Superovulatory responses using pregnant mare serum gonadotropin hormone in Murrah buffalo cows

Superovulation plays an important role in embryo transfer program. A preliminary study carried out in the Research Institute for Animal Production evaluated superovulatory responses in Murrah buffalo cows using pregnant mare serum gonadotropin (PMSG) hormone. The three buffalo cows were estrus synchronized using 5 ml prostaglandin (PGF) twice, with an interval of 11 days. PMSG was injected intra-muscularly 3000 IU on day-10 after estrus. Prostaglandins were administered 48 hours after PMSG injection. Fixed-Time artificial insemination (FTAI) was carried out at 72 hours after the last PGF treatment. Administration of hCG 2 ml/head was given at the time of FTAI. A non-surgery flushing was performed on day 6 after FTAI. Parameters observed using ultrasonography (USG) were diameter of follicle (DFL), total follicle (TFL), and number embryos (NE). Data were analyzed descriptively. The mean of DFL before PMSG treatment was 8.2 mm and after PMSG treatment was 12.5 mm. The mean of TFL before PMSG treatment was 7.7 and after PMSG treatment was 16.1. The NE obtained was one degenerative embryo. Superovulation using PMSG increased TCL and DFL. It can be concluded that the Murrah buffalo cows superovulated by PMSG showed a good response but no transferable embryo was found.

Evaluation of Serum Gonadotropin and Prolactin Level among Sudanese Patients with Chronic Renal Failure

Background: Generally, patients on hemodialysis for chronic renal failure also have endocrine defects and sexual function disorders. In this study, we aimed to assess the serum prolactin (PRL), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in patients with chronic renal failure. Methods: This hospital-based case–control study was conducted at Jabal Aulia Teaching Hospital, Khartoum, Sudan. The study was carried out between August 2019 and February 2020. A total of 100 subjects were enrolled – 50 chronic renal failure patients and 50 as controls. The serum hormones were estimated using Tosoh 360. SPSS version 25 was used to analyze the results. Results: The serum PRL, LH, and FSH were significantly increased among chronic renal failure patients than their healthy counterparts (p-value = 0.000). The age of patients was positive correlated with plasma hormones, PRL (r = 0.332, p = 0.001), LH (r = 0.387, p = 0.000), and FSH (r = 0.320, p = 0.001). No correlation was found between the duration of the disease and serum hormones. Conclusion: Patients with chronic renal failure had a highly significant increase of serum PRL, LH, and FSH and also the age of the patients was positively correlated with serum hormones. Keywords: chronic renal failure, prolactin, gonadotropin, hemodialysis

Peroxiredoxin 1 Controls Ovulation and Ovulated Cumulus–Oocyte Complex Activity through TLR4-Derived ERK1/2 Signaling in Mice

Peroxiredoxins (PRDXs) are expressed in the ovary and during ovulation. PRDX1 activity related to the immuno-like response during ovulation is unknown. We investigated the roles of Prdx1 on TLR4 and ERK1/2 signaling from the ovulated cumulus–oocyte complex (COC) using Prdx1-knockout (K/O) and wild-type (WT) mice. Ovulated COCs were collected 12 and 16 h after pregnant mare serum gonadotropin/hCG injection. PRDX1 protein expression and COC secretion factors (Il-6, Tnfaip6, and Ptgs2) increased 16 h after ovulated COCs of the WT mice were obtained. We treated the ovulated COCs in mice with LPS (0.5 μg/mL) or hyaluronidase (Hya) (10 units/mL) to induce TLR4 activity. Intracellular reactive oxygen species (ROS), cumulus cell apoptosis, PRDX1, TLR4/P38/ERK1/2 protein expression, and COC secretion factors’ mRNA levels increased in LPS- and Hya-treated COCs. The ERK inhibitor (U0126) and Prdx1 siRNA affected TLR4/ERK1/2 expression. The number and cumulus expansion of ovulated COCs by ROS were impaired in Prdx1 K/O mice but not in WT ones. Prdx1 gene deletion induced TLR4/P38/ERK1/2 expression and cumulus expansion genes. These results show the controlling roles of PRDX1 for TLR4/P38/ERK1/2 signaling activity in ovulated mice and the interlink of COCs with ovulation.

Determining the Effects of Serial Injections of Pregnant Mare Serum Gonadotropin on Plasma Testosterone Concentrations, Testicular Dynamics, and Semen Production in Leopard Geckos (Eublepharis macularius)

Reptiles are highly susceptible to anthropogenic activities as a result of their narrow geographical ranges and habitat specialization, making them a conservation concern. Geckos represent one of the mega-diverse reptile lineages under pressure; however, limited assisted reproductive technologies currently exist for these animals. Exogenous pregnant mare serum gonadotropin (PMSG) has been found to exhibit follicle stimulating hormone-like action and has been routinely used to alter reproductive hormones of vertebrates in assisted reproductive protocols. The purpose of this study was to determine the effects of serial injections of 20 IU and 50 IU PMSG on circulating testosterone concentrations, testicular dynamics, and semen production in a model species of gecko. Twenty-four captive-bred, adult, male leopard geckos (Eublepharis macularius) were divided into three treatment groups and administered a once-weekly injection of either PMSG or saline for a total of nine weeks. Ultrasonographic testicular measurements, electrostimulation for semen collection, and venipuncture were performed on days 0, 21, 42, and 63. Right unilateral orchidectomies and epididymectomies were performed in all animals on day 63; tissues were submitted for histopathology. PMSG treated geckos had significantly higher testicular volumes and weights, spermatozoa motility, and spermatozoa concentrations compared with controls. However, there were no significant differences in testosterone concentrations by treatment or time. Under the conditions outlined, PMSG is effective at stimulating spermatogenesis and increasing testicular size, but not effective at increasing testosterone concentrations in the leopard gecko between October–December in the Northern hemisphere.

Endocrine and molecular milieus of ovarian follicles are diversely affected by human chorionic gonadotropin and gonadotropin-releasing hormone in prepubertal and mature gilts

Different strategies are used to meet optimal reproductive performance or manage reproductive health. Although exogenous human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) agonists (A) are commonly used to trigger ovulation in estrous cycle synchronization, little is known about their effect on the ovarian follicle. Here, we explored whether hCG- and GnRH-A-induced native luteinizing hormone (LH) can affect the endocrine and molecular milieus of ovarian preovulatory follicles in pigs at different stages of sexual development. We collected ovaries 30 h after hCG/GnRH-A administration from altrenogest and pregnant mare serum gonadotropin (eCG)-primed prepubertal and sexually mature gilts. Several endocrine and molecular alternations were indicated, including broad hormonal trigger-induced changes in follicular fluid steroid hormones and prostaglandin levels. However, sexual maturity affected only estradiol levels. Trigger- and/or maturity-dependent changes in the abundance of hormone receptors (FSHR and LHCGR) and proteins associated with lipid metabolism and steroidogenesis (e.g., STAR, HSD3B1, and CYP11A1), prostaglandin synthesis (PTGS2 and PTGFS), extracellular matrix remodeling (MMP1 and TIMP1), protein folding (HSPs), molecular transport (TF), and cell function and survival (e.g., VIM) were observed. These data revealed different endocrine properties of exogenous and endogenous gonadotropins, with a potent progestational/androgenic role of hCG and estrogenic/pro-developmental function of LH.

Clinical significance of serum gonadotropin and androgen levels among Egyptian overweight/obese pubertal girls

Objectives Evaluate the association between overweight/obesity with serum gonadotropin and androgen levels in Egyptian pubertal girls. Subjects and methods A case-control study carried out in “Obesity Clinic” of “Diabetes, Endocrine and Metabolism Pediatric Unit (DEMPU)”, Pediatric Hospital, Cairo University. It included 40 overweight and obese girls and 40 age-matching normal weight (control) ones, aged 12–18 years. Anthropometric assessment (weight, height and hip and waist circumferences) was done, and waist/hip and BMI were calculated. Laboratory investigations: lipid profile, serum gonadotropin (LH, FSH), androgen (free and total testosterone), estradiol, insulin, and FBG were quantified, while insulin resistance (IR) was calculated. Results Hypogonadotropins (FSH and LH) and hyperandrogenaemia (total and free testosterone) were significantly prominent among obese girls. Correlation between gonadotropin, androgen and all of the studied variables, for the three studied groups (obese, overweight and control) revealed constant relations. Gonadotropin and androgens showed opposing correlations. Gonadotropin had significant negativ e correlations with the anthropometric parameters of obesity (BMI, Waist C, and W/H ratio), insulin, insulin resistance and lipid profile (triglycerides, total cholesterol and LDL), whereas androgens had significant positiv e ones. In addition, gonadotropin showed significant positiv e correlations with estradiol and HDL, while androgens showed significant negative ones. Conclusions Overweight/obesity had no effect on the correlations between gonadotropin and androgen on one side, with the anthropometric measurements and laboratory investigations on the other one. Alterations in androgen levels occur at earlier ages than gonadotropin, among both overweight and obese girls.

Recovery of Male Reproductive Endocrine Function Following Prolonged Injectable Testosterone Undecanoate Treatment

Background: Exogenous androgen treatment suppresses the hypothalamo-pituitary testicular (HPT) axis causing reduced serum LH, FSH and testosterone (T). Recovery of male reproductive endocrine function in past androgen abusers takes 9-18 months with persistent mild lowering of serum T. The natural history of recovery of HPT axis following prolonged injectable testosterone undecanoate (TU) treatment at standard dose is not known. Therefore, the Runoff Study investigated the rate and extent of reproductive hormone recovery over 12 months following cessation of 2 years of TU treatment in the Testosterone for Diabetes Mellitus (T4DM) Study, while men remain blinded to treatment allocation. Methods: T4DM participants without pathological hypogonadism (n=1007) were randomised to TU or Placebo (P) injections every 3 months for 2 years with 303 subsequently volunteering to enter the Runoff study at 12 weeks after last injection. Before T4DM study unblinding, they provided blood samples and validated sexual function questionnaires (PDQ, IIEF-15) at entry (3 months after last injection), 6, 12, 18, 24, 40 and 52 weeks later. Serum steroid profile (T, DHT, E2, E1) was measured batchwise by LCMS and serum LH, FSH and SHBG by immunoassays. Results: Runoff study participants in both groups were similar and did not differ from all T4DM participants. As expected, at entry to Runoff serum T was higher in TU-treated men but at all timepoints from 12 weeks onwards serum T and SHBG remained consistently 11% and 13%, respectively, lower in TU-treated than in P-treated men. Similarly, at entry sexual function scores were higher in TU-treated men but subsequently no different from P-treated men. Serum LH and FSH recovered slowly with the median time to reach their own pre-treatment baseline of serum LH was 51.1 weeks [95% CI 50.4 – 53.0 weeks] and for serum FSH was 52.7 weeks [51.0 – 60.9 weeks]. Conclusion: After stopping 2 years of standard dose injectable TU treatment in men without pathological hypogonadism, recovery of testicular endocrine function is eventually complete but slow with serum gonadotropin recovery taking on 12 months since the last dose. Persistent mild, proportionate reduction in serum SHBG and T reflects lasting exogenous T effects on hepatic SHBG secretion rather than signifying androgen deficiency. This suggests that recovery from androgen-induced HPT axis suppression depends primarily on time since cessation rather than dose or duration of androgen exposure.

C57BL/6J mouse superovulation: schedule and age optimization to increase oocyte yield and reduce animal use

Superovulation protocols have been described for different mouse strains, however the numbers of animals used are still high and still little information is known about hormone administration schedules and estrous cycle phases. In this study, we aimed to optimize a superovulation protocol by injecting 5 IU of pregnant mare serum gonadotropin followed by 5 IU of hCG 48 h later, using three different schedules related to the beginning of the dark cycle (3, 5 and 7 pm) in a light cycle of 7 am to 7 pm, with light on at 7 am. C57BL/6J mice at 3, 4 and 5 weeks of age were used and the estrous cycle phase for times of PMSG and hCG injections was also analyzed. Total oocyte number was counted in the morning after hCG injection. Hormones given at 3 weeks of age at 3 pm (59 ± 15 oocytes) and 7 pm (61 ± 10 oocytes) produced a significantly higher oocyte number compared with oocytes numbers collected from females at the same age at 5 pm (P = 0.0004 and <0.0001 respectively). Females at 4 and 5 weeks of age produced higher numbers of oocytes when superovulated at 7 pm. No statistical differences between females at different phases of the estrous cycle were found. These results showed that in C57BL/6J mice, hormones should be given at 3 or 7 pm for females at 3 weeks of age, however older females should be superovulated closer to the beginning of the dark cycle to reduce female mouse use and increase the numbers of oocytes produced per female.

Histochemical Study of the Rat Uterine Glycoconjugate Alteration following Treatment with Exogenous Gonadotropic Hormones during the Implantation Period

One of the female causes of infertility is anovulation which is treatable with gonadotropin hormones. These hormones affect the molecular organization of the uterus such as glycoconjugates that are the first site of contact between the blastocyst and the uterus. The objective of this project was to study the alteration of glycoconjugates on the uterine apical, Golgi zone, and basement membrane of epithelial cells and the uterine gland after hyperstimulation with pregnant mare serum gonadotropin (PMSG) (4, 8, 16, 24, and 40 IU), during the implantation period. Injection of PMSG (in experimental groups) and injection of distilled water (in the control group) were followed by HCG administration (10 IU), mating, isolation of positive vaginal plug rats, and killing at 5.5 days of pregnancy. Histochemistry was done on the pregnant uterine horns with the use of WGA, DBA, PNA, ConA, SBA, and UEA lectins. The intensity of the immunohistochemical staining was scored, and quantitative data were generated. 4 IU did not show any significant differences with the control, 8 IU had less effect on the alteration of the Golgi zone, and apical and basement membrane glycoconjugates and 40 IU had the least effects on the alteration of uterine gland glycoconjugates. Also, 24 IU had the most effect on the alteration of uterine glycoconjugates. Understanding of the effects of gonadotropin hormones at the uterine level in implantation time helps to optimize hormonal manipulation for improving the outcome of assisted reproductive procedures. It seems that the optimal dose for superovulation and less alteration in uterine glycoconjugates of rats at implantation time were induced by the administration of 8 IU PMSG.