• 한국동물학회:학술대회논문집
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• 한국동물학회 2003년도 한국생물과학협회 학술발표대회
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• pp.166.3-166
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• 2003

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• Asian-Australasian Journal of Animal Sciences
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• 제13권5호
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• pp.595-599
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• 2000

Fine wool sheep (n=18) maintained in a tropical environment were allocated to three treatment groups. Estrus was induced with two injections of $PGF_{2{\alpha}}$ (10 mg. im) at 10 days interval. Superovulation treatment started 2 days prior to the second injection of $PGF_{2{\alpha}}$. Each ewe was treated with a total dose of 25 units FSH (Super-OV) i.m. every 12 hover 3 days; Group 2 were also injected i.m. with 200 IU PMSG at the first injection of FSH; Group 3 was treated as in Group 2 and also with GnRH ($4{\mu}g$ Buserelin) at the onset of estrus. The ewes in estrus were mated with a fertile ram. Ovarian examination and recovery of embryo and ova were performed at laparoscopy and laparotomy on day 3 or 4 after mating. Data for onset of estrus, duration of estrus, number of corpora lutea (CL), number of unnovulated large follicle (LF), embryo recovery rate, embryo quality and fertilization recorded for the 3 groups. Ewes in the Group 1 set in estrus later (p<0.05; $50.0{\pm}7.29h$) than the ewes in Group 2 ($24.5{\pm}3.58$) and 3 ($32.5{\pm}3.58h$). The duration of estrus, ovarian size and ovarian response (number of CL and LF) did not differ significantly (p>0.05) among the 3 groups. The proportion of ewes with a superovulatory response (${\geq}2$ CL) was the lowest (50%) in Group 1 treated with FSH alone but ova/embryo recovery (100%) and fertilization (100%) was significantly (p<0.05) higher than Group 2 (58.3 and 85.7%, respectively) and Group 3 (48.6 and 50%, respectively). It is concluded that in tropical fine wool sheep, there is no difference in the 3 treatments for yield of good quality embryos but ovarian response and ovulation rate increased on additional use of PMSG and GnRH respectively to FSH alone.

• Clinical and Experimental Reproductive Medicine
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• 제18권2호
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• pp.201-208
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• 1991

Recently the application of gonadotropin-releasing hormone (GnRH) agonist to superovulation in previous poor responders has resulted in the improved outcomes after in vitro fertilization (IVF) outcome. However, poor responders with poor estradiol $(E_2)$ rise or single dominant follicle are a particularly challenging group. Recent reports have also shown that patients with higher basal serum follicle stimulating hormone (FSH) level, result in poorer ovarian response and lower pregnancy rate. Analysis of the differences of superovulation outcomes according to the different protocols of GnRH agonist, long (L, n = 18) and short (S, n = 16) protocols, in patients with high basal FSH levels (>20mIU/ml) were undertaken at Seoul National University Hospital from June to October 1990. The administration of GnRH agonist was begun on day 21 of the cycle in long protocol, and on day 2 in short protocol. Ages of patients and husbands, basal FSH and luteinizing hormone (LH) levels and FSH/LH ratio did not differ significantly. Types and causes of infertility were evenly distributed. Whereas the duration of stimulation and the amounts of gonadotropins administered were significantly reduced in short protocol, the numbers of oocytes retrieved and cleaved, the cleavage rate and the number of embryos transferred were higher in long protocol without statistical signifieance. The pregnancy rate per ET was 16.7% (2/12) in short protocol, and 17.6% (3/17) in long protocol. These data suggest that both protocols result in the similar superovulation outcomes in patients with higher basal serum FSH levels.

• Animal Bioscience
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• 제35권12호
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• pp.1850-1859
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• 2022

Objective: The quantitative reverse transcription polymerase chain reaction (qPCR) is the most accurate and reliable technique for analysis of gene expression. Endogenous reference genes (RGs) have been used to normalize qPCR data, although their expression may vary in different tissues and experimental conditions. Verification of the stability of RGs in selected samples is a prerequisite for reliable results. Therefore, we attempted to identify the most stable RGs in the hypothalamic-pituitary-gonadal (HPG) axis in sows. Methods: The cycle threshold values of nine commonly used RGs (18S, HPRT1, GAPDH, RPL4, PPIA, B2M, YWHAZ, ACTB, and SDHA) from HPG axis-related tissues in the domestic sows in the different stages of estrus cycle were analyzed using two RG-finding programs, geNorm and Normfinder, to rank the stability of the pool of RGs. In addition, the effect of the most and least stable RGs was examined by normalization of the target gene, gonadotropin-releasing hormone (GnRH), in the hypothalamus. Results: PPIA, HPRT1, and YWHAZ were the most stable RGs in the HPG axis-related tissues in sows regardless of the stages of estrus cycle. In contrast, traditional RGs, including 18S and ACTB, were found to be the least stable under these experimental conditions. In particular, in the normalization of GnRH expression in the hypothalamus against several stable RGs, PPIA, HPRT1, and YWHAZ, could generate significant (p<0.05) elevation of GnRH in the preovulatory phase compared to the luteal phase, but the traditional RGs with the least stability (18S and ACTB) did not show a significant difference between groups. Conclusion: These results indicate the importance of verifying RG stability prior to commencing research and may contribute to experimental design in the field of animal reproductive physiology as reference data.

• 한국발생생물학회지:발생과생식
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• 제20권1호
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• pp.51-61
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• 2016

Neurokinin B (NKB) and neurokinin B related peptide (NKBRP) belong to tachykinin peptide family. They act as a neurotransmitter and/or neuromodulator. Mutation of NKB and/or its cognate receptor, NK3R resulted in hypogonadotropic hypogonadism in mammals, implying a strong involvement of NKB/NK3R system in controlling mammalian reproduction. Teleosts possess NKBRP as well as NKB, but their roles in fish reproduction need to be clarified. In this study, NKB and NKBRP coding gene (tac3) was cloned from Nile tilapia and sequenced. Based on the sequence, Nile tilapia NKB and NKBRP peptide were synthesized and their biological potencies were tested in vitro pituitary culture. The synthetic NKBRP showed direct inhibitory effect on the expression of GTH subunits at the pituitary level. This inhibitory effect was confirmed in vivo by means of intraperitoneal (ip) injection of synthetic NKB and NKBRP to mature female tilapia (20 pmol/g body weight [BW]). Both NKB and NKBRP had no effect on the plasma level of sex steroids, E2 and 11-KT. However, NKBRP caused declines of expression level of GnRH I, Kiss2 and tac3 mRNAs in the brain while NKB seemed to have no distinct effect. These results indicate some inhibitory roles of NKBRP in reproduction of mature female Nile tilapia, although their exact functions are not clear at the moment.

• Clinical and Experimental Reproductive Medicine
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• 제42권2호
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• pp.67-71
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• 2015

Objective: Elevated serum progesterone (P) levels on triggering day have been known to affect the pregnancy rate of in vitro fertilization (IVF). This study aimed to identify the possible factors influencing serum P levels on triggering day in stimulated IVF cycles. Methods: Three hundred and thirty consecutive fresh IVF cycles were included in the study. All cycles were first attempts and were performed in a single infertility center. The indications for IVF were male factor infertility (n=114), ovulatory infertility (n=84), endometriosis (n=61), tubal infertility (n=59), unexplained infertility (n=41), and uterine factor infertility (n=39). A luteal long protocol of a gonadotropin-releasing hormone (GnRH) agonist (n=184) or a GnRH antagonist protocol (n=146) was used for pituitary suppression. Ovarian sensitivity was defined as the serum estradiol level on triggering day per 500 IU of administered gonadotropins (OS[a]) or the retrieved oocyte number per 500 IU of administered gonadotropins (OS[b]). Results: Univariate analysis revealed that the serum P level on triggering day was associated with the serum estradiol level on triggering day (r=0.379, p<0.001), the number of follicles ${\geq}14mm$ (r=0.247, p<0.001), the number of retrieved oocytes (r=0.384, p<0.001), and ovarian sensitivity (OS[a]: r=0.245, p<0.001; OS[b]: r=0.170, p=0.002). The woman's age, body mass index, antral follicle count, and basal serum follicle stimulating hormone and estradiol levels were not associated with serum P level on triggering day. The serum P level on triggering day did not show significant variation depending on the type or cause of infertility, pituitary suppression protocol, or the type of gonadotropins used. Conclusion: The serum P level on triggering day was closely related to the response to ovarian stimulation.

• 한국발생생물학회지:발생과생식
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• 제16권2호
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• pp.121-128
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• 2012

최근 여러 포유류 종에서 Kisspeptin이 그 수용체인 GPR54와 함께 GnRH 분비를 자극하여 성 성숙과 최종배란에 영향을 미친다고 보고되고 있다. 이러한 보고들은 KiSS-GPR54 system이 번식과 밀접한 연관이 있음을 제시하고 있지만, 어류에 관한 연구는 미비한 실정이다. 이 연구는 어류에서의 KiSS-GPR54 system의 생리적 기능과 번식내분비적 기능을 탐색하기 위해 홍바리 Epinephelus fasciatus 뇌에서 KiSS1, KiSS2, GPR54 mRNA의 부분 염기서열을 클로닝 하였으며, KiSS1, KiSS2, GPR54 mRNA의 각 조직별 발현을 RT-PCR을 이용하여 확인하였다. 성 성숙과 KiSS-GPR54 system과의 연관성을 확인하기 위해 생식소 발달 상태(성숙과 미성숙)에 따른 발현을 qRT-PCR을 이용하여 분석하였다. KiSS1, KiSS2, GPR54의 부분 염기서열은 각각 232 bp, 304 bp, 613 bp 였으며, 조직별 발현을 조사한 결과 공통적으로 뇌에서 발현되었다. 생식소발달에 따른 발현 양상은 KiSS1, KiSS2 mRNA의 경우 성숙 상태에서가 미성숙 상태에 비해 유의적으로 높게 발현되었다. 하지만 GPR54 mRNA의 경우 미성숙 상태에서 더 높게 발현되었다. 이러한 결과들로 미루어 보아, KiSS-GPR54 system은 홍바리에 있어 번식과 밀접한 관련이 있을 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.33-39
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• 2017

Objective: The aim of this study was to assess the changes of follicular fluid (FF) and serum levels of cerebellin precursor protein 1 (cbln1) and betatrophin in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotropin-releasing hormone (GnRH) antagonist protocol. Methods: Twenty infertile women with PCOS and 20 control women diagnosed as poor responders undergoing ovarian stimulation with a GnRH antagonist were included. Blood samples were obtained during ovum pick-up. Follicular fluid from a dominant follicle was collected from the subjects. Using enzyme-linked immunosorbent assays, FF and serum levels of cbln1 and betatrophin were measured in both groups of participants. Metabolic and hormonal parameters were also determined and correlated with each other. Results: Both groups of women had similar serum and FF betatrophin levels ($55.0{\pm}8.9ng/mL$ vs. $53.1{\pm}10.3ng/mL$, p=0.11). The serum and FF betatrophin levels of poor responders were found to be similar ($49.9{\pm}5.9ng/mL$ vs. $48.9{\pm}10.7ng/mL$, p=0.22). Conversely, the FF cbln1 levels of PCOS women were found to be significantly higher than the serum cbln1 levels ($589.1{\pm}147.6ng/L$ vs. $531.7{\pm}74.3ng/L$, p<0.02). The FF cbln1 levels of control participants without PCOS were significantly higher than their serum cbln1 levels ($599.3{\pm}211.5ng/L$ vs. $525.3{\pm}87.0ng/L$, p=0.01). Positive correlations were detected among body mass index, insulin resistance, serum insulin, total testosterone, and betatrophin levels in the PCOS group. Conclusion: Follicular fluid betatrophin and cbln1 concentrations may play a pivotal role on follicular growth in PCOS subjects undergoing IVF/ICSI with an antagonist protocol.

• 한국동물생명공학회지
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• 제36권1호
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• pp.51-58
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• 2021

Kisspeptin, a neuropeptide and the master controller of reproductive axis upstream to GnRH neurons, and its receptor are also expressed in extra-hypothalamic tissues, such as ovaries. As systemic kisspeptin has been shown to modulate follicular dynamics in cattle, we hypothesized that kisspeptin has direct actions on the ovarian follicular development. We also hypothesized that kisspeptin regulation of primordial follicle development is via modulation of VEGF expression. In order to test these hypotheses, we cultured caprine ovarian cortical strips in vitro for 7 days with supplementation of kisspeptin at 1, 10 and 100 µM concentration and observed the development of primordial follicles into intermediate, primary and secondary follicles. We also studied the alteration in the expression profile of VEGF and VEGF transcript variant 2 mRNA during follicular development in the presence of kisspeptin. We confirmed the presence of GPR54 in goat ovaries in our preliminary studies. Supplementation of kisspeptin at 1 and 10 µM concentration facilitated the development of primordial follicles into intermediate, primary and secondary follicles with less number of degenerated follicles while the same at 100 µM resulted in degeneration of follicles. We observed a drastic increase in the expression profile of VEGF and VEGF transcript variant 2 mRNA upon culture which was independent of kisspeptin treatment. In conclusion, our studies show that kisspeptin facilitates ovarian primordial development in vitro.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2005년도 제48차 춘계 학술대회
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• pp.110-111
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• 2005