Injection Of Follicle-stimulating Hormone Research Articles

Cumulative outcome of pre-implantation genetic diagnosis for sickle cell disease: a 5-year review.

To review the cumulative outcome of pre-implantation genetic diagnosis (PGD) cycles performed for prevention of sickle cell disease (SCD). Couples referred for PGD for SCD between April 2012 and October 2017 were included. Ovarian stimulation was performed using a short gonadotrophin-releasing hormone (GnRH) antagonist protocol and follicle-stimulating hormone injections. The GnRH agonist was used to trigger oocyte maturation. Oocytes were fertilised using intracytoplasmic sperm injection. Trophectoderm biopsy was performed on day 5 or 6 followed by vitrification. Genetic testing was done using pre-implantation genetic haplotyping. A total of 60 couples started 70 fresh PGD cycles (mean 1·2cycles/couple) and underwent a total of 74 frozen-embryo-transfer (FET) cycles (mean 1·3FET/couple). The mean (SD) female age was 33(4·4)years and the mean (SD) anti-müllerian hormone level was 22·9(2·8)pmol/l. The cumulative live-birth rate was 54%/PGD cycle started and 63%/couple embarking on PGD. The rate of multiple births was 8%. The cumulative outcome of PGD treatment for prevention of SCD transmission is high and PGD treatment should be offered to all at-risk couples.

Real-time assessment of the superovulatory effect of FSH and eCG with laparoscopy at different seasons in Akkaraman ewes.

Conventional methods for determining the reproductive performance of sheep bred either after estrus synchronization during the breeding season or after induction of estrus/ovulation during the non-breeding season take a long time and may give misleading results due to the effect of environmental factors. Laparoscopic observations allow real-time monitoring of ovarian activity around estrus or ovulation. This study was aimed at assessing the superovulatory effects of follicle-stimulating hormone (FSH) and equine chorionic gonadotropin (eCG) treatments by laparoscopy during breeding (September-November, n=12) and non-breeding (April-June, n=12) seasons in Akkaraman sheep. In both seasons, after CIDR withdrawal, the ewes were injected either with 600 IU eCG or 300 μl (20 mg/ml) FSH twice at 12 hour intervals. Plasma P4, E2 and LH concentrations were determined at the time of intra-vaginal CIDR insertion (day 0) and then at its withdrawal (day 12), followed by 3 and 6 days of eCG or FSH injections. After 3 (first observation) and 6 (second observation) days of hormone injections, laparoscopy was performed to record ovarian activity in both seasons. The eCG increased (p⟨0.05) the numbers of large follicles (first observation) and CL (first and second observations) in the breeding season compared to FSH treatment. CL, small-moderate and large follicle numbers of eCG treated ewes were higher (p⟨0.05) than those of FSH at both observations in the non-breeding season. In the breeding season, eCG treated ewes had higher (p⟨0.05) plasma P4 (3 and 6 days after hormones injections) and E2 (3 days after hormones injections) concentrations than those of FSH. In conclusion, the results of the present study indicate that treatment with eCG during the non-breeding season can support ovarian activity, and thus increase ovulation rate and plasma hormone concentrations around induced estrus/ovulation in Akkaraman ewes.

The hypothalamic-pituitary-gonadal axis and thyroid hormone regulation interact to influence seasonal breeding in green anole lizards (Anolis carolinensis)

Reproductive physiology and behavior is mainly regulated by the hypothalamus-pituitary-gonad (HPG) axis, although abnormal thyroid hormone (TH) levels alter HPG axis activity. Seasonally breeding animals, such as green anole lizards (Anolis carolinensis), undergo drastic hormonal and behavioral changes between breeding and non-breeding seasons, with increased sex steroid hormones, larger gonads and increased reproductive behaviors during the breeding compared to non-breeding seasons. Relatively less is known regarding the regulation of gonadal TH in seasonal reproduction. We examined whether the gonadal expression of enzymes involved in TH activation are altered in concert with seasonal reproduction. Type 2 deiodinase (Dio2) mRNA, the TH activating enzyme, was upregulated in breeding compared to non-breeding testes, while type 3 deiodinase (Dio3) mRNA, the TH deactivating enzyme, was upregulated in breeding ovaries. To study the association between the HPG axis and local activation of TH, we manipulated the HPG axis during the non-breeding season by subcutaneously injecting luteinizing hormone (LH) and follicle stimulating hormone (FSH) in male lizards. We found that acute LH and FSH injections induced many aspects of breeding, with increased testes size and testosterone levels. Surprisingly, Dio3 was upregulated in the testes after LH and FSH injections, while Dio2 mRNA levels were unchanged. These results suggest that there might be different roles for local TH activation in developing and maintaining fully mature and functional gonads. Our findings continue to support the role for TH in regulating reproduction.

Follicle Stimulating Hormone and Gonadotropin Releasing Hormone Administration to the Superovulation of Buffalo (Bubalus bubalis)

This study aims to determine the response of superovulation, corpus luteum number, onset and duration of estrus in Swamp Buffalo after administration of Follicle Stimulating Hormone (FSH) and Gonadotropin Releasing Hormone (GnRH). The material used is 16 mother Swamp Buffaloes with an average body weight of 500 kg and age between 3-5 years. The dairy performance index such as the lactation period was 127.5±104.6 days, the milk production accounted for 2.0±0.9 kg/cow/day and the milk yield was 255±209 kg/lactation. The experimental design used was a Completely Randomized Design (CRD) consisting of four replications. Swamp buffaloes are synchronized with 5 mL of PGF2α injection. GnRH hormone injection on the 8th day at a dose of 500 µg with 2 times in the morning and evening. FSH injection begins on the 10th day with 2 times a day i.e., morning and night (12 h interval) with a dose decreased for 3 days intra-muscularly. Artificial insemination was carried out after signs appear estrus. The parameters observed include the superovulation response, the number of corpus luteum, the onset and duration of estrus in the Swamp Buffalo. The results obtained are the superovulation response in 56.25% Swamp Buffalo. The average number of corpus luteum, speed and duration of swamp buffalo estrus at different FSH doses and addition of 500 μg GnRH were 1.75, 20.25 and 145.75 h respectively. It was concluded that superovulation by administering a dose of 16 ml FSH + 500 μg GnRH has shown the best superovulation response and a long duration of estrus. The administration of 22 mL FSH dose and the addition of 500 μg GnRH showed the fastest onset of estrus.

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.

Generation of cynomolgus monkey fetuses with intracytoplasmic sperm injection based on the MII-stage oocytes acquired by personalized superovulation protocol.

BackgroundMature oocytes at the metaphase II status (MII-stage oocytes) played an important role in assisted reproductive technology in non-human primates.ObjectivesIn order to improve the proportion of MII-stage oocytes retrieval, three different superovulation protocols were performed on 24 female cynomolgus monkeys.MethodsAll the monkeys received once-daily injection of follicle-stimulating hormone (25 international unit [IU]) on day 3 of the menstruation, 3-day intervals, twice daily for 8–12 days until the time of human chorionic gonadotropin (1,500 IU) injection, on the 14–17th day of menstruation collecting oocytes. The difference between protocol I and protocol II was that 0.1 mg the gonadotropin-releasing hormone agonist was injected on day 1 of the menstruation, while the difference between personalized superovulation protocol and protocol II was that oocytes could be collected on the 14–17th day of menstrual cycle according to the length of each monkey.ResultsThe total number of oocytes harvested using the personalized superovulation protocol was much higher than that using protocol I (p < 0.05), and the proportion of MII-stage oocytes was significantly greater than that from either superovulation protocol I or II (p < 0.001 and p < 0.01 respectively), while the proportion of immature oocytes at the germinal vesicle was less than that from superovulation protocol I (p < 0.05).ConclusionsThe personalized superovulation protocol could increase the rate of MII-stage oocytes acquired, and successfully develop into embryos after intracytoplasmic sperm injection, and eventually generated fetus.

Ultrasound guidance versus the blind method for intrauterine catheter insemination: A randomized controlled trial.

ObjectiveThe primary objective of this study was to compare clinical pregnancy rates in intrauterine insemination (IUI) treatment cycles with transabdominal ultrasound guidance during intrauterine catheter insemination (US-IUI) versus the “blind method” IUI without ultrasound guidance (BM-IUI). The secondary objective was to compare whether US-IUI had better patient tolerability and whether US-IUI made the insemination procedure easier for the clinician to perform compared to BM-IUI.MethodsThis was a randomized controlled trial done at the Reproductive Medicine Unit of General Hospital Kuala Lumpur, Malaysia. We included women aged between 25 and 40 years who underwent an IUI treatment cycle with follicle-stimulating hormone injections for controlled ovarian stimulation.ResultsA total of 130 patients were recruited for our study. The US-IUI group had 70 patients and the BM-IUI group had 60 patients. The clinical pregnancy rate was 10% in both groups (p> 0.995) and there were no significant difference between the groups for patient tolerability assessed by scores on a pain visual analog scale (p= 0.175) or level of difficulty for the clinician (p> 0.995). The multivariate analysis further showed no significant increase in the clinical pregnancy rate (adjusted odds ratio, 1.07; 95% confidence interval, 0.85–1.34; p= 0.558) in the US-IUI group compared to the BM-IUI group even after adjusting for potential covariates.ConclusionThe conventional blind method for intrauterine catheter insemination is recommended for patients undergoing IUI treatment. The use of ultrasound during the insemination procedure increased the need for trained personnel to perform ultrasonography and increased the cost, but added no extra benefits for patients or clinicians.

New Insights Into the Role of Follicle-Stimulating Hormone in Sex Differentiation of the Protogynous Orange-Spotted Grouper, Epinephelus coioides.

Follicle-stimulating hormone (FSH) signaling is considered to be essential for early gametogenesis in teleosts, but its functional roles during sex differentiation are largely unknown. In this study, we investigated the effects of long-term and short-term FSH injection on sex differentiation in the protogynous orange-spotted grouper (Epinephelus coioides). Long-term FSH treatment initially promoted the formation of ovaries but subsequently induced a male fate. The expression of female pathway genes was initially increased but then decreased, whereas the expression of male pathway genes was up-regulated only during long-term FSH treatment. The genes related to the synthesis of sex steroid hormones, as well as serum 11-ketotestosterone and estradiol, were also up-regulated during long-term FSH treatment. Short-term FSH treatment activated genes in the female pathway (especially cyp19a1a) at low doses but caused inhibition at high doses. Genes in the male pathway were up-regulated by high concentrations of FSH over the short term. Finally, we found that low, but not high, concentrations of FSH treatment activated cyp19a1a promoter activities in human embryonic kidney (HEK) 293 cells. Overall, our data suggested that FSH may induce ovarian differentiation or a change to a male sex fate in the protogynous orange-spotted grouper, and that these processes occurred in an FSH concentration-dependent manner.

OR33-2 Blocking FSH Inhibits Hepatic Cholesterol Biosynthesis and Reduces Serum Cholesterol

Menopause is associated with dyslipidemia and an increased risk of cardio-cerebrovascular disease. The classic view assumes that the underlying mechanism of dyslipidemia is attributed to an insufficiency of estrogen. In addition to a decrease in estrogen, circulating follicle-stimulating hormone (FSH) levels become elevated at menopause. In this study, we found that blocking FSH reduced serum cholesterol via inhibiting hepatic cholesterol biosynthesis. First, the epidemiological results showed that the serum FSH levels were positively correlated with the serum total cholesterol levels, even after excluding serum estrogen effects. In addition, the prevalence of hypercholesterolemia was significantly higher in peri-menopausal women than that in pre-menopausal women. Furthermore, we generated an FSH-elevated mouse model by intraperitoneal injection of exogenous FSH in ovariectomized (OVX) mice, in which a normal level of estrogen (E2) was maintained by exogenous supplementation. Consistently, the results indicated that FSH, independent of estrogen, increased the serum cholesterol level in the FSH-elevated mouse model. Moreover, blocking FSH by FSHβ-antibody or ablating the FSH receptor (FSHR) gene could effectively prevent hypercholesterolemia induced by FSH injection or high-cholesterol diet feeding. Mechanistically, FSH, binding with hepatic FSHRs, activated the Gi2a/β-arrestin-2/Akt pathway and subsequently inhibited the binding of FoxO1 with the SREBP-2 promoter, thus preventing FoxO1 from repressing SREBP-2 gene transcription. This effect, in turn, resulted in the upregulation of SREBP-2, which drove HMGCR nascent transcription and de novo cholesterol biosynthesis, leading to the increase of cholesterol accumulation. This study uncovers a new opportunity for treating hypercholesterolemia during menopause by blocking FSH signaling, particularly for women in peri-menopause characterized by only FSH elevation.

Effects of follicle-stimulating hormone followed by gonadotropin-releasing hormone on embryo production by ovum pick-up and in vitro fertilization in the river buffalo (Bubalus bubalis).

In this study, we examined the effects of superstimulation using follicle‐stimulating hormone (FSH) followed by gonadotropin‐releasing hormone (GnRH) on buffalo embryo production by ultrasound‐guided ovum pick‐up (OPU) and in vitro fertilization (IVF). Nine Murrah buffaloes were subjected to OPU‐IVF without superstimulation (control). The morphologies of the oocytes collected were evaluated, and oocytes were then submitted to in vitro maturation (IVM). Two days after OPU, same nine buffaloes were treated with twice‐daily injections of FSH for 3 days for superstimulation followed by a GnRH injection. Oocytes were collected by OPU 23–24 hr after the GnRH injection and submitted to IVM (the superstimulated group). The total number of follicles, number of follicles with a diameter > 8 mm, and number of oocytes surrounded by multi‐layered cumulus cells were higher in the superstimulated group than in the control group (p ≤ 0.05). After IVF, the percentages of cleavage and development to blastocysts were higher in the superstimulated group than in the control group (p < 0.05). In conclusion, superstimulation improved the quality of oocytes and the embryo productivity of OPU‐IVF in river buffaloes.

Corifollitropin- compared to daily r-FSH in for patients undergoing intracytoplasmic sperm injection: Clinical trial study.

BackgroundThe current treatment regimen for ovarian stimulation in Intracytoplasmic sperm injection (ICSI) patients is daily injections of Gonadotropins. Recombinant DNA technologies have produced a new recombinant molecule that is a long-acting Follicle Stimulating Hormone (FSH), named corifollitropin alfa. A single injection of long-acting FSH can replace seven daily FSH injections during the first week of controlled ovarian stimulation (COS) and can make assisted reproduction more patients-friendly. There is limited data with different results in this area.ObjectiveTo compare the effectiveness of long-acting FSH vs. daily r-FSH in terms of pregnancy and safety outcomes in women undergoing ICSI cycles.Materials and MethodsIn this clinical trial study, 109 women who were the candidates for ICSI at azzahra hospital were divided in two groups. The first group received 150 units of daily Gonal-f from second or third day of menstruation. The second group received a 150IU corifollitropin alfa on the second or third day of mensuration, and the treatment continued from day eighth of stimulation with Gonal-f based on the ultrasound finding. Both the groups received GnRH antagonist from fifth day of stimulation. Two groups were compared in terms of number of dominant follicles, number of oocytes, stimulation duration, total number of embryos, number of transferred embryos, and success rate of pregnancy.ResultsNo significant difference was found between the two groups in terms of stimulation duration, number of follicles, number of oocytes, total number of embryos, and number of transferred embryos. Moreover, pregnancy outcomes including chemical pregnancy rate (positive pregnancy test), clinical pregnancy rate (detection of fetal heart), the rate of ovarian hyper-stimulation syndrome, multiple-pregnancy, ectopic pregnancy, and miscarriage didn't have a significant difference between the two groups.ConclusionAs corifollitropin alfa was as effective as r-FSH, it could be used as an alternative to ovulation stimulation method in patients undergoing ICSI.

Response of Superovulation by Using FSH (Follicle Stimulating Hormone) and Sex Determination of Embryos Using PCR in Pesisir Cows of West Sumatra

This study was conducted to determine the response of superovulation by giving 16 ml dosage of FSH hormone to female Pesisir cattle. The estrus schedule of 15 Pesisir cows was set by inserting CIDR (Controlled Internal Drug Release) into the vagina for 12 days. At day 10, all cattles were injected with FSH for three consequent days but with decreasing dosage. On the 3rd day, FSH injection was accompanied by PGF2α injection and CIDR was removed. The detection of estrus was performed at day 13. Natural mating was proceeded after the estrus signs visible. Collection of donor embryos was done on the 6th and 8th day after mating. The variables measured were the response of superovulation, total number of corpus luteum, number of embryos and sex ratio. The results obtained were all Pesisir cows responded to superovulation. The average number of of corpus luteum and embryoes per cow were 5.93±3.17 and -----, respectively, while the total of transferable embryoes were 90, with an average of 6.00 or 61.64%. The sexing of embryoes obtained in this study from 146 embryoes awere 76.03% males (111 embryoes) and 23.97% females (35 embryoes). Based on total of transferable embryoes, there were 51.37% male embryos and 11.28% of females embryos. The result of this study showed that the sex ratio of male embryos was higher than female embryos.

EFEKTIVITAS PENYUNTIKAN FSH SECARA SUBKUTAN DAN INTRAMUSKULAR TERHADAP RESPON SUPEROVULASI SAPI SIMENTAL

Superovulation is a necessary technique to produce large number of embryos for embryo` transfer. Hormonal treatment is superovulation methode can be done by implant CIDR and injection of Follicle Stimulating Hormone (FSH). Experiment were carried out to observe the effectiveness of subcutaneous and intramuscular FSH injections on superovulation response in simmental cattle. All animal (n=10) were treated with intravaginal CIDR implant before FSH injection. Studies were devided into two experiment ie: P1 (400 mg FSH diluted in 4 ml sterile diluent) injected in five simmental cattle by single subcutan injection and P2 (400 mg FSH diluted in 20 ml sterile diluent) injected in five simmental cattle by twice daily intramuscular injection over 4 day in decreasing doses. The number of corposa luteal (CL), total embryos collected, and total transferable embryos were observed in this experiment. Data were analyzed by T-test method. The result showed that effectiveness of single subcutan FSH injection were significanthy different (P &lt; 0,05) than intramuscular FSH injection superovulation with single subcutan FSH injetcion is easier than twice daily intramuscular injection in decreasy dose. In conclusion the average of CL (21,4 ± 3,6) and number of tranferable embryo (71,96 %) of the single subcutan FSH injection tended to be better than intramuscular FSH injection. Single subcutan FSH injection more efficient than intramuscular FSH injection. Single subcutan injection can decreasing sterss level in the cattle and be easier in handling the cattle during the experiment.Keywords: Simmental cattle, FSH, superovulation, single subcutan injection, intramuscular injection.

Analysis of patient and nurse preferences for self-administered FSH injection devices in select European markets.

PurposeThe purpose of this study was to assess product-specific features for a variety of self-administered injection devices and identify key factors that patients and nurses in select European markets find most important when selecting injection devices for self-administration of recombinant human follicle-stimulating hormone and urinary human follicle-stimulating hormone for fertility/reproductive therapy.Patients and methodsPatients (N=402) in France, Italy, Spain, Germany, the UK, the Netherlands, and Belgium, as well as reproductive/fertility nurses (N=40) in Germany, Italy, France, Spain, and the Czech Republic were surveyed. All patients were previously prescribed a follicle-stimulating hormone (FSH) treatment for either in vitro fertilization or ovulation induction. Patient and nurse preferences for attributes across all injection devices in the market were obtained via an online questionnaire and evaluated using the maximum differential scaling (MaxDiff) and conjoint analyses, which captured the relative importance of the selected FSH injection device attributes to determine specific qualities in overall product preference.ResultsBoth the MaxDiff and conjoint analyses indicated that, for patients and nurses, the ideal FSH injection device would be a highly accurate, multi-use reusable pen injector with a dial-back function that would be easy for both use and education/instruction. Patients and nurses each selected attributes pertinent to their own experiences with the FSH injection device. Categorically, patients valued factors that resulted in minimal impact on daily life, including reduced injection volume to minimize injection-site pain, as well as a reusable device that would be easy to use; nurses placed greater value on a device that would be easy to teach in order to instruct the greatest number of patients while minimizing risk.ConclusionPatient and nurse preferences were aligned on certain selected attributes of the FSH products. Although this study was an unbranded examination of attributes across all injection devices currently in the market, results demonstrated that the preferred product attributes were all characteristics of the Ovaleap® Pen.

Blocking FSH inhibits hepatic cholesterol biosynthesis and reduces serum cholesterol.

Menopause is associated with dyslipidemia and an increased risk of cardio-cerebrovascular disease. The classic view assumes that the underlying mechanism of dyslipidemia is attributed to an insufficiency of estrogen. In addition to a decrease in estrogen, circulating follicle-stimulating hormone (FSH) levels become elevated at menopause. In this study, we find that blocking FSH reduces serum cholesterol via inhibiting hepatic cholesterol biosynthesis. First, epidemiological results show that the serum FSH levels are positively correlated with the serum total cholesterol levels, even afteradjustment by considering the effects of serum estrogen. In addition, the prevalence of hypercholesterolemia is significantly higher in peri-menopausal women than that in pre-menopausal women. Furthermore, we generated a mouse model of FSH elevation by intraperitoneally injecting exogenous FSH into ovariectomized (OVX) mice, in which a normal level of estrogen (E2) was maintained by exogenous supplementation. Consistently, the results indicate that FSH, independent of estrogen, increases the serum cholesterol level in this mouse model. Moreover, blocking FSH signaling by anti-FSHβ antibody or ablating the FSH receptor (FSHR) gene could effectively prevent hypercholesterolemia induced by FSH injection or high-cholesterol diet feeding. Mechanistically, FSH, via binding to hepatic FSHRs, activates the Gi2α/β-arrestin-2/Akt pathway and subsequently inhibits the binding of FoxO1 with the SREBP-2 promoter, thus preventing FoxO1 from repressing SREBP-2 gene transcription. This effect, in turn, results in the upregulation of SREBP-2, which drives HMGCR nascent transcription and de novo cholesterol biosynthesis, leading to the increase of cholesterol accumulation. This study uncovers that blocking FSH signaling might be a new strategy for treating hypercholesterolemia during menopause, particularly for women in peri-menopause characterized by FSH elevation only.

Follicle capacitation: a meta-analysis to investigate the transcriptome dynamics following follicle-stimulating hormone decline in bovine granulosa cells.

In recent years, exciting progress was made to improve the embryo outcome after ovarian stimulation in domestic animals. The practice of follicle-stimulating hormone (FSH) withdrawal, which is defined as the period of time between the last injection of FSH and oocyte retrieval, resulted in embryo yields significantly superior. Since then, specific changes in the transcriptome of granulosa cells were associated with the increase and also the decline in oocyte developmental competence following the FSH decline. In this study, we integrated large datasets from a public depository using a meta-analysis in order to elucidate the molecular changes occurring in granulosa cells following FSH decline in association with oocyte developmental competence. The meta-analysis revealed that the gene expression patterns observed during this period resulted from the downregulation of proliferative signals, and the upregulation of differentiation signals and early apoptotic signals. Additionally, FSH decline induced cellular hypoxia and triggered the expression of proinflammatory molecules which resulted in early atresia and mimicked the luteinizing hormone (LH) surge signaling to ovulation. To characterize this unique differentiation period, we suggest using the term "follicle capacitation" to refer to the functional changes occurring within the follicle in order to prepare the molecular machinery for the LH surge and ovulation following FSH decline. During this period, the follicle confers the oocyte with developmental competence to become a viable embryo. However, if this period is not rapidly followed by an LH surge, apoptosis signals are increased to generate follicular atresia and decrease oocyte quality.

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.

Quality of common marmoset (Callithrix jacchus) oocytes collected after ovarian stimulation

The common marmoset (Callithrix jacchus) is an experimental animal that is considered suitable for the creation of next-generation human disease models. It has recently been used in the reproductive technology field. Oocytes can be effectively collected from female marmosets via ovarian stimulation with injections of follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG). The oocytes, collected about 28 h after the hCG injection, include both premature oocytes and postmature (in vivo matured; IVO) oocytes, and the premature oocytes can be matured by in vitro culture (in vitro matured; IVM). Although IVM and IVO oocytes are equivalent in appearance at the MII stage, it remains unclear whether there are differences in their properties. Therefore, we investigated their in vitro fertilization and developmental capacities and cytoskeletal statuses. Our findings revealed that the IVM and IVO oocytes had similar fertilization rates but that no IVO oocytes could develop to the blastocyst stage. Additionally, IVO oocytes showed abnormal cytoskeletal formation. It is concluded that IVM oocytes maintain normal function, whereas IVO oocytes would be affected by aging and other factors when they remain for a long time in the ovary.

Lipid droplets in cultured luteal cells in non-pregnant sheep fed different planes of nutrition

Accumulation of lipid droplets (LD) in luteal cells likely is important for energy storage and steroidogenesis in the highly metabolically active corpus luteum (CL). The objective of this study was to determine the effect of plane of nutrition on progesterone (P4) secretion, and lipid droplet number and size in cultured ovine luteal cells. Ewes were randomly assigned to one of three nutritional groups: control (C; 100% NRC requirements, n=9), overfed (O; 2×C, n=12), or underfed (U; 0.6×C, n=10). Superovulation was induced by follicle stimulating hormone injections. At the early and mid-luteal phases of the estrous cycle, CL were dissected from ovaries, and luteal cells isolated enzymatically. Luteal cells were incubated overnight in medium containing serum in chamber slides. Media were then changed to serum-free and after 24h incubation, media were collected for P4 analysis, and cells were fixed in formalin and stained with BODIPY followed by DAPI staining. Z-stacks of optical sections of large and small luteal cells (LLC and SLC, respectively) were obtained using a laser-scanning microscope. Rendered 3D images of individual LLC and SLC were analyzed for cell volume, and total and individual LD volume, number and percentage of cellular volume occupied by LD by using Imaris software. Concentrations of P4 in serum and media were greater (P<0.05) at the mid than early-luteal phase, and were not affected by nutritional plane. LD total volume and number were greater (P<0.001) in LLC than SLC; however, mean volume of individual LD was greater (P<0.02) in SLC than LLC. In LLC, total LD volume was greater (P<0.02) in O than C and U ewes. In SLC, total LD volume and number was greater (P<0.003) at the mid than early-luteal phase, and percentage of cell volume occupied by LD was greater (P<0.002) in U than C and O ewes. These data demonstrate that both stage of luteal development and nutritional plane affect selected LD measurements and thus may affect luteal functions. Furthermore, these data confirm that LD dynamics differ among parenchymal steroidogenic luteal cell types.