• 한국수정란이식학회지
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• 제28권3호
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• pp.169-175
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• 2013

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{\mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{\mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

• 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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• 제25권3호
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• pp.293-297
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• 1998

Hypogonadotropic hypogonadism, or the lack of function of the testis or ovary secondary to the lack of pituitary and or hypothalamic trophic hormones, is also sometimes generally termed Kallmann's syndrome. Whether such deficiencies arise from an inborn error of hypothalamic organization and pituitary connection or damage to the hypothalamic pituitary system in prepubertal life, the manifestations of a eunuchoid or apubertal individual with potentially competent pituitary and gonadal function will result. Beyond the achievement of puberty, a similar situation can be recreated by the administration of a long-acting GnRH analog or by conditions of secondary hypothalamic dysfunction such as anorexia nervosa where shutdown of GnRH and its resultant effects cause cessation of gonadal function and even a regression of secondary sexual characteristics. Technically, these conditions are not Kallmann's syndrome but one must recognize the similarities. We have experienced a case of isolated gonadotropin deficiency which showed a negative KALIG-1 gene in infertile patient with primary amenorrhea. So we report this case with a brief review of literatures.

• 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.

• 한국발생생물학회지:발생과생식
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• 제9권1호
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• pp.1-5
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• 2005

시상하부-뇌하수체-생식소(HPG) 호르몬 축은 유아기와 아동기에는 작동하지 않다가 사춘기 개시 직전에 활성화되는 흥분성 및 억제성 신호들의 복잡한 중추성 조절 네트워크에 의해 조절된다. 최근 주목받고 있는 kisspeptin은 KiSS-1 유전자의 펩타이드 산물로, 최초 orphan receptor로 클로닝된 G protein-coupled receptor 54(GPR54)의 내인성 리간드이다. KiSS-l은 본래 종양전이억제 유전자로 알려졌으나, 최근의 연구들은 KiSS-1/GPR54 시스템이 생식계의 주요한 조절자임을 시사하고 있다. 비록 생식 관련 호르몬 분비의 신경내분비적 조절에서 KiSS-1/GPR54 시스템의 정확한 역할은 아직 자세히 모르지만, 이 시스템은 생식호르몬 축에서의 일차적인 연결 고리일 수 있다. 중추적(icv) 또는 말초적(sc 또는 ip)으로 kisspeptin을 주사할 경우 시상하부-뇌하수체-생식소 축이 자극되어 혈중 LH 수준이 증가함이 설치류, 양, 원숭이 그리고 인간을 대상으로 한 실험들에서 관찰되었다. 이러한 kisspeptin의 효과는 시상하부 GnRH 신경계를 경유하여 나타나는 것으로 보이지만, 뇌하수체 전엽에 직접 작용할 수도 있다. GPR54 녹아웃 생쥐에서는 사춘기 개시의 소실과 생식소자극호르몬 저하성 생식소 기능저하증(hypogonadotropic hypogonadism, HH)이 나타났다. 따라서 kisspeptin 주사는 인간의 생식계 이상을 치료하기 위한 HPG 축을 활성화시키는 치료에 응용할 수 있을 것이다.

• 한국발생생물학회지:발생과생식
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• 제26권3호
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• pp.107-115
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• 2022

Kisspeptins, products of KISS1 gene, are ligands of the G-protein coupled receptor (GPR54), and the kisspeptin-GPR54 signaling has an important role as an upstream regulator of gonadotropin releasing hormone (GnRH) neurons. Interestingly, extrahypothalamic expressions of kisspeptin/GPR-54 in gonads have been found in primates and experimental rodents such as rats and mice. Hamsters, another potent experimental rodent, also have a kisspeptin-GPR54 system in their ovaries. The presence of testicular kisspeptin-GPR54 system, however, remains to be solved. The present study was undertaken to determine whether the kisspeptin is expressed in hamster testis. To do this, reverse transcription-polymerase chain reactions (RT-PCRs) and immunohistochemistry (IHC) were employed. After the nest PCR, two cDNA products (320 and 280 bp, respectively) were detected by 3% agarose gel electrophoresis, and sequencing analysis revealed that the 320 bp product was correctly amplified from hamster kisspeptin cDNA. Modest immunoreactive (IR) kisspeptins were detected in Leydig-interstitial cells, and the weak signals were detected in germ cells, mostly in round spermatids and residual bodies of elongated spermatids. In the present study, we found the kisspeptin expression in the testis of Syrian hamster. Further studies on the local role(s) of testicular kisspeptin are expected for a better understanding the physiology of hamster testis, including photoperiodic gonadal regression specifically occurred in hamster gonads.

• 한국해양생명과학회지
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• 제1권1호
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• pp.41-49
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• 2016

Kisspeptin (Kiss) and its cognate receptor, kisspeptin receptor (KissR; G protein coupled receptor 54, GPR54), have recently been recognized as potent regulators of reproduction in teleosts. Additionally, leptin plays an important role in energy homeostasis and reproductive function in teleosts. The purpose of this study was to examine differences in the concentration of the hormones of the Kiss/KissR system and leptin and the expression of their underlying genes, all of which are involved in the sexual maturation of female goldfish, Carassius auratus, following treatment with Kiss. The expression levels of KissR increased after the Kiss injection. Furthermore, the peptide hormone leptin also increased after the injection (in vivo and in vitro). Additionally, the expression of GnRH and GTHs (GTHα, FSHβ, and LHβ) increased in the brain and pituitary (in vitro and in vitro). These results support the hypothesis that Kiss plays important roles in the direct regulation of the hypothalamus-pituitary-gonad axis and leptin in goldfish. Therefore, we suggest that Kiss system gene expression is correlated with energy balance and reproduction.

• 한국발생생물학회지:발생과생식
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• 제14권3호
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• pp.185-197
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• 2010

The reproduction of animals is a means to maintain their species. The golden hamsters are seasonal breeders, and their reproduction is regulated by the photoperiod. The shifts of the sexual functions imply the changes of the protein expression, and those are reflected by the proteome. Thus the present study was to examine via two dimensional polyacrylamide gel electrophoresis (2d PAGE), the physiological changes and the alterations of protein expressions in hypothalamus upon the reproductive situation related to the pineal gland. Among the pineal intact animals, the reproductive functions were sustained in animals housed in long photoperiod (LP) but regressed in animals housed in short photoperiod. And those pinealectomized animals showed high sexual activities regardless of photoperiod. Ultimately they were branched into dichotomy, sexually active and inactive animals. Apart from the changes of physiological parameters upon the reproductive conditions, there were obvious differences in proteins extracted from the hypothalamus. The hypothalamus of LP animals presented high levels of enzymes which are involved in the production of energy, glycolysis and Krebs cycle. The increased energy might be related to the GnRH synthesis in hypothalamus and indirectly to the constant cell divisions in spermatogenesis. Taken together, the impacts of the photoperiodic information controlling reproduction could be observed through 2d PAGE. Therefore, the present results suggest the potential of biomarkers collectively to diagnose the fertility and the infertility by way of proteomics in organs with regard to the reproductive system, further could be applied to diagnose various diseases.

• 한국발생생물학회지:발생과생식
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• 제19권1호
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• pp.11-17
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• 2015

This study investigated possible involvement of photoperiodic regulation in reproductive endocrine system of female olive flounder. To investigate the influence on brain-pituitary axis in endocrine system by regulating photoperiod, compared expression level of Kisspeptin and sbGnRH mRNA in brain and FSH-${\beta}$, LH-${\beta}$ and GH mRNA in pituitary before and after spawning. Photoperiod was treated natural photoperiod and long photoperiod (15L:9D) conditions from Aug. 2013 to Jun. 2014. Continuous long photoperiod treatment from Aug. (post-spawning phase) was inhibited gonadal development of female olive flounder. In natural photoperiod group, the Kiss2 expression level a significant declined in Mar. (spawning period). And also, FSH-${\beta}$, LH-${\beta}$ and GH mRNA expression levels were increasing at this period. However, in long photoperiod group, hypothalamic Kiss2, FSH-${\beta}$, LH-${\beta}$ and GH mRNA expression levels did not show any significant fluctuation. These results suggest that expression of hypothalamic Kiss2, GtH and GH in the pituitary would change in response to photoperiod and their possible involvement of photoperiodic regulation in reproductive endocrine system of the BPG axis.

• Journal of Ginseng Research
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• 제32권1호
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• pp.8-14
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• 2008

스트레스 반응은 위급 상황에서만 활성화 되는 것이 아니라 우리 주변을 둘러싼 모든 사회적, 물리적 환경에 의해 끊임없이 기인된다. 예를 들어 소음, 강한 빛과 열, 주거의 공간 등과 같은 물리적 환경과 타인과의 관계, 부당한 대우, 규칙, 형식과 같은 사회적 관계 이 그것이다. 고려 인삼은 항스트레스 작용을 한다고 알려져 있으나 현재까지 대부분의 연구는 인삼 엑기스 또는 총 사포닌을 투여 시 나타나는 물리화학적인 척도 (수영 시간, 뜨거움에 대한 반응, 포도당 및호르몬 수준 등)를 측정하였으며 뇌에서 ginsenoside의 기능연구는 매우 미흡한 수준이다. 특히 신호전달, 전사체 (transcriptome),proteomics, system biology와 같은 분자생물학적인 방법을 이용한 기전 연구는 전혀 수행된 바 없다. 따라서 인삼의 항 스트레스 반응 기전을 이해하기 위해서는 인삼투여 후 뇌에서 나타나는 분자적 변화에 대한 연구가 앞으로 요구된다. 또한 microarray 등의 분자생물학적 기법을 이용하여 인삼 투여에 의해 반응하는 유전자를 발굴함으로써 이러한 인자들이 인삼 효능 시험 등에 지표로 응용될 수 있을것이다.