Administration Of Follicle-stimulating Hormone Research Articles

Enhancing effect of FSH on follicular development through yolk formation and deposition in the low-yield laying chickens

Healthy and efficient development of ovarian follicles largely determines poultry laying performance. In low-yield laying chickens, retarded follicle progression resulted in decreased prehierarchical follicles. In this study the extenuating effect of follicle-stimulating hormone (FSH) on delayed follicular development was investigated in the low-yield chickens. Results showed that FSH administration in vivo accelerated development of prehierarchical follicles, with increased expression of steroidogenic enzymes and follicular angiogenesis through elevating plasma levels of 17β-estradiol, progesterone, luteinizing hormone and the expression of vascular endothelial growth factor and its receptor as well as angiopoietins. Furthermore, treatment with FSH raised expression of lipid uptake and adipogenesis-related proteins and decreased tight junctions between granulosa cells. Meanwhile, the results of the in vivo studies were confirmed by the in vitro studies as FSH promoted development of the cultured prehierarchical follicles with increased angiogenesis, cell proliferation, steroid hormones synthesis and yolk deposition. These results indicated FSH enhanced follicular development in the low-yield laying chickens involving increased follicular angiogenesis.

Follicle-Stimulating Hormone Treatment and Male Idiopathic Infertility: Effects on Sperm Parameters and Oxidative Stress Indices according to FSHR c. 2039 A/G and c. -29 G/A Genotypes.

Scientific evidence shows that the administration of follicle-stimulating hormone (FSH) to infertile patients with normal serum FSH concentrations improves sperm parameters in oligozoospermic men. The aim of this study was to evaluate the effects of highly purified urofollitropin (hpFSH) on conventional and bio-functional sperm parameters and on oxidative stress indices in patients with idiopathic infertility. We also evaluated the response to hpFSH on these parameters in relationship to FSHR c. 2039 A/G and FSHR c. -29 G/A genotypes. A prospective longitudinal study was conducted on 42 patients with idiopathic male infertility, 23 of whom underwent to FSHR c. 2039 A/G and FSHR c. -29 G/A genotyping. Each patient was asked to collect two semen samples before and after administration of 150 IU hpFSH three times a week for 16 weeks. Patients were divided into responders or non-responders based on whether their total sperm count had at least doubled or was less than double at the end of treatment, respectively. Responders showed a significantly higher semen volume, sperm concentration, spermatids, and leukocytes. Non-responders had a significant decrease of the percentage of spermatozoa in early apoptosis after hpFSH administration. Oxidative stress indexes did not differ significantly after FSH administration in both groups. Conventional and bio-functional sperm parameters did not differ in patients with FSHR c. 2039 GG and AA genotypes, and FSHR c. -29 GG genotype both before and after FSH administration. The FSHR c. 2039 and FSHR -29 G/A genotypes and allelic distribution did not differ between responders and non-responders. FSH showed to be capable of ameliorating sperm parameters in about half patients treated, therefore it may be helpful in patients with idiopathic infertility.

Prospects for FSH Treatment of Male Infertility.

Despite the new opportunities provided by assisted reproductive technology (ART), male infertility treatment is far from being optimized. One possibility, based on pathophysiological evidence, is to stimulate spermatogenesis with gonadotropins. We conducted a comprehensive systematic PubMed literature review, up to January 2020, of studies evaluating the genetic basis of follicle-stimulating hormone (FSH) action, the role of FSH in spermatogenesis, and the effects of its administration in male infertility. Manuscripts evaluating the role of genetic polymorphisms and FSH administration in women undergoing ART were considered whenever relevant. FSH treatment has been successfully used in hypogonadotropic hypogonadism, but with questionable results in idiopathic male infertility. A limitation of this approach is that treatment plans for male infertility have been borrowed from hypogonadism, without daring to overstimulate, as is done in women undergoing ART. FSH effectiveness depends not only on its serum levels, but also on individual genetic variants able to determine hormonal levels, activity, and receptor response. Single-nucleotide polymorphisms in the follicle-stimulating hormone subunit beta (FSHB) and follicle-stimulating hormone receptor (FSHR) genes have been described, with some of them affecting testicular volume and sperm output. The FSHR p.N680S and the FSHB -211G>T variants could be genetic markers to predict FSH response. FSH may be helpful to increase sperm production in infertile men, even if the evidence to recommend the use of FSH in this setting is weak. Placebo-controlled clinical trials, considering the FSHB-FSHR haplotype, are needed to define the most effective dosage, the best treatment length, and the criteria to select candidate responder patients.

Follicle-stimulating Hormone (FSH) Action on Spermatogenesis: A Focus on Physiological and Therapeutic Roles.

Background: Human reproduction is regulated by the combined action of the follicle-stimulating hormone (FSH) and the luteinizing hormone (LH) on the gonads. Although FSH is largely used in female reproduction, in particular in women attending assisted reproductive techniques to stimulate multi-follicular growth, its efficacy in men with idiopathic infertility is not clearly demonstrated. Indeed, whether FSH administration improves fertility in patients with hypogonadotropic hypogonadism, the therapeutic benefit in men presenting alterations in sperm production despite normal FSH serum levels is still unclear. In the present review, we evaluate the potential pharmacological benefits of FSH administration in clinical practice. Methods: This is a narrative review, describing the FSH physiological role in spermatogenesis and its potential therapeutic action in men. Results: The FSH role on male fertility is reviewed starting from the physiological control of spermatogenesis, throughout its mechanism of action in Sertoli cells, the genetic regulation of its action on spermatogenesis, until the therapeutic options available to improve sperm production. Conclusion: FSH administration in infertile men has potential benefits, although its action should be considered by evaluating its synergic action with testosterone, and well-controlled, powerful trials are required. Prospective studies and new compounds could be developed in the near future.

Use of follicle-stimulating hormone for the male partner of idiopathic infertile couples in Italy: Results from a multicentre, observational, clinical practice survey.

The management of male idiopathic infertility is heterogeneous. Although meta-analyses reported the effectiveness on pregnancy rate, the real clinical impact of follicle-stimulating hormone (FSH) was not evaluated so far. In Italy, FSH is approved by the National Medicines Agency (AIFA) for idiopathic infertile patients with FSH<8IU/L, independently of semen parameters. Primary endpoint was to record the therapeutic approach to the male partner of infertile couples. Secondary aim was to assess changes of semen parameters during FSH treatment. A multicentre, prospective, observational, clinical practice survey was carried out, enrolling the male partner of infertile couples attending ten Italian participating centres. Inclusion criteria were as follows: couple infertility, age >18years and FSH serum levels <8IU/L. Thus, all men in which AIFA allowed the FSH prescription were enrolled. Primary endpoint was the number of infertile patients treated with FSH. Secondary outcomes were semen parameters. The treating physician decided whether to offer FSH therapy and whether to re-evaluate the male partner. A total of 718 infertile couples were enrolled, and 241 patients were re-evaluated (median follow-up: 4.5months). In 64.9% (466 patients), a treatment was prescribed. FSH was prescribed in 397 patients (85.2% of treated men). Sperm concentration (P=.002) and normal form percentage (P<.001) significantly improved during FSH administration. No correlation was found between these parameters and FSH duration (P=.545 and P=.627, respectively) or dosage (P=.455 and P=.533, respectively). Among patients treated with FSH, the incidence of oligozoospermia decreased from 73.0% to 56.0% (P<.001) and teratozoospermia from 43.6% to 27.7% (P<.001). This first nation-wide survey reveals a FSH prescription rate of 55% in patients qualifying for treatment according to AIFA. Although the study was not designed to highlight FSH efficacy in male infertility, a slight increase in semen parameters was demonstrated in about half of the treated men without adverse events.

The Effect of Follicle-Stimulating Hormone Administration on the Day of Human Chorionic Gonadotropin Trigger on Assisted Reproductive Technique Outcomes in Patients Undergoing In vitro Fertilization-Embryo Transfer: A Retrospective Cohort Study.

Aim:The aim is to study the effect of follicle-stimulating hormone (FSH) administration on the day of human chorionic gonadotropin (hCG) trigger on the assisted reproductive technique (ART) outcomes in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles.Settings and Design:Retrospective cohort study was conducted in the ART center of our hospital.Materials and Methods:Two hundred and ninety IVF/ICSI cycles performed between September 2012 and August 2017 were included in the study. Patients who received 375 IU of FSH on the day of hCG trigger (149 cycles) were compared with those who did not receive FSH on the day of trigger (141 cycles).Statistical Analysis Used:Chi-square test and Student's t-test were used.Results:The FSH co-administered group had a significantly higher number of oocytes retrieved, mature oocytes, and fertilization rate compared to those who did not receive FSH on the day of trigger (p < 0.001). The total number of embryos, the number of grade 1 embryos and the number of embryos available for cryopreservation were also significantly higher in the FSH administered group (p < 0.001). Implantation rate, clinical pregnancy rate, and live birth rate were not significantly different between the two groups.Conclusions:This study has shown that FSH administration on the day of the trigger may be considered in IVF cycles receiving hCG trigger to improve the oocyte recovery and maturity if the patient is not at increased risk of ovarian hyperstimulation and serum estradiol on the day of the trigger is <4500 pg/ml. However, there is only an increase in the total number of oocytes retrieved and the number of mature oocytes but no significant change in the implantation, clinical pregnancy, and live birth rates.

Consequences of twinning induction to Noemi ewes by a recombinant human follicle-stimulating hormone compared with pituitary-derived porcine follicle-stimulating hormone on follicular dynamics, maternal biochemical attributes, and neonatal traits.

Aim:The aim of this study was to investigate the effectiveness of using recombinant human follicle-stimulating hormone (FSH) compared with pituitary-derived porcine FSH given as one dose or multiple doses on the neonatal traits, follicular dynamics, and maternal blood biochemical constituents in Noemi ewes.Materials and Methods:A 3×2 factorial arrangement was designed utilizing 60 adults Noemi ewes to test the effects of using two sources of FSH (human vs. porcine) in addition to control, either given as a single total dose or six descending doses to provoke twinning. Six treatments (T) were tested (n=10 ewes/T). C1 and C6 served as control ewes given saline as one dose and six doses, respectively; H1 and H6 ewes were given human FSH as one and six doses; and P1 and P6 ewes were given porcine FSH similar to the above treatments. Saline and/or FSH administration were administered at days 8, 9, and 10 of the 10-day controlled internal drug release (CIDR) implant. At CIDR removal, fertile rams were used for natural mating. Blood samples for the assessment of serum metabolites were collected.Results:Twinning increased in FSH-treated ewes than control. However, giving FSH of either source as a single dose resulted in a higher incidence of stillbirths. Pregnancy rates were 30, 40, 50, 60, 70, and 80% in C1, C6, P1, P6, H1, and H6, respectively. Respective percent of ewes delivering twins/multiple birth was 0, 0, 80, 66.7, 71.4, and 87.5%. FSH of human source was more efficient for folliculogenesis than porcine FSH. Administration of FSH increased blood cholesterol, decreased high-density lipoprotein; however, low-density lipoprotein levels were not different than control. Moreover, an interaction (p<0.05) exists between source and type of FSH administration on blood glucose. Six doses of FSH elevated blood protein. Blood albumin decreased by porcine-FSH but not affected by human-FSH. Blood globulins were not different due to source of FSH, whereas giving FSH as six doses increased globulins than in single-dose protocol. Contrariwise, an interaction was found between source and type of FSH administration on elevating the activity of alanine aminotransferase and reducing the activity of aspartate aminotransferase.Conclusion:Administration of human FSH at 180 IU in six descending doses resulted in the best neonatal outcomes and maternal health in Noemi ewes.

Porcine embryo collection using single subcutaneous administration of follicle-stimulating hormone in a large volume of saline.

The effects of a single subcutaneous or intramuscular injection of follicle-stimulating hormone (FSH) on follicular growth and expression of estrous behavior and its single subcutaneous administration on the number of corpora lutea (CL) and embryos were investigated in pigs. All four sows that were subcutaneously administered 5 AU FSH expressed normal estrus and had no ovarian cysts. Two of the four sows that were administered 5 AU FSH intramuscularly did not exhibit estrus, and another sow had a short estrus period. All four sows had ovarian cysts. The mean numbers of CL, embryos, and blastocysts following the subcutaneous administration of 5 AU FSH (16.8, 16.0, and 13.8, respectively) did not differ significantly from those for the control animals treated intramuscularly with 1000 IU equine chorionic gonadotropin (18.5, 16.5, and 14.3, respectively). In conclusion, embryo recovery was possible using a single subcutaneous administration of FSH.

Effect of single dose follicle stimulating hormone on follicular aspiration, in vitro fertilization and pregnancy rate

The present study evaluated Brangus cows treated with single doses of follicle stimulating hormone (FSH) subjected to follicular aspiration after 24 h to assess oocyte recovery, in vitro fertilization and pregnancy rate. Follicles exceeding 3 millimeters in diameter were aspirated, 200 mg of FSH was administered 2 days later, and a new ovum pickup was performed 24 h afterward. These methods were performed 3 times every 3 days. In control, follicular aspirations occurred at intervals of 1-week without FSH administration o. The aspirated oocytes were evaluated, submitted to in vitro fertilization and the embryos were transferred to the recipients. The average recovery of oocytes was higher (p&lt;0.05) in control cows (12.4±1.8) than in treated cows (9.4±1.3). There was no difference (p&gt;0.05) in the mean percentage of viable oocytes (52.0±3.9 and 62.7±4.7%) or the mean percentage of embryos (41.4±4.8 and 41.5±4.2%) among control and treated cows, respectively. The mean percentage of pregnancy did not differ (p&gt;0.05) for control cows (43.8±2.7%), and treated cows (40.9±6.8%). In conclusion, FSH treatment did not improve oocyte recovery, in vitro fertilization, and pregnancy percentage. However, there is possibility of several consecutive ovum pickup every t3 days, concentrating the in vitro fertilization and the pregnancy percentage.

The effect of follicle stimulating hormone administration at the time of human gonadotropin triggering, is it improve the oocyte/embryo profile in in vitro fertilization cycles?

The effect of follicle stimulating hormone administration at the time of human gonadotropin triggering, is it improve the oocyte/embryo profile in in vitro fertilization cycles?

FSH and estradiol benzoate administration recover spermatogenesis and sexual hormone levels in a busulfan-injured rat model

Busulfan induces azoospermia and testicular atrophy, while follicle stimulating hormone (FSH) and estradiol benzoate (EB) may improve spermatogenesis. Our objective was to determine the effects of FSH and EB on the recovery of spermatogenesis, histology, sexual hormones, and testicular gene expression in the testis of a busulfan-injured rat model. Forty adult rats were assigned into eight groups. Half received a single intraperitoneal saline or 30 mg/kg busulfan injection, and 42 days after, the animals received intramuscular injections for 10 days: 1 ml saline, 7.5 IU FSH, and 12 μg/kg EB or FSH+EB. One day after, blood plasma was obtained for hormone analysis, sperm was recovered from epididymis, and testes were processed for histology and RNA extraction. Busulfan decreased sperm count, motility, and viability; induced seminiferous tubular atrophy; and decreased blood testosterone, FSH, and luteinizing hormone. Hormonal treatments partially recovered spermatogenesis, decreasing tubular atrophy. They also decreased caspase 3 and iNOS expression while increasing Bcl-2 expression, compared with the busulfan treatment. FSH+EB was the most efficient treatment, also increasing the concentration of sexual hormones close to no-busulfan levels. Thus, busulfan has a clear negative effect in spermatogenesis in the rat model, and FSH and EB, especially combined, could help to reverse these effects.

Follicle-stimulating hormone administration affects amino acid metabolism in mammalian oocytes†.

Culture media used in assisted reproduction are commonly supplemented with gonadotropin hormones to support the nuclear and cytoplasmic maturation of in vitro matured oocytes. However, the effect of gonadotropins on protein synthesis in oocytes is yet to be fully understood. As published data have previously documented a positive in vitro effect of follicle-stimulating hormone (FSH) on cytoplasmic maturation, we exposed mouse denuded oocytes to FSH in order to evaluate the changes in global protein synthesis. We found that dose-dependent administration of FSH resulted in a decrease of methionine incorporation into de novo synthesized proteins in denuded mouse oocytes and oocytes cultured in cumulus-oocyte complexes. Similarly, FSH influenced methionine incorporation in additional mammalian species including human. Furthermore, we showed the expression of FSH-receptor protein in oocytes. We found that major translational regulators were not affected by FSH treatment; however, the amino acid uptake became impaired. We propose that the effect of FSH treatment on amino acid uptake is influenced by FSH receptor with the effect on oocyte metabolism and physiology.

FSH dosage effect on conventional sperm parameters: a meta-analysis of randomized controlled studies

Follicle-stimulating hormone (FSH) represents a therapeutic option in normogonadotropic patients with idiopathic oligozoospermia. The aim of this review was to evaluate the possible dose- and drug-dependent efficacy of FSH treatment on conventional sperm parameters. We performed a comprehensive systematic review via a meta-analysis of all available randomized controlled trials, in which FSH administration was compared with placebo or no treatment when administered to normogonadotropic patients with idiopathic oligozoospermia. Of the 971 articles that were retrieved, 5 were finally included, including a total of 372 patients and 294 controls. Overall, FSH treatment was effective in ameliorating the sperm concentration, total count, progressive motility, but not normal forms. On the basis of the weekly dosage, the studies were classified into those using low (175–262.5 IU per week), intermediate (350–525 IU per week), and high (700–1050 IU per week) doses. At low doses, FSH improved only sperm motility. At intermediate doses, FSH ameliorated sperm concentration and morphology. Total sperm count and progressive motility showed a trend toward the increase. At high doses, FSH increased sperm concentration, total sperm count, and progressive motility. Sperm morphology showed a trend toward the increase. Finally, both highly purified FSH (hpFSH) and recombinant human FSH (rhFSH) improved sperm concentration, total sperm count, progressive motility, but not morphology. No different efficacy was observed between these two preparations. This meta-analysis provides evidence in favor of high FSH doses. The FSH efficacy was not related to the preparation type (recombinant vs highly purified). Further studies are needed to evaluate the effectiveness of long-standing treatment regimes.

Effects of Luteinizing Hormone Supplementation on Ovarian Response and Assisted Reproductive Technology Outcomes in Antagonist In vitro Fertilization/Intracytoplasmic Sperm Injection Cycles: A Meta-analysis

Objective: The study objective was to investigate the effects of luteinizing hormone (LH) supplementation on ovarian response and assisted reproductive technology (ART) outcomes in in vitro fertilization/intracytoplasmic sperm injection cycles with a gonadotropin (Gn)-releasing hormone antagonist protocol. Methods: This is a meta-analysis, and nine published randomized controlled trials (1,685 patients) were included. Continuous data were extracted in the form of mean ± standard deviation and population size, whereas dichotomous data were extracted in the form of odds ratio. Results: The total amount of follicle-stimulating hormone (FSH) used, the duration of stimulation (DOS), the number of eggs in MII stage, the total number of formed embryos, the clinical pregnancy rate, and live birth rates were similar between groups, but the estrogen level on the day of human chorionic Gn (hCG) administration was slightly higher in the LH supplementation group. On subgroup analysis, it was reported that the addition of LH could significantly increase estrogen levels on the day of hCG administration in patients older than 35 years, and LH supplementation starting on the day of FSH administration may slightly extend the DOS. Moreover, regardless of the timing of LH supplementation, an increase in estrogen levels was found on the day of hCG administration. Conclusions: LH supplementation of an antagonist protocol increases estrogen levels on the day of hCG administration, but does not increase the number of mature oocytes retrieved, and also fails to improve ART outcomes.

Effects of FSH on Sperm DNA Fragmentation: Review of Clinical Studies and Possible Mechanisms of Action.

Sperm DNA fragmentation (sDF) is an important reproductive problem, associated to an increased time-to-pregnancy and a reduced success rate in natural and in vitro fertilization. sDF may virtually originate at any time of sperm's life: in the testis, in the epididymis, during transit in the ejaculatory ducts and even following ejaculation. Studies demonstrate that an apoptotic pathway, mainly occurring in the testis, and oxidative stress, likely acting in the male genital tract, are responsible for provoking the DNA strand breaks present in ejaculated spermatozoa. Although several pharmacological anti-oxidants tools have been used to reduce sDF, the efficacy of this type of therapies is questioned. Clearly, anti-apoptotic agents cannot be used because of the ubiquitous role of the apoptotic process in the body. A notable exception is represented by Follicle-stimulating hormone (FSH), which regulates testis development and function and has been demonstrated to exert anti-apoptotic actions on germ cells. Here, we review the existing clinical studies evaluating the effect of FSH administration on sDF and discuss the possible mechanisms through which the hormone may reduce sDF levels in infertile subjects. Although there is evidence for a beneficial effect of the hormone on sDF, further studies with clear and univocal patient inclusion criteria, including sDF cut-off levels and considering the use of a pharmacogenetic approach for patients selection are warranted to draw firm conclusions.

Recombinant FSH Improves Sperm DNA Damage in Male Infertility: A Phase II Clinical Trial.

Background and objectives: Male infertility is a global health dilemma and Follicle-Stimulating Hormone (FSH) administration has shown promising results. Several studies showed that infertile men with normal semen parameters have low levels of DNA damage while infertile men with abnormal semen parameters have more damage at the DNA level. Sperm DNA damage may affect the reproductive outcome and has been associated with failure in the achievement of competent embryos and pregnancy fulfillment. The aim of this study was to evaluate whether the administration of recombinant FSH (Gonal-f® PEN 900 IU) could improve sperm DNA fragmentation in men with infertility. The secondary endpoints of this study were to evaluate the FSH effects on sperm parameters and hormonal assets.Methods: A longitudinal, prospective, multicenter, open-label clinical trial was carried out. Infertile couples were recruited from six Italian Reproductive Medical Centers and 115 infertile men were enrolled for this study. All participants were treated with subcutaneous injections of Gonal-f® 150 IU every other day, within a 3 month-time frame. The semen samples were examined in accordance to the 2010 World Health Organization criteria. Sperm DNA Fragmentation (DFI) was determined by fluorescence microscopy using terminal deoxynucleotidyl transferase-mediated d-UTP Nick-end Labeling (TUNEL) assay. Statistical analysis was performed using both the t-test for paired samples and the Wilcoxon signed-rank test.Results: FSH administration improved DFI in 67% of patients, with an average decrease of 35.4% compared to the baseline. This improvement is more evident in men with basal DFI lower than 17% and in those with FSH basal levels between 2.16 and 4.27 IU/L. In addition, FSH enhanced the gonadal function, increasing the hormones AMH and Inhibin B and semen parameters. Limitation of these results are represented by the absence of a placebo group and of FSHR genotype stratification sub-analysis.Conclusion: Recombinant FSH 150 IU is well tolerated and effective in eliciting a significant DFI reduction as well as in improving gonadal function.Trial Registration: EUDRACT Number 2010-020196-23. Registred 14 April 2011.

Response: Commentary: Efficacy of Follicle-Stimulating Hormone (FSH) Alone, FSH + Luteinizing Hormone, Human Menopausal Gonadotropin or FSH + Human Chorionic Gonadotropin on Assisted Reproductive Technology Outcomes in the "Personalized" Medicine Era: A Meta-analysis.

Setting: Luteinizing hormone(LH) and human chorionic gonadotropin (hCG) act on the same receptor, activating different signal transduction pathways. The role of LH or hCG addition to follicle stimulating hormone (FSH) as well as menopausal gonadotropins (hMG) in controlled ovarian stimulation (COS) is debated. Objective: To compare FSH+LH, or FSH+hCG or hMG vs FSH alone on COS outcomes. Design: A meta-analysis according to PRISMA statement and Cochrane Collaboration was performed, including prospective, controlled clinical trials published until July 2016, enrolling women treated with FSH alone or combined with other gonadotropins. Trials enrolling women with polycystic ovarian syndrome were excluded (PROSPERO registration n. CRD42016048404). Results: Considering 70 studies, the administration of FSH alone resulted in higher number of oocytes retrieved than FSH+LH or hMG. The MII oocytes number did not change when FSH alone was compared to FSH+LH, FSH+hCG or hMG. Embryo number and implantation rate were higher when hMG was used instead of FSH alone. Pregnancy rate was significantly higher in FSH+LH-treated group versus others. Only twelve studies reported live birth rate, not providing protocol-dependent differences. Patients’ stratification by GnRH agonist/antagonist identified patient subgroups benefiting from specific drug combinations. Conclusion: In COS, FSH alone results in higher oocyte number. HMG improves the collection of mature oocytes, embryos and increases implantation rate. On the other hand, LH addition leads to higher pregnancy rate. This study supports the concept of a different clinical action of gonadotropins in COS, reflecting previous in vitro data.

Effect of pre-in vitro maturation with cAMP modulators on the acquisition of oocyte developmental competence in cattle.

The administration of follicle-stimulating hormone (FSH) prior to oocyte retrieval improves oocyte developmental competence. During bovine embryo production in vitro, however, oocytes are typically derived from FSH-unprimed animals. In the current study, we examined the effect of pre-in vitro maturation (IVM) with cAMP modulators, also known as the second messengers of FSH, on the developmental competence of oocytes derived from small antral follicles (2–4 mm) of FSH-unprimed animals. Pre-IVM with N6,2ʹ-O-dibutyryladenosine 3′,5′-cyclicmonophosphate (dbcAMP) and 3-isobutyl-1-methylxanthine (IBMX) for 2 h improved the blastocyst formation in oocytes stimulated by FSH or amphiregulin (AREG). Furthermore, pre-IVM enhanced the expression of the FSH- or AREG-stimulated extracellular matrix-related genes HAS2, TNFAIP6, and PTGS2, and epidermal growth factor (EGF)-like peptide-related genes AREG and EREG. Additionally, pre-IVM with dbcAMP and IBMX enhanced the expression of EGFR, and also increased and prolonged cumulus cell-oocyte gap junctional communication. The improved oocyte development observed using the pre-IVM protocol was ablated by an EGF receptor phosphorylation inhibitor. These results indicate that pre-IVM with cAMP modulators could contribute to the acquisition of developmental competence by bovine oocytes from small antral follicles through the modulation of EGF receptor signaling and oocyte-cumulus/cumulus-cumulus gap junctional communication.

Effect of a single epidural administration of follicle-stimulating hormone via caudal vertebrae on superstimulation for in vivo and in vitro embryo production in Japanese black cows.

Here, we describe a simplified procedure for embryo production in the Japanese black cow that uses a single caudal epidural injection of follicle-stimulating hormone (FSH). First, we compared the efficiency of superovulation for in vivo embryo production between conventional multiple FSH treatment (control, n = 10) and single epidural administration (epidural, n = 5). The number of transferable blastocysts was similar between control and epidural groups (4.7 ± 3.5 and 9.0 ± 6.0, respectively). Next, we compared in vitro embryo production by ovum pick-up and in vitro fertilization (OPU-IVF) between control (n = 12) and epidural groups (n = 12). The rate of development to transferable blastocysts was higher in the epidural group than in the control (23.3 vs. 10.5%, P < 0.001). In conclusion, a single epidural administration of FSH can induce follicular development comparable to that of the conventional superovulation protocol and may improve the productivity of OPU-IVF.

193 Comparison of Superovulatory Responses to Single Subcutaneous FSH Injection and Twice-Daily Intramuscular FSH Injection Protocols in Japanese Black Cows

The conventional superovulation treatment using follicle-stimulating hormone (FSH) comprises 2 intramuscular injections per day for 3 to 4 days. This conventional method requires frequent handling of donors—which may cause increasing stress in donors—and high labour costs. Therefore, to alleviate these issues, a treatment protocol using a single FSH injection under on-farm conditions is needed. The objective of this study was to evaluate the feasibility of the single subcutaneous FSH injection protocol (Sugawara et al. 2017 Reprod. Fertil. Dev. 29, 209), compared with with the traditional twice-daily intramuscular FSH injection protocol, in Japanese Black cows. Japanese Black cows (n = 13) were superovulated using 2 treatments in a crossover design. All cows received an intravaginal progesterone device (CIDR) at random stages of the oestrous cycle (Day 0), and 2 mg of oestradiol benzoate on Day 1 (24 h after CIDR insertion). On Day 6, for the group receiving a single subcutaneous FSH injection (single-FSH), 20 armor units (AU) of pFSH dissolved in 30 mL of saline was subcutaneously injected in the neck region and 0.5 mg of prostaglandin F2α was intramuscularly injected simultaneously. For the control group, FSH administration was initiated at Day 6. A total dose of 20 AU of FSH was administered in 3 days (with decreasing doses of 5, 3, and 2 AU, respectively, per injection) with 2 intramuscular injections per day; 0.5 mg of prostaglandin F2α was injected 48 h after the first FSH injection. For both groups, CIDR was removed 60 h after FSH injection and AI was done 42 to 48 h after CIDR removal. Embryo collections were performed 7 days after AI. Data were analysed using t-test for the mean numbers of collected ova/embryos and transferable embryos, and chi-squared test for the proportion of transferable embryos. There were no significant differences in the mean numbers (single-FSH v. control) of ova/embryos collected (17.1 ± 12.1 v. 17.8 ± 9.3), or transferable embryos (8.4 ± 9.3 v. 10.3 ± 8.7), and proportion of transferable embryos (45.0 ± 34.0 v. 56.6 ± 36.5). These results showed that although a slightly higher number and proportion of transferable embryos were obtained in the control group, a superovulation treatment protocol involving a single subcutaneous FSH injection can be effectively used for Japanese Black cows, under on-farm conditions.