbit does before and immediately after a suckling stimulus from 7-9 or 10-12 kits were significanltly different, as we only detected basal levels, with a rise after weaning in both groups. More studies are necessary, delaying blood sampling to later periods of time after the suckling stimulus, in order to conclude whether the peak release of this hormone is altered or not. There were also no differences in plasma LH and progesterone levels after artificial insemination, or in productive performance of these females after their second pregnancy. In conclusion, the litter size adjustment of prolific primiparous rabbits with 7 to 12 kits determines adequate pituitary, ovarian and reproductive responses at second parturition if the does are inseminated after weaning.

Characteristics of Neurokinin-3 Receptor and Its Binding Sites by Mutational Analysis.

Simple SummaryThis study presents in silico models of neurokinin B receptor tiTac3Ra (tilapia Tachykinin 3 receptor a) and its potential binding sites, as well as docking native tilapia Neurokinin F (tiNKF) and tilapia Neurokinin B (tiNKB) to theses orthosteric binding sites. For a better understanding of the binding confirmation and interaction of the structures, we compared the conformation between peptide docking and induced fit docking results. We have tried to analyze the affinity of binding and binding site interactions parallel to in-vitro results of receptor activity. NKB antagonists inhibit male tilapia gonadal development and gonadotropin release. We further verified the in-vivo effect of the antagonists on gonadal development in males. Studying the receptor activity of variants with alanine mutations at Phe2516.44 and Met2897.43 respectively, we found that, while both variants completely underperformed, there was no direct interaction with the ligand in the binding process indicating their role in post binding receptor activation rather than the binding process itself.NKB (Neurokinin B) is already known to play a crucial role in fish reproduction, but little is known about the structure and function of NKB receptors. Based on an in silico model of the tilapia NKB receptor Tachykinin 3 receptor a (tiTac3Ra) found in the current study, we determined the key residues involved in binding to tilapia NKB and its functional homologue NKF (Neurokinin F). Despite studies in humans suggesting the crucial role of F2516.44 and M2897.43 in NKB binding, no direct peptide interaction was observed in tilapia homologs. In-silico, Ala mutations on residues F2516.44 and M2897.43 did not influence binding affinity, but significantly affected the stability of tiTac3Ra. Moreover, in vitro studies indicated them to be critical to tiNKB/tiNKF-induced receptor activity. The binding of NKB antagonists to tiTac3Ra both in-vitro and in vivo inhibits FSH (follicle stimulating hormone) and LH (luteinizing hormone) release and sperm production in mature tilapia males. Non-peptide NKB antagonist SB-222200 had a strong inhibitory effect on the Tac3Ra activation. SB-222200 also decreased LH plasma levels; two hours post intraperitoneal injection, changed sperm volume and the ratios of the different stages along the spermatogenesis in tilapia testes.

Effects of neurokinin 3 receptor antagonist fezolinetant on hot flash-like symptoms in ovariectomized rats

The majority of women experience vasomotor symptoms (VMS), such as hot flashes and night sweats, during the menopausal transition. Recent evidence strongly suggests a connection between neurokinin 3 (NK3) receptor signaling and VMS associated with menopause. The NK3 receptor antagonist fezolinetant is currently in phase 3 development for treatment of moderate to severe VMS associated with menopause. We investigated the pharmacological effects of repeated administration of fezolinetant on levels of sex hormones and gonadotropins, neuronal activity in the hypothalamus, and skin temperature as an index of hot flash-like symptoms in ovariectomized rats as a model of menopause. Ovariectomized rats exhibited several typical menopausal symptoms: hyperphagia, increased body weight, significantly decreased plasma estradiol levels, increased luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels, and significantly increased skin temperature. Increased c-Fos expression (an indirect marker of neuronal activity) in median preoptic nucleus (MnPO) hypothalamic neurons was also observed in ovariectomized rats. Repeated oral administration of fezolinetant (1–10 mg/kg, twice daily) for 1 week dose-dependently reduced plasma LH levels without affecting estradiol or FSH levels, inhibited the activation of MnPO neurons, and attenuated hot flash-like symptoms. In addition, fezolinetant dose-dependently reduced hyperphagia and weight gain in ovariectomized rats. These preclinical findings suggest that fezolinetant attenuates hot flash-like symptoms via inhibition of neuronal activity in the MnPO of ovariectomized rats and provides further support for the ongoing clinical development of fezolinetant for the treatment of VMS associated with menopause.

Effects of Diet-Induced Obesity on Hypothalamic Kisspeptin-Neurokinin-Dynorphin (KNDy) Neurons and Luteinizing Hormone Secretion in Sex Hormone-Primed Male and Female Rats

Metabolic stress resulting from a nutrient excess causes infertility in both sexes. Kisspeptin-neurokinin B-dynorphin (KNDy) neurons in the arcuate nucleus (ARC) have been suggested to be key players in reproduction via direct stimulation of gonadotropin-releasing hormone (GnRH) and subsequent gonadotropin release in mammalian species. In this study, we investigated the sex differences in the effects of a high-fat diet (HFD) on KNDy-associated gene expression in the ARC to determine the pathogenic mechanism underlying obesity-induced infertility. Wistar-Imamichi strain male and female rats (7 weeks of age) were fed either a standard diet (10% calories from fat) or high-fat diet (45% calories from fat) for 4 months. In male rats, the HFD caused a significant suppression of Kiss1(encoding kisspeptin), Tac3(encoding neurokinin B), and Pdyn(encoding dynorphin A) gene expression in the ARC, resulting in a decrease in plasma luteinizing hormone (LH) levels. In female rats, 58% of the HFD-fed female rats exhibited irregular estrous cycles, while the other rats showed regular cycles. LH pulses were found, and the numbers of ARC Kiss1-,Tac3-, and Pdyn-expressing cells were high in control animals and almost allHFD-fed female rats, but two out of 10 rats showed profound HFD-induced suppression of LH pulse frequency and reduction in these cells. No statistical differences in LH secretion or ARC KNDy gene expression were observed between HFD-fed and control female rats. Additionally, the number of Gnrh1-expressing cells in the preoptic area was comparable between the groups in both sexes. Our findings revealed that HFD-fed male rats showed KNDy-dependent infertility, while irregular menstruation was mainly induced by KNDy-independent pathways during the incipient stage of obese infertility in female rats. Taken together, hypothalamic kisspeptin neurons in male rats may be susceptible to HFD-induced obesity compared with those in female rats.