Highlights • Expression of estrus in varied intensities (mild, moderate, and strong) among buffaloes was observed. • A Significant fold variationwas observed only between estrus phase and baseline ...value. • LH surge window in urine was extended for several days due to lingering excretion. • Small LH humps were spotted besides the major excretory LH surge.
Mammalian ovulation is induced by a luteinizing hormone surge, which is triggered by elevated plasma estrogen levels; however, chronic exposure to high levels of estradiol is known to inhibit ...luteinizing hormone secretion. In the present study, we hypothesized that the inhibition of the luteinizing hormone surge by chronic estradiol exposure is due to the downregulation of the estrogen receptor alpha in kisspeptin neurons at hypothalamic anteroventral periventricular nucleus, which is known as the gonadotropin-releasing hormone/luteinizing hormone surge generator. Animals exposed to estradiol for 2 days showed an luteinizing hormone surge, whereas those exposed for 14 days showed a significant suppression of luteinizing hormone. Chronic estradiol exposure did not affect the number of kisspeptin neurons and the percentage of kisspeptin neurons with estrogen receptor alpha or c-Fos in anteroventral periventricular nucleus, but it did affect the number of kisspeptin neurons in arcuate nucleus. Furthermore, chronic estradiol exposure did not affect gonadotropin-releasing hormone neurons. In the pituitary, 14-day estradiol exposure significantly reduced the expression of Lhb mRNA and LHβ-immunoreactive areas. Gonadotropin-releasing hormone-induced luteinizing hormone release was also reduced significantly by 14-day estradiol exposure. We revealed that the suppression of an luteinizing hormone surge by chronic estradiol exposure was induced in association with the significant reduction in kisspeptin neurons in arcuate nucleus, luteinizing hormone expression in the pituitary, and pituitary responsiveness to gonadotropin-releasing hormone, and this was not caused by changes in the estrogen receptor alpha-expressing kisspeptin neurons in anteroventral periventricular nucleus and gonadotropin-releasing hormone neurons, which are responsible for estradiol positive feedback. Summary Sentence Suppression of luteinizing hormone surge by chronic estradiol exposure was induced in association with alterations of arcuate nucleus kisspeptin, pituitary luteinizing hormone, and pituitary responsiveness and was not caused by the changes in anteroventral periventricular nucleus kisspeptin neurons, which are responsible for estradiol positive feedback. Graphical Abstract
The aim of the present study was to determine the direct cause of the mutation-induced, increased ovulation rate in Booroola Merino (BB) sheep. Granulosa cells were removed from antral follicles ...before ovulation and post-ovulation from BB (n=5) and WT (n=12) Merino ewes. Direct immunofluorescence measurement of mature cell surface receptors using flow cytometry demonstrated a significant up-regulation of FSH receptor (FSHR), transforming growth factor beta type 1, bone morphogenetic protein receptor (BMPR1B), and LH receptor (LHR) in BB sheep. The increased density of FSHR and LHR provide novel evidence of a mechanism for increasing the number of follicles that are recruited during dominant follicle selection. The compounding increase in receptors with increasing follicle size maintained the multiple follicles and reduced the apoptosis, which contributed to a high ovulation rate in BB sheep. In addition, we report a mutation-independent mechanism of down-regulation to reduce receptor density of the leading dominant follicle in sheep. The suppression of receptor density coincides with the cessation of mitogenic growth and steroidogenic differentiation as part of the luteinization of the follicle. The BB mutation-induced attenuation of BMPR1B signaling led to an increased density of the FSHR and LHR and a concurrent reduction in apoptosis to increase the ovulation rate. The role of BMPs in receptor modulation is implicated in the development of multiple ovulations.
Objective To test by genomic analysis whether empty follicle syndrome (EFS) in a family with two affected sisters has a genetic basis. Design Whole-exome sequencing in the context of clinical ...genetics. Setting University hospital. Patient(s) Two women (36 and 32 years old at the time of the study) with EFS. Intervention(s) Genetic counseling based on autosomal recessive inheritance. Main Outcome Measure(s) Discovery of a mutation in the LH/choriogonadotropin receptor ( LHCGR ) as the cause of EFS. Result(s) A novel missense mutation in LHCGR , p.N400S, was homozygous in sisters with EFS and/or infertility, but not in their unaffected siblings or parents. The mutation was not present in 500 ancestry-matched control subjects. Asparagine at residue 400 is highly conserved and its substitution by serine predicted to alter critical interactions that stabilize LHCGR. Conclusion(s) We describe a genetic basis for EFS and provide strong evidence for the existence of genuine EFS in some patients. A mutation impairing the function of LHCGR explains the lack of response of these patients to repeated administration of β-hCG.
Polycystic ovarian syndrome (PCOS) is a well-known endocrinopathy and one of the most frequent endocrine-reproductive-metabolic syndromes in women, which can result in reduced fertility. While the ...actual cause is unknown, PCOS is regarded as a complicated genetic characteristic with a great degree of variability. Moreover, hormones and immune cells, including both innate and acquired immune cells, are thought to interact in PCOS. Chronic low-grade inflammation raises the risk of autoimmune disease. The study's purpose is to investigate the chemokine monocyte chemoattractant protein-1 (MCP-1) and fertility hormones in samples of women patients with polycystic ovary syndrome (PCOS) in the City of Medicine. Sixty PCOS women comprise 30 healthy control women; their average age was 20–40 years, and their weight ranged from 60 to 100 kg. The results showed an increase in the level of MCP1 in PCOS patients, but this increase was not significant (P<0.05), which was not influenced by BMI or fertility hormones. As well as elevated fertility hormones, this study, when compared to controls as well as patients with PCOS, showed a significant increase in the level of testosterone (14.63 ±2.30 nmol/L) while in control women (0.627 ±0.04), LH hormone in patients and control group (6.54 ±0.51 mIU/mL), and 2.93 ±0.18, respectively. Prolactin hormone was increased in PCOS patients (16.27 ±1.25 ng/mL) when compared to the control group \ (12.85 ±0.62). There was no significant difference in FSH hormone in women with PCOS (5.27 ±0.28 mIU/mL) compared with the control group (5.59 ±0.18).
The blood luteinizing hormone (LH) surge in cows is well studied. However, little is known about urinary LH in cows. This study examined urinary LH concentrations after administration of ...gonadotropin-releasing hormone (GnRH) in six Japanese black cows to induce LH secretion from the pituitary gland into the bloodstream. Abrupt rises in plasma and urinary LH were observed after GnRH administration. Plasma and urinary LH peaked at 2 and 5 hr, respectively. A positive correlation was observed between plasma LH concentrations and urinary LH amounts. Ovulation was confirmed in the cows after 48 hr of GnRH administration. These data strongly suggest that urinary LH is derived from plasma LH, which triggers ovulation in cows.
Safe, reliable, and economic hydrogen storage is a bottleneck for large-scale hydrogen utilization. In this paper, hydrogen storage methods based on the ambient temperature compressed gaseous ...hydrogen (CGH2), liquid hydrogen (LH2) and cryo-compressed hydrogen (CcH2) are analyzed. There exists the optimal states, defined by temperature and pressure, for hydrogen storage in CcH2 method. The ratio of the hydrogen density obtained to the electrical energy consumed exhibits a maximum value at the pressures above 15 MPa. The electrical energy consumed consists of compression and cooling down processes from 0.1 MPa at 300 K to the optimal states. The recommended parameters for hydrogen storage are at 35–110 K and 5–70 MPa regardless of ortho-to parahydrogen conversion. The corresponding hydrogen density at the optimal states range from 60.0 to 71.5 kg m−3 and the ratio of the hydrogen density obtained to the electrical energy consumed ranges from 1.50 to 2.30 kg m−3 kW−1. While the ortho-to para-hydrogen conversion is considered, the optimal states move to a slightly higher temperatures comparing to calculations without ortho-to para-hydrogen conversion.
•Optimal states are proposed for cryo-compressed hydrogen storage.•Ratio of hydrogen density obtained to electrical energy consumed exhibits a maximum.•With ortho-to para-hydrogen conversion, optimal states move to higher temperatures.
The glycoprotein receptors, members of the large G protein-coupled receptor family, are characterized by a large extracellular domains responsible for binding their glycoprotein hormones. ...Hormone-receptor interactions are traditionally analyzed by ligand-binding assays, most often using radiolabeling but also by thermal shift assays. Despite their high sensitivity, these assays require appropriate laboratory conditions and, often, purified plasma cell membranes, which do not provide information on receptor localization or activity because the assays typically focus on measuring binding only. Here, we apply bioluminescence resonance energy transfer in living cells to determine hormone-receptor interactions between a Gaussia luciferase (Gluc)-luteinizing hormone/chorionic gonadotropin receptor (LHCGR) fusion and its ligands (human chorionic gonadotropin or LH) fused to the enhanced green fluorescent protein. The Gluc-LHCGR, as well as other Gluc-G protein-coupled receptors such as the somatostatin and the C-X-C motif chemokine receptors, is expressed on the plasma membrane, where luminescence activity is equal to membrane receptor expression, and is fully functional. The chimeric enhanced green fluorescent protein-ligands are properly secreted from cells and able to bind and activate the wild-type LHCGR as well as the Gluc-LHCGR. Finally, bioluminescence resonance energy transfer was used to determine the interactions between clinically relevant mutations of the hormones and the LHCGR that show that this bioassay provides a fast and effective, safe, and cost-efficient tool to assist the molecular characterization of mutations in either the receptor or ligand and that it is compatible with downstream cellular assays to determine receptor activation/function.
Wilms tumor gene, Wt1, is abundantly expressed in testis Sertoli cells. Our recent study demonstrated that Wt1 is involved in spermatogenesis by regulating Sertoli cell polarity. In the present ...study, we found that Wt1 is also required for steroidogenesis in Leydig cells and that deletion of the Wt1 gene resulted in defects in testosterone biosynthesis and downregulation of steroidogenic gene expression, including cytochrome P450 side-chain cleavage (P450scc), steroidogenic acute regulatory protein (StAR), 3beta-hydroxysteroid dehydrogenase I (3beta-HSD), and cytochrome P450 17A1 (Cyp17a1). The expression of LHR was significantly decreased in Wt1(-/flox); Cre-ER(TM) testes after tamoxifen induction, whereas the luteinizing hormone level in serum was unchanged. Further studies revealed that desert hedgehog (Dhh) expression was regulated by Wt1 in Sertoli cells and that its expression was significantly reduced in Wt1-deficient testes. In vitro study demonstrated that the defect in testosterone production and decreased expression of several steroidogenic genes in Wt1-deficient testis explants was partially rescued by smoothened agonist (SAG), a hedgehog pathway agonist. These results indicate that Wt1 is most likely involved in Leydig cell steroidogenesis by regulating the expression of paracrine factors in seminiferous tubules. Dhh probably had important roles in this process, but we could not exclude the possibility that other factors were also required for Leydig cell steroidogenesis. Loss of Wt1 leads to downregulation of paracrine factors, which in turn causes a decrease in steroidogenic enzyme expression and reduces testosterone production in Leydig cells. The results of this study further confirm that the cross talk between Sertoli cells and Leydig cells has important roles in Leydig cell steroidogenesis.
Objective To investigate the biological role of growth differentiation factor 8 (GDF8) in the regulation of steroidogenesis in human granulosa-lutein (hGL) cells. Design Experimental study. Setting ...Academic medical center. Patient(s) In vitro fertilization patients who provided hGL cells. Intervention(s) Cultured hGL cells treated with recombinant human GDF8 for 24 hours. Main Outcome Measure(s) Expression of steroidogenic enzymes and steroid production in primary cultures of hGL cells used to investigate the effects of GDF8 via specific mRNA and protein levels examined using real time-quantitative polymerase chain reaction and Western blot analysis, respectively, and levels of estradiol and progesterone measured by enzyme immunoassays. Result(s) Extracts were prepared from cultured hGL cells after exposure to GDF8. The levels of cytochrome P450 aromatase (aromatase), the FSH receptor, and estradiol were increased, whereas steroidogenic acute regulatory protein (StAR), luteinizing hormone (LH) receptor, and progesterone levels were decreased after treatment with GDF8. In addition, follicle-stimulating hormone (FSH) stimulated the production of aromatase/estradiol, and LH induced the production of StAR/progesterone. Furthermore, pretreatment with GDF8 for 24 hours enhanced the effects of FSH on aromatase/estradiol induction, whereas GDF8 suppressed the effects of LH on StAR/progesterone stimulation. Conclusion(s) In human granulosa cells, GDF8 may play an important role in the modulation of cellular responsiveness to gonadotropins and in the regulation of ovarian steroid production, most likely as a luteinization inhibitor.
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