• Asian-Australasian Journal of Animal Sciences
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• 제27권9호
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• pp.1270-1275
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• 2014

An experiment was conducted to evaluate the efficacy of porcine follicle stimulating hormone (pFSH) dosage based on body weight (BW) on ovarian responses of crossbred does. Thirty donor does were divided into 3 groups getting pFSH dosages of 3, 5, and 8 mg pFSH per kg BW, respectively, and were named as pFSH-3, pFSH-5 and pFSH-8, respectively. Estrus was synchronized by inserting a controlled internal drug release (CIDR) device and a single injection of prostaglandin $F2{\alpha}$ ($PGF2{\alpha}$). The pFSH treatments were administered twice a day through 6 decreasing dosages (25, 25, 15, 15, 10, and 10% of total pFSH amount; decreasing daily). Ovarian responses were evaluated on Day 7 after CIDR removal. After CIDR removal, estrus was observed 3 times in a day and pFSH treatments were initiated at 2 days before the CIDR removal. All does in pFSH-5 and pFSH-8 showed estrus signs while half of the does in pFSH-3 showed estrus signs. No differences (p>0.05) were observed on the corpus luteum and total ovarian stimulation among the treatment groups, while total and transferable embryos were higher (p<0.05) in pFSH-5 (7.00 and 6.71) than pFSH-3 (3.00 and 2.80) and pFSH-8 (2.00 and 1.50), respectively. In conclusion, 5 mg pFSH per kg BW dosage gave a higher number of embryos than 3 and 8 mg pFSH per kg BW dosages. The results indicated that the dosage of pFSH based on BW is an important consideration for superovulation in goats.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.433-440
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• 1999

Objective: To evaluate the effectiveness of CC+FSH or CC+hMG in intrauterine insemination (IUI) cycles for the treatment of infertility. Method: Patients received daily 100 mg of clomiphene citrate (CC) for 5 days followed by hMG or FSH. A single IUI was performed at 36 h after hCG. Clinical pregnancy was classified if a gestational sac or fetal cardiac activity was seen on ultrasound. Results: The overall clinical pregnancy rate was 19.1% per cycle (17/89) and 21.5% per patient (17/79). More clinical pregnancies were recorded in CC+FSH (23.1%, 6/26) than CC+ hMG cycles (17.5%, 11/63), but this difference was not statistically significant. No differences were found in age, duration of infertility, follicle size, levels of estradiol ($E_2$) on the day of hCG injection and total motile sperm counts between pregnant and non-pregnant groups. However, more ampules of gonadotropins were used in pregnant group than non-pregnant group (p<0.05). Conclusion: Combination of CC and hMG may economically be more effective to induce ovulation for IUI compared to CC and FSH.

• 한국수정란이식학회지
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• 제19권2호
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• pp.81-87
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• 2004

본 연구는 재래산양에 있어서 과배란 처리시 공란 산양의 반복사용에 따른 oocyte의 회수방법과 회수란의 질적 개선방법을 확립하기 위하여 실시하였으며, 성선자극호르몬의 1회 처리와 반복처리, 호르몬제의 종류에 따른 oocyte의 회수율과 등급을 조사하였다. 공시동물은 체중 15~25kg 전${\cdot}$후의 성숙한 미경산 재래산양으로서 발정 동기화를 위하여 CIDR를 10일간 질내에 삽입하고 과배란 처리는 70 mg의 FSH 와 1,000 IU의 PMSG를 투여하여 과배란을 유기하였다. ${PGF_{2\alpha}$ 는 8일째에 투여하여 과배란을 유기하였으며, CIDR는 10일째 제거와 동시에 hCG 400 IU를 투여하였다. 난자의 회수는 hCG 투여 후 35~40시간째에 외과적인 방법으로 실시하였으며, 회수한 난자는 난구세포와 세포질의 부착정도에 따라 4등급으로 분류하여 회수율을 조사하였다. FSH와 PMSG를 투여하여 공란 산양으로 처음 사용하였을 때 49두에서 324(6.63${\pm}$0.63) 개와 27두에서 305(11.30${\pm}$1.29)개의 배란점을 확인하였으며, 이 중 43.8%(142/324)와 69.2%(211/305)가 회수되어 두당 회수율은 2.92${\pm}$0.52 개와 7.81${\pm}$1.14개로서 FSH와 PMSG간에 전체 회수율과 두당 회수율은 PMSG 처리구가 유의적(P<0.05)으로 높았다. 반복 사용하였을 때 FSH와 PMSG 처리구의 두당 배란수는 2.88${\pm}$0.69 개와 7.56${\pm}$1.11 개였으며, 회수율은 각각 17.4%(4/23) 및 44.9%(61/136) 로 PMSG 처리구가 유의적(P<0.05)으로 높았다. 두당 회수율은 0.50${\pm}$0.27개 및 3.39${\pm}$0.86개로서 PMSG 처리구가 유의적으로 높았다. 회수한 난포란의 1 및 2등급에 있어서 공란 산양으로 처음 사용하였을 때 FSH 투여구는 각각 20.6%(GI) 및 14.9%(GII)였으며, PMSG 투여구는 12.9%(GI) 및 14.1%(GII)로 나타났다. 반복 사용하였을 때, 회수한 난포란의 등급은 FSH 투여구는 각각 31.3%(GI) 및 28.1%(GII)였으며, PMSG 투여구는 36.4%(GI), 5.1%(GII)로 나타났다. 이상의 결과로 볼 때 배란된 난자의 회수율은 PMSG 가 FSH보다 높았으며, 난포내 난자의 회수율은 투여 호르몬 및 반복사용 여부에 따른 차이는 없었다.

• Reproductive and Developmental Biology
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• 제34권1호
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• pp.33-40
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• 2010

Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.

• Clinical and Experimental Reproductive Medicine
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• 제22권2호
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• pp.123-130
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• 1995

Many types of medication regimens have been used for controlled ovarian hyperstimulation for assisted reproductive technique(ART). Questions are now being raised regarding how to lower the escalating costs of assisted reproduction and decrease the extent of patient discomfort and disruption of life style without sacrificing success rates. In this investigation, from January 1994 through August 1994 patients presenting to the Chung-Ang university hospital, infertility clinic were offered the option of the clomiphene citrate (CC)/single Human Menopausal Gonadotropin(HMG) combination and conventional GnRH-agonist combination method. 60 patients (78 cycles) were given CC/single HMG combination as a study group, and 78 patients (102 cycles) were given conventional GnRH-a combined ultrashort protocol as a control group for IVF-ET program and the resulting number of oocyte retrieved, embryo produced, and pregnancy initiated were compared. There were no differences between the two groups in mean age, serum $E_2$, LH and FSH level on menstrual cycle day 2. HMG requirement was 2 ampules in study group and $24.2{\pm}6.8$ ampules in control group. On the day of HCG injection, serum LH and FSH levels were not significantly different, but serum $E_2$, was significantly higher in control group(p<0.001). There was relatively well endometrial quality in control group but not significant compare to study group. In control group, numbers of retrieved oocyte and transferred embryo were significantly more than study group(p<0.001). Fertilization rate was not significantly different in the two groups and pregnancy rates were 20.2% in study group 28.4% in control group(p<0.001). CC/single HMG protocol for IVF-ET is less expensive than GnRH-a combined ultrashort protocol and minimizes patients discomfort. In addition, CC/single HMG protocol produces acceptable pregnancy rate and represents an attractive alternative to select patients undergoing IVF-ET.

• The Korean Journal of Physiology and Pharmacology
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• 제27권2호
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• pp.167-176
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• 2023

This study aims to explore the impact of Rehmannioside D (RD) on ovarian functions of rats with diminished ovarian reserve (DOR) and its underlying mechanisms of action. A single injection of cyclophosphamide was performed to establish a DOR rat model, and fourteen days after the injection, the rats were intragastrically administrated with RD for two weeks. Rat estrus cycles were tested using vaginal smears. Ovarian tissues were histologically evaluated, the number of primordial, mature, and atretic follicles was calculated, and the apoptotic rate of granulosa cells. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E₂) levels were determined by ELISA assays. Protein levels of Forkhead Box O1 (FOXO1), KLOTHO, Bcl-2, and Bax were investigated in ovarian tissues of DOR rats. The binding between FOXO1 and KLOTHO was verified by ChIP assay. High-dose administration of RD into DOR rats improved their estrus cycles, increased ovarian index, enhanced the number of primordial and mature follicles, reduced the number of atretic follicle number, and ovarian granulosa cell apoptosis in addition to inhibiting FSH and LH levels and upregulating E₂ expression. FOXO1 and KLOTHO were significantly suppressed in DOR rats. FOXO1 knockdown partially suppressed the protective effects of RD on DOR rats, and KLOTHO overexpression could restore RD-induced blockade of DOR development despite knocking down FOXO1. FOXO1 antibody enriched KLOTHO promoter, and the binding between them was reduced in DOR group compared to that in sham group. RD improved ovarian functions in DOR rats and diminished granulosa cell apoptosis via the FOXO1/KLOTHO axis.

• 한국발생생물학회지:발생과생식
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• 제11권1호
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• pp.49-54
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• 2007

Ethane 1,2-dimethane sulfonate(EDS)은 Leydig cells(LC)만을 선별적 사멸을 유도하는 약물로서 가역적인 테스토스테론(testosterone, T) 결핍 흰쥐 모델을 만드는데 널리 사용된다. 본 연구에서는 수컷 흰쥐 뇌하수체의 생식소자극호르몬인 LH와 FSH의 발현에 미치는 EDS 투여 효과를 조사하였다. 성숙한 수컷 흰쥐(SD strain, $300{\sim}350\;g$ B.W.)에 EDS(75 mg/kg, i.p.)를 1회 복강주사하고 주사 후 0, 1, 2, 3, 4, 5, 6 그리고 7주가 경과한 날 희생시켰다. 뇌하수체로부터 total RNA를 추출한 후 뇌하수체 glycoprotein hormone common alpha subunit($C{\alpha}$), LH beta subunit($LH{\beta}$), FSH beta subunit($FSH{\beta}$) 그리고 GnRH 수용체(GnRH-R)의 발현 변화를 semi-quantitative RT-PCR로 측정하였다. 그 결과, $C{\alpha}$ 전사수준은 주사 후 1주부터 급격히 상승하여 주사 후 4주까지 유의하게 높게 유지되다가 5주 후부터 control 수준으로 회귀하였다. $LH{\beta}$ 전사 수준은 주사 후 2주부터 유의하게 상승하여 주사 후 4주에 최고 수준에 도달하였으며, 5주 후부터 control 수준으로 감소하였다. $FSH{\beta}$ 전사수준은 주사 후 2주부터 유의하게 상승하여 주사 후 3주에 최고 수준에 도달하였으며, 4주 후부터 감소하여 5주 후에 최소치를 보였다. 유사하게, GnRH-R 전사 수준도 주사 후 2주부터 유의하게 상승하여 주사 후 3주에 최고 수준에 도달하였으며, 5주 후부터 control 수준으로 감소하였다. 본 연구는 EDS 주사에 의해 수컷 흰쥐 뇌하수체 전엽에서의 생식소 자극호르몬 subunit들과 GnRH-R의 발현 변화가 가역적으로 유도될 수 있음을 보여준 것이다. EDS 주사 모델은 수컷 흰쥐에서의 시상하부-뇌하수체 신경내분비 축의 호르몬 조절에 대한 기작을 이해하는데 도움이 될 것이다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 1997년도 추계학술대회 및 워크숍
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• pp.47-47
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• 1997

• Asian-Australasian Journal of Animal Sciences
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• 제14권4호
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• pp.455-461
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• 2001

The effect of exogenous gonadotrophins on superovulation in rabbits was examined. One hundred and sixteen sexually mature California, Chinchilla and New Zealand White rabbits were randomly allocated to control (100 IU hCG), PMSG-treated (100 IU HCG following 150 IU PMSG) and FSH-treated groups (0.3 mg/head /12 h for 3 days followed by 100 IU hCG). All does were mated after hCG injection and were sacrificed or laparotomized within 1 to 4 days postcoitus for counting the number of ovulation points. The number of ovulations was higher in FSH-treated animals than in the control and PMSG-treated groups (37.2 vs. 10.4 and 14.5, p<0.05). Follicle haemorrhagicum was observed in many cases in the PMSG-treated group. No significant difference in ovulation number was observed between left and right ovaries regardless of gonadotropin treatment. In another experiment, 2-cell stage embryos were collected at 26 h postmating and blastomeres were separated by mechanical pipetting or gentle pressure with a fine glass needle. Aggregated or chimeric embryos were produced from two single blastomeres from two breeds, New Zealand White and Chinchlla, with different coat colors. All the embryos were cultured in Ham's F-10 medium supplemented with 1.5% BSA (bovine serum albumin fraction V) and 10% PRS (pregnant rabbit serum), and incubated in a humidified atmosphere with 5% $CO_2$ at $38^{\circ}C$. After development to morula or early blastocyst, the embryos were transferred into the oviducts of recipient does. Results showed that 7 out of 10 does (70%) receiving intact embryos (control) became pregnant and 41 kits were delivered. However, no pregnancy was obtained from the recipient of either denuded demi- or aggregated embryos. It is suggested that embryos without zona pellucida could not develop to term in rabbits.

• 한국발생생물학회지:발생과생식
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• 제14권1호
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• pp.35-41
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• 2010

Mammalian reproduction is regulated by a feedback circuit of the key reproductive hormones such as GnRH, gonadotropin and sex steroids on the hypothalamic-pituitary-gonadal axis. In particular, the onset of female puberty is triggered by gain of a pulsatile pattern and increment of GnRH secretion from hypothalamus. Previous studies including our own clearly demonstrated that genistein (GS), a phytoestrogenic isoflavone, altered the timing of puberty onset in female rats. However, the brain-specific actions of GS in female rats has not been explored yet. The present study was performed to examine the changes in the activities of GnRH neurons and their neural circuits by GS in female rats. Concerning the drug delivery route, intracerebroventricular (ICV) injection technique was employed to eliminate the unwanted actions on the extrabrain tissues which can be occurred if the testing drug is systemically administered. Adult female rats (PND 100, 210-230 g BW) were anaesthetized, treated with single dose of GS ($3.4{\mu}g$/animal), and sacrificed at 3 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly raised the transcriptional activities of enhanced at puberty1 (EAP-1, p<0.05), glutamic acid decarboxylase (GAD67, p<0.01) which are known to modulate GnRH secretion in the hypothalamus. However, GS infusion could not change the mRNA level of nitric oxide synthase 2 (NOS-2). GS administration significantly increased the mRNA levels of KiSS-1 (p<0.001), GPR54 (p<0.001), and GnRH (p<0.01) in the hypothalami, but decreased the mRNA levels of LH-$\beta$ (p<0.01) and FSH-$\beta$ (p<0.05) in the pituitaries. Taken together, the present study indicated that the acute exposure to GS could directly activate the hypothalamic GnRH modulating system, suggesting the GS's disrupting effects such as the early onset of puberty in immature female rats might be derived from premature activation of key reproduction related genes in hypothalamus-pituitary neuroendocrine circuit.