Gonadotropin Research Articles

Характер психічних відхилень та їх гормональний супровід у дівчат-підлітків з розладами менструальної функції

The pubertal period is accompanied by the tension of the adaptation system during the hormonal adjustment. Purpose - to define the hormonal determinants of stress-related menstrual disorders in adolescent girls with comorbid mental disorders and the peculiarities of adaptation reactions. Materials and methods. 150 adolescent girls with menstrual cycle disorders were examined. The levels of luteotropic hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), estradiol (E2), testosterone (T), cortisol (K), dehydroepiandrosterone sulfate (DHEA-S) were determined. They were divided into three groups depending on the examination by a psychiatrist: without mental status disorders, with anxiety-phobic disorders and depressive states. The control group included 35 girls of similar age with a normal menstrual cycle who were examined in the early follicular phase. Results. There were no significant differences in the content of gonadotropic hormones in girls with menstrual function disorders in view of the state of mental health. However, in girls with oligomenorrhea (OM), in relation to patients with abnormal uterine bleeding (AUB), elevated LH values are significantly more often registered (p<0.03). Levels of peripheral T, as well as its precursor DHEA-S and metabolite (E2), were associated with mental health status. The lowering of E2 significantly elevated in adolescents with comorbid psychopathology. The number of girls with high T values increased in the presence of psychopathology in AUB and remained unchanged in OM. Cortisol in psychopathology, regardless of the type of menstrual function disorders, was reduced in relation to the control group, however, in girls with OM, its content was significantly higher, especially in depressive states (p˂0.05). The specific gravity of girls with a reduced level of DHEA-S in patients with AUB increased with comorbid psychopathology. In case of OM, a reduced and increased content of DHEA-S was registered with the same frequency. An increase in the C/DHEA-S indicator of the functioning of the hypothalamic-pituitary-adrenal system in patients with OM was noted much less frequently than in patients with AUB (p˂0.006). Conclusions. It is believed that the hormonal relationship between cortisol and DHEA-S indicates the body’s response to stressors and can be used as one of the biomarkers of neurotic and depressive states. An increase in the C/DHEA-S ratio suggests a possible dissociation of their secretion. Adolescents with AUB, unlike girls with OM, have a decrease in the “protective” hormone DHEA-S, which can affect the deterioration of mental health. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Effect of Aqueous Leaves Extract of Datura alba ness (Thorn Apple) on Gonadotropic Hormones (FSH, LH) and Prolactin Following Administration of MSG and Clomiphene Citrate in Female Wistar Rats

Phytomedicine has become popular among and majority of women depends on herbal medicine when seeking healthcare. Several women opted for herbal medicine when looking for fertility and most the women that hormonal imbalance depends on herbal medicine for correction. Hormones is substance that must be required and must be in the right values for reproduction to occur. The aim is this study is to ascertain the Effect of aqueous leave extract of D. alba ness (Thorn Apple) on gonadotropic hormones (FSH, LH) Treated with monosodium glutamate (MSG) and clomiphene citrate in female wistar rats. The animals were randomly selected into 7 with five animals per group. Each group received the following: group 1 10mls/kg of distil water, group 2 low dose of extract (1000mg/kg), group 3 high dose of extract(2000mg/kg), group 4 MSG 800mg/kg, group 5 clomid 50mg/kg, group 6 MSG + high dose of extract and group 7 clomid+ high dose of extract. Administration of extract was done for 21 days, and on the 22nd day, the animals were sacrificed and blood samples were collected for hormonal analysis. The results revealed significant decreased in the serum levels of follicle stimulating hormones (FSH) in all the treatment groups (extract 1000mg/kg, extract 2000mg/kg, MSG 800mg/kg, MSG 800mg/kg + extract 2000mg/kg, and clomid 50mg/kg + extract 2000mg/kg) when compared with control and significant increase in serum levels FSH in clomid 50mg/kg treated group. Luteinizing hormone (LH) serum levels significantly decreased in all the treatment groups when compared with the control group. LH levels in the clomid group significantly increased when compared with other treatment groups. The statistical analysis was carried out using Statistical Package for Social Science (SPSS) version 23.

Effect of Aqueous Leaves Extract of Datura alba ness (Thorn Apple) on Gonadotropic Hormones (FSH, LH) Following Administration of Monosodium Glutamate and Clomiphene Citrate in Male Wistar Rats

Reproductive process cannot be complete without hormone and if these hormones are not in the right proportion, then reproductive process could fail. Hormones are chemical substances produced by the endocrine gland and its physiological action is exerted on a target organ different from the secretion’s site. The aim is this study is to evaluate Effect of aqueous leave extract of D. alba ness (Thorn Apple) on gonadotropic hormones (FSH, LH) and prolactin following administration of MSG and clomid citrate in male wistar rats. The study involves 35 male wistar rats weighing between 130g to 160g. The animals were randomly selected into 7 with five animals per group. Each group received the following: group 1 10mls/kg of distil water, group 2 low dose of extract (1000mg/kg), group 3 high dose of extract(2000mg/kg), group 4 MSG 800mg/kg, group 5 clomid 50mg/kg, group 6 MSG + high dose of extract and group 7 clomid+ high dose of extract. Administration of extract was done through oral gavage for 21 days, thereafter, the animals were sacrificed on the 22nd day, and blood samples were collected for hormonal analysis. The results showed significant increase in serum levels of FSH and LH in the MSG group and decreased in FHS levels in all others treated groups when compared with control. The serum levels of FSH and LH rises when extract was co-administered with MSG. The serum levels of LH slightly increased when extract was co-administered with clomid serum levels of FSH do not increased when extract was co-administered with clomid. The statistical analysis was carried out using Statistical Package for Social Science (SPSS) version 23.

Adverse effects of type 2 diabetes mellitus on ovarian reserve and pregnancy outcomes during the assisted reproductive technology process.

To study the effect of type 2 diabetes mellitus(T2DM)on overall ovarian reserve and pregnancy outcomes during assisted reproductive technology (ART) among childbearing infertile women. Retrospective cohort study. The Reproductive Medicine Special Hospital, The First Hospital of Lanzhou University, between January 2019 and December 2022. A total of 265 infertile female patients aged 20-45 years who underwent in vitro fertilization-embryo transfer (IVF-ET), intracytoplasmic sperm injection-embryo transfer (ICSI-ET), or rescue intracytoplasmic sperm injection-embryo transfer (RICSI-ET) in the first fresh cycle. None. Serum Anti-Müllerian Hormone (AMH) levels, clinical pregnancy rate (CPR), live birth rate (LBR), and abortion rate (AR) in the T2DM group and non-T2DM group. Patients with T2DM showed statistically decreased levels of AMH compared to the non-T2DM group. During ovarian stimulation, those with T2DM required significantly higher total and initial doses of gonadotropin (GN), although they had fewer retrieved oocytes and worse pregnancy outcomes than the non-T2DM group. Multivariate logistic regression analysis adjusting for confounding factors showed that T2DM alone was an independent risk factor for CPR and LBR (adjusted odds ratio [a OR], 0.458, adjusted 95% confidence interval [CI], 0.235-0.891, P = 0.022; a OR, 0.227, 95% CI, 0.101-0.513, P<0.001; respectively), and the abortion rate in the T2DM group was 3.316 times higher than the non-T2DM group(a OR, 3.316, 95%CI, 1.248-8.811, P = 0.016). Infertile patients with T2DM have decreased ovarian reserve, and T2DM has a deleterious impact on clinical pregnancy outcomes during the ART process compared with non-T2DM infertile women. Infertile women with T2DM have decreased ovarian reserve and pregnancy outcomes during the assisted reproductive technology process compared with non-T2DM infertile women.

26 The Kisspeptin Knockout Pig: a New Model for Dissecting Mechanisms Controlling Reproduction

Abstract The biological role of kisspeptin in the reproduction of swine has been far less studied than in other species. In the porcine hypothalamus, kisspeptin is coexpressed with neurokinin B and dynorphin in KNDy neurons. Although kisspeptin-stimulated secretion of gonadotropin hormones in gilts was established early, increased expression of hypothalamic KISS1 was only recently confirmed to be associated with puberty and cyclicity in pigs. Gene knockout (KO) models to study kisspeptin in livestock did not exist until TALEN gene editing technology was used to knockout KISS1R in pigs, resulting in a hypogonadotropic-hypogonadal (HH) phenotype that caused pubertal failure. Little is known about the reproductive physiology of KISS1R KO pigs, except that exogenous follicle-stimulating hormone and gonadotropin-releasing hormone only partially restored gonadal development in boars. To further explore the biological function of kisspeptin and its receptor in swine, we recently used CRISPR/cas9-ribonucleoproteins to knockout KISS1 in pigs (10.3389/fgene.2022.1078991). A total of 684 zygotes were injected resulting in 69 piglets with 58% bearing KISS1 KO alleles (KISS1-/-). The proportion of KISS1-/- alleles did not affect BW or ADG in these mosaic founder pigs. Two boars and 2 gilts with complete KISS1 KO exhibited HH and failed to become pubertal. Germ and Sertoli cells were disorganized in small seminiferous tubules. Mating mosaic F0 pigs (frequency of KISS1-/-, 35 to 88%) resulted in 170 F1 piglets with KISS1-/- alleles observed in a Mendelian frequency. These KISS1-/- pigs displayed normal BW growth during development but were also HH. Ovarian development of KISS1-/- gilts was arrested at a juvenile state, although antral follicles were present. The KISS1-/- boars lacked testicular development and had dramatically reduced concentrations of androgens and skatole, suggesting the lack of boar taint in their meat. Importantly, KISS1 and KISS1R are expressed in ovarian, uterine and testicular tissue; however, the direct role of kisspeptin in these reproductive organs remains unknown in livestock. These KISS1-/- pigs provide a unique animal model that opens new avenues for understanding the role of kisspeptin and gonadotropin hormones in regulation of reproductive development and function in pigs as well as livestock in general. These animals facilitate our ability to explore questions such as: 1) how KNDy neurons function and control gonadotropin pulses or 2) what role does KISS1 play in the development and function of the ovary, uterus and testis in domestic animals. Some of these questions have heretofore only been addressable with difficult surgical modifications, and therefore, KISS1-/- pigs represent an alternative to painful procedures in livestock research. Pursuing these fundamental studies is expected to lead to methods that restore fertility in KISS1-/- pigs. This, in turn, could provide new possibilities for the elimination of surgical castration from pork production. The USDA is an equal opportunity provider and employer.

Gonadotropin-Releasing Hormone Receptor (GnRHR) and Hypogonadotropic Hypogonadism

Human sexual and reproductive development is regulated by the hypothalamic-pituitary-gonadal (HPG) axis, which is primarily controlled by the gonadotropin-releasing hormone (GnRH) acting on its receptor (GnRHR). Dysregulation of the axis leads to conditions such as congenital hypogonadotropic hypogonadism (CHH) and delayed puberty. The pathophysiology of GnRHR makes it a potential target for treatments in several reproductive diseases and in congenital adrenal hyperplasia. GnRHR belongs to the G protein-coupled receptor family and its GnRH ligand, when bound, activates several complex and tissue-specific signaling pathways. In the pituitary gonadotrope cells, it triggers the G protein subunit dissociation and initiates a cascade of events that lead to the production and secretion of the luteinizing hormone (LH) and follicle-stimulating hormone (FSH) accompanied with the phospholipase C, inositol phosphate production, and protein kinase C activation. Pharmacologically, GnRHR can be modulated by synthetic analogues. Such analogues include the agonists, antagonists, and the pharmacoperones. The agonists stimulate the gonadotropin release and lead to receptor desensitization with prolonged use while the antagonists directly block the GnRHR and rapidly reduce the sex hormone production. Pharmacoperones include the most recent GnRHR therapeutic approaches that directly correct the misfolded GnRHRs, which are caused by genetic mutations and hold serious promise for CHH treatment. Understanding of the GnRHR’s genomic and protein structure is crucial for the most appropriate assessing of the mutation impact. Such mutations in the GNRHR are linked to normosmic hypogonadotropic hypogonadism and lead to various clinical symptoms, including delayed puberty, infertility, and impaired sexual development. These mutations vary regarding their mode of inheritance and can be found in the homozygous, compound heterozygous, or in the digenic state. GnRHR expression extends beyond the pituitary gland, and is found in reproductive tissues such as ovaries, uterus, and prostate and non-reproductive tissues such as heart, muscles, liver and melanoma cells. This comprehensive review explores GnRHR’s multifaceted role in human reproduction and its clinical implications for reproductive disorders.

Chemical Induce Polycystic Ovarian Syndrome-Preclinical Animal Models

Polycystic ovarian syndrome is a complex endocrine disturbance that leads to hyperandrogenism, disruption in the functioning of the Hypothalamic-Pituitary-Ovary (HPO) axis and multiple cysts in ovaries. To understand and study different treatment approaches of polycystic ovarian syndrome, there are several chemical-induced animal models available that mimic polycystic ovarian syndrome. These animal models are designed to closely resemble the characteristic symptoms. Polycystic Ovarian Syndrome’s key characteristics are changes in gonadotropin and sex steroid hormone, ovarian morphology, and metabolic characteristics. Direct hormone-regulated animal models are frequently utilized to study PCOS. Rodent animal model is often used which aims to replicate the key feature of human PCOS. Various endocrine-disrupting chemicals also makes a major role in the development of PCOS. In order to bridge the gap between basic research and clinical application in the field of PCOS, PCOS-induced models are essential tools for improving our understanding of the illness and evaluating innovative therapies. The review discusses various animal models used to induced PCOS by various inducers such as aromatase inhibitor inducer (letrozole), androgen excess inducer (dihydrotestosterone, dehydroepiandrosterone, testosterone), estrogen-induced (estradiol valerate), antiprogesterone (mifepristone), monosodium-L-glutamate, bisphenol-A and tributyltin chloride. This article contributed to underlying the current understanding and provides you a complete review that overall covers various aspects, including the impact of chemical-induced models, which also includes changes in the morphology of ovaries, gonadotropin as well as, and alterations in the level of various sex steroid hormone profile. Additionally it explores the metabolic abnormalities caused by various chemical-inducers used to induce PCOS in animal. The objective of this review is to provide a comprehensive review about various chemical inducers which are responsible for the development of PCOS.

Potential Significance of Serum Autoantibodies to Endometrial Antigens, α-Enolase and Hormones in Non-Invasive Diagnosis and Pathogenesis of Endometriosis.

The objective of the study was to evaluate the profile of serum autoantibodies and their diagnostic and pathogenetic significance in ovarian endometrioma (OEM) and deep infiltrative endometriosis (DIE). The study enrolled 74 patients with endometriosis (Group 1), including 53 patients with OEM (Subgroup 1a); 21 patients with DIE without ovarian lesions (Subgroup 1b); and 27 patients without endometriosis (Group 2). The diagnosis was confirmed by laparoscopic surgery and histologic examination of resected tissues. Antibodies (M, G) to tropomyosin 3 (TPM), tropomodulin 3 (TMOD), α-enolase (ENO), estradiol (E2), progesterone (PG), and human chorionic gonadotropin (hCG) were identified in blood serum using modified ELISA. In endometriosis, antibodies to endometrial antigens, hormones, and ENO were detected more often than antiphospholipid and antinuclear antibodies. Higher levels of IgM to TPM, hCG, E2, and PG and IgG to TMOD, ENO, E2, and hCG were found in Subgroup 1a compared to Group 2. IgM to TPM, hCG, E2, PG, and IgG to E2 and ENO had a high diagnostic value for OEM (AUC > 0.7), with antibodies to TPM having the highest sensitivity and specificity (73.6% and 81.5%). In Subgroup 1b, only the levels of IgM to TPM and hCG were higher than in Group 2. These antibodies had a high diagnostic value for DIE. Thus, endometriosis is associated with autoantibodies to endometrial antigens, α-enolase, steroid, and gonadotropic hormones. A wider spectrum of antibodies is detected in OEM than in DIE. These antibodies have a high diagnostic value for OEM and DIE and potential pathogenetic significance for endometriosis and associated infertility.

The hyperthyroidism in gestational trophoblastic disease: a case report

Gestational trophoblastic disease, also referred to as GTD, is an extremely rare form of pregnancy-related complication that can arise. It covers a wide range of conditions, from choriocarcinoma to molar pregnancy. Patients who are diagnosed with gestational trophoblastic disease have an increased likelihood of developing hyperthyroidism. It is speculated that this occurs as a result of molecular mimicry that exists between human chorionic gonadotrophin (hCG) and thyroid-stimulating hormone (TSH), which results in cross-reactivity with the TSH receptor. This would explain why the two molecules would react similarly to each other. The gestational trophoblastic disease-induced thyroid storm is an extremely rare but potentially fatal complication that can arise during pregnancy. It is necessary to detect and treat this complication as soon as possible in order to prevent additional complications from occurring. Once the patient's hemodynamic status has been stabilized, the removal of the mole is the primary component of the definitive treatment that will be administered. In most cases, hyperthyroidism usually resolves once the gestational trophoblastic disease has been successfully treated and the hCG levels have been brought back to normal.

Evaluation of the effect of extrasystolic arrhythmia in pregnant women on fetal condition

Relevance much attention is paid to the problem of the effect of extrasystolic arrhythmia in pregnant women on the condition of the fetus. The aim of the study is to study the effect of extrasystolic arrhythmia in pregnant women on the condition of the fetus by cardiotocography. Material and methods. 32 pregnant women with extrasystole (group 1) and 30 without arrhythmia (group 2) were examined in the 3rd trimester from the 32nd week of pregnancy. All pregnant women of both groups underwent daily registration of electrocardiography (ECG), ultrasound examination of the heart, fetal cardiotocography (CTG of the fetus), determination of the level of human chorionic gonadotropin (hCG), prolactin and thyroid hormones. Results. The correlation analysis revealed a close relationship between the number of fetal cardiotocography scores and the level of thyroid-stimulating hormone (TSH) (r = 0,917, p &lt;0,01), free thyroxine (T4c) (r = -0,877, p &lt; 0,01) and the number of extrasystoles (r = -0,872, p &lt; 0,01) in pregnant women. Conclusion. In pregnant women with and without extrasystole, there are differences between reproductive hormones and electrocardiography indicators within the reference values. It was found that in pregnant women with and without extrasystole, fetal cardiotocography indicators do not significantly differ. A quantitative threshold value of ventricular extrasystole was revealed to reduce the number of fetal cardiotocography scores in the third trimester from week32. The threshold value of extrasystole was 1500 per day, AUC (area under curve) – 0,792, sensitivity – 81,3 %, specificity – 100,0 %.

Gonadotropin Versus Gonadotropin/Letrozole Protocol in Previously Failed Antagonist Cycles in Patients with Low Prognosis

Objective: Letrozole is an aromatase inhibitor which has been used for ovulation induction. Our aim is to evaluate the effectiveness of adding letrozole to gonadotropins in the early follicular phase of gonadotropin-releasing hormone antagonist (GnRH-a) protocol in patients with poor ovarian response who had failed in the preceding GnRH-a cycle.&#x0D; Materials and Method: Ninety-eight patients with poor ovarian response who had previously failed GnRH-a cycle were included. Patients (n; 58) who were treated with letrozole plus gonadotropins (LzGA) were compared with patients (n; 40) who received gonadotropins (GT). The number of total oocytes retrieved, the number of MII oocytes and fertilized oocytes, fertilization and implantation ratios, the rate of cycle cancellation as well as clinical pregnancy and live birth rates&#x0D; Results: The LzGA group had a significantly shorter duration of GnRH-a stimulation and higher progesterone level at trigger day (p=0.005, p=0.049 respectively). Although letrozole administration demonstrated lower estradiol levels at trigger day and the total dose of gonadotropins used were lower in LzGA group, this was statistically non-significant (p=0.13, p=0.13 respectively).&#x0D; Conclusion: Adding letrozole to gonadotropins in GnRH-a protocol in patients with poor prognosis did not improve pregnancy outcomes. But it seems to decrease IVF costs by reducing the GnRH-antagonist and gonadotropin dosage.

Quantification of hCG Hormone Using Tapered Optical Fiber Decorated with Gold Nanoparticles.

In this study, a novel technique for the quantification of the human chorionic gonadotropin (hCG) hormone using localized surface plasmons and a tapered optical fiber decorated with gold nanoparticles (Au-NPs) is reported. The tapered optical fiber fabrication process involves stretching a single-mode optical fiber using the flame-brushing system. The waist of the tapered optical fiber reaches a diameter of 3 μm. Decoration of the taper is achieved through the photodeposition of 100 nm Au-NPs using the drop-casting technique and a radiation source emitting at 1550 nm. The presence of the hCG hormone in the sample solutions is verified by Fourier-transform infrared spectroscopy (FTIR), revealing the presence of bands related to functional groups, such as C=O (1630 cm-1), which are associated with proteins and lipids, components of the hCG hormone. Quantification tests for hormone concentrations were carried out by measuring the optical power response of the tapered optical fiber with Au-NPs under the influence of hCG hormone concentrations, ranging from 1 mIU/mL to 100,000 mIU/mL. In the waist of the tapered optical fiber, the evanescent field is amplified because of localized surface plasmons generated by the nanoparticles and the laser radiation through the optical fiber. Experimental results demonstrated a proportional relationship between measured radiation power and hCG concentration, with the optical power response decreasing from 4.45 mW down to 2.5 mW, as the hCG hormone concentration increased from 1 mIU/mL up to 100,000 mIU/mL. Furthermore, the spectral analysis demonstrated a spectral shift of 14.2 nm with the increment of the hCG hormone concentration. The measurement system exhibits promising potential as a sensor for applications in the biomedical and industrial fields.

AXL receptor tyrosine kinase modulates gonadotropin-releasing hormone receptor signaling

BackgroundGonadotropin-releasing hormone (GnRH) receptors are essential for reproduction and are expressed in numerous urogenital, reproductive, and non-reproductive cancers. In addition to canonical G protein-coupled receptor signaling, GnRH receptors functionally interact with several receptor tyrosine kinases. AXL is a receptor tyrosine kinase expressed in numerous tissues as well as multiple tumors. Here we tested the hypothesis that AXL, along with its endogenous ligand Gas6, impacts GnRH receptor signaling.MethodsWe used clonal murine pituitary αT3-1 and LβT2 gonadotrope cell lines to examine the effect of AXL activation on GnRH receptor-dependent signaling outcomes. ELISA and immunofluorescence were used to observe AXL and GnRH receptor expression in αT3-1 and LβT2 cells, as well as in murine and human pituitary sections. We also used ELISA to measure changes in ERK phosphorylation, pro-MMP9 production, and release of LHβ. Digital droplet PCR was used to measure the abundance of Egr-1 transcripts. A transwell migration assay was used to measure αT3-1 and LβT2 migration responses to GnRH and AXL.ResultsWe observed AXL, along with the GnRH receptor, expression in αT3-1 and LβT2 gonadotrope cell lines, as well as in murine and human pituitary sections. Consistent with a potentiating role of AXL, Gas6 enhanced GnRH-dependent ERK phosphorylation in αT3-1 and LβT2 cells. Further, and consistent with enhanced post-transcriptional GnRH receptor responses, we found that Gas6 increased the abundance of Egr-1 transcripts. Suggesting functional significance, in LβT2 cells, Gas6/AXL signaling stimulated LHβ production and enhanced GnRH receptor-dependent generation of pro-MMP9 protein and promoted cell migration.ConclusionsAltogether, these data describe a novel role for AXL as a modulator of GnRH receptor signaling.5LYCavpivq5eiBQqEyV1BjVideo

Plurihormonal PIT1-lineage pituitary neuroendocrine tumors: a clinicopathological study

Objective: To investigate the clinicopathological characteristics of plurihormonal PIT1-lineage pituitary neuroendocrine tumors. Methods: Forty-eight plurihormonal PIT1-lineage tumors were collected between January 2018 and April 2022 from the pathological database of Sanbo Brain Hospital, Capital Medical University. The related clinical and imaging data were retrieved. H&E, immunohistochemical and special stains were performed. Results: Out of the 48 plurihormonal PIT1-lineage tumors included, 13 cases were mature PIT1-lineage tumors and 35 cases were immature PIT1-lineage tumors. There were some obvious clinicopathological differences between the two groups. Clinically, the mature plurihormonal PIT1-lineage tumor mostly had endocrine symptoms due to increased hormone production, while a small number of immature PIT1-lineage tumors had endocrine symptoms accompanied by low-level increased serum pituitary hormone; patients with the immature PIT1-lineage tumors were younger than the mature PIT1-lineage tumors; the immature PIT1-lineage tumors were larger in size and more likely invasive in imaging. Histopathologically, the mature PIT1-lineage tumors were composed of large eosinophilic cells with high proportion of growth hormone expression, while the immature PIT1-lineage tumors consisted of chromophobe cells with a relatively higher expression of prolactin; the mature PIT1-lineage tumors had consistently diffuse cytoplasmic positive staining for keratin, while the immature PIT1-lineage tumors had various expression for keratin; the immature PIT1-lineage tumors showed more mitotic figures and higher Ki-67 proliferation index; in addition, 25.0% (12/48) of PIT1-positive plurihormonal tumors showed abnormal positive staining for gonadotropin hormones. There was no significant difference in the progression-free survival between the two groups (P=0.648) by Kaplan-Meier analysis. Conclusions: Plurihormonal PIT1-lineage tumor belongs to a rare type of PIT1-lineage pituitary neuroendocrine tumors, most of which are of immature lineage. Clinically increased symptoms owing to pituitary hormone secretion, histopathologically increased number of eosinophilic tumor cells with high proportion of growth hormone expression, diffusely cytoplasmic keratin staining and low proliferative activity can help differentiate the mature plurihormonal PIT1-lineage tumors from the immature PIT1-lineage tumors. The immature PIT1-lineage tumors have more complicated clinicopathological characteristics.

SAT481 Metastatic Choriocarcinoma, A Rare Cause of hCG-Mediated Hyperthyroidism

Abstract Disclosure: S. Jain: None. M. Crabtree: None. C. Houston: None. Background: Structural homology between human chorionic gonadotropin (hCG) and thyrotropin stimulating hormone (TSH) allows high levels of hCG to exert thyrotropic effects via TSH receptors. Although hCG-mediated hyperthyroidism is most frequently encountered in the context of pregnancy, it also occurs rarely as a paraneoplastic syndrome. We present a case of hCG-mediated hyperthyroidism due to metastatic choriocarcinoma in a postmenopausal woman. Case Presentation: A previously healthy 59-year-old woman was evaluated for shortness of breath and found to have multiple lung nodules concerning for metastatic malignancy. Further imaging demonstrated an enlarged heterogeneous uterus with an enlarged endometrial cavity. Endometrial biopsy revealed poorly differentiated carcinoma with some areas of possible squamous differentiation. Subspecialty cytopathology review was requested. One week after biopsy, the patient was brought to the emergency department with acute hypoxic respiratory failure following a seizure. She was found to be febrile, tachycardic, hypertensive, and minimally responsive. Head CT revealed a left frontal brain mass with surrounding vasogenic edema, concerning for metastasis. Laboratory evaluation revealed TSH of 0.03 mIU/L (normal 0.35-4.94) and free T4 of 2.30 ng/dL (normal 0.70-1.48). Thyroid stimulating immunoglobulin was negative. Beta-hCG was found to be significantly elevated at 982,093 mIU/mL after 1:100 dilution. She was started on treatment with propranolol, dexamethasone, and chemotherapy with etoposide and cisplatin. Meanwhile, cytopathology review identified scattered cells with features of choriocarcinoma and positive hCG immunostaining, felt to represent a diagnosis of poorly differentiated choriocarcinoma. After one cycle of chemotherapy, the patient’s beta-hCG decreased to 477,668 mIU/mL and free T4 decreased to 1.05 ng/dL. Due to progressive worsening of her clinical status, she was transitioned to palliative care and passed away on hospital day 14. Discussion: Hyperthyroidism mediated by hCG is most often seen in patients with hyperemesis gravidarum, gestational transient thyrotoxicosis, or gestational trophoblastic disease. Our case underscores the importance of considering hCG-mediated hyperthyroidism outside the context of pregnancy. Rare paraneoplastic sources of hCG include choriocarcinoma, seminoma, and other germ cell tumors. Hyperthyroidism in these cases is usually subclinical or mild, but instances of thyroid storm have been reported with metastatic disease. Beta blockers, glucocorticoids, and thionamides can be used as temporizing measures, but definitive therapy requires treatment of the causative malignancy. Presentation Date: Saturday, June 17, 2023

FRI288 Single-Cell Transcriptomics Identifies Pituitary Gland Cell Type Population Changes In Diet Induced Obesity

Abstract Disclosure: R.E. Ruggiero-Ruff: None. B. Le: None. N. Lainez: None. D. Coss: None. Obesity is a worldwide growing public health concern, with a current prevalence of 43% in the United States and projected future increase. Obesity is associated with type 2 diabetes, cardiovascular disease, stroke, and endocrine abnormalities. The pituitary gland is the master gland that responds to metabolic perturbations and regulates homeostasis via hormone secretion. Thyrotrophic, gonadotropic, somatotropic, lactotropic and corticotropic dysfunctions may occur as a consequence of obesity and perpetuate pathophysiologic conditions since pituitary hormones regulate metabolic function in return. Previous work from our lab determined that mice fed high fat diet have reduced levels of gonadotropin hormones, indicating that diet-induced obesity may play a role in pituitary hormone production. We postulate that diet-induced obesity alters the pituitary transcriptome and/or select population changes. To determine the effect of diet-induced obesity on pituitary gland population plasticity, we performed single-cell RNA-sequencing (scRNA-seq) of pituitary glands from control and high-fat diet-fed mice and characterized population changes and gene expression changes in each population. While gonadotrope population changes levels of gene expression that results in the lower gonadotropin levels, somatotrope and lactotrope populations, on the other hand, change at the population level with alterations in the cell numbers that result in altered levels of growth hormone and prolactin in the circulation. In summary, we defined diet-induced changes in pituitary hormone production and population shifts that may play a role in altered hormone production in obese patients and consequently dysregulation of homeostasis. Presentation: Friday, June 16, 2023

OR09-01 Mini-Puberty Could Be A Key Phase Regulating Reproductive Function In Female Mice

Abstract Disclosure: M. Chester: None. M. Devillers: None. L. Naule: None. F. Souaré: None. R. Corre: None. D. Quintas: None. C. Petrovic: None. J. Cohen-Tannoudji: None. S. Mhaouty-Kodja: None. C.J. Guigon: None. After birth, there is a transient activation of the hypothalamic-pituitary-ovary (HPO) axis, with high secretion of LH and FSH acting on the ovary to promote the secretion of sex steroids and in particular that of estradiol (E2). This period, called mini-puberty, occurs between 10-17 days postnatal (dpn) in mice. During reproductive life, E2 plays an important role on the HPO axis by regulating GnRH pulsatility and, thus, the cyclic fluctuations of gonadotropin hormone levels required for follicular development and ovulation. E2 acts on hypothalamic kisspeptin neurons of the arcuate (ARC) and the anteroventral periventricular nucleus (AVPV), which project to GnRH neurons and regulate their activity by negative and positive feedbacks, respectively. The possibility that E2 contributes, during mini-puberty, to the maturation of hypothalamic structures involved in puberty onset and reproductive function, has never been investigated. The aim of this study was to elucidate the putative roles of mini-puberty on reproduction. We developed a mouse model injected with a GnRH receptor antagonist (GonadoSTOP mice) suppressing LH and FSH surges to cause a 50% decrease in E2 levels during mini-puberty. Our results show that GonadoSTOP mice exhibited later first diestrus 2 (D2) (31.0 dpn versus 29.9 dpn in CTR; n=55-56 mice/group; P=0.0285), suggesting a possible delay in the timing of puberty. During reproductive life, GonadoSTOP mice spent a longer % of time in D2 than control mice, with no alteration in total estrous cycle time (64.4% versus 56% in CTR; n=18-19 mice/group; P=0.0350). Immunohistochemical analysis of kisspeptin neurons in ARC and AVPV of ovariectomized and hormonally-primed mice showed that their numbers were significantly lower in AVPV from GonadoSTOP mice than in control mice (n=5-6 brains/group; P=0.038). In fertility studies, we observed no difference in the time to conception or % of pregnant mice between the two groups at early reproductive life (3, 4 and 5 months). Contrary to these results, at the time of reproductive decline in control mice (11 months), a substantial % of GonadoSTOP mice could still be pregnant (47% versus 18% of CTR mice; P=0.0017). At this age, GonadoSTOP mice had lower LH levels than control mice (P=0.0074), but similar levels to young control mice at 4 months (P&amp;gt;0.05). RT-qPCR analyses of markers reflecting ovarian activity (Cyp19a1, Inhbb, Amh, Gdf9) at 4 and 11 months revealed that ovarian aging occurred similarly in both groups, suggesting that reproductive longevity in GonadoSTOP mice does not have an ovarian origin. Taken together these data suggest that transient and sustained activation of the HPO axis at mini-puberty may regulate key aspects of reproductive function with long-term impact on fertility, possibly by acting on the hypothalamus. Presentation: Friday, June 16, 2023

FRI275 Glutamate Receptor Agonists Stimulate Kisspeptin Neurons In The Arcuate Nucleus Of The Hypothalamus

Abstract Disclosure: R. Bearss: None. R. Piet: None. KNDy neurons co-express kisspeptin, neurokinin B (NKB), dynorphin A (Dyn A), and glutamate. These neurons, located in the arcuate nucleus of the hypothalamus, drive pulsatile gonadotropin releasing hormone (GnRH) secretion. This eventually promotes pulsatile gonadotropin hormone release from the anterior pituitary, important for fertility in both sexes. The current hypothesized mechanism for pulse generation involves KNDy neuron interconnectivity and reciprocal regulation through the combined actions of NKB, DynA and glutamate to coordinate KNDy neuron activity. There is evidence that KNDy neurons express NKB, Dyn A and glutamate receptors and that NKB stimulates KNDy neuron electrical activity while Dyn A suppresses it. There is also evidence that ionotropic glutamate receptor antagonists can inhibit KNDy neuron coordinated activity and pulsatile gonadotropin hormone release, indicating that glutamatergic neurotransmission plays a significant role in activity coordination. The purpose of the following experiments was to investigate the response of KNDy neurons to varied glutamate receptor activation. We used calcium imaging to monitor the activity of KNDy neurons in brain slices. We crossed Kiss1-Cre mice—expressing Cre recombinase (Cre) in kisspeptin neurons—with Cre-dependent GCaMP6f mice to generate Kiss1-Cre x GCaMP6f mice that express GCaMP6f in kisspeptin neurons. GCaMP6f fluorescence increases when binding to intracellular calcium and can, thus, be used as an inferential marker of electrical activity. Coronal brain slices (200 µm thickness) obtained from male Kiss1-cre x GCaMP6f mice and containing the arcuate nucleus were placed in a recording chamber under an epifluorescence microscope. KNDy neurons were exposed for 2 minutes to a bath application of either the ionotropic glutamate receptor agonist α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA, 10µM), the ionotropic glutamate receptor agonist N-methyl-D-aspartate (NMDA, 50µM), or the group 1 metabotropic glutamate receptor (mGluR) agonist (S)-3,5-dihydroxyphenylglycine (DHPG, 50µM). AMPA, NMDA, and DHPG increased KNDy neuron fluorescence by 7.8±0.5% (n=113 in 7 slices from 6 mice), 7.5±0.5% (n=154 in 6 slices from 5 mice), and 6.1±0.7% (n=159 in 7 slices from 5 mice) respectively. All cells that were excited by NMDA or DHPG were also excited by AMPA. These experiments provide evidence that activation of ionotropic AMPA and NMDA receptors as well as group 1 mGluRs stimulate KNDy neuron activity. Further investigation into the precise role of individual glutamate receptor subtypes on KNDy neuron coordinated activity is needed. Presentation: Friday, June 16, 2023

OR09-03 Loss of Inhibin Negative Feedback In The Pituitary Leads To Enhanced Ovulation But Pregnancy Failure In Mice

Abstract Disclosure: Y. Lin: None. E. Brule: None. M. Cowan: None. U. Boehm: None. D.J. Bernard: None. Loss of inhibin negative feedback in the pituitary leads to enhanced ovulation but pregnancy failure in mice Follicle-stimulating hormone (FSH) is an essential regulator of gonadal function, particularly in females. Inhibins are TGFβ family ligands made in the gonads that suppress FSH synthesis. Inhibins require a co-receptor, betaglycan or TGFBR3L, to mediate their actions in pituitary gonadotrope cells. Female mice with a gonadotrope-specific knockout of betaglycan or global deletion of Tgfbr3l exhibit increased ovarian follicle development, numbers of ovulated eggs, and litter sizes compared to controls. Females with both co-receptors knocked out (hereafter dKO) show dramatic increases in FSH levels, numbers of antral follicles and corpora lutea, serum progesterone levels, and ovarian aromatase expression. They ovulate 3-4 times as many eggs in natural cycles compared to control females. dKO females can become pregnant and show an increased number of implanted embryos at 7.5 days post coitum (7.5 dpc). Nevertheless, dKO females do not give birth to live offspring. By 10.5 dpc, the weight of the fetoplacental unit is decreased in dKO females, and many embryos display morphological abnormalities. By 14.5 dpc, most embryos in dKO females are either dead or resorbing. Wild-type surrogates give birth to live young following transplantation of embryos from control or dKO females. Conversely, control but not dKO females carry wild-type embryos to term. These data suggest that the maternal environment in dKO mice cannot support full-term pregnancies. Based on the hormonal profile of the cycling females, we suspected a deleterious effect of elevated estrogens. Consistent with this idea, treatment with the aromatase inhibitor anastrozole increased fetal survival in some dKO mice, suggesting a role for estrogens in embryo death. We are currently more thoroughly characterizing the maternal hormonal environment throughout gestation and investigating placental structure/function to gain more insight into the precise nature of the pregnancy failure. Our results will show how loss of inhibin action in the pituitary impede embryo survival and whether pregnancy loss is linked to increases in FSH. Presentation: Friday, June 16, 2023

OR2801 Novel Cis-Regulatory Mechanisms Of Lhb Transcription By Non-Coding RNAs And Non-Canonical DNA Structures

Abstract Disclosure: T. Refael: None. H. Gruber: None. G. Golan: None. L. Pnueli: None. P. Melamed: None. Expression of the Lhb gene was shown previously to be directed by synergistic actions of lineage-specific transcription factors at conserved regions of the proximal promoter. However extensive genomic analysis during the last few decades has expanded the repertoire of factors we now know to be involved in transcriptional regulation, revealing roles for distal transcriptional enhancers and long non-coding RNAs (lncRNAs), whose modes of action are only beginning to be elucidated1. Recently it has also become evident that regulatory regions of the genome are enriched with non-canonical DNA structures. These include G-quadruplexes (G4s) which form on guanine-rich stretches of single-stranded DNA and iMotifs (iMs) on the complementary strand, and are reported to bind various proteins which presumably regulate and mediate their actions. We identified in available mice pituitary scATAC-seq data a region of gonadotrope-specific open chromatin upstream of the Lhb gene, and chromatin conformation capture (3C) in murine gonadotrope cell lines indicated the distal sites that are in physical proximity to the Lhb promoter. Further, we show by chromatin immunoprecipitation (ChIP) a peak of H3K4 monomethylation (H3K4me1) ∼3 kbp upstream of the Lhb TSS, and transcription of two bi-directional non-coding enhancer RNAs (eRNAs), both of which are characteristic of transcriptional enhancers. One of the eRNAs was found to direct open chromatin at the Lhb promoter and is required for Lhb transcription, while guide RNA-mediated recruitment of dCas9- KRAB or dCas9-VP64 to this locus altered Lhb mRNA levels accordingly, confirming its functional enhancer activity. However this enhancer activity is not sufficient for optimal Lhb transcription, and we found that splicing in situ of a distinct downstream lncRNA is also required. The DNA at the central non-coding region of the enhancer, and at the promoter of this lncRNA, forms G4 and iM structures, seen by circular dichroism and ChIP. These DNA structures are affected differentially by local conditions including changes in pH or hormonal treatments, and appear to have diverse effects. HMGB2, which is known to bind structured DNA, was found to bind the enhancer iM in cells and in vitro. Further, HMGB2 knockdown reduced the iM signal and Lhb mRNA, eRNA and lncRNA expression levels quite dramatically. We thus present novel elements involved in the regulation of Lhb transcription through various classes of non-coding RNAs and DNA structures, while emphasizing the complexity of transcriptional regulation via context-dependent cis-acting elements.