• Journal of Embryo Transfer
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• v.13 no.3
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• pp.261-275
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• 1998

본 연구에서는 GnRH가 과배란 처치된 래트의 초기 난포기와 후기 난포기에서 난소기능에 어떠한 영향을 미치는지를 이해하기 위해서, 30IU PMSG와 10IU hCG로 전처치된 미성숙 래트에 있어서 배란반응, 배란 난자의 형태학적 이상 유무 및 핵 성숙도, 난소 중량, 난소의 조직학적인 변화 및 혈중 스테로이드 호르몬 (17$\beta$-estradiol, progesterone 및 testosterone) 농도에 대하여 GnRH agonist의 효과를 검사하였다. GnRH agonist는 PMSG 전처치 후 초기 난포기 (PMSG 투여 후 6시간부터) 또는 후기 난포기(PMSG 투여 후 54시간부터)에 4시간 동안 20분 간격으로 경정맥 카테타를 통해 혈관내로 투여하였다. 각 실험동물은 혈중 스테로이드 호르몬의 변화를 측정하기 위하여 PMSG 투여 후 54시간, 72시간에 혈액을 채취하고 72시간에 희생시켰다. PMSG로 전처치한 미성숙 래트의 초기 난포기에 GnRH agonist의 투여는 GnRH agonist를 투여하지 않은 군(대조군)에 비해 과배란 억제, 형태학적 비정상 배란난자의 증가, 난소 중량의 감소, 난포폐쇄의 증가 및 혈중 스테로이드 호르몬의 농도 감소가 보였다. 한편 후기 난포기에 GnRH agonist의 투여는 대조군에서의 반응과 전반적으로 유사하였다. 이상의 결과, PMSG 및 hCG 처치로 과배란된 래트의 초기 난포기에 GnRH agonist의 투여는 난소기능을 전반적으로 억제하지만, 후기 난포기에 GnRH agonist의 투여는 난소기능에 영향을 미치지 않았다.

• Development and Reproduction
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• v.13 no.4
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• pp.353-362
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• 2009

GnRH and its receptor are known to express locally in the ovary and to regulate the ovarian function by affecting on granulosa and lutein cells. It has been reported that GnRH directly causes apoptosis in the granulosa and lutein cells of the ovary. However, whether the apoptosis of the cells by GnRH is recovered by FSH as an anti-apoptotic factor is not yet known. In this study, we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ after treatment with 5, 50, and 100 ng/$m\ell$ GnRH and 1 IU/ml FSH in the granulosa-lutein cells that are obtained during oocyte-retrieval for IVF-ET. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in the cells treated with 100 ng/$m\ell$ GnRH. In addition, we found that FSH suppresses the apoptosis of the cells induced by GnRH. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by GnRH treatment, whereas $E_2$ production was not changed. We also demonstrated that FSH inhibits the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present results suggest that GnRH agonist using in ovarian hyperstimulation protocol might induce the dysfunction of the ovary, but its function could be recovered by FSH. These results also will be expected to use as the basic data to elucidate the physiological role of GnRH and to develop new ovarian hyperstimulation protocols for IVF-ET.

• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.384-394
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• 2004

This study was carried out to monitor the response of ovaries and cyst according to treatment with GnRH or combination of GnRH and $PGF_2{\alpha}$ in dairy cows with ovarian follicular cysts. Thirty cows were diagnosed as having follicular cysts by rectal palpation, ultrasonography and progesterone (P4) assays. Ten cows were treated with GnRH (control), and the other twenty were treated with $PGF_2{\alpha}$ at 10 days after GnRH treatment. All the animals were re-examined by ultrasonography and blood was collected for the measurement of plasma P4 concentration at day 0 (the day of treatment), day 7, day 10, day 13, day 24 and day 34, respectively. In 30 cows that were diagnosed with follicular cysts, mean plasma P 4 concentrations on day -II and day -I were 0.3 ng/ml and 0.4 ng/ml. On day 10 increased as 2.7$\pm$0.2 ng/ml. Mean cystic wall thickness by ultrasonography on day -11 and day -I were 2.1 mm and 2.2 mm. In 9 cows responded on luteinization of cystic wall, cystic wall thickness was 3.9$\pm$0.5 mm at day 10 after GnRH treatment. The responses of ovaries until day 10 after GnRH treatment included development of corpus luteum in the ovary bearing the cyst or in the contralateral ovary (12 cows), luteinization of cystic wall (6 cows) and clouding of the anechoic antrum of cysts (2 cows). The ovarian responses according to the combination of GnRH and $PGF_2{\alpha}$ included regression of the corpus luteum (12 cows), increase (1 cow) and no change (1 cow) of cyst size until last examination, and complete disappearance on day 13 (6 cows), 23 (6 cows) and 34 (4 cows). Combination treatment group of GnRH and $PGF_2{\alpha}$ showed a higher pregnancy rate within 100 days after initial treatment (40.0 vs 65.0%) and shorter intervals from the treatment to conception (45.4$\pm$25.8 vs 53.5$\pm$31.4 days) compared with control. It was concluded that the administration of $PGF_2{\alpha}$ following GnRH treatment is effective in shortening the interval from treatment to conception in cows with follicular cyst. Also, this study suggested that the response of the cyst according to treatment revealed various types. Therefore, veterinarians should pay attention to monitor of the response of cystic ovaries after treatment, specially no change, slowly decrease or increasement of cyst size after treatment.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.4
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• pp.265-270
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• 2001

Objective : The aim of this study was to evaluate the outcome the GnRH antagonist (Cetrotide) short protocol in controlled ovarian hyperstimulation comparing with GnRH agonist long protocol. Materials and Method: From July 2000 to November 2001, 26 patients, 28 cycles were performed in controlled ovarian hyperstimulation by GnRH antagonist and GnRH agonist. GnRH antagonist (Cetrotide) was administered in 12 patients (14 cycles, Group 1) and GnRH agonist (Lucrin, Sub Q, Group 2) in 14 patients (14 cycles). Ovulation induction was performed by hMG (Pergonal) in group 1, and by Combo (Metrodine HP + Pergonal) in group 2. We compared the fertilization rate, good quality embryo, and clinical pregnancy rate between the two groups. Student-t test and Chi-square were used to determine statistical significance. Statistical significance was defined as p<0.05. Results: Ovarian hyperstimulation syndrome did not occurred in which estradiol (E2) level was $3874{\pm}809\;pg/ml$ and the number of retrieved oocytes was $18.4{\pm}2.4$. The number of used gonadotropin ampules was significantly decreased in Group 1 (26.0 vs. 33.1, p<0.04). There were no significant difference in the number of preovulatory oocyte ($10.6{\pm}6.9$ vs. $10.0{\pm}6.1$), fertilization rate ($74.8{\pm}23.4$ vs. $72.2{\pm}21.8$), good quality embryo ($58.7{\pm}23.6$ vs. $38.7{\pm}36.6$), and embryo transfer ($4.3{\pm}1.6$ vs. $4.4{\pm}1.6$). Although the age of the group 1 was older than the group 2 (34.4 vs. 30.8), there was no significant difference in clinical pregnancy rate (50.0% vs. 57.1%). Conclusions: We suggest that GnRH antagonist was a safe, effective, and alternative method in the controlled ovarian hyperstimulation, especially in PCOD patients who will be develop the ovarian hyperstimulation syndrome.

• Journal of Aquaculture
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• v.9 no.1
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• pp.3-10
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• 1996

Ovulation of maturing female yellow puffer, Takifugu obscrus, was induced by using single injection of human chorionic gonadotropin (HCG) or gonadotropin releasing hormone-analogue (GnRH-A) $des-Gly^{10}[D-Ala^6]$ GnRH-ethylamide plus pimozide. The response was evaluated using the fertilization and embryo-formation rate after insemination and the gonadotropin (GTH) level in blood plasma using radioimmunoassay. In the fertilization and embryo-formation, maximal effects were recorded by using 1,000 IU/kg HCG or $10\;{mu}g/kg$ GnRH-A plus 5 mg/kr pimozide. Pimozide (1, 5 mg/kg) or GnRH-A treatment alone was not effective in elevation of GTH level, however combinations of these treatments were particularly effective. Injection of dopamine blocked the rapid elevation of plasma GTH levels of blood. These data suggest that yellow puffer secrete GnRH and gonadotropin-releasing-inhibiting factor during the spawning or the other period.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.3
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• pp.191-200
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• 2004

Objective: To compare the clinical efficacy of double intrauterine insemination with single intrauterine insemination in GnRH antagonist combined ovarian hyperstimulation (Mild ovarian hyperstimulation) Materials and Methods: From Jan. 2001 to Jul. 2004, a retrospective clinical analysis was done of a total of 295 cycles in 170 patients who underwent ovarian hyperstimulation for ART (assisted reproductive technique). Subjects were divided into three groups; only clomiphene citrate ovarian hyperstimulation (n=55, 95cycles), GnRH antagonist combined ovarian hyperstimulation (soft ovarian hyperstimulation) (n=66 99cycles), and GnRH agonist combined ovarian hyperstimulation (short protocol) (n=49, 101cycles) Each group were randomly devided into two subgroups. One group underwent single IUI and the other group underwent double IUI. Results: GnRH antagonist group and GnRH agonist group had similar pregnancy rate. In GnRH antagonist Group, pregnancy rate was 36.1% in single IUI subgroup and was 36.6% in double IUI subgroup. These finding were not statistically significant. And Pregnancy rate was 20.8% in single IUI subgroup and was 19.3% in double IUI subgroup in single clomiphene citrate group, and 36.3% in single IUI subgroup and was 33.3% in double IUI subgroup in GnRH agonist group. These finding were not statistically significant, too. Conclusion: Pregnancy rate of GnRH antagonist was high and complication rate such as OHSS and multiple pregnancy was lower. In GnRH antagonist group, to compare with single IUI and double IUI, the result do not statistically differ. So GnRH antagonist single injection with single IUI was relatively comparable ART in infertiliry patient.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.3
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• pp.261-268
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• 2005

Objective: To compare the clinical results and pregnancy outcomes of in vitro fertilization (IVF) between GnRH antagonist cycles and GnRH agonist (GnRH-a) cycles including flare-up and long protocol in women with advanced age. Materials and Methods: Retrospective clinical study. From January 2001 to September 2003, IVF cycles of female patient 37 years over were included in this study. GnRH-a long protocol (62 cycles, 61 patients) and GnRH antagonist multi-dose flexible protocol (66 cycles, 51 patients) were compared with the control group of GnRH-a flare-up protocol (151 cycles, 138 patients). IVF cycles for non-obstructive azoospermia (NOA), endometriosis III, IV and polycystic ovarian syndrome (PCOS) were excluded in this study. Clinical results such as total gonadotropin dose, serum E2 on hCG administration, the number of retrieved oocytes and the pregnancy outcomes - clinical pregnancy rate (CPR), implantation rate (IR) and live birth rate (LBR) per embryo transfer - were compared. Results: There were significant differences in the total dose of gonadotropin (GnRH-a flare-up vs. GnRH-a long vs. GnRH-antagonist; 41.8 vs. 54.7 vs. 24.8), serum E2 on hCG administration (1787.2 vs. 1881.6 vs. 788.0), the numbers of retrieved oocytes (8.1 vs. 11.1 vs. 4.5) and endometrial thickness (9.1 vs. 10.4 vs. 8.0) which were significantly lower in GnRH-antagonist cycles. But pregnancy outcomes shows no significant differenced in CPR (25.0% vs. 35.8% vs. 24.5%), IR (11.7% vs. 12.3% vs. 10.1%) and LBR (15.8% vs. 28.3% vs. 15.1%) Conclusion: In women with advanced age, GnRH-antagonist cycles can result in comparable pregnancy outcomes to GnRH-a cycles including flare-up and long protocol. GnRH-a long protocol show higher CPR, IR and LBR than GnRH antagonist multi-dose flexible protocol and flare-up protocol without significant differences.

• Journal of Aquaculture
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• v.9 no.3
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• pp.205-213
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• 1996

Experiments were carried out to investigate the effect of GnRH-analogue (GnRH-a) on the induction of ovulation in catfish, S. asotus. Fully matured female catfish ($250\~600\;g$) received a single intraperitoneal injection of GnRH-a ($50\~200\;{\mu}g/kg{\cdot}body$ weight) showed the successful induction of ovulation. More than $86\%$ of treated females were ovulated after injection of GnRH-a ($90\;{\mu}g/kg$) at $25{\pm}1^{\circ}C$. The majority of spawning took place within 22 to 25 hours after the injection. The gonadosomatic index (GSI) and pseudo-GSI in the group treated with $120\;{\mu}g/kg$ GnRH-a were $23-30\%$ and $18-21\%$, respectively. Average fertilization and hatching rates were $94\%\;and\;81\%$, respectively. Electron microscopically, gonadotrophs of maturing female catfish were characterized by the presence of numerous small, electron-dense granules of approximately $150\~300$ nm in diameter and a few larger, less electron-dense granules of approximately $800\~1000$ nm in size in their cytoplasm. Gonadotrophs of GnRH-a treated catfish showed that their was a distinct decrease in number of small and large granules. The rough endoplasmic reticulum was composed of numerous cisternae conspicuosly dilated to various degrees.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.2
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• pp.115-123
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• 2006

Objective: To investigate new signal transduction cascade through integrin, FAK and ERK in the suppressed cell proliferation by GnRH-I and -II. Method: Human endometrial cancer cells (HEC1A) were cultured under the following condition: DMEM/F12 (10% FBS). GnRH-I and -II were treated time (0, 5, 10, 15, 20, 30 min; 100 nM) and dose (10 nM or 100 nM; 20 min) dependent manner according to experimental purposes. Cell proliferation was measured using [$^3H$] thymidine incorporation assay. Immunoblotting was utilized to detect proteins. Results: GnRH-I and -II inhibited proliferation of HEC1A cells and induced expression of integrin ${\beta}3$. Phosphorylation of FAK and ERK were induced by GnRH-I and -II. Conclusion: GnRH inhibited cell proliferation via the expression of integrin and FAK, ERK phosphorylation.

• Molecules and Cells
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• v.25 no.1
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• pp.91-98
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• 2008

The Glu/$Asp^{7.32}$ residue in extracellular loop 3 of the mammalian type-I gonadotropin-releasing hormone receptor (GnRHR) interacts with $Arg^8$ of GnRH-I, conferring preferential ligand selectivity for GnRH-I over GnRH-II. Previously, we demonstrated that the residues (Ser and Pro) flanking Glu/$Asp^{7.32}$ also play a role in the differential agonist selectivity of mammalian and non-mammalian GnRHRs. In this study, we examined the differential antagonist selectivity of wild type and mutant GnRHRs in which the Ser and Pro residues were changed. Cetrorelix, a GnRH-I antagonist, and Trptorelix-2, a GnRH-II antagonist, exhibited high selectivity for mammalian type-I and non-mammalian GnRHRs, respectively. The inhibitory activities of the antagonists were dependent on agonist concentration and subtype. Rat GnRHR in which the Ser-Glu-Pro (SEP) motif was changed to Pro-Glu-Val (PEV) or Pro-Glu-Ser (PES) had increased sensitivity to Trptorelix-2 but decreased sensitivity to Cetrorelix. Mutant bullfrog GnRHR-1 with the SEP motif had the reverse antagonist selectivity, with reduced sensitivity to Trptorelix-2 but increased sensitivity to Cetrorelix. These findings indicate that the residues flanking $Glu^{7.32}$ are important for antagonist as well as agonist selectivity.