IU Human Chorionic Gonadotropin Research Articles

The Effect of Sulpiride Treatment During the Periovulatory Period on Prolactin Concentration and Ovulation in the Mare

Sixteen estrous cycles from 10 cyclic mares were randomly assigned to a control or sulpiride group (n = 8 each). All mares received 1,500 IU of human chorionic gonadotropin (hCG) (hour 0) during estrus with a follicular diameter ≥32 mm. Mares were scanned every 12 hours until ovulation. In the treatment group, beginning at hour 0, each mare received 1.5 mg/kg of sulpiride every 12 hours intra-muscularly until ovulation or formation of a luteinized unruptured follicle (LUF). Concentrations of luteinizing hormone (LH) and prolactin (PRL) were measured by radioimmunoassay. In each group, there were 10 preovulatory follicles for the eight cycles. The ovulation rate (9/10, 90%) was similar in the control and sulpiride groups. Two mares formed an LUF, which was first detected at hours 48 and 72 for the sulpiride and control mares, respectively. The interval from hCG to ovulation was 49.5 ± 11.1 and 43.5 ± 5.8 hours, for the control and sulpiride groups, respectively (P > .5). LH followed the typical preovulatory surge pattern, with no difference between groups (P > .5). Sulpiride administration increased PRL concentration in treated mares at 24 (P < .1), 36, and 48 hours (P < .05) after treatment. In conclusion, sulpiride administration every 12 hours increased PRL concentration in treated mares after 24 hours of the beginning of treatment. However, at this time window and concentration, PRL did not have any effect on ovulation. The control mare that developed an LUF had a PRL concentration similar to other ovulatory control mares (always ≤10 ng/mL).

Effect of bovine blastocyst size at embryo transfer on day 7 on conceptus length on day 14: Can supplementary progesterone rescue small embryos?

Conceptus size on Day 14 after multiple embryo transfer of Day 7 in vitro–produced blastocysts varies greatly within animal. One explanation for this variation may be related to blastocyst cell number at the time of transfer. The aim of this study was to examine the effect of Day 7 blastocyst cell number on Day 14 conceptus size and to examine the effect of progesterone (P4) supplementation on embryo development after the transfer of Day 7 blastocysts containing a low total cell number. The estrous cycles of crossbred beef heifers were synchronized using an 8-day progesterone (P4)–releasing intravaginal device (PRID) with the administration of a prostaglandin F2α analog on the day before device removal. Only those heifers recorded in standing estrus (Day 0) were used. Heifers were randomly assigned to one of four treatment groups: (1) control: large blastocysts (high total cell number), (2) control: small blastocysts (low total cell number), (3) small blastocysts plus a single intramuscular injection of 3000 IU human chorionic gonadotropin (hCG) on Day 2 after estrus, or (4) small blastocysts plus insertion of a vaginal P4 insert (PRID, 1.55 g P4) between Days 3 and 5 after estrus. In vitro–produced blastocysts were transferred to each heifer on Day 7 (n = 10 blastocysts per heifer), and conceptuses were recovered at slaughter on Day 14. Daily blood samples were collected from Day 0 to 14 to measure serum P4 concentrations. Data were analyzed using the PROC MIXED procedure of SAS. Total cell number on Day 7 was significantly lower in small versus large blastocysts (72.4 ± 3.93 vs. 144.8 ± 3.90, P < 0.05). Conceptus recovery rate was 53.8% overall (140 of 260) and was highest in the large blastocyst group (68.3%, 41 of 60) compared with the other groups (45.7%–55.0%). Concentrations of serum P4 were similar in the two unmanipulated recipient groups but were significantly elevated (P < 0.05) by Day 8 in the hCG-treated heifers and on Days 4 and 5 in the PRID group (P < 0.003). In the absence of supplemental P4, Day 14 conceptuses resulting from the transfer of small blastocysts (2.48 ± 0.54 mm) were smaller than those from large blastocysts (3.32 ± 0.52 mm). Administration of hCG on Day 2 approximately doubled conceptus length on Day 14 (4.94 ± 1.15 mm; P < 0.05), whereas insertion of a PRID from Days 3 to 5 increased conceptus length approximately fivefold (13.09 ± 2.11 mm; P < 0.05) compared with controls. In conclusion, results indicate that supplemental P4 is capable of “rescuing” poor-quality blastocysts, presumably via the now well-described actions on the endometrium and consequent effects on uterine lumen fluid composition.

Identification of inadequate maternal progesterone concentrations in nulliparous dairy heifers and treatment with human chorionic gonadotrophin

Plasma progesterone concentrations were estimated for 381 nulliparous dairy heifers from eight farms on days 4–6 after insemination. Heifers were synchronised using two treatments of prostaglandin F2α (PGF2α) 11 days...

The role of midluteal progesterone for the prediction of pregnancy after ovulation induction with clomiphene citrate for unexplained infertility

To investigate the role of midluteal progesterone (MLP) for the prediction of pregnancy after ovulation induction with clomiphene citrate (CC) for unexplained infertility. prospective cohort. A total of 271 ovulation induction cycles with CC in 140 patients aged between 18-40 years diagnosed with unexplained infertility were recruited in the study between April and December 2012. The women were given a 5 day course of 50-150 mg of CC starting on day 2 -5 of the menstrual cycle and ovulation was triggered with 10 000 IU of human chorionic gonadotropin (hCG) when the dominant folicule reached 18mms. We performed intrauterine insemination (IUI) 36 hours after the hCG injection. We collected venous blood samples on day 21-23 of the cycle for the measurement of MLP levels. We performed a pregnancy test in women who missed their menstrual periods and clinical pregnancy was documented with detection of cardiac activity on ultrasonographic examination. Statistical analysis was performed by the SPSS package 20.0 version. Paired t test was used to compare baseline and biological characteristics. ROC analysis was performed to determine a cut off value for MLP for the prediction of pregnancy. Clinical pregnancy rate was 10,3 % (28/271). Age and BMI of the patients with a positive pregnancy test and the ones who failed to achieve pregnancy were similar (p=0.81and p=0.15, repectively). Midluteal progesterone levels were significantly elevated in patients who achieved pregnancy than the ones who failed (31,72 ±12,58 ng/ml versus 18,59±8,91ng/ml, p=0.00). A treshold value of 22,5 ng/ml was established for MLP following analyses repeated with balanced distributions. MLP values over 22,5 ng/ml had a positive predictive value of 67,8 % and those below 22,5 ng/ml had a negative predictive value of 81,2% for the prediction of pregnancy. Midluteal progesterone levels may be useful for the prediction of pregnancy in CC cycles in patients with unexplained infertility.

EMBRYONIC DEVELOPMENT AND SALINITY TOLERANCE OF MUDSKIPPER (PERIOPHTHALMUS MAGNUSPINNATUS)

Periophthalmus magnuspinnatus inhabit in China which was previously mistaken as Periophthalmus modestus and recently reported from the coasts of Bohai Sea,Yellow Sea,East China Sea,and South China Sea.For the taxonomically new P.magnuspinnatus,there is only one study on the skin structure and investigating the life history Lifestyle of Korean mudskipper.This paper was studied the fecundity and effects of salinity on embryonic development of the P.magnuspinnatus in Qiantang River estuary.The fertilized eggs were observed through a semi-dry artificial fertilization method,which were cultured and analysed in constant temperature of sea water with weak aeration in different salinities.The embryonic development of mudskipper was characterized by continual viviperception.The results indicated that the individual absolute fecundity(F) ranged from 2249 to 3472 eggs,the average was 2746 eggs,and the standard error of mean was 230.The individual relative fecundity(FL) ranged from 32 to 44 eggs per millimeter,the average was 35 eggs,and the standard error of mean was 2.The individual relative fecundity(FW) ranged from 643 to 1320 eggs per gram,the average was 698 eggs,and the standard error of mean was 71.From the injection of 0.2 μg of LHRH-A3(luteinizing hormone releasing hormone-analogue) then after 24 hours injection of 20 IU of HCG(Human Chorionic Gonadotropin) into female fish and 20 IU of HCG into male fish,each male fish received one injection at the same time as female fish received the second injection.Most of experimental fish was able to develop to mature.With the water temperature of 27 ℃,salinity of 10‰,and pH of 7.8±0.3,the embryonic development required 158 hours and 46 minutes,while the accumulated temperature was up to 4286.70 h.℃.The development time from heart beating to hatch larva was relatively longer which need 127 hours and 52 minutes under 27 ℃ and 10‰ salinity condition,accounting for 80.5% of the total embryonic development time.The development time from air bladder formation(the last development stage) to hatch larva was 88 hours and 52 minutes,accounting for 56.0% of the total embryonic development time.Before the eggs were hatched,the organs had differentiated normally in shape already,such as distinct gut,visible anus,air bladder elliptical,and melanophore of eyes per minute.The time series of embryonic development was slightly different from other Gobioidei species which experienced intestinal canal formation,blood circulation and pectoral fin appearance successively.The heart rate in heart beating stage of P.magnuspinnatus was ranged from 52 to 78 times per minute,and then sped up which could reach to 283 times per minute until hatch larva stage.The optimum salinity for embryo development was 5‰-15‰.The hatchability under 5‰,10‰ and 15‰ salinity was 38.5%,47.3% and 42.8% respectively.Experimental observation results indicated that salinity showed no influence on embryonic development of P.magnuspinnatus,but the development time and development accumulative temperature shortened as the decrease of salinity.Under high salinity concentrations of 35‰,30‰,25‰,and 20‰,the fish larvaes were not hatched from the fertilized eggs.

Anti-mullerian hormone and antral follicle count as predictors of ovarian response in assisted reproduction.

OBJECTIVE:The objective of this study was to test the hypothesis that AMH and antral follicle count (AFC) are good predictors of ovarian response to controlled ovarian stimulation and to compare them.MATERIALS AND METHODS:This observational cross-sectional study included 56 subjects aged between 25 and 42 years who were enrolled between 1st January and 31st December 2010 for their first intracytoplasmic sperm injection (ICSI) program. Baseline hormone profiles including serum levels of Estradiol (E2), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), and Anti-mullerian Hormone (AMH) were determined on day 3 of the previous cycle. The antral follicle count measurements were performed on days 3-5 of the same menstrual cycle. Antral follicles within the bilateral ovaries between 2-6 mm were recorded. The subjects were treated with long protocol for ovarian stimulation. Ovulation was induced with 10,000 IU of human chorionic gonadotropin (hCG) when at least 3 follicles attained the size of more than 17 mm. Transvaginal oocyte retrieval was performed under ultrasound guidance 36 hours after hCG administration. An oocyte count less than 4 and absence of follicular growth with controlled ovarian hyper stimulation was considered as poor ovarian response. Oocyte count of 4 or more was considered as normal ovarian response.RESULTS:Statistical analysis was performed using SPSS software trail version 16.0. Subjects were divided into 2 groups, depending on the ovarian response. The mean oocyte counts were 12.27 ± 6.06 and 2.22 ± 1.24 in normal and poor responders, respectively, (P = 001). Multiple regression analysis revealed AMH and antral follicle count as predictors of ovarian response (β coefficient ± SE for AMH was 1.618 ± 0.602 (P = 0.01) and for AFC, it was, 0.528 ± 0.175 (P = 0.004). AFC was found to be a better predictor of ovarian response compared to AMH in controlled ovarian hyper stimulation.CONCLUSION:The observations made in this study revealed that both AMH and AFC are good predictors of ovarian response; AFC being a better predictor compared to AMH.

Penile growth in response to hormone treatment in children with micropenis

Introduction:Micropenis is defined as a stretched penile length 2.5 standard deviations less than the mean for age without the presence of any other penile anomalies, such as hypospadias. The term refers to a specific disorder that has a known set of causative factors and defined treatment modalities. The purpose of this study was to determine the effect of hormonal therapy on the gonadal response and penile growth in children who presented with micropenis.Materials and Methods:Children (<18 years) who met the criteria for micropenis were included in this study. Children more than 11 years old were treated using a standard protocol of 1,500 to 2,000 IU human chorionic gonadotrophin administrated intramuscularly, once per week, for 6 weeks. Children less than 11 years old were treated with parenteral testosterone enanthate 25 mg once a month for 3 months. Response was evaluated in terms of change in testosterone levels and size of penis.Results:Serum testosterone levels at baseline and after 8 weeks of hormonal treatment were <20 and 449.4 ng/mL, respectively (P < 0.0001) in all children more than 11 years old. Stretched penile length after hormonal treatment increased from 15.54 to 37.18 mm in children less than 11 years old and from 26.42 to 64.28 mm in children more than 11 years old (P < 0.001).Conclusions:Management of isolated micropenis revolves around testosterone (direct administration or encouraging the patient's body to make its own), and results with respect to increase in penile length are promising.

Ovarian Stimulation Affects the Population of Mouse Uterine NK Cells at Early Pregnancy

The aim of this study was to determine the influence of ovarian stimulation on endometrial mouse NK cell population. For superovulation, the female adult NMRI mice were injected i.p. with 10 IU of the pregnant mare serum gonadotropin followed 48 h later by an i.p. injection of 10 IU human chorionic gonadotropin hormone. Ovarian stimulated and nonstimulated mice were mated with fertile male. The presence of vaginal plug proved natural pregnancy, and this day was considered as day one of pregnancy. Tissue samples were prepared from the uterine horn and spleen of both groups of study on 7th day of pregnancy. Serum estradiol-17β and progesterone were measured at the same time. The tissue cryosections were prepared and double stained for CD 161 and CD3 markers, and NK cells population was analyzed. Relative frequency of NK cells was significantly lower in stroma and myometrium in hyperstimulated mice compared with the control group. However, no difference was seen in percentage of NK cells in spleen. The ovarian stimulation influences the proportion of uterine NK cells and may affect the embryo implantation.

197 EXPRESSION OF MELATONIN-RELATED GENES IN RAT CUMULUS–OOCYTE COMPLEXES

Melatonin is a hormone usually associated with the modulation of circadian rhythms and the regulation of seasonal reproductive function. There is evidence that melatonin acts directly on the regulation of ovary function. The mRNA expression of melatonin membrane receptors genes was detected in mammalian and non-mammalian ovaries. In spite of melatonin receptors and Asmt (limiting enzyme in melatonin biosynthesis) genes being present in cattle cumulus–oocyte complexes (COC), no information regarding rat COC has been reported. The aim of this study was to investigate the expression of melatonin receptor (Mt2) and melatonin synthesis enzyme (Asmt) genes at different meiotic cell cycle stages in COC from 27-day-old female rats. All the procedures involving animals were approved by the Animal Care Committee of the Institute of Biomedical Sciences. To obtain germinal vesicle (GV) immature COC from ovarian follicles, rats were treated with 20 IU equine chorionic gonadotropin (eCG) for induction of follicular development and killed by euthanasia 48 h later. To obtain COC at MII oocyte stage from oviducts, rats were injected with 20 IU of eCG and 48 h later with 20 IU of human chorionic gonadotropin (hCG), and then killed after 14 to 16 h. The oocytes in COC were separated from their cumulus cells by repeated pipetting through a narrow-bore pipette in culture medium. Oocytes and cumulus cells total RNA were isolated using a Trizol reagent (Invitrogen Corp., Carlsbad, CA, USA). Pools of 80 COC per cDNA sample were used. Quantitative real-time PCR (qRT-PCR) was performed (7500 Real-Time PCR System; Applied Biosystems Inc., Foster City, CA, USA) with 25-µL reactions containing 2 µL of cDNA (10 ng µL–1), SYBR green (Power SYBR Green; Applied Biosystems Inc.), and 400 nM specific intron-spanning primers. A set of 10-fold serial dilutions of each internal standard (102 to 106 copies/2 µL) was used to generate a standard curve. Transcript numbers were determined by the system software and normalized using the geometric mean calculated from the reference genes: Actb and Rpl37a. All the results were plotted as the mean ± SEM, and 4 replicates were performed. Unpaired t-test was used to evaluate the cell type (oocyte v. cumulus cells) differences. The qRT-PCR analyses revealed the presence of transcripts of the Mt2 gene only in oocytes from immature (GV) and mature (MII) COC. At both maturational stages, the copy numbers for Asmt in oocytes were significantly higher than in cumulus cells. The results confirm the presence of the Asmt gene in rat COC and suggest the possible involvement of these cells on melatonin biosynthesis. The presence of Mt2 transcript in immature and mature oocytes also suggests the potentially important role of melatonin in regulating the rat meiotic cellular cycle. Research supported by FAPESP.

Improvement of ovarian response and oocyte quality of aged female by administration of bone morphogenetic protein-6 in a mouse model

BackgroundAdvancing female age remains a difficult problem in infertility treatment. Ovarian angiogenesis plays an important role in follicular development and the activation of ovarian angiogenesis has been emerged as a new strategy for the improvement of age-related decline of oocyte quality. BMP-6 affect gonadotropin signals in granulosa cells and it promotes normal fertility by enabling appropriate response to LH and normal oocyte quality. BMP-6 has a potential role in regulation of angiogenesis and regulates the expression of inhibitor of DNA-binding proteins (Ids). Ids involved in the control and timing of follicle selection and granulosa cells differentiation. Especially, Id-1 is well-characterized target of BMP-6 signaling. Therefore, this study investigated whether co-administration of BMP-6 during superovulation process improves ovarian response, oocyte quality and expression of Id-1 and vascular endothelial growth factor (VEGF) in the ovary of aged female using a mouse model.MethodsAged C57BL/6 female mice (26–31 weeks old) were superovulated by injection with 0.1 mL of 5 IU equine chorionic gonadotropin (eCG) containing recombinant mouse BMP-6 at various doses (0, 0.01, 0.1, 1, and 10 ng), followed by injection with 5 IU human chorionic gonadotropin (hCG) 48 h later. Then, the mice were immediately paired with an individual male. The aged control group was superovulated without BMP-6. Young mice of 6–9 weeks old were superovulated without BMP-6 as a positive control for superovulation and in vitro culture of embryos. Eighteen hours after hCG injection, zygotes were retrieved and cultured for 4 days. Both ovaries of each mouse were provided in the examination of ovarian expression of Id-1 and VEGF by reverse transcriptase-polymerase chain reaction, western blot, and immunohistochemistry.ResultsAdministration of 0.1 ng BMP-6 significantly increased the number and blastocyst formation rate of oocytes ovulated and ovarian expression of Id-1 and VEGF compared to aged control mice. These increased levels were comparable to those of young control mice.ConclusionsThis result suggests that BMP-6 during ovulation induction plays an important role in improvement of oocyte quality and ovarian response of aged female, possibly by regulating of ovarian Id-1 and VEGF expression.

Metabolism of energy substrates of in vitro and in vivo derived embryos from ewes synchronized and super ovulated with norgestomet and porcine follicle stimulating hormone.

The synchronization and ovulatory responses of Sangsari cross bred ewes and metabolism of energy substrates in 8-cell stage embryos to hatched blastocysts stage produced in vitro or in vivo were investigated. Ewes were assigned randomly to receive 37.5 IU of porcine follicle stimulating hormone (FSH-P) daily for the 3 days preceding implant removal (Day 0). Synchronization of estrus was carried out using a 1.5 mg norgestomet (Crestar) ear implant for 12 days. Ewes in estrus were mated two to three times with rams of proven fertility. At the time of first mating each ewe was administered 1000 IU of human chorionic gonadotrophin (hCG) to induce ovulation. Surgical embryo recovery was performed on Days 4 and 6 after onset of estrus (Day 0) and recovered embryos were subjected to comparative metabolism studies with in vitro derived embryos at the same stage of development. The number of corpora lutea (CL), unovulated follicles and overall ovarian activity were recorded for each ewe during the breeding and non-breeding seasons. While the pattern of oxidation was similar among in vitro and in vivo derived embryos, a low pyruvate to lactate ratio was the preferred substrate of embryos derived in vitro. A high level of production of CO2 and lactate resulted from a stress response to the suboptimal culture environment. The first marked increase in the metabolism of glucose by ovine embryos was detected in compact morula stage, but there was no significant increase in the oxidation of glucose after the morula stage. Two different concentrations of glucose were compared, but this did not affect metabolism. However, the rate of incorporation and metabolism of glucose tended to be higher at the 0.56 mmol/L glucose dosage.

The Effect of n-Hexane on the Gonad Toxicity of Female Mice

ObjectiveTo investigate the toxic effects of n-hexane on the Ganod of female mice. Methodsn-Hexane was administered to four groups of mice by inhalation at doses of 0, 3.0, 15.1, and 75.8 mL/m3 respectivelyfor five weeks. Each group consisted of 10 mice, of which half were injected in first with 10 IU of pregnant mare serum gonadotrophin (PMSG) on the 33rd days, and then with 10 IU of human chorionic gonadotrophin (HCG) 48 hrs later. After the treatment, mouse sera were sampled and ovulating hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), and progesterone (P4) levels were measured by electrochemiluminescence immunoassays (ECLIA). In each group, the right ovaries of the non-super-ovulated mice were stained with hematoxylin and eosin while ovaries on the left side were prepared with the TUNEL method in order to detect apoptotic cells. ResultsThe duration of the diestrus stage decreased significantly (P<0.05) in the 75.8 mL/m3 group. All super-ovulated mice in each treatment group produced fewer eggs than those in the control group (P<0.05). The number of follicles in ovaries in the 75.8 mL/m3 group was smaller compared with the control group (P<0.05). The serum P4 levels in each treatment group were lower than those in the control group (F=6.196, P<0.01). The cell apoptotic rate in the 75.8 mL/m3 group was higher (P<0.05). Conclusionsn-Hexane may have directly mediated via alterations hormone secretion and promoted granulosal cell apoptotic, which may be one of the important mechanisms for n-hexane induced mouse ovary impairment.

Ovarian reaction and estrus manifestation in delayed puberty gilts after treatment with equine chorionic gonadotropin

BackgroundProlonged pre-insemination anestrus (i.e. delayed puberty) is a major contributing factor for culling up to 30% of the replacement gilts at large breeding farm units in Vojvodina. It is imperative to determine if these gilts are acyclic (prepubertal) or cyclic, but just fail to exhibit behavioural estrus. Recent investigations demonstrate that treatment with equine chorionic gonadotropin (eCG) can increase the diestrous phase duration in sexually mature gilts. Based on these finding, the aim of the present studies was to determine the reproductive status of delayed puberty gilts following injection with eCG.MethodsTwo experiments were conducted on a swine breeding farm in Vojvodina. In Exp. 1, 20 prepubertal (acyclic) gilts, and 120 sexually mature (cyclic) gilts were injected with a single injection of 400 IU eCG + 200 IU human chorionic gonadotropin (hCG) or with 1000 IU eCG (cyclic gilts), at d5, d11 or d17 after spontaneous estrus detection, to determine their ovarian reaction and induced estrus manifestation. In Exp. 2, sixty delayed puberty gilts (estrus not detected until 8 month of age, av. 258 days) were culled from breeding herd and slaughtered to determine their reproductive status based on ovarian anatomical features. The second group of gilts (n = 60) was treated with a single 1000 IU eCG injection to determine their reproductive status, based on the interval between eCG injection to estrus detection and duration. The data were analyzed by descriptive statistics, t-test, analysis of variance and Duncan’s test in the software package Statistics 10th.ResultsOvulations were induced in 90% of acyclic (sexually immature) and, on average, 93.3% of cyclic (sexually mature) gilts after the eCG injection. On average, 4 days after the eCG injection, estrus was detected in 85% of the treated acyclic (sexually immature) gilts and in 95% (19/20) of the cyclic (sexually mature) gilts, treated with eCG on day 17 after spontaneous estrus detection. The interval from eCG to induced estrus detection was prolonged (av. 25 days) in 95% (19/20) of the sexually mature gilts treated with eCG on day 5 and in 90% (18/20) of gilts treated on day 11 after spontaneous estrus detection (Exp. 1). Forty anestrous gilts reached cyclic pubertal ovarian activity. Estrus manifestation was detected in 56 gilts (93.3% of the total 60 treated prolonged anestrous gilts, av. 259 days of age), after a single 1000 IU eCG injection. Thirty-four gilts (60.7% of the total gilts in estrus) with prolonged eCG to estrus interval (av. 24.7 days) were considered spontaneously cyclic (sexually mature), but behaviourally anestrous before treatment. The remaining 22 (39.3% of the total gilts in estrus) were considered truly sexually immature (acyclic) before the treatment or were eCG injected in the late luteal or proestrous phase of spontaneous estrous cycle (Exp. 2).ConclusionsIn 66.7% of the delayed puberty gilts, pre-ovulatory follicles (PoF), corpora hemorrhagica (CH), corpora lutea (CL), or corpora albicantia (CA) were found on the ovaries upon post mortem examination. These gilts were considered as sexually mature before slaughtering. In 60.7% of the delayed puberty gilts, behavioural estrus was detected an average of 24.7 days following eCG injections. These gilts were considered as eCG treated during the luteal phase (diestrus) of the spontaneous estrus cycle. Both findings suggest that delayed puberty gilts actually reached cyclic pubertal ovarian activity (sexual maturity) before culling from the breeding herd.

Letrozole versus clomiphene citrate for superovulation in Egyptian women with unexplained infertility: a randomized controlled trial

To compare the efficacy of letrozole with clomiphene citrate (CC) in Egyptian women with unexplained infertility. This randomized controlled trial was conducted at Ain Shams University Hospital and Dar Al Hekma hospital between February 2010 and May 2011. Two hundreds and seventy women with unexplained infertility were randomized into two groups using computer-generated randomization plan, and were given letrozole 2.5 mg/day from cycle day 3 to 7 (Letrozole group, n=136) or CC 100 mg/day from cycle day 3 to 7 (CC group, n=134). On day 9, the participants underwent a transvaginal sonography (TVS) every other day to monitor their ovulation and measure both endometrial thickness and Doppler flow indices of uterine and subendometrial vessels. Single injection of 10,000 IU of human chorionic gonadotropin (hCG) was given when the mean diameter of at least one ovarian follicle was ≥18 mm, quantitative βhCG was done 2 weeks after hCG injection to diagnose chemical pregnancy. Clinical pregnancy was confirmed by observing a gestational sac with fetal echoes and pulsation 4 weeks after positive pregnancy test by TVS. Both groups were comparable with regard to the day of hCG administration (letrozole 13.4±5 vs CC 12.1±4.9, respectively, p=0.06). The mean number of mature follicles was significantly higher in CC group (2±0.9 vs 1±0.0, P=0.02). Serum estradiol was significantly greater in CC group (817±299 vs 364±149 pg/ml, P<0.001). The clinical pregnancy rate was significantly higher in letrozole group (23.07 vs 10.68%, P<0.001). There was statistically significant increase in endometrial receptivity in letrozole group as assessed by endometrial thickness and Doppler flow indices of uterine and subendometrial vessels. No serious side effects were reported in either group. Letrozole has beneficial effect on endometrium, an action that may improve the implantation and pregnancy rates in women with unexplained infertility.

Microarray analysis of mRNA from cumulus cells following in vivo or in vitro maturation of mouse cumulus–oocyte complexes

The IVM of mammalian cumulus-oocyte complexes (COCs) yields reduced oocyte developmental competence compared with oocytes matured in vivo. Altered cumulus cell function during IVM is implicated as one cause for this difference. We have conducted a microarray analysis of cumulus cell mRNA following IVM or in vivo maturation (IVV). Mouse COCs were sourced from ovaries of 21-day-old CBAB6F1 mice 46h after equine chorionic gonadotrophin (5IU, i.p.) or from oviducts following treatment with 5IU eCG (61h) and 5IU human chorionic gonadotrophin (13h). IVM was performed in α-Minimal Essential Medium with 50 mIU FSH for 17h. Three independent RNA samples were assessed using the Affymetrix Gene Chip Mouse Genome 430 2.0 array (Affymetrix, Santa Clara, CA, USA). In total, 1593 genes were differentially expressed, with 811 genes upregulated and 782 genes downregulated in IVM compared with IVV cumulus cells; selected genes were validated by real-time reverse transcription-polymerase chain reaction (RT-PCR). Surprisingly, haemoglobin α (Hba-a1) was highly expressed in IVV relative to IVM cumulus cells, which was verified by both RT-PCR and western blot analysis. Because haemoglobin regulates O2 and/or nitric oxide availability, we postulate that it may contribute to regulation of these gases during the ovulatory period in vivo. These data will provide a useful resource to determine differences in cumulus cell function that are possibly linked to oocyte competence.

Administration of visfatin during superovulation improves developmental competency of oocytes and fertility potential in aged female mice

To examine whether visfatin administration during superovulation improves ovarian response, developmental competence of oocytes, and fertility in aged female mice. Controlled experimental study. University hospital. Two groups of differently aged C57BL female mice (6-11 and 26-31 weeks). Female mice were coinjected intraperitoneally with 5 IU pregnant mare's serum gonadotropin (PMSG) and visfatin of various doses (0-500 ng/mL), followed by 5 IU human chorionic gonadotropin (hCG) injection 48 hours later. Then the mice were immediately mated with an individual male. After 18 hours zygotes were cultured, and expression of ovarian visfatin and vascular endothelial growth factor (VEGF) was examined. Potential pregnancies of visfatin-administered aged female mice were monitored for delivery of offspring. Number of zygotes retrieved, embryo developmental competency, fertility potential, ovarian visfatin and VEGF expression. Ovarian visfatin expression was significantly decreased in the aged mice group compared with the young. Visfatin administration significantly increased embryo developmental rate and ovarian visfatin and VEGF expressions in the aged mice. Visfatin-administered aged mice delivered significantly higher numbers of offspring than controls. This study suggests that visfatin administration during superovulation plays an important role in regulating oocyte quality and can improve oocyte quality and fertility of aged female mice.

A case report: successful pregnancy after controlled ovarian hyperstimulation by using short-acting gonadotropin releasing hormone agonist only

Controlled ovarian hyperstimulation (COH) is widely used in in vitro fertilization and embryo transfer (IVF-ET) to achieve multiple follicle development during one treatment cycle. COH is usually initiated by gonadotropin releasing hormone agonist (GnRHa) to down-regulate pituitary secretion of gonadotropins, suppress endogenous LH surge and synchronize multiple follicles development, which is followed by stepwise exogenous gonadotropins stimulation [1, 2]. Here we report a case of successful pregnancy after inducing multiple follicles development by using short-acting GnRHa only. A 31-year-old woman, 50 kg weight, was administrated in our unit for four-year primary infertility. Her menstrual cycle was 28 days, lasting 6–7 days. The basal serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) level was 5.1 mIU/mL, 3.56 mIU/mL, and 38.81 pg/mL, respectively. The numbers of antral follicles were eight per ovary. In the mid-luteal phase, the patient was administrated of short-acting GnRHa (Decapeptyl, Ferring, Germany) 0.1 mg/day for pituitary down-regulation] to initiate an IVF treatment cycle. After 14 days administration, the serum FSH was 4.58 mIU/mL, LH was 29.23 mIU/mL, and E2 was 26.79 pg/mL. Transvaginal ultrasound examination showed that there were five follicles with average diameter more than 10 mm, and the largest one was 14 mm. 0.1 mg Decapeptyl was continuously given for one more day and the LH level maintained 29.96 mIU/mL. Then, we decreased the dose of GnRHa to 0.05 mg/day and continuously given for four more days. Ultrasound examination showed five follicles with average diameter more than 20 mm, the largest one was 28 mm. The patients’ serum E2 increased to 1,270 pg/mL, LH was 13.51 mIU/mL. 0.05 mg Decapeptyl was continuously given in the morning and 10,000 IU human chorionic gonadotropin (hCG) was administered to trigger egg maturation in the evening. The patient was scheduled for oocyte retrieval 34 h after the hCG injection. Five oocytes were obtained and three fertilized successfully. Two grade I embryos at the 8-cell stage were transferred at d3, and a surplus grade I embryos at the 7-cell stage were frozen. Routine luteum support (UTROGESTAN 200 mg, Bid, Laboratoires Besins-Iscovesco, France) was given after embryo transfer (ET). Serum β-hCG level examined 14 days after ET was 613.9 mIU/mL, indicating a successful biochemical pregnancy. A transvaginal ultrasound examination 4 months after ET confirmed an ongoing intrauterine twin pregnancy.

A superovulation protocol for the spiny mouse (Acomys cahirinus)

This study aimed to develop a superovulation protocol for the spiny mouse (Acomys cahirinus). The spiny mouse is a desert-adapted rodent species, with a long oestrus cycle (11 days) compared with rat and mouse, and gives birth to few (mean litter size is 3) precocial offspring after a relatively long gestation (39 days). We successfully optimised a superovulation protocol that elicited a 5-fold increase in the normal ovulation rate of this species. To induce superovulation in the spiny mouse 2 injections of equine chorionic gonadotrophin (eCG, 10 IU each), 9h apart, were required, followed by 20 IU of human chorionic gonadotrophin (hCG). This protocol was successful in 100% of females trialed and at 33 h post-hCG an average of 14.7 ± 1.5, 1-2 cell embryos were recovered. Histological analysis of ovaries following superovulation revealed large corpus lutea and post-ovulatory follicles occupying a large part of the ovary. Ovulation commenced 6-12 h after the hCG injection and continued until 24-33 h post-hCG as indicated by both histological analysis of ovaries and the presence of oocytes/embryos in the oviduct. This superovulation protocol will facilitate the development of an in vitro culture system for spiny mouse embryos.

Identification of variables contributing to superovulation efficiency for production of transgenic prairie voles (Microtus ochrogaster).

BackgroundThe prairie vole (Microtus ochrogaster) is an emerging animal model for biomedical research because of its rich sociobehavioral repertoire. Recently, lentiviral transgenic technology has been used to introduce the gene encoding the green fluorescent protein (GFP) into the prairie vole germline. However, the efficiency of transgenesis in this species is limited by the inability to reliably produce large numbers of fertilized embryos. Here we examined several factors that may contribute to variability in superovulation success including, age and parentage of the female, and latency to mating after being placed with the male.MethodsFemales produced from 5 genetically distinct breeder lines were treated with 100 IU of pregnant mare serum gonadotrophin (PMSG) and immediately housed with a male separated by a perforated Plexiglas divider. Ovulation was induced 72 hr later with 30 IU of human chorionic gonadotropin (hCG) and 2 hrs later mating was allowed.ResultsSuperovulation was most efficient in young females. For example, females aged 6-11 weeks produced more embryos (14 +/- 1.4 embryos) as compared to females aged 12-20 weeks (4 +/- 1.6 embryos). Females aged 4-5 weeks did not produce embryos. Further, females that mated within 15 min of male exposure produced significantly more embryos than those that did not. Interestingly, there was a significant effect of parentage. For example, 12 out of 12 females from one breeder pair superovulated (defined as producing 5 or more embryos), while only 2 out of 10 females for other lines superovulated.ConclusionsThe results of this work suggest that age and genetic background of the female are the most important factors contributing to superovulation success and that latency to mating is a good predictor of the number of embryos to be recovered. Surprisingly we found that cohabitation with the male prior to mating is not necessary for the recovery of embryos but is necessary to recover oocytes. This information will dramatically reduce the number of females required to generate embryos for transgenesis in this species.

Reinsemination intervals after timed artificial insemination or estrus-detected inseminations (2012)

The objective was to quantify the reinsemination intervals of lactating dairy cows that were either inseminated at estrus or received a timed AI (TAI) at first service. Cows in Experiment 1 were enrolled in a TAI program before first AI after calving. Cows detected in estrus after 50 days in milk (DIM) were inseminated, whereas the remainder continued in the TAI program and were inseminated as scheduled. Cows in Experiment 2 also were enrolled in a TAI program and were inseminated accordingly at first service after calving. On day 7 after TAI, cows were assigned randomly to receive either saline (control) or 1,000 IU human chorionic gonadotropin (hCG) to induce accessory luteal structures (corpora lutea) in an attempt to improve pregnancy outcome. First-repeat insemination dates were recorded for all cows after the initial AI and grouped as 25 days since first AI. More cows in Experiment 1 that were inseminated at estrus returned to estrus before 25 days than TAI cows and during summer months had shorter average return intervals by 1.7 days. More cows in Experiment 2 that received saline and had no accessory luteal structures also returned to estrus before 25 days than cows receiving hCG. Equal proportions of saline and hCG-treated cows (25%) in Experiment 2 had retained at least one of their original luteal structures until day 28 after TAI, but were not pregnant at day 32. Of those nonpregnant cows that retained luteal structures, average concentrations of pregnancy-specific protein B (BioPRYN test) concentrations were slightly elevated, but failed to retain the embryos to day 32 after AI. Furthermore, progesterone concentrations of these cows that lost their embryos were compromised compared with pregnant cows by day 21 after AI. Regardless of the number of luteal structures after first insemination, 25% were retained up to 28 days after AI, indicating pregnancy had occurred but embryo loss occurred between pregnancy recognition (day 15) and days 28 to 32 after insemination. Cows receiving TAI also had longer reinsemination intervals than cows inseminated at estrus, a phenomenon that is exaggerated during summer heat stress.