Follicle-stimulating Hormone Release Research Articles

Effect of kisspeptin (KISS) and RFamide-related peptide-3 (RFRP-3) on the synthesis and secretion of FSH in vitro by pituitary cells in pigs

Kisspeptins (KISSs) and RFamide-related peptide-3 (RFRP-3) affect the synthesis and secretion of luteinizing hormone (LH) and modulate female reproductive processes. The presence of KISS and RFRP-3 in the porcine pituitary gland and their contribution to the regulation of follicle-stimulating hormone (FSH) synthesis and secretion is unknown. This study analyzed the presence of KISS and RFRP-3 in the pituitary of estrous-cyclic pigs on days 2 to 3, 10 to 11, 12 to 13, 15 to 16 and 19 to 20 and early pregnant pigs on days 10 to 11, 12 to 13 and 15 to 16, and evaluated the effect of KISS and RFRP-3 on β-Fsh mRNA expression and FSH secretion in vitro by pituitary cells collected on selected days of the estrous cycle. The cells were cultured in vitro and treated with KISS (10−6 M, 10−7 M) and RFRP-3 (10−6 M, 10−7 M) or gonadotropin-releasing hormone (GnRH; 100 ng/mL) alone and in combinations (4 h or 24 h). The relative abundance of Kiss and Rfrp-3 and their receptor mRNA transcripts, as well as the KISS and RFRP-3 proteins, were found in the pituitaries of estrous-cyclic and early pregnant pigs. KISS after 4 h increased the secretion of FSH in estrous cyclic pigs mostly during the early-luteal phase and luteolysis. RFRP-3 inhibited the synthesis and secretion of FSH in estrous-cyclic pigs on days 19 to 20 and the secretion of FSH on days 2 to 3 and 10 to 12 of the estrous cycle compared with GnRH-treated cells. KISS in co-treatment with GnRH after 24 h enhanced FSH release on days 2 to 3 and 15 to 16 of the estrous cycle. In conclusion, KISS and RFRP-3 systems are present in the pituitary of estrous-cyclic and pregnant pigs. In estrous-cyclic pigs, KISS and RFRP-3 may affect the synthesis and secretion of FSH by pituitary cells.

Distribution of GnRH and Kisspeptin Immunoreactivity in the Female Llama Hypothalamus.

Llamas are induced non-reflex ovulators, which ovulate in response to the hormonal stimulus of the male protein beta-nerve growth factor (β-NGF) that is present in the seminal plasma; this response is dependent on the preovulatory gonadotrophin-releasing hormone (GnRH) release from the hypothalamus. GnRH neurones are vital for reproduction, as these provide the input that controls the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the pituitary gland. However, in spontaneous ovulators, the activity of GnRH cells is regulated by kisspeptin neurones that relay the oestrogen signal arising from the periphery. Here, we investigated the organisation of GnRH and kisspeptin systems in the hypothalamus of receptive adult female llamas. We found that GnRH cells exhibiting different shapes were distributed throughout the ventral forebrain and some of these were located in proximity to blood vessels; sections of the mediobasal hypothalamus (MBH) displayed the highest number of cells. GnRH fibres were observed in both the organum vasculosum laminae terminalis (OVLT) and median eminence (ME). We also detected abundant kisspeptin fibres in the MBH and ME; kisspeptin cells were found in the arcuate nucleus (ARC), but not in rostral areas of the hypothalamus. Quantitative analysis of GnRH and kisspeptin fibres in the ME revealed a higher innervation density of kisspeptin than of GnRH fibres. The physiological significance of the anatomical findings reported here for the ovulatory mechanism in llamas is still to be determined.

Treatment of uterine bleeding in pubertal girls without application of hormonal therapy on the background of related thyroid pathology

Pubertal uterine bleeding occupies an important place in the structure of gynecological pathology since, in the years, it can lead to persistent and irreversible changes in the process of formation of the reproductive system.The purpose of the work is to investigate the effectiveness of non-hormonal therapy in the treatment of uterine bleeding in adolescent girls against the background of concomitant thyroid pathology.Material and methods. Thirty adolescent girls with pubertal uterine bleeding were examined and treated against the background of concomitant thyroid pathology, which were treated in the gynecological department of the city clinical maternity hospital No. 1 in Chernivtsi. All examined patients also underwent a comprehensive examination to determine the level of hormones in the blood serum and to determine individual indicators of hemostasis.Results. It was established that the complex therapy used in adolescent girls suffering from pubertal uterine bleeding against the background of thyroid pathology is effective, as indicated by the positive feedback mechanism, namely the release of follicle-stimulating hormone in response to an increase in the level of estradiol in peripheral blood.Conclusion. The comprehensive treatment technique developed and used by us in adolescent girls with uterine bleeding with concomitant thyroid pathology contributes to the gradual normalization of the level of sex and thyroid hormones in the blood serum, shortens the treatment period in the hospital, and helps to normalize the overall hormonal profile of the examined girls with concomitant thyroid pathology.

Osteoprotective effect of bread enriched with protein, dietary fiber, calcium, iron and iodine in hypoestrogen-induced osteoporosis among rats

The search for new strategies for the prevention and control of osteoporosis is an urgent task. Functional foodstuffs and their components are of particular interest in this regard. The aim was to study the effect of bread enriched with protein, dietary fiber, calcium, iron and iodine on the state of the bone tissue of rats in a model of postmenopausal osteoporosis. Material and methods. The experiment was performed on sexually mature female Wistar rats divided into groups: K - control (sham-operated rats, not ovariectomized); О30 - osteoporosis model (animals were sacrificed 30 days after ovariectomy); groups О120 and О120+ - a model of osteoporosis (rats were sacrificed 120 days after ovariectomy). All animals were fed a standard vivary diet. For rats of the О120+ group, from the 40th to the 120th day, enriched bread was included in the diet in an amount of 6 g per 100 g of body weight per day. The bread was fortified with protein (whey protein, blood plasma proteins from farm animals), dietary fiber, calcium (eggshell), iron (purified hemoglobin) and iodized whey protein. Animals of groups K and О120 received unfortified bread in the same amount. Blood levels of total calcium (by colorimetric method), gonadotropins, testosterone, and estradiol (by enzyme-linked immunosorbent assay) were analyzed. Microtomographic evaluation of the architecture and mineral density of the trabecular part of the femur and lumbar vertebrae was performed. Histomorphological analysis of the uterus and femur of animals was performed. Results and discussion. In animals of the О120+ group, in comparison with the О120 sample, there was a decrease in blood testosterone and a marked compensatory release of follicle-stimulating hormone, while no changes were detected in the concentration of estradiol and the state of the uterus atrophied against the background of ovariectomy. There was an increase in the trabecular mineral density of the femur and lumbar vertebrae. The proportion of bone trabeculae in the total volume of the femoral metaphysis (BV/TV) in animals of the О120+ sample was 12.5±0.66% compared to 10.4±0.52% in the О120 group. The values of the structural model index (SMI) reflecting the loss of bone strength and the trabecularity coefficient (TbPf) in О120+ rats (1.44±0.07 and 5.96±0.29 1/mm) were significantly lower than these parameters in the О120 group (1.74±0.08; 9.13±0.46 1/mm, р<0.05). The micro-architectural structure of the femur in the О120+ group of rats was close to that of the О30 sample, which serves as a model of the early stage of osteoporosis (SMI 1.42±0.07; TbPf 5.55±0.28 1/mm). The percentage of bone resorption perimeter and the number of osteoclasts in the О120+ femoral trabeculae were lower than in the О120 group. In the О120+ group, active osteoblasts were observed in a significant part of the resorption cavities. Cell differentiation more was observed in the osteogenic direction than in the adipogenic direction. Conclusion. Bread enriched with protein, fiber, calcium, iron and iodine, effectively weakens osteoporosis induced by ovariectomy in rats. Its inclusion in the diet may be beneficial for the prevention and treatment of systemic postmenopausal osteoporosis.

Enzymatic preparation of Crassostrea oyster peptides and their promoting effect on male hormone production

Ethnopharmacological relevanceCrassostrea gigas Thunberg and other oysters have been traditionally used in China as folk remedies to invigorate the kidney and as natural aphrodisiacs to combat male impotence. Aim of the studyErectile dysfunction (ED) has become a major health problem for the global ageing population. The aim of this study is therefore to evaluate the effect of peptide-rich preparations from C. gigas oysters on ED and related conditions as increasing evidence suggests that peptides are important bioactive components of marine remedies and seafood. Materials and methodsCrassostrea oyster peptide (COP) preparations COP1, COP2 and COP3 were obtained from C. gigas oysters by trypsin, papain or sequential trypsin-papain digestion, respectively. The contents of testosterone, cyclic adenosine monophosphate (cAMP) and nitric oxide (NO) and the activity of nitric oxide synthase (NOS) in mice and/or cells were measured by enzyme-linked immunosorbent assays. Real-time PCR was used to assess the expression of genes associated with sex hormone secretion pathways. The model animal Caenorhabditis elegans was also used to analyze the gene expression of a conserved steroidogenic enzyme. In silico analysis of constituent peptides was performed using bioinformatic tools based on public databases. ResultsThe peptide-rich preparation COP3, in which >95% peptides were <3000 Da, was found to increase the contents of male mouse serum testosterone and cAMP, both of which are known to play important roles in erectile function, and to increase the activity of mouse penile NOS, which is closely associated with ED. Further investigation using mouse Leydig-derived TM3 cells demonstrates that COP3 was able to stimulate the production of testosterone as well as NO, a pivotal mediator of penile erection. Real-time PCR analysis reveals that COP3 up-regulated the expression of Areg and Acvr2b, the genes known to promote sex hormone secretion, but not Fst, a gene involved in suppressing follicle-stimulating hormone release. Furthermore, COP3 was also shown to up-regulate the expression of let-767, a well-conserved C. elegans gene encoding a protein homologous to human 17-β-hydroxysteroid dehydrogenases. Preliminary bioinformatic analysis using the peptide sequences in COP3 cryptome identified 19 prospective motifs, each of which occurred in more than 10 peptides. ConclusionsIn this paper, Crassostrea oyster peptides were prepared by enzymatic hydrolysis and were found for the first time to increase ED-associated biochemical as well as molecular biology parameters. These results may help to explain the ethnopharmacological use of oysters and provide an important insight into the potentials of oyster peptides in overcoming ED-related health issues.

Relative abundance of chemerin mRNA transcript and protein in pituitaries of pigs during the estrous cycle and early pregnancy and associations with LH and FSH secretion during the estrous cycle

Adipokines such as chemerin affect metabolic status and reproductive function in many species. The hypothesis in the present study was that there were chemerin mRNA transcript and protein in the pituitary of pigs and that relative abundances fluctuate during the estrous cycle and early pregnancy. Chemerin is thought to modulate luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion during the estrous cycle. Changes in the relative abundance of chemerin mRNA transcript and protein in anterior (AP) and posterior (PP) pituitaries of pigs were investigated, for the first time in the present study, during four phases of the estrous cycle and four periods of early pregnancy. Chemerin protein was localized in gonadotrophs, thyrotrophs and somatotrophs during the estrous cycle and early gestation. Chemerin treatments affected both basal, GnRH- and/or insulin-induced LH and FSH production, with there being variations with phase of the estrous cycle when tissues were collected. These findings indicate chemerin may be produced locally in the pituitary and may affect female reproductive function by controlling the release of LH and FSH from AP cells.

Effect of Wubi Shanyao Pills on sexual dysfunction in rats with adenine-induced kidney-Yang deficiency

The aim of this paper was to study the effect of Wubi Shanyao Pills on sexual dysfunction in rats with kidney-Yang deficiency and to investigate its possible mechanism. Adenine(100 mg·kg~(-1)) was administered to male SD rats for 8 weeks to establish kidney-Yang deficiency model, and at the same time, Wubi Shanyao Pills(2, 1, 0.5 g·kg~(-1)) were administered to rats for 8 weeks. The syndrome manifestation of kidney-Yang deficiency was observed in rats and the scores of symptoms were evaluated. Sexual behavior indexes(incubation period and times of capture, straddle and ejaculation) were measured by mating experiment. The levels of serum testosterone(T), estradiol(E_2), follicle stimulating hormone(FSH), luteinizing hormone(LH), and gonadotropin releasing hormone(GnRH) were measured by radioimmunoassay. The wet weights of testis and seminal vesicle were measured. The content of fructose in seminal plasma was detected by UV spectrophotometry. The pathological changes of testis and epididymis were observed by HE staining. The expression levels of transforming growth factor(TGF-β1) and cytochrome P450 aromatase(CYP19) in testis were detected by immunohistochemistry and Western blot. The results showed that Wubi Shanyao Pills could significantly reduce the score of kidney-Yang deficiency syndrome, improve the symptoms of kidney-Yang deficiency syndrome, shorten capture, straddle and ejaculation latency, increase capture and straddle times, increase serum T, LH, FSH, E_2 and GnRH levels, increase the wet weight of testis and seminal vesicle and fructose content in seminal plasma, improve the pathological structure of testis and epididymis, and inhibit the expression of TGF-β1 and increase CYP19 in testis of the model rats. Therefore, Wubi Shanyao Pills can significantly improve sexual dysfunction in rats with kidney-Yang deficiency, and its mechanism may be related to regulating the low function of hypothalamus pituitary gonad(HPG) axis and improving the disorder of sex hormone secretion. In addition, it may be also related to inhibiting the expression of testicular TGF-β1, increasing the expression of CYP19 protein, and then regulating the amount of T converted to E_2.

Interactions of circulating estradiol and progesterone on changes in endometrial area and pituitary responsiveness to GnRH†.

Changes in circulating progesterone (P4) and estradiol (E2) during proestrus produce dynamic changes in endometrial function and pituitary release of gonadotropins. Independent and combined effects of P4 and E2 on endometrium and pituitary were evaluated. In a preliminary study, an exogenous hormone model of proestrus was created by removal of corpus luteum and follicles ≥5mm followed by gradual removal of intravaginal P4 implants during 18h and treatment with increasing doses of estradiol benzoate during 48h to mimic proestrus using high E2 (n=9) or low E2 (n=9). Decreased P4, increased E2, and increased endometrial area (EA) simulated proestrus in high-E2 cows and this was used subsequently. The main experiment used a 2×2 factorial design with: high E2 and low P4 (n=11); high E2 and high P4 (n=11); low E2 and high P4 (n=11); low E2 and low P4 (n=10). At 48h, gonadotropin-releasing hormone (GnRH)-induced luteinizing hormone (LH) and follicle stimulating hormone (FSH) release was determined. Variables were analyzed using PROCMIXED of Statistical Analysis System. The EA increased dramatically during 48h only in high-E2 and low-P4 cows. For FSH, high-E2 cows had greater area under the curve (AUC) and FSH peak after GnRH than low E2, with mild negative effects of high P4. For LH, concentration at peak and AUC were 2-fold greater in high E2 compared to low-E2 groups, with low P4 also 2-fold greater than high-P4 groups. Thus, maximal changes in uterus and pituitary during proestrus depend on both low P4 and high E2, but different physiologic responses are regulated differently by E2 and P4. Changes in endometrium depend on low P4 and high E2, whereas GnRH-induced FSH secretion primarily depends on high E2, and GnRH-induced LH secretion is independently increased by high E2 or reduced by high P4.

When male seahorses take the female contraceptive pill ...

17α-ethinylestradiol (EE2), the female contraceptive pill, has been detected in mediterranean coasts where seahorse populations, Hippocampus guttulatus, live. Low environmental concentrations have the potential to disrupt growth but also endocrine metabolism, and this imbalance is all the more critical in early life stage. To investigate the impact of EE2 in reared seahorses, we exposed aged 2months and sexually undifferentiated seahorses to an environmental concentration of 21ng/L of EE2 for a period of 30days. EE2 exposure led to a 19% reduction in weight, but also a mortality rate of 27%. This exposure predicted demasculinization of male individuals with a late onset of secondary sexual characteristics. EE2 exposure led to an increase of the free androgen index, but significant reductions of estradiol and testosterone in males were observed. This low estrogen concentration seemed to impact the positive feedback on luteinizing hormone (LH) with a decrease in LH production. Added to this, synthetic estrogen had a negative impact on the production and the release of follicle-stimulating hormone. Contrary to all expectations, females demonstrated a significant decrease in vitellogenin, following exposure to EE2 at 21ng/L, while no changes were detected in males. This first study on the European long-snouted seahorses confirmed the deleterious impact of the female contraceptive pill with a real impact on growth, sexual differentiation, and maturation in young immature seahorses.

Follistatin expression in the central nervous system of the adult rat

Follistatin was initially cloned as a monomeric polypeptide that inhibits the release of follicle-stimulating hormone. Although follistatin also plays pivotal roles in skeletal muscle hypertrophy and immunoregulation in the epididymis, little information is available regarding follistatin function in the adult central nervous system (CNS). Hence, we investigated follistatin expression in the adult rat CNS using immunohistochemistry. Follistatin was intensely expressed in most neurons and their axons. Furthermore, oligodendrocytes, ependymal cells, and some astrocytes also expressed follistatin protein. These data indicate that follistatin is widely expressed throughout the adult CNS. The abundant expression of follistatin in the adult brain suggests that this protein plays important roles in the CNS.

Contraception and CKD.

In general, nephrologists give little thought to contraception, leaving this task to other practitioners, but the significant risks associated with drugs routinely prescribed to treat kidney disease; the potential for pregnancy-associated adverse outcomes in women with CKD; and the effect that

Transcriptome analysis during photostimulated recrudescence reveals distinct patterns of gene regulation in Siberian hamster ovaries†.

In Siberian hamsters, exposure to short days (SDs, 8h light:16h dark) reduces reproductive function centrally by decreasing gonadotropin secretion, whereas subsequent transfer of photoinhibited hamsters to stimulatory long days (LDs, 16L:8D) promotes follicle stimulating hormone (FSH) release inducing ovarian recrudescence. Although differences between SD and LD ovaries have been investigated, a systematic investigation of the ovarian transcriptome across photoperiod groups to identify potentially novel factors that contribute to photostimulated restoration of ovarian function had not been conducted. Hamsters were assigned to one of four photoperiod groups: LD to maintain ovarian cyclicity, SD to induce ovarian regression, or post transfer (PT), where females housed in SD for 14-weeks were transferred to LD for 2-days or 1-week to reflect photostimulated ovaries prior to (PTd2) and following (PTw1) the return of systemic FSH. Ovarian RNA was extracted to create RNA-sequencing libraries and short-read sequencing Illumina assays that mapped and quantified the ovarian transcriptomes (n=4/group). Ovarian and uterine masses, plasma FSH, and numbers of antral follicles and corpora lutea decreased in SD as compared to LD ovaries (P<0.05). When reads were aligned to the mouse genome, 18 548 genes were sufficiently quantified. Most of the differentially expressed genes noted between functional LD ovaries and regressed SD ovaries; however, five main expression patterns were identified across photoperiod groups. These results, generally corroborated by select protein immunostaining, provide a map of photoregulated ovary function and identify novel genes that may contribute to the photostimulated resumption of ovarian activity.

Effects of central orexin A on gonadotropins and progesterone secretion in ewes during the luteal phase of the estrous cycle and during anestrus

Prior studies, mainly on rodents demonstrated that orexin A (OXA) plays an important role in regulating the hypothalamo-pituitary-gonadal (HPG) axis. However, the results of those works have indicated that the central impact of OXA on the HPG axis may vary depending on the species or sex of the animal, the phase of the estrous cycle, or the experimental model system (in vivo vs. in vitro). The aim of the present study was to further investigate the role of OXA in the regulation of reproduction in seasonally breeding, intact, non-steroid-primed ewes during the luteal phase of the estrous cycle and during anestrus. We examined the effect of fourth-ventricle injections of OXA (0.3 μg/kg body weight) on plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH) and progesterone concentrations and on the level of gonadotropin-releasing hormone receptor (GnRH-R) protein in the anterior pituitary (AP). Radioimmunoassays revealed that OXA treatments inhibited (P < 0.001) LH and FSH secretion during both examined periods and progesterone (P < 0.001) during the luteal phase. The influence of OXA on LH and FSH release may be partially attributable to the additional response of the pituitary to OXA, as, under the influence of OXA, the concentration of GnRH-R protein in the AP was downregulated during the luteal phase (P < 0.01) and during anestrus (P < 0.05). We conclude that OXA may be a negative regulator of gonadotropin secretion in intact, non-steroid-primed ewes in periods when the HPG axis is suppressed by negative feedback through steroid sex hormones.

Acute restraint stress triggers progesterone withdrawal and endometrial breakdown and shedding through corticosterone stimulation in mouse menstrual-like model.

Stress impacts the reproductive axis at the level of the hypothalamus and the pituitary gland, which exert an effect on the ovary. Menstruation is regulated by the hypothalamic-pituitary-ovary (HPO) axis. However, the role of stress in menstruation remains unclear. The objective of this study was to explore the role of stress in endometrial breakdown and shedding, using the pseudopregnant mouse menstrual-like model. Female mice were mated with vasectomized males and labeled day 0.5, upon observation of a vaginal seminal plug. On day 3.5, decidualization was induced in pseudopregnant mice using arachis oil. On day 5.5, pseudopregnant mice with artificial decidualization were placed in restraint tubes for 3 h. The findings indicated that acute restraint stress resulted in the disintegration of the endometrium. While corticosterone concentration in the serum increased significantly due to restraint stress, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and progesterone (P4) levels in the serum decreased significantly. An endometrial histology examination indicated that progesterone implants may rescue P4 decline caused by acute stress and block endometrium breakdown and shedding. In addition, mice were treated with metyrapone, an inhibitor of corticosterone synthesis, 1 h prior to being subjected to restraint stress. Interestingly, metyrapone not only inhibited stress-induced endometrium breakdown and shedding, but also prevented stress-induced reduction of P4, LH and FSH. Furthermore, real-time PCR and western blot showed that mRNA and protein expression of CYP11A1 (cytochrome P450, family 11, subfamily A, polypeptide 1) and steroidogenic acute regulatory protein (StAR), the two rate-limiting enzymes for progesterone synthesis in the ovary, decreased following acute stress. But metyrapone prevented the reduction of StAR expression induced by restraint stress. Overall, this study revealed that acute stress results in an increase in corticosterone, which may inhibit LH and FSH release in the serum and CYP11A1 and StAR expression in the ovary, which finally leads to the breakdown and shedding of the endometrium. These experimental findings, based on the mouse model, may enable further understanding of the effects of stress on menstruation regulation and determine the potential factors affecting stress-associated menstrual disorders.

Activin A in Mammalian Physiology.

Activins are dimeric glycoproteins belonging to the transforming growth factor beta superfamily and resulting from the assembly of two beta subunits, which may also be combined with alpha subunits to form inhibins. Activins were discovered in 1986 following the isolation of inhibins from porcine follicular fluid, and were characterized as ovarian hormones that stimulate follicle stimulating hormone (FSH) release by the pituitary gland. In particular, activin A was shown to be the isoform of greater physiological importance in humans. The current understanding of activin A surpasses the reproductive system and allows its classification as a hormone, a growth factor, and a cytokine. In more than 30 yr of intense research, activin A was localized in female and male reproductive organs but also in other organs and systems as diverse as the brain, liver, lung, bone, and gut. Moreover, its roles include embryonic differentiation, trophoblast invasion of the uterine wall in early pregnancy, and fetal/neonate brain protection in hypoxic conditions. It is now recognized that activin A overexpression may be either cytostatic or mitogenic, depending on the cell type, with important implications for tumor biology. Activin A also regulates bone formation and regeneration, enhances joint inflammation in rheumatoid arthritis, and triggers pathogenic mechanisms in the respiratory system. In this 30-yr review, we analyze the evidence for physiological roles of activin A and the potential use of activin agonists and antagonists as therapeutic agents.

A small population of hypothalamic neurons govern fertility: the critical role of VAX1 in GnRH neuron development and fertility maintenance

Fertility depends on the correct maturation and function of approximately 800 gonadotropin-releasing hormone (GnRH) neurons in the brain. GnRH neurons are at the apex of the hypothalamic-pituitary-gonadal axis that regulates fertility. In adulthood, GnRH neurons are scattered throughout the anterior hypothalamic area and project to the median eminence, where GnRH is released into the portal vasculature to stimulate release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the pituitary. LH and FSH then regulate gonadal steroidogenesis and gametogenesis. Absence of GnRH neurons or inappropriate GnRH release leads to infertility. Despite the critical role of GnRH neurons in fertility, we still have a limited understanding of the genes responsible for proper GnRH neuron development and function in adulthood. GnRH neurons originate in the olfactory placode then migrate into the brain. Homeodomain transcription factors expressed within GnRH neurons or along their migratory path are candidate genes for inherited infertility. Using a combined in vitro and in vivo approach, we have identified Ventral Anterior Homeobox 1 (Vax1) as a novel homeodomain transcription factor responsible for GnRH neuron maturation and fertility. GnRH neuron counts in Vax1 knock-out embryos revealed Vax1 to be required for the presence of GnRH-expressing cells at embryonic day 17.5 (E17.5), but not at E13.5. To localize the effects of Vax1 on fertility, we generated Vax1flox mice and crossed them with Gnrhcre mice to specifically delete Vax1 within GnRH neurons. GnRH staining in Vax1flox/flox:GnRHcre mice show a total absence of GnRH expression in the adult. We performed lineage tracing in Vax1flox/flox:GnRHcre:RosaLacZ mice which proved GnRH neurons to be alive, but incapable of expressing GnRH. The absence of GnRH leads to delayed puberty, hypogonadism and complete infertility in both sexes. Finally, using the immortalized model GnRH neuron cell lines, GN11 and GT1-7, we show that VAX1 is a direct regulator of Gnrh1 transcription by binding key ATTA sites within the Gnrh1 promoter. This study identifies VAX1 as a key transcription factor regulating GnRH expression and establishes VAX1 as a novel candidate gene implicated in heritable infertility.

Protein adsorption onto polysaccharides: Comparison of chitosan and chitin polymers

Chitosan (CHS) and chitin (CHT) biopolymers have found many applications in the field of controlled-release drug delivery systems. Herein, molecular dynamics (MD) simulation and binding free energy calculations were used to investigate the potentials of CHS and CHT polymers for the controlled release of follicle-stimulating hormone (FSH). The results indicated that FSH conformation did not change in the presence of CHS and CHT. In addition, FSH-polymer interactions caused stability of the 310-helix structure of the alpha subunits of FSH (FSHα). Both the biopolymers interacted with the protein mainly through the hydrophobic forces. CHS has more affinity for FSH when compared with CHT. Furthermore, in both systems, the affinity of polymers for FSHα was more than that for beta subunits of FSH (FSHβ). The results suggested that the polysaccharides might improve the controlled-release FSH delivery.

Herb Formula ZhenRongDan Balances Sex Hormones, Modulates Organ Atrophy, and Restores ERα and ERβ Expressions in Ovariectomized Rats.

Herb mixtures are widely used for treatment of the menopausal syndrome long before the hormonal therapy. However, there is insufficient data for herb remedies in treating menopausal syndromes. Here we aim to investigate the effect of ZhenRongDan (ZRD) in balancing female hormones, regulating expression of estrogen receptors (ERs), and preventing organ atrophy in menopausal rats. Rats that underwent bilateral ovariectomy were used in the experiments; the effects of ZRD on serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and estradiol (E2) levels were observed. Histology of vagina and ERs expression in vagina, uterus, and adrenal gland were also examined. ELISAs were used to analyze the changes of FSH, LH, PRL, and E2 in serum, and the morphological changes of the cervical epithelium cells were observed by Hematoxylin & Eosin (H&E) staining. Immunohistochemistry and western blot were applied to detect estrogen receptors subtypes alpha (ERα) and beta (ERβ) expression in vagina, uterus, and adrenal gland. We found that ZRD could significantly reduce the weight of the adrenal gland and increase the weight of the uterus. It could decrease the release of FSH and LH as well as increasing E2 and PRL levels. Furthermore, ZRD could improve the number of cervical vaginal epithelial cells and increase the thickness of the vaginal wall. And the altered expressions of ERα and ERβ are also restored by ZRD. ZRD could obviously relieve the endocrine disorders, modulate organ atrophy, and restore ERα and ERβ expression in the ovariectomized rat model.

A second successful pregnancy using oocytes from a woman with premature ovarian failure with embryos transferred to a gestational carrier

Purpose: To describe a second successful pregnancy in a woman with premature ovarian failure who was successfully able to induce ovulation by restoring down-regulated follicle stimulating hormone (FSH) receptors, by lowering elevated FSH by inhibiting FSH release from the pituitary, by using ethinyl estradiol with subsequent stockpiling of embryos following in vitro fertilization (IVF) for subsequent future transfer to a gestational carrier. Materials and Methods: A day 3 eight-cell embryo obtained from a fertilized oocyte 10.5 years before was transferred to a gestational carrier. Results: The gestational carrier conceived and she successfully carried to six months to date. Conclusions: This is the first case of two successful pregnancies from the same woman with premature ovarian failure. Also, this is the longest an embryo derived from a woman in ovarian failure has been frozen leading to a successful pregnancy.

Final follicular maturation by administration of GnRH agonist plus HCG versus HCG in normal responders in ART cycles: An RCT

Gonadotropin-releasing hormone agonists (GnRH-a) was increasingly used for triggering oocyte maturationfor the prevention of ovarian hyperstimulation syndrome. Studies suggest that GnRH-a might be used as a better trigger agent since it causes both Luteinizing hormone and follicle stimulating hormone release from a physiologic natural cycle. The aim of this study was to evaluate the effect of dual-triggering in assisted reproductive technology outcomes. 192 normal responder women aged ≤42 years and 18< Body Mass Index <30 kg/m2 enrolled in this single-blind randomized controlled trial. All participants received antagonist protocol. For final triggering, women randomly were divided into two groups. Group, I was triggered by 6500 IU human chorionic gonadotropin (hCG) alone, and group II by 6500 IU hCG plus 0.2 mg of triptorelin. The implantation, chemical, clinical and ongoing pregnancy, and abortion rates were measured. The mean of retrieved oocytes and obtained embryos were statistically higher in the dual-trigger group (group I), but the implantation and pregnancy rates were similar in two groups. The results of our study did not confirm the favorable effect of dual-triggered oocyte maturation with a GnRH-a and a standard dosage of hCG as an effective strategy to optimize pregnancy outcome for normal responders in GnRH-antagonist cycles. We think that this new concept requires more studies before becoming a universal controlled ovarian hyperstimulation protocol in in vitro fertilization practice.