• 한국수정란이식학회지
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• 제26권4호
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• pp.229-235
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• 2011

Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea (CL) formation to predict the possible involvement of Ski in luteinization. In addition, we performed to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

• 한국수정란이식학회지
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• 제7권2호
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• pp.117-124
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• 1992

미성숙 래트의 외경정맥에 카테타를 장착하고, 다음날 (28일령) 대조군에는 4IU, 다배란 처치 군에는 20IU의 PMSG를 피하주사하였다. 각 실험동물은 혈중의 LH농도 변화를 측정하기 위하여 PMSG 투여 직전 (0시간), 투여후 12시간, 그 이후 6시간 간격으로 혈액을 채취하고 72시간에 희생시켰다. 그 결과 다배란 용량의 PMSG 투여는 먼저 배란반응 및 난소중량을 대조군에 비하여 각각 4,7배 및 2.1배나 현저하게 (P<0.05) 증가시켰다. 그리고 혈청 LH농도는 Radioimmunoassay(RIA)에 의하여 결정되었는데, 먼저 두 군 모두 두 개의 분명한 peak을 가진 경시적 변화관계를 보였다. 즉 이들 두 군네 있어서 LH농도 변화는 0-18시간대에 처음으로 완만한 증가와 54-60시간대에 두번째는 급격한 증가(surge)를 보였다. 그러나 두 군간에 LH농도의 크기는 현저하게 달라, 다배란처치군의 동물에 있어서는 두번째의 LH peak에 앞서 전반적인 LH농도가 대조군보다 현저하게 (P<0.001) 높았으며, 반대로 PMSG 투여후 60시간에 일어나는 peak에 있어서는 LH농도가 대조군보다 현저하게 (P<0.001) 54%나 낮았다. 덧붙여 두 peak간의 증가폭은 대조근에 비하여 다배란 처치군에서 훨씬 낮았다. 본 열구 결과는 PMSG 처치된 래트에 있어서 두 가지의 분명한LH peak의 존재를 정의하며, 다배란 처치에 따른 난소과잉 자극과 내인성 LH surge의 감소와의 연관성을 밝힌다.

• Clinical and Experimental Reproductive Medicine
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• 제20권1호
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• pp.37-43
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• 1993

In 27 patients with the past history of poor response to the gonadotropin superovulation induction due to poor follicular growth or permature surge of endogenous luteinizing hormone, the effectiveness of pituitary supperssion with the gonadotropin releasing hormone agonist(GnRH-a) in in vitro fertilization(IVF) program was evaluated in 43 cycles using a combination regimen of D-Trp-6 LHRH(Decapeptyl, Ferring)and FSH/hMG from June, 1989 to August, 1990 at Korea University Hospital IVF Clinic. At midluteal phase of menstrual cycle, Decapeptyl-CR was administered by long-term protocol to minimize initial agonistic effect of endogenous gonadotropins. After the confirmation of pituitary suppression, about 2-3 weeks after GNRH-a administration, ovarian follicle growth was stimulated with FSH/hMG and followed by transvaginal ultrasonic measurement of follicle size and by monitoring of serm E2 and LH if necessary. When compared with the control group stimulated with gonadotropin regimen only, the cancellation rate and occurrence rate of premature LH surge during gonadotropin treatment were significantly lower in study group(11.6% and 2.4%, respectively). There is no significant differences in the mean number of aspirated oocytes, fertilization/cleavage rate, embryo transfer(ET) rate, and mean number of embryos transferred between the two groups. The pregnancy rate per treatment cycle, 16.3%, and per ET cycle, 23.3%, were significantly higher in the study group compared with those of control group. These data suggest that GnRH-a therapy is effective for previous poor responder In gonadotropin superovulation induction for IVF.

• Clinical and Experimental Reproductive Medicine
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• 제40권2호
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• pp.90-94
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• 2013

Objective: To evaluate the efficacy of earlier oocyte retrieval in IVF patients with a premature LH surge on hCG day. Methods: One hundred forty IVF patients (164 cycles) with premature LH surge on hCG day were included, retrospectively. We divided them into 2 study groups: LH surge with timed ovum pick-up (OPU) 36 hours after hCG injection (group B, 129 premature cycles), and LH surge with earlier OPU within 36 hours after hCG injection (group C, 35 cycles). Control groups were tubal factor infertility without premature LH surge (group A, 143 cycles). Results: The mean age (year) was statistically higher in group C than in groups A or B ($38.2{\pm}5.4$ vs. $36.2{\pm}4.2$ vs. $36.8{\pm}4.9$, respectively; p=0.012). The serum LH levels (mIU/mL) on hCG day were significantly higher in group B and C than in group A ($22.7{\pm}14.9$ vs. $30.3{\pm}15.9$ vs. $3.2{\pm}2.9$, respectively; p>0.001). Among groups A, B, and C, 4.9%, 31.7%, and 51.4% of the cycles, respectively, had no oocytes, and the overall rates of cycle cancellation (OPU cancellation, no oocyte, or no embryos transferrable) were 15.4%, 65.9%, and 74.3%, respectively. The fertilization rate (%) was significantly higher in group B than in group C ($73.2{\pm}38.9$ vs. $47.8{\pm}42.9$, p=0.024). The clinical pregnancy rate was significantly higher in group C than in groups A and B (44.4% vs. 27.3% vs. 9.1%, respectively, p=0.021). However, the miscarriage rate was also higher in group C than in group B (22% vs. 0%, respectively, p=0.026). Conclusion: Earlier OPU may not be effective in reducing the risk of cycle cancellation in patients with premature LH surge on hCG day. A larger scale study will be required to reveal the effectiveness of earlier ovum retrieval with premature LH surge.

• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.199-205
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• 1998

흰쥐 난소에서의 Luteinizing hormonoe (LH) subunit 유전자 발현과 LH polypeptide의 존재를 조사하였다. 이를 위해 LH subunit들에 대한 역전사 중합효소연쇄반응 (RT-PCR)을 시행하고, 난소내 LH 함량을 방사면역측정법으로 정량하였다. 뇌하수체와 정소에서 공통적으로 존재하는 LH-$\beta$ subunit$(LH-\beta})$의 exon에 해당되는 primer를 사용하여 RT-PCR을 시행한 결과 흰쥐 난소에서도 뇌하수체, 정소와 같이 306 bp band가 확인되었고, 정소특이적인 exon에 해당되는 primer를 사용한 결과 정소와 난소에서 예상대로 428 bp band가 검출되었다. 또한 LH, FSH, TSH 그리고 hCG에서 공통적으로 발현되는 common $\alpha$-subunit $(C_\alpha)$의 전사물질도 PCRdml 의해 증폭되었다. 방사면역측정법에서는 LH standard curve와 난소추출물을 사용한 curve가 동일하게 sigmoid 형태를 보임으로서 흰쥐 난소내에 immunoreactive LH가 존재함이 증명되었다. 인위적으로 성적인 성숙을 유도한 PMSG 주사 동물에서 혈중 LH 수준은 주사 후 48시간에 preovulatory LH surge와 유사한 최고 수준을 나타냈으나, 난소내 LH 함량의 경우 주사 24시간 후부터 급격히 감소하여 주사 48, 72시간군까지도 낮은 수준이 유지되었다. 이 결과는 흰쥐 난소의 LH가 생리적으로 조절되고 그 조절방식이 뇌하수체에서와는 다를 가능성을 시사하는 것이다. 본 연구는 흰쥐의 난소에서 LH가 유전자가 발현됨을 최초로 보고한 것이며, LH의 경우 내분비적 경로 (endocrine; 뇌하수체로 부터의 LH)외에도 국부적 경로 (autocrine이나 paracrine; 난소내에서 합성되는 LH)를 통해 난소의 생리와 기능 조절을 담당함을 시사한다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.355-361
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• 2011

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to locate Ski protein in the rat ovary during luteinization to predict the possible role of Ski. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadotropin to immature female rat, and luteinization was induced by human chorionic gonadotropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally.

• 한국임상수의학회지
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• 제31권1호
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• pp.19-24
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• 2014

본 연구의 목적은 LH surge 형태의 분비가 선행적으로 투여되는 각각 다른 Progesterone (P) 농도에 의해 어떻게 조절되는지에 대한 탐구이다. 선행된 연구에서 LH surge는 난포기(follicular phase)에서 보이는 P 농도에서만 발현되었고 황체기(luteal phase)와 아황체기(subluteal phase) P 농도에서는 완전히 억제되었다. 이런 결과를 토대로 본 연구의 가설은 아황체기 P농도(스트레스, 황체형성부전)와 황체기 P 농도를 선행 처치 후 외인성 estradiol에 의한 LH surge 발현은 동시에 발생할 것으로 예상한다. 이 가설을 증명하기 위해 난소가 제거된 Shiba 염소를 이용하여 인위적으로 난포기(non-P), 아황체기(low-P), 기능성황체기(high-P)의 P 농도 및 Estradiol (E) 농도를 유도하였다 (Day 0: P와 E 이식일). 스테로이드 호르몬의 농도를 알아보기 위해 매일 혈액을 채혈하였으며, Day 7에 각 그룹의 P 농도를 유지하기 위한 packet를 제거하고 E 처치용 capsule은 그대로 유지시켰다. 또한 Day 7에 P와 E의 농도변화를 세밀하게 조사하기 위해 12시간 동안 1시간 간격으로 채혈 후 검사하였다. LH surge 분비패턴은 P packet 제거 후 13시간부터 E를 3 ${\mu}g/h$ 농도로 36시간 동안 주입하고, 52시간 동안(E 주입 4시간 전부터 주입 후 48시간) 2시간 간격으로 채혈을 실시했다. 그 결과, 모든 3그룹의 염소는 LH surge가 발현되었지만 그 발현시간은 각 그룹마다 유의성있는 발현시간 차를 나타내었다. 즉, 각각의 선행된 P 농도 (non-P, low-P, high-p) 그룹은 외인성 Estradiol에 의한 LH surge 발현시간에 농도의존성으로 영향을 미치는 것으로 판단되었다.

• 한국임상수의학회지
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• 제31권1호
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• pp.25-30
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• 2014

본 연구의 목적은 외인성 Estradiol(E)에 의해 발현되는 LH surge가 다른 농도의 Progesterone (P)에 억제되고, 그 기전이 E의 존재에 의해 더욱 강화된다는 것을 증명하는 것이다. 선행된 연구에서 E의 존재하에 LH surge는 P의 농도에 의해 LH surge 발현 시간은 점진적/유의성 있게 억제된다는 것을 확인했다. 이런 결과를 토대로 난소가 제거된 Shiba 염소를 이용하여 인위적으로 난포기(Non-P), 아황체기(Low-P), 기능성황체기(High-P)의 P 농도 및 Estradiol (E) 농도를 유도하였다 (Day 0: P와 E 이식일). 스테로이드 호르몬의 농도를 알아보기 위해 매일 혈액을 채혈하였으며, Day 7에 각 그룹의 P 농도를 유지하기 위한 packet과 E 처치용 capsule를 동시에 제거하였다. 또한 Day 7에 P와 E의 농도변화를 세밀하게 조사하기 위해 12시간 동안 1시간 간격으로 채혈 후 검사하였다. LH surge 분비패턴은 P packet 제거 후 13시간부터 E를 3-6 ${\mu}g/h$ 농도로 36시간 동안 주입하고, 52시간 동안(E 주입 4시간 전부터 주입 후 48시간) 2시간 간격으로 채혈을 실시했다. 그 결과, 모든 3그룹의 염소는 LH surge가 발현되었지만 그 발현시간은 Low-P와 High-P 그룹에서 유의성 없는 발현시간 차를 나타내었다. 즉, 각각의 선행된 P 농도 (Non-P, Low-P, High-p) 그룹은 외인성 Estradiol에 의한 LH surge 발현을 조절하는데 있어서 E의 존재가 중요하며, E는 GnRH/LH surge-generating 시스템에 작용하는 P의 억제적 효과를 더욱 강화시키는 것으로 생각되었다.

• 한국수정란이식학회지
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• 제26권4호
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• pp.237-244
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• 2011

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vitro models. RT-PCR and real time PCR analysis respectively revealed that LH had no effect on c-Ski mRNA expression in the cultured granulosa cells regardless of LH treatment. Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally. Moreover, expression of mRNA of Arkadia, an E3 ubiquitin ligases, in luteinizing granulosa cells in vivo was assessed by realtime-PCR. The levels of Arkadia mRNA expression were unchanged during follicular growth and postovulatory luteinization. These findings suggest that Ski protein level may be regulated during luteinization at translational and/or post-translational level but not by Arkadia.

• Clinical and Experimental Reproductive Medicine
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• 제15권2호
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• pp.103-117
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• 1988

Ovulation induction was done with 3 different regimens as clomid combined with HMG, HMG only, and FSH combined with HMG in 28 menstrual cycles for IVF-ET and GIFT program. The appearance of endogenous LH surge, estradiol plateau, atypical LH surge, and time from initiation to peak of LH surge in serum and urine were observed and compared in 3 groups. 1. The estradiol concentration of serum LH surge day was similar in three groups but 1st group (Clomiphene Citrate+Sequential HMG) was slightly higher at $1924.0{\pm}865.1\;pg/ml$. In regards to the existence of serum estradiol plateau, 3rd group (FSH+Sequential HMG) was highest at 60%, and 1st group and 2nd group (HMG only) were similar at 33% and 44% respectively. 2. The number of ovarian of ovarian follicle which was more than 18mm in diameter was $4.1{\pm}2.0$, $4.2{\pm}2.1$ respectibely for 2nd group and 3rd group. Although the numbers were slightly higher thean 1st group for each ovarian follicle, serum estradiol value per follicle was higher for 1st group at $583.0{\pm}261.2pg/ml$. 3. When measuring the urine LH surge according to Hi-Gonavi and according to the standard set by three different types of surge, simultameous satisfaction for 1st group, 2nd group, 3rd group was two cases, five cases, four cases respectively at 40%, and the remained cases were composed of numorous type combination which satisfied the two definition, simultaneously in this study, the LH surge starting time was determined only in the cases tow combination were satisfied simultaneously at first, but there are something to study more. In one case of the 3rd group. 4. The concentration of LH surge start in urine and serum of 2nd group was highest at306. $0{\pm}287.2IU/l$ and $34.0{\pm}9.9mIU/ml$ and 1st group was low at $116.6{\pm}66.1IU/l$ and 7.4mIU/ml. The urine and serum value of LH was highest at $1644.4{\pm}988.8IU/l$, $65.9{\pm}15.0mIU/ml$ for 2nd group, 1st group was low at urine, and 3rd group was low of serum. With pregnancy established, the LH concentration of urine was relatively high but on the contrary the LH concentration of serum was low compared to urine concentration. 5. Time from LH surge start to the maximun of urine and serum value was highest at 15. $7{\pm}9.1$ hrs and $10.8{\pm}4.9$ hrs for 1st group and 3rd group. With pregnancy established, time was shortened for urine but on the contrary serum showed an increase in time. 6. The concentration of LH which increases with time on urine was highest at 2nd group ($194.6{\pm}76.8\;IU/hour$). The lowest increase for serum was at 3rd group (2.1mIU/hour). With pregnancy established, urine showed more increase than control group ($266.5{\pm}47.4\;IU/hour$) and for serum there was similar increase ($3.4{\pm}0.8\;mIU/hour$). 7. There were two examples of non-typical surge from 1st group and 3rd group each. Among these three cases showed a reestablishment of LH surge after 10-24 hours. 8. Endogenous spontaneous Lh surge occurs once for each 2nd group and 3rd group.