Regulation Of Reproductive Function Research Articles

The saccus vasculosus of the neotropical cichlid fish Cichlasoma dimerus: characterization, developmental studies and its response to photoperiod.

The saccus vasculosus is an organ present in gnathostome fishes, located ventral to the hypothalamus and posterior to the pituitary gland, whose structure is highly variable among species. In some fishes, this organ is well-developed; however, its physiological function is still under debate. Recently, it has been proposed that this organ is a seasonal regulator of reproduction. In the present work, we examined the histology, ultrastructure, and development of the saccus vasculosus in Cichlasoma dimerus. In addition, immunohistochemical studies of proteins related to reproductive function were performed. Finally, the potential response of this organ to different photoperiods was explored. C. dimerus presented a well-developed saccus vasculosus consisting of a highly folded epithelium, composed of coronet and supporting cells, closely associated with blood vessels, and a highly branched lumen connected to the third ventricle. Coronet cells showed all the major characteristics described in other fish species. In addition, some of the vesicles of the globules were positive for thyrotropin beta subunit, while luteinizing hormone beta subunit immunostaining was observed at the edge of the apical processes of some coronet cells. Furthermore, neuropeptide Y and gonadotropin inhibitory hormone innervation in the saccus vasculosus of C. dimerus were shown. Finally, animals exposed to the long photoperiod showed lower levels of thyrotropin beta and common alpha subunits expression in the saccus compared to those of animals exposed to short photoperiod. All these results support the hypothesis that the saccus vasculosus is involved in the regulation of reproductive function in fish.

Response of the serine/threonine kinase AKT and phosphoinositide-dependent kinase PDK in Frankliniella occidentalis (Thysanoptera: Thripidae) to three kinds of foods and their regulation of reproductive function.

Frankliniella occidentalis (Pergande) is a typical omnivorous insect that feeds on host plants, pollens and mite eggs, and poses a threat to crops worldwide. The insulin signalling pathway (ISP) is a typical nutrient-sensitive pathway that participates in the regulation of various functions in insects. Serine/threonine kinases (AKTs) and phosphoinositide-dependent kinases (PDKs) are key components of the ISP. In this study, the FoAKT and FoPDK genes in F. occidentalis were cloned, and the effects of three foods on their expression were determined. The expression of FoAKT and FoPDK in the thrips fed on kidney bean leaves supplemented with pine pollen or mite eggs was higher than in those primarily fed on leaves alone. Meanwhile, the fecundity of thrips fed on leaves supplemented with pine pollen was highest. In addition, RNA interference-mediated knockdown of FoAKT and FoPDK decreased vitellogenin (Vg) content and Vg expression in females, shortened ovariole length, delayed egg development and reduced fecundity and offspring hatching rates. Furthermore, the synthesis of juvenile hormone (JH) was reduced, and the contents of glucose, trehalose, glycogen and trehalase were affected. These results suggest that FoAKT and FoPDK regulate the reproduction of F. occidentalis by regulating Vg and JH production as well as carbohydrate metabolism.

Neuropeptide expression and action in the reproductive system of the starfish Asterias rubens.

Reproductive processes are regulated by a variety of neuropeptides in vertebrates and invertebrates. In starfish (phylum Echinodermata), relaxin-like gonad-stimulating peptide triggers oocyte maturation and spawning. However, little is known about other neuropeptides as potential regulators of reproduction in starfish. To address this issue, here, we used histology and immunohistochemistry to analyze the reproductive system of the starfish Asterias rubens at four stages of the seasonal reproductive cycle in male and female animals, investigating the expression of eight neuropeptides: the corticotropin-releasing hormone-type neuropeptide ArCRH, the calcitonin-type neuropeptide ArCT, the pedal peptide-type neuropeptides ArPPLN1b and ArPPLN2h, the vasopressin/ocytocin-type neuropeptide asterotocin, the gonadotropin-releasing hormone-type neuropeptide ArGnRH, and the somatostatin/allatostatin-C-type neuropeptides ArSS1 and ArSS2. The expression of five neuropeptides, ArCRH, ArCT, ArPPLN1b, ArPPLN2h, and asterotocin, was detected in the gonoducts and/or gonads. For example, extensive ArPPLN2h expression was revealed in the coelomic epithelial layer of the gonads throughout the seasonal reproductive cycle in both males and females. However, seasonal and/or sexual differences in the patterns of neuropeptide expression were also observed. Informed by these findings, the in vitro pharmacological effects of neuropeptides on gonad preparations from male and female starfish were investigated. This revealed that ArSS1 causes gonadal contraction and that ArPPLN2h causes gonadal relaxation, with both neuropeptides being more effective on ovaries than testes. Collectively, these findings indicate that multiple neuropeptide signaling systems are involved in the regulation of reproductive function in starfish, with some neuropeptides exerting excitatory or inhibitory effects on gonad contractility that may be physiologically relevant when gametes are expelled during spawning.

Implications of lifestyle factors on male reproductive health.

In recent decades, there has been a substantial decline in sperm quality in humans, with lifestyle factors playing a major role in this trend. There are several lifestyle factors which are contributing to male infertility. This review, however, discusses factors such as obesity, diet/nutrition, psychological stress, radiation exposure, cigarette smoking, and alcohol use with reference to male infertility. Sperm count, motility, morphology and sperm DNA may be adversely affected by lifestyle factors, which may also affect the endocrine regulation of reproductive function. The decline in male fertility has a significant impact on fertility rates, and the resulting implications for the human population make this a serious public health concern in the twenty-first century. Thus, lifestyle interventions through a specific framework of educational, environmental, nutritional/physical exercise, and psychological support coupled with the use of nutritional antioxidants supplements can help couples achieve better health and well-being and improve their fertility prospects or increase their chances of conception.

Statistical assessment of the concentration of anti-Muller hormone and biochemical parameters of the blood of donor cows in the luteal phase of the cycle

Increasing the productivity of cows in the modern world is a key aspect of animal husbandry. Embryo transplantation in cows makes it possible to increase the production of genetically valuable animals and improve the breeding qualities of the herd. One of the methods for determining the activity and reserve of the ovaries is the determination of anti-Muller hormone in the blood. AMH plays an important role in the regulation of reproductive function in animals, including cows. High levels of AMH may indicate a good reserve capacity of the ovaries, which may be useful when choosing animals for breeding. In the presented study, the bio-chemical parameters of the blood of 9 Holstein cows in the luteal phase were analyzed. A direct correlation of a high degree (0.89 and 0.89, respectively) was found between the AMH level indicator and urea and urea nitrogen indicators. There was a direct noticeable correlation between the AMH level and the serum creatinine level (0.63). Also, when assessing the indicators of mineral metabolism, strong correlations were revealed: negative when comparing the level of AMH with the level of calcium (-0.71) and positive when comparing the level of AMH with the level of phosphorus (0.78). The regulation of the synthesis of AMH itself and its metabolic effects have been studied little, there is only a limited amount of data on the relationship of the hormone with other factors of autocrine and paracrine regulation. For a more accurate understanding of this correlation and its significance, additional research is needed.

Biotechnology of regulation of reproductive functions of Bos primigenius taurus

Since cattle are biologically capable of giving birth to only one calf per year, they are considered a single-fruited animal species. One of the cardinal methods of solving the problem of accelerated reproduction of livestock is biotechnological regulation of reproductive function in cows and synchronisation of their sexual activity. The aim of the study is to investigate the peculiarities of the reproductive function of Red Steppe cows under the influence of various methods of its regulation using modern hormonal agents. To evaluate the reproductive function of cows, generally accepted zootechnical methods were used, methods of variation statistics were used to calculate data, and biotechnological methods were used to regulate sexual activity in cows. Based on the analysis of the state of reproduction of the cattle herd, it was established that 164-187 cows and 85-96 heifers are artificially inseminated annually, which ensures the yield of calves per 100 cows of 89-91 heads and the annual introduction of 36 to 40 first-born calves per 100 cows. The shortened service period contributes to both a higher calf yield and an increase in milk yield, which is a desirable production effect. However, calving to conception interval, necessary to prepare the cow’s body for a new insemination, can be reduced to at least 30 days. Insemination of heifers with artificial insemination devices should be started when they reach a live weight of 314±6.5 kg, which will allow to obtain fertility from the first insemination at the level of 73.7-77.8% and thereby prevent obesity of heifers, which mainly reduces the percentage of fertility from the first insemination. It is necessary to establish obstetric and gynaecological examinations and treatment of problematic replacement heifers and cows, and it is advisable to use prostaglandin F2α analogues in combination with vitamins or biologically active drugs to stimulate the sexual function of cows and heifers. The use of prophylactic drugs will help to synchronise the heifers' sexual desire and increase their fertility during insemination. And the organisation of cattle reproduction and the proposed biotechnological ways to improve artificial insemination of the red steppe breed breeding stock will significantly improve their reproductive status, which will increase the overall economic efficiency of the industry

Fibroblast Growth Factor 21 Has a Diverse Role in Energetic and Reproductive Physiological Functions of Female Beef Cattle.

Fibroblast growth factor 21 (FGF21) has been identified in multiple mammalian species as a molecular marker of energy metabolism while also providing negative feedback to the gonads. However, the role of FGF21 in regulating the energetic and reproductive physiology of beef heifers and cows has yet to be characterized. Herein, we investigated the temporal concentrations of FGF21 in female beef cattle from the prepubertal period to early lactation. Circulating concentrations of FGF21, non-esterified fatty acids, plasma urea nitrogen, glucose, and progesterone were assessed. Ultrasonography was employed to determine the onset of puberty and resumption of postpartum ovarian cyclicity as well as to measure backfat thickness. Finally, cows and calves underwent the weigh-suckle-weigh technique to estimate rate of milk production. We have revealed that FGF21 has an expansive role in the physiology of female beef cattle, including pubertal onset, adaptation to nutritional transition, rate of body weight gain, circulating markers of metabolism, and rate of milk production. In conclusion, FGF21 plays a role in physiological functions in beef cattle that can be applied to advance the understanding of basic scientific processes governing the nutritional regulation of reproductive function but also provides a novel means for beef cattle producers to select parameters of financial interest.

Pathogenesis of the crosstalk between reproductive function and stress in animals - part 2: Prolactin, thyroid, inflammation and oxidative stress.

Stress has a significant impact on reproductive health and fertility in both humans as well as various animal species. In particular, chronic stress can disrupt the delicate balance of the hormonal system that regulates reproductive function, leading to a variety of reproductive disorders and fertility issues. Beside the action of the hypothalamic-pituitary-adrenal (HPA) system and the sympatho-adrenomedullary system (SAM), other subsequent mechanisms have been incriminated. Thus, stress has also been associated with increased prolactin level, resulting in an inhibition of the hypothalamo-pituitary-gonadal (HPG) system leading to several reproductive disorders. Thyroid function is inhibited during chronic stress, and therefore considered an important regulator of reproductive function. Thus, and in particular by interfering with the HPA system, stress-induced immune dysregulation can have adverse effects on reproduction. In addition, oxidative stress and inflammation have been proposed as potential mechanisms by which chronic stress affects reproductive function. This is caused by an increase in reactive oxygen species (ROS) production that has a harmful effect on cells. Furthermore, inflammation can lead to tissue damage and scarring, which can affect fertility. The present review completes the complex mechanism linking stress and reproduction through the current knowledge in various animal species in a comparative point of view.

The influence of the opioid pentapeptide methionine-enkephalin on seasonal and FSH-induced ovarian recrudescence in the gecko Hemidactylus frenatus

Although methionine-enkephalin (M-ENK) is implicated in the regulation of reproductive functions in vertebrates, its function in reptiles is little understood. This study aims to elucidate the role of M-ENK on seasonal and follicle stimulating hormone (FSH)-induced ovarian recrudescence in the gecko Hemidactylus frenatus. In the first experiment, administration of 5 µg M-ENK did not affect germinal bed activity or follicular developmental stages I, II, and III (previtellogenic) and IV (vitellogenic), but there were no stage V (vitellogenic) follicles in the ovary. However, there was a significant decrease in the mean numbers of oogonia and primary oocytes in the germinal bed associated with the complete absence of stage IV and V follicles in 25 µg M-ENK-treated lizards in contrast to experimental controls. Furthermore, there was a significant decrease in gonadotropin-releasing hormone - immunoreactive (GnRH-ir) content in the median eminence (ME) and pars distalis (PD) of the pituitary gland and sparse labelling of hypothalamic GnRH-ir neurons in 25 µg M-ENK-treated lizards. In the second experiment, treatment with FSH during the regression phase of the ovarian cycle resulted in the appearance of stage IV and V follicles, in contrast to their absence in the initial controls and treatment controls. However, treatment with 25 µg M-ENK + FSH did not result in the appearance of these follicles, indicating the inhibitory effect of M-ENK on FSH-induced ovarian recrudescence. These findings suggest that M-ENK inhibits the germinal bed and vitellogenic follicular growth in a dose-dependent manner, possibly mediated through the suppression of GnRH release in the ME and PD. In addition, M-ENK may also act at the level of the ovary in the gecko.

Discovery of Paralogous GnRH and Corazonin Signaling Systems in an Invertebrate Chordate.

Gonadotropin-releasing hormone (GnRH) is a key regulator of reproductive function in vertebrates. GnRH is related to the corazonin (CRZ) neuropeptide which influences metabolism and stress responses in insects. Recent evidence suggests that GnRH and CRZ are paralogous and arose by a gene duplication in a common ancestor of bilaterians. Here, we report the identification and complete characterization of the GnRH and CRZ signaling systems in the amphioxus Branchiostoma floridae. We have identified a novel GnRH peptide (YSYSYGFAP-NH2) that specifically activates two GnRH receptors and a CRZ peptide (FTYTHTW-NH2) that activates three CRZ receptors in B. floridae. The latter appear to be promiscuous, as two CRZ receptors can also be activated by GnRH in the physiological range. Hence, there is a potential for cross-talk between these closely related signaling systems. Discovery of both the GnRH and CRZ signaling systems in one of the closest living relatives of vertebrates provides a framework to discover their roles at the transition from invertebrates to vertebrates.

Physiological action of vitamin D3 in health and disease

Vitamin D is the common molecules for every steroid that has the biological effects of cholecalciferol. Vitamin D can be synthesized in the mammalian skin after exposed to ultraviolet (UV) waves and this process occur endogenously. Amongst diverse Vitamins, VitD has powerful effects on the immune system. As major components of the diet, vitamins have essential effects on the innate and acquired immune system. The active form of VitD is calcitriol (1,25(OH)2VitD3). Calcitriol (1,25(OH)2VitD3) is the active form of VitD. Calcitriol regulates antimicrobial peptides productions, comprising defensin and cathelicidin, that controller the natural intestine microbiota floor and supports intestinal barriers. VitD in controlling the immune response in infectious and autoimmune diseases. There is a theory that VitD complements could be beneficial for treatment of COVID-19. Vit D has an important anti-inflammatory function on the immune system by reducing the production of pro-inflammatory cytokines and increasing anti-inflammatory cytokines in immune cells. Also, vitamin D deficiency is closely related to chronic diseases such as osteoporosis, type 1 and type 2 diabetes, hypertension, cardiovascular disease and cancer. In addition, recently it was revealed that vitamin D receptors (VDR) are expressed in many organs such as the testes, and vitamin D may be a adjustable regulator of reproductive function and fertility.

Enkephalin-δ Opioid Receptor Signaling Mediates Glucoprivic Suppression of LH Pulse and Gluconeogenesis in Female Rats.

Energy availability is an important regulator of reproductive function at various reproductive phases in mammals. Glucoprivation induced by 2-deoxy-D-glucose (2DG), an inhibitor of glucose utilization, as an experimental model of malnutrition suppresses the pulsatile release of GnRH/LH and induces gluconeogenesis. The present study was performed with the aim of examining whether enkephalin-δ-opioid receptor (DOR) signaling mediates the suppression of pulsatile GnRH/LH release and gluconeogenesis during malnutrition. The administration of naltrindole hydrochloride (NTI), a selective DOR antagonist, into the third ventricle blocked the suppression of LH pulses and part of gluconeogenesis induced by IV 2DG administration in ovariectomized rats treated with a negative feedback level of estradiol-17 β (OVX + low E2). The IV 2DG administration significantly increased the number of Penk (enkephalin gene)-positive cells coexpressing fos (neuronal activation marker gene) in the paraventricular nucleus (PVN), but not in the arcuate nucleus (ARC) in OVX + low E2 rats. Furthermore, double in situ hybridization for Penk/Pdyn (dynorphin gene) in the PVN revealed that approximately 35% of the PVN Penk-expressing cells coexpressed Pdyn. Double in situ hybridization for Penk/Crh (corticotropin-releasing hormone gene) in the PVN and Penk/Kiss1 (kisspeptin gene) in the ARC revealed that few Penk-expressing cells coexpressed Crh and Kiss1. Taken together, these results suggest that central enkephalin-DOR signaling mediates the suppression of pulsatile LH release during malnutrition. Moreover, the current study suggests that central enkephalin-DOR signaling is also involved in gluconeogenesis during malnutrition in female rats.

Socially Induced Infertility in Naked and Damaraland Mole-Rats: A Tale of Two Mechanisms of Social Suppression.

The naked mole-rat (Heterocephalus glaber) and the Damaraland mole-rat (Fukomys damarensis) possess extreme reproductive skew with a single reproductive female responsible for reproduction. In this review, we synthesize advances made into African mole-rat reproductive patterns and physiology within the context of the social control of reproduction. Non-reproductive female colony members have low concentrations of luteinising hormone (LH) and a reduced response of the pituitary to a challenge with gonadotropin releasing hormone (GnRH). If the reproductive female is removed from the colony, an increase in the basal plasma LH and increased pituitary response to a GnRH challenge arises in the non-reproductive females, suggesting the reproductive female controls reproduction. Non-reproductive male Damaraland mole-rats have basal LH concentrations and elevated LH concentrations in response to a GnRH challenge comparable to the breeding male, but in non-breeding male naked mole-rats, the basal LH concentrations are low and there is a muted response to a GnRH challenge. This renders these two species ideal models to investigate physiological, behavioural and neuroendocrine mechanisms regulating the hypothalamic-pituitary-gonadal axis. The recently discovered neuropeptides kisspeptin and RFamide-related peptide-3 are likely candidates to play an important role in the regulation of reproductive functions in the two mole-rat species.

Saliva and plasma steroidome in mare during reproductive stages: A GC-MS/MS study

Precision livestock farming using omics approach to acquire precise and real-time data can help farmers in individual animal management and decision making. Since steroid hormones play a key role for the regulation of reproductive functions, reproduction management could be improved by characterizing the steroidome during reproductive stages. Moreover, saliva collection is a non-invasive, painless, inexpensive and easy sampling method. Thus, this prospective study proposes a steroidomic analysis in mare saliva during reproductive stages, that could help to identify potential biomarkers to accurately detect their reproductive stage in a welfare friendly production system, for real-time decision making at the individual animal level. Correlation between saliva and plasma steroidome was also investigated. Saliva and blood samples from 6 mares were collected in anestrus, in the follicular phase 3 days, 2 days and 1 day before ovulation and the day when ovulation was detected, in the luteal phase 6 days after ovulation and in gestation 18 days after ovulation and insemination. Steroidome analysis was performed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). We characterized 25 and 36 steroids in saliva and plasma respectively. Pregnenolone concentrations in saliva during gestation were significantly higher than during anestrus or follicular phase and tended to be higher than during luteal phase. Most of the 5α-reduced metabolites of progesterone showed higher salivary concentrations during the luteal phase 6 days post-ovulation and during gestation 18 days post-ovulation compared to anestrus and follicular phase. These steroids could be potential salivary biomarkers of the reproductive stage of the mare. Further studies with a greater number of animals are in progress to confirm the reliability of these potential candidate biomarkers and to develop field-friendly assays.

The Kisspeptin and Kisspeptin receptor in follicular microenvironment: is that really necessary for oocyte maturation and fertilisation?

The aim of this study was to determine whether Kisspeptin and Kisspeptin receptor in the follicular microenvironment is necessary for human oocyte maturation and fertilisation. The cumulus cell (CC) and follicle fluids (FF) obtained from the first aspirated follicles (n = 52) from 32 patients were divided into three groups considering nuclear maturation and fertilisation results of oocytes: (1) Metaphase I or germinal vesicle stage oocytes (incomplete nuclear maturation, n = 10), (2) unfertilised metaphase II oocytes (incomplete cytoplasmic maturation, n = 16), and (3) fertilised metaphase II oocytes (completed nuclear-cytoplasmic maturation, n = 26). The gene expression levels were assessed by RT-PCR. The levels of Kisspeptin (KISS1) and Kisspeptin receptor (KISS1R) were measured by ELISA. There were no significant efficacy KISS1 and KISS1R gene expressions in cumulus cells in terms of oocyte nuclear maturation stage (Group 1, vs Group 2 + Group 3) (respectively p = .49; p = .45). In terms of the cytoplasmic maturation stage (Group 2, vs Group 3); KISS1 and KISS1R expressions in CCs were comparable (respectively p = .07; p = .08). In FFs, KISS1 and KISS1R concentrations were similar between all groups (respectively p = .86; p = .26). In conclusion, the relative KISS1 and KISS1R expressions in CC and also KISS1 and KISS1R level of FF were independent of oocytes nuclear and/or cytoplasmic maturation. Impact statement What is already known on this subject? It has been demonstrated that Kisspeptin is an essential regulator of reproductive function and plays a key role in the modulation of GnRH secretion and gonadotropin release. Still, no information is available about the link between gene expression or concentration in the follicular microenvironment and oocyte development. What do the results of this study add? The study has shown that the relative Kisspeptin (KISS1) and Kisspeptin receptor (KISS1R) and expressions in cumulus cell (CC) and also KISS1 and KISS1R levels of follicle fluids (FF) were independent of oocytes nuclear and/or cytoplasmic maturation. What are the implications of these findings for clinical practice and/or further research? Based on the findings, it is difficult to establish a concept that kisspeptin can directly induce oocyte maturation. Nevertheless, to confirm these findings, further studies with a larger sample size are needed.

Kisspeptin Modulation of Reproductive Function

Kisspeptin is a peptide expressed mainly in the infundibular nucleus of the hypothalamus. Kisspeptin plays a crucial role in the regulation of reproductive functions. It is regarded as the most important factor responsible for the control of the hypothalamic–pituitary–gonadal axis, the onset of puberty, and the regulation of menstruation and fertility. Kisspeptin activity influences numerous processes such as steroidogenesis, follicular maturation, ovulation, and ovarian senescence. The identification of kisspeptin receptor mutations that cause hypogonadotropic hypogonadism has initiated studies on the role of kisspeptin in puberty. Pathologies affecting the neurons secreting kisspeptin play a major role in the development of PCOS, functional hypothalamic amenorrhea, and perimenopausal vasomotor symptoms. Kisspeptin analogs (both agonists and antagonists), therefore, may be beneficial as therapy in those afflicted with such pathologies. The aim of this review is to summarize the influence of kisspeptin in the physiology and pathology of the reproductive system in humans, as well as its potential use in therapy.

Comparison of Reproductive Function Between Normal and Hyperandrogenemia Conditions in FemaleMice With Deletion of Hepatic Androgen Receptor.

Obesity, altered glucose homeostasis, hyperinsulinism, and reproductive dysfunction develops in female humans and mammals with hyperandrogenism. We previously reported that low dose dihydrotestosterone (DHT) administration results in metabolic and reproductive dysfunction in the absence of obesity in female mice, and conditional knock-out of the androgen receptor (Ar) in the liver (LivARKO) protects female mice from DHT-induced glucose intolerance and hyperinsulinemia. Since altered metabolic function will regulate reproduction, and liver plays a pivotal role in the reversible regulation of reproductive function, we sought to determine the reproductive phenotype of LivARKO mice under normal and hyperandrogenemic conditions. Using Cre/Lox technology, we deleted the Ar in the liver, and we observed LivARKO female mice have normal puberty timing, cyclicity and reproductive function. After DHT treatment, like control mice, LivARKO experience altered estrous cycling, reduced numbers of corpus lutea, and infertility. Liver Ar is not involved in hyperandrogenemia-induced reproductive dysfunction. The reproductive dysfunction in the DHT-treated LivARKO lean females with normal glucose homeostasis indicates that androgen-induced reproductive dysfunction is independent from metabolic dysfunction.

Highlights of neuroanatomical discoveries of the mammalian gonadotropin-releasing hormone system.

The anatomy and morphology of gonadotropin‐releasing hormone (GnRH) neurons makes them both a joy and a challenge to investigate. They are a highly unique population of neurons given their developmental migration into the brain from the olfactory placode, their relatively small number, their largely scattered distribution within the rostral forebrain, and, in some species, their highly varied individual anatomical characteristics. These unique features have posed technological hurdles to overcome and promoted fertile ground for the establishment and use of creative approaches. Historical and more contemporary discoveries defining GnRH neuron anatomy remain critical in shaping and challenging our views of GnRH neuron function in the regulation of reproductive function. We begin this review with a historical overview of anatomical discoveries and developing methodologies that have shaped our understanding of the reproductive axis. We then highlight significant discoveries across specific groups of mammalian species to address some of the important comparative aspects of GnRH neuroanatomy. Lastly, we touch on unresolved questions and opportunities for future neuroanatomical research on this fascinating and important population of neurons.

Human pluripotent stem cell-derived cells endogenously expressing follicle-stimulating hormone receptors: modeling the function of an inactivating receptor mutation.

Follicle-stimulating hormone (FSH) is crucial in the development and regulation of reproductive functions. The actions of human FSH and its receptor (FSHR) and mutations therein have mainly been studied using in vivo models, primary cells, cancer cells and cell lines ectopically expressing the FSHR. To allow studies of endogenous FSHR function in vitro, we differentiated FSHR-expressing cells from human pluripotent stem cells. FSH stimulation of the wild-type (WT), but not the inactivating Finnish founder mutant (A189V) receptor, activated the canonical cyclic adenosine monophosphate (cAMP)-dependent signaling pathway and downstream mediators. To investigate protein–protein interaction partners of FSHR at resting state and upon FSH stimulation, we expressed FSHR in HEK293 cells followed by affinity purification mass spectrometry analyses. We found 19 specific high-confidence interacting proteins for WT FSHR and 14 for A189V FSHR, several of which have been linked to infertility. Interestingly, while only WT FSHR interacted with FSH, insulin-like growth factor 1 receptor (IGF1R), for example, interacted with both WT and A189V FSHR upon FSH stimulation. In conclusion, our protocol allows detailed studies of FSH action and disease modeling in human cells endogenously expressing FSHR.

Interplay between nitric oxide and gonadotrophin-releasing hormone in the neuromodulation of the corpus luteum during late pregnancy in the rat

BackgroundNitric oxide and GnRH are biological factors that participate in the regulation of reproductive functions. To our knowledge, there are no studies that link NO and GnRH in the sympathetic ganglia. Thus, the aim of the present work was to investigate the influence of NO on GnRH release from the coeliac ganglion and its effect on luteal regression at the end of pregnancy in the rat.MethodsThe ex vivo system composed by the coeliac ganglion, the superior ovarian nerve, and the ovary of rats on day 21 of pregnancy was incubated for 180 min with the addition, into the ganglionic compartment, of L-NG-nitro arginine methyl ester (L-NAME), a non-selective NO synthase inhibitor. The control group consisted in untreated organ systems.ResultsThe addition of L-NAME in the coeliac ganglion compartment decreased NO as well as GnRH release from the coeliac ganglion. In the ovarian compartment, and with respect to the control group, we observed a reduced release of GnRH, NO, and noradrenaline, but an increased production of progesterone, estradiol, and expression of their limiting biosynthetic enzymes, 3β-HSD and P450 aromatase, respectively. The inhibition of NO production by L-NAME in the coeliac ganglion compartment also reduced luteal apoptosis, lipid peroxidation, and nitrotyrosine, whereas it increased the total antioxidant capacity within the corpora lutea.ConclusionCollectively, the results indicate that NO production by the coeliac ganglion modulates the physiology of the ovary and luteal regression during late pregnancy in rats.