간행물

한국발생생물학회 학술대회논문집

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한국발생생물학회 2015년도 추계학술대회 (2015년 9월) 53

심포지움 I – 초청강연

1.
2015.09 서비스 종료(열람 제한)
한국발생생물학회가 1995년 창립된 지 벌써 20년! 회고담을 준비하게 되니 기쁘고 벅찬 가슴에 힘이 솟습니다. 그 동안 학회를 이끌어 주신 회장님들과 더불어 임원진들 그리고 발생학회의 발전을 위해 많은 노력을 해주신 회원님들의 노력의 결과이기에 여러분들께 충심으로 감사와 치하를 드립니다. 우리 학회의 최초 10년은 기초를 다지는 시기라고 생각됩니다. 우리 한국발생생물학회가 창립된 초기 상황을 약간 소개하려 합니다. 다음 10년은 여러분이 같이 학회의 발전을 만들어온 시기이고, 앞으로 10년은 우리의 운명이 스스로의 결정과 노력으로 우수한 학회, 우수 논문을 싣는 학회지를 가진 학회로 자리잡는 시기가 될 것으로 생각됩니다.
2.
2015.09 서비스 종료(열람 제한)
Somatic cell nuclear transfer (SCNT) has long been envisioned as a means for generating patient-specific stem cells to treat a range of age-related diseases. Until now, only three research groups have reported the successful derivation of SCNT-derived pluripotent stem cells (SCNT-PSCs). Our group has shown for the first time that human SCNT-PSCs can be successfully generated using dermal fibroblasts from 35 and 75 year-old males, and also recently established another SCNT-PSC from a patient with disease. However, despite cloning success in these groups, the derivation of stem cell lines from cloned human embryos has proven elusive. So, several approaches for the optimization of SCNT conditions, such as the use of protein phosphatase inhibitors, oocyte activation method and epigenetic regulation have been applied in order to overcome the obstacle. This study reveals mechanistic insights and establishes a promising method for improving human SCNT for regenerative medicine.
3.
2015.09 서비스 종료(열람 제한)
Autophagy means “self-eating” and it is a major catabolic pathway within cells. A basal level of autophagy is required for survival of cells or organisms, but prolonged activation of autophagy may have an adverse effect. In mammalian systems, autophagy is stimulated by nutrient starvation or deprivation of growth factors. Ovariectomy on day 4 of pregnancy in mice to deprive blastocysts of estrogen induces “dormancy” in blastocysts and delay the process of implantation until estrogen is given. Dormant blastocysts maintain a state of low metabolism in utero and survive for many days without initiating implantation under the unfavorable condition of estrogen deficiency. We tested the hypothesis if an autophagic response is operative in dormant blastocysts for prolonged survival in utero during the delayed implantation. We observed that autophagy is highly activated in dormant blastocysts. Interestingly, autophagic activation is more prominent in trophectoderm than in inner cell mass. Activation of blastocysts by estrogen supplementation induces formation of multivesicular bodies and exosomes in the trophectoderm. Dormant blastocysts with longer period of autophagic activation show compromised development after implantation. Thus, autophagy may be a critical cellular mechanism to provide energy source during extended survival of dormant blastocysts. However, prolonged activation of autophagy may compromise developmental outcome of blastocysts with irreparable cellular damage.
4.
2015.09 서비스 종료(열람 제한)
Dynamic reorganization of actin filaments is essential for various stages of mammalian oocyte maturation, including spindle migration, actin cap formation, polar body extrusion, and cytokinesis. Various actin binding proteins (ABPs) have been known to be involved in the regulation of actin filament remodeling. We elucidate roles of three different actin binding proteins in mouse oocyte maturation. The heterodimeric actin-capping protein (CP) binds to the fast-growing(barbed) ends of actin filaments and plays essential roles in various actin-mediated cellular processes. When CP is knockdowned or inhibitory component was overexpressed, asymmetric division of oocyte have been compromised. It turns out that knockdown or inhibition of CP deplete cytoplasmic actin mesh level, which have been known to be essential for maintain cytoplasmic actin mesh. Another actin binding proteins, tropomodulin 3 (Tmod3), binds to the slow-growing end of actin filaments and knockdown or expression deletion mutant of Tmod3 also decrease actin mesh level in maturing oocyte and it severely ablated asymmetric division of oocyte. Finally, tropomyosin 3, actin filament binding proteins protect actin filament from depolymerization, is also important to maintain cortex integrity in maturing oocyte. Taken together, these finding showed the essential roles of actin binding proteins in remodeling of actin filaments in mammalian oocyte development.
5.
2015.09 서비스 종료(열람 제한)
Although decapod crustaceans are one of the most important fisheries resources with high market value, we still have only limited knowledge about their basic physiology related to growth, development, and reproduction. This is mainly due to the lack of tools to manipulate genetic information leading the phenotype changes. Recently physiological study for decapod crustaceans changed dramatically by both the development of next-generation sequencing (NGS) technology and RNA interference (RNAi). Significant decrease in the cost for reading genome or transcriptome allowed the even single lab can manage the omics-level study about the non-model system, decapod crustaceans. As genomic and transcriptomic data increased, we are able to screen novel genes in decapod crustaceans related to growth and development. As another useful tool, gene silencing through RNAi is gaining momentum for decapod crustaceans. RNAi has proven instrumental in a growing number of crustacean species, revealing the functionality of novel crustacean genes. Major research topics in decapod crustaceans include immune response, reproduction, development, homeostasis, molting and growth, and environmental stress. In addition to any changes in phenotype, tanscriptomic analysis induced by the specific gene knockdown by RNAi extended our knowledge of physiological responses of novel crustacean genes. Those new techniques extend our knowledge about crustacean physiology providing the basis for increasing productivity of decapod crustaceans.

심포지엄 II - 내분비계장애물질 최신 연구동향 및 대체물질 개발

6.
2015.09 서비스 종료(열람 제한)
스테로이드 호르몬의 하나인 여성호르몬 에스트로젠은 난소에서 생성되어 혈관을 통해 표적 세포에 도달하여 단순확산에 의해 표적세포 내로 들어가, 표적세포의 핵 내에 존재하는 전사인자인 에스트로젠 수용체를 통하여 표적 유전자의 전사 활성도에 영향을 미침으로써 그 기능을 나타낸다. 1960년대 후반에 에스트로젠 수용체 알파가 분리되고, 1996년에 스웨덴의 Gustafsson 그룹에 의해 에스트로젠 수용체 베타가 발견되면서 연구에 많은 진척을 가져왔다. 에스트로젠 수용체 연구분야에 에스트로젠에 대한 연구는 생식기능 이외에도 심혈관 질환, 뇌질환, 골다공증 등과 같은 만성 퇴행성 질환 및 유아 청소년의 발달과도 연관이 되어 있어 스테로이드 호르몬 수용체 중에서도 연구가 가장 활발하게 이루어지고 있는 분야 중 하나이다. 분자적 수준에서의 에스트로젠 수용체의 작용기전의 이해는 에스트로젠에 의한 많은 질병의 예방과 치료에 큰 역할을 하고 있으며, 사회적으로 문제가 되고 있는 환경 호르몬에 의한 건강 관리 문제에도 사회적으로 큰 기여를 하고 있다. 에스트로젠 수용체 전사 조절 기전과 에스트로젠과 구조가 유사한 환경 호르몬의 작용 기전에 대한 이해를 바탕으로 환경 호르몬의 스크리닝 기법과 국소적인 여성호르몬 합성을 통한 내분비 교란 등을 통하여 환경호르몬이 유방암, 전립선암, 조기폐경, 성조숙증에 미치는 최근 연구 결과를 발표하고자 한다.
7.
2015.09 서비스 종료(열람 제한)
양서류의 변태(metamorphosis)는 갑상선호르몬에 의해 유발되며, 이러한 특성을 이용해 시상하부-뇌하수체-갑상선 축(hypothalamic-pituitary-thyroid axis; HPT axis)을 교란하는 화학물질을 스크리닝하는 기술이 개발되어왔다. 1998년 OECD는 Endocrine Disrupters Testing and Assessment 팀을 구축하고, 이 과정에서 ‘Amphibian Metamorphosis Assay(AMA)’를 갑상선교란물질 스크리닝법으로 제안하였다. 현재는 OECD test guideline No. 231로서 아프리카발톱개구리(Xenopus laevis) 유생을 이용한 In-vivo 시험법이 공개되어 있으며, 이에 관한 다양한 실증 및 활용 연구가 학계에 보고되고 있다. 본 시험법은 갑상선호르몬 작용제 또는 길항제를 스크리닝하기 위해 특정 화학물질을 양서류 유생이 있는 탱크에 처리하는 방식으로 진행되며, 21일간 진행되는 시험법으로서 현존하는 갑상선교란물질 스크리닝 방법 중 독성종말점이 가장 뚜렷하며, 분석비용이 저렴하다는 장점을 갖는 것으로 평가되고 있다. 특히, 모델생물로서 이용되는 X. laevis는 수생양서류로서 어류 사육시설에서도 쉽게 유지가 가능하며, 호르몬(human chorionic gonadotropin; hCG)을 이용해 쉽게 배아획득이 가능하다. 독성종말점(toxicological end-point)으로 제시되는 항목은 유생의 발생단계, 뒷다리 길이, 전장, 무게가 있으며, 추가적으로 갑상선의 조직학적 변화를 확인한다. 본 시험법 또는 변형된 유사 시험법을 통해 현재까지 다양한 제초제, 살충제, 중금속, 알킬페놀류 등을 비롯한 다양한 물질들의 갑상선교란효과가 밝혀졌으며, X. laevis 외의 양서류에서도 본 시험법이 적용되는 사례가 보고되고 있다. 최근에는 세대주기가 1~2년이며, 4배체(allotetraploid)인 X. laevis보다 세대주기가 짧고(6 month) 이배체(diploid)인 X. tropicalis를 활용한 연구가 증가하는 추세이며, 자국에 서식하는 양서류에 적용하기 위한 노력도 이루어지고 있다. 국내에서 내분비계 교란물질이 검출되는 젖병, 수액튜브 등이 이슈화되면서 내분비계 교란 활성이 없는 계면활성제, 가소제 등을 개발하기 위한 연구가 활발하다. 향후, 이들의 신물질 안전성 평가에 있어 AMA는 갑상선교란성을 파악하기 위해 매우 유용하게 사용될 것이다.
8.
2015.09 서비스 종료(열람 제한)
세칭 환경호르몬인 내분비계장애물질(endocrine disrupting chemical, EDC)에 노출될 경우, 내분비계가 교란되어 항상성이 무너지고, 심지어는 후손에 까지 유해성이 전달될 수 있음이 속속 드러나고 있다. 따라서 EDC는 인간의 건강은 물론 생태계의 안정성에 심대한 영향을 끼치는 요인으로 지목되어 학계는 물론 일반인들의 관심이 날로 커지고 있다. EDC는 단기 노출 시 치사에 이르는 급성 독성보다는 비교적 장기간 노출 시 발생독성, 생식독성, 대사독성, 면역독성 등이 나타나는 특징을 가지고 있는데, 본 발표에서는 생식독성에 초점을 맞추고자 한다. EDC의 유해성 조사에 있어서 가장 큰 문제는 잠재적으로 EDC 활성을 지닌 것으로 추정되는 물질이 많은데 비해, 연구의 속도는 그에 미치지 못함이다. 또한 대부분의 EDC 또는 EDC-like 물질이 산업적으로 중요하며, 일상생활에서도 널리 사용되고 있다는 점이다. 따라서 EDC 추정 물질의 유해성을 신속, 정확하게 조사하는 기술 개발과 함께, 생산량-소비량-인체 노출량을 고려한 조사 우선순위를 정하고, 궁극적으로는 유해성이 적은 대체 물질을 개발함이 중요하다. 주로 설치류를 대상으로 한 포유동물 사춘기 개시에 미치는 EDC 유해성 연구는 생식독성 조사의 일환으로 여러 연구자들에 의해 수행되어 왔다. 특히 암컷 사춘기 개시는 수컷보다 빠르게 일어나고, 수컷에 비해 질구개방(vaginal opening)과 같은 뚜렷한 지표와 더불어 신경내분비 활성에 대한 자료가 많이 축적되어 있어 연구 수행에 용이한 장점들이 있다. 사춘기 개시는 궁극적으로 시상하부-뇌하수체-난소 호르몬 축의 활성이 증진되어야만 하는데, 따라서 pro-estrogenic EDC에 대한 연구가 주종을 이루며, anti-androgenic EDC에 대해서는 일부 수행되었다. 본 발표에서는 먼저 정상 사춘기 개시에 있어서의 신경내분비 조절에 대한 현재의 지식을 알아보고, 다음 본 연구자 그룹이 수행해온 연구내용을 소개하고자 한다. EDC 노출에 따른 사춘기 개시 조절의 교란 가능성은 의학적으로도 중요한 의미를 가지고 있다. 최근 관심이 고조되는 분야는 직접적인 유전적인 요인이 없이도 10세 전후에 나타나는 조기 사춘기로써, 이는 심신이 미숙한 상태의 조기 임신, 조산이나 기형아 출산 같은 심각한 가정문제를 유발할 수 있다. 또한 골성장 역시 조기에 개시된 후 정상인에 비해 상대적으로 일찍 종료되어 해당자에게 육체적인 측면은 물론 삶의 질을 저하시킬 수 있다. 이러한 측면에서 볼 때, EDC 연구는 앞으로도 계속 중요한 연구 분야가 될 것으로 예상한다.
9.
2015.09 서비스 종료(열람 제한)
‘환경호르몬’(내분비계 교란물질)은 생체 외부에서 들어와 인간의 내분비기관에서 호르몬의 생리작용을 교란시키는 화합물을 뜻한다. 현재 환경호르몬은 오존층 파괴, 지구 온난화와 함께 세계 3대 환경문제 중의 하나로 대두되고 있다. 환경호르몬은 생체 내 호르몬의 합성, 방출, 수송 등 다양한 과정에 관여해 각종 형태의 교란을 일으킴으로써 야생동물 및 인간의 발생과 생식기능에 변화를 초래할 수 있으며, 성장(변태) 억제, 면역저해, 발암 가능성을 증가시킬 수 있다. 환경호르몬은 다수의 플라스틱 및 세제에서 검출되고 있어, 유해 환경호르몬을 대체할 수 있는 물질의 개발과 이를 이용한 제품생산이 필요하게 되었다. 현재 플라스틱 가소제 부분에서 비스페놀A, 프탈레이트를 대체할 수 있는 다양한 대체가소재가 개발중에 있고, 노닐페놀을 대체할 수 있는 계면활성제 개발 또한 활발하다. 그러나 일부 제품화되어 안전한 것으로 선전되고 있는 신물질들에 대한 내분비학적 안정성에 대해 여전히 논란의 여지가 남아있다. 한양대학교는 미래창조과학부가 2015년부터 3년간 지원하는 ‘환경호르몬으로부터 국민의 건강을 보호하기 위한 기술개발 연구’의 주관사업단으로 선정되어 환경호르몬 대체물질 개발 및 국민 생활에 필요한 기술 연구 활동에 들어간다. 연구내용은 신체에 직접적인 악영향을 미치는 노닐페놀, 프탈레이트, 비스페놀A를 대체할 저독성 환경호르몬 개발, 환경호르몬 감지센서기술을 개발한다. 특히 기술 개발을 넘어서, 대체물질을 이용한 취약계층 생활용품 제작 및 보급형 제품 실용화 단계까지 목표로 하고 있다. 또한, 대체물질의 내분비계교란 활성에 대한 정밀 평가, 환경호르몬 관리 법제도 연구 및 대국민홍보체계 수립을 위한 연구도 예정되어 있다. 본 사업은 통해 현재 사용되고 있는 계면활성제, 플라스틱 가소제 및 새롭게 개발되고 있는 주요 대체물질들의 세포독성, 유전독성, 후성유전학적독성, 에스로젠성, 안드로젠성, 항안드로젠성, 갑상선호르몬성에 대한 in vitro 및 in vivo 독성모니터링 및 최종후보 물질들에 대한 생태동태(ADME)을 통해 대체후보물질을 선발하게 된다. 이러한 연구개발 노력은 환경호르몬 노출로 인한 국민 불안감을 해소하고, 대체물질을 이용한 제품 개발로 새로운 시장개척 효과도 있을 것으로도 예상된다.
10.
2015.09 서비스 종료(열람 제한)
Spermatogonial stem cells (SSCs) possess the unique capacity of self-renewal and differentiation and thereby can transmit genetic information to the next generation. Combination with techniques such as isolation, culture, preservation, and transplantation of the SSC has facilitated the efficient method for production of transgenic animals, and preservation of livestock and endangered species. The purpose of this study was to genetically modify enriched populations of pre-pubertal germ cells using lentiviral transduction and to develop an efficient in vitro culture system and cryopreservation technique for bovine SSCs. To maximize the efficiency of genetic modification of bovine SSCs, effective enrichment techniques need to be developed. Selection of bovine SSCs using a combination of laminin and gelatin was resulted in a 8-fold enrichment. Selected cells were then transduced using a lentiviral vector containing the transgene for the enhanced green fluorescent protein. Transduction efficiency was 17%. Next, to enhance the efficiency of proliferation for in vitro culture, the effects of various culture conditions and growth factors on bovine cell proliferation were evaluated. Based on the results, we developed the optimal culture conditions [2× rat sertum free medium (rSFM) containing 0.1% FBS together with GDNF, GFRα1, bFGF, EGF, LIF, and CSF-1] for maintaining bovine SSCs over 3 months without any alteration of stem cell characteristics and functions. Also, to develop an effective cryopreservation technique for bovine SSCs, the effects of different freezing methods and various cryoprotective agents were tested. The recovery rate, and proliferation capacity of bovine SSCs were significantly greater for germ cells frozen using tissue freezing methods compared to cell freezing methods. Cryopreservation in the presence of 200 mM trehalose resulted in significantly greater recovery rate, and proliferation capacity of germ cells compared to control. As a results, cryopreservation using tissue freezing methods in the presence of 200 mM trehalose is an efficient cryopreservation protocol for bovine SSCs. Collectively, these findings can serve as a model for comprehensively understanding the biology of SSCs and the factors that regulate male fertility. Furthermore, results of this study will be integral for the continued refinement of techniques to manipulate bovine SSCs.

한국발생생물학회 젊은과학자상

구두 발표

14.
2015.09 서비스 종료(열람 제한)
Cryopreservation has become a powerful method of the assisted reproduction technology and supports fertility preservation of cancer and other indication patients. After controlled ovarian hyperstimulation, surplus oocytes and embryos were recommended to store using cryopreservation. Recently, vitrification is replaced with traditional slow freezing protocol, because of improved survival rates and clinical outcomes. Vitrification requires a high concentration of CPAs that may induce significant osmotic and metabolic damage to cells including oocytes even in a short exposure of a few minutes. Generally, MPF plays a crucial role in the cell cycle regulation and maintaining the meiotic arrest of oocytes. In fact, it has been observed to decline in MII ovine oocytes after vitrification and would be suggested that one of the main causes of low fertilization rate and developmental competence derived from cryoinjury during vitrification. Therefore, the aim of this study was to evaluate the effect of caffeine treatment on the activity of MPF, MAPK level in vitrified/warmed mouse mature eggs. Caffeine, Phosphataseinhibitor, may maintain active form of MPF. We evaluated their survival after warming procedure, fertilization, cleavage, and developmental rates. Ovulated MII eggs were retrieved from 6 weeks old B6D2F1 female mouse at 14hr post hCG injection. Collected MII eggs were maintained in HTF medium containing 10% KSR with or without caffeine for 1hr. Eggs were vitrified in 7.5%EG +7.5%DMSO equilibrium solution, 15%EG + 15%DMSO + 0.5M sucrose vitrification solution with or without caffeine. Also warming solution contained sucrose (0.5M, 0.25M, 0.125M, and 0M) with or without caffeine. After warming, eggs were cultured in HTF medium with or without caffeine for 2 hr then fertilized with epididymal sperm in vitro and cultured in KSOM for 5 days to analyze embryonic development. Survival rates were similar in all experimental groups. However, fertilization rate was higher in with caffeine group compare to without caffeine significantly (80% vs. 85%, p<0.05). 2-cell and blastocyst formation were increased in caffeine group (p<0.05). MPF activity and MAP kinase activity were recovered in with caffeine group after vitrification/warming process. In conclusion, Caffeine may maintain MPF and MAPK level in vitrified/warmed MII eggs, and enhance fertilization and further embryonic development.
15.
2015.09 서비스 종료(열람 제한)
DGCR8 is a RNA-binding protein working with DROSHA to produce pre-microRNA in the nucleus, while DICER does not only mature microRNA but also endogenous siRNAs in the cytoplasm. Here, we have produced Dgcr8 conditional knock-out mice using progesterone receptor (PR)-Cre (Dgcr8flox/flox; PRcre/+ mice, Dgcr8d/d) and demonstrated that canonical microRNAs dependent of DROSHA-DGCR8 complex are required for uterine development as well as female fertility in mice. Adult Dgcr8d/d females did not undergo regular reproductive cycle and produce any pups when housed with fertile males, whereas administration of exogenous gonadotropins induced normal ovulation with corpus luteal formation in these mice. Ovulated oocytes from Dgcr8d/d mice had comparable fertilization potentials and were normally developed to the blastocyst after fertilization as compared to those in control Dgcr8f/f mice. Interestingly, PR-Cre-dependent Dgcr8 deletion showed aberrant infiltration of acute inflammatory immune cells to female reproductive organs only when Dgcr8d/d mice were mated with male mice. With respect to uterine development, gross morphology, histology, and weight of Dgcr8d/d uterus were similar to those of control at 3-week-old age. However, multiple uterine abnormalities were noticeable at 4-week-old age when PR expression is significantly increased, and these deformities became severe onwards. Gland formation and myometrial layers were significantly reduced, and stromal cell compartment did not expand and became atrophic during uterine development in these mice. These results were consistent with aberrantly reduced cell proliferation in stromal cell compartments of Dgcr8d/d mice. Collectively, our results suggest that DGCR8 dependent-canonical microRNAs are essential for development and physiology of the uterus with respect to morphogenesis, proper immune modulation, reproductive cycle, and steroid hormone responsiveness in mice.
16.
2015.09 서비스 종료(열람 제한)
Primary oocytes that are arrested in first meiotic prophase for years enter maturation process to meet a critical precondition for successful fertilization. During maturation, oocyte finishes meiosis I and progresses to the metaphase II stage, achieving meiotic maturity. Although importance of oocyte maturation for oocyte quality has been recognized, it is not fully understood for molecular mechanisms underlying oocyte maturation. Here, we found that dexamethasone-induced Ras-related protein 1 (RASD1), a member of RAS superfamily of small GTPases, was expressed in the mouse ovary. Immunohistochemical analysis revealed that Rasd1 expression was dominant in oocyte cytoplasm. Real-time PCR and RT-PCR analyses showed that Rasd1 mRNA was steadily expressed in germinal vesicle (GV), germinal vesicle break down (GVBD), metaphase I (MI) oocytes, but decreased in metaphase II(MII) oocytes during oocyte maturation. Konckdown of Rasd1 using RNAi system in the GV oocytes suppressed oocyte maturation through disruption of meiotic spindle and formation of misarranged chromosomes. Taken together, Rasd1 is a critical factor for MI-MII transition of oocyte and is involved in the regulation of spindle formation during oocyte maturation. Further study is needed to examine relationship between Rasd1 and spindle formation in MI-MII transition.
18.
2015.09 서비스 종료(열람 제한)
Accurate chromosome segregation is critical to ensure genomic integrity during cell division. This process is facilitated by the kinetochore, a multiprotein structure that is assembled on centromeric regions of chromosomes. The kinetochore establishes a mechanical link between the chromosomes and spindle microtubules and modulates cell cycle progression by regulating spindle assembly checkpoint (SAC). Defects in this process result in an aneuploidy, leading to miscarriages, infertility and various genetic disorder such as Down’s syndrome. Although the numerous kinetochore proteins have been identified and studied, the mechanisms that engaged in kinetochore assembly and chromosome segregation are poorly understood. Here we investigated the function of kinetochore protein Zwint-1 on homologous chromosome segregation during oocyte meiotic maturation. We found that Zwint-1 was localized at the kinetochore during meiotic maturation. Knockdown of Zwint-1 caused premature polar body extrusion, indicating acceleration of meiosis I. Interestingly, Zwint-1 knockdown impaired the recruitment of Mad2 at the kinetochores. However, BubR1 localization at the kinetochores was not affected by Zwint-1 knockdown, suggesting that Zwint-1 selectively regulates the recruitment of SAC components into the kinetochores. We also found that Zwint-1 knockdown abrogated chromosome alignment and segregation, thereby resulting in a high incidence of aneuploidy. These chromosomal defects were mostly due to the abnormal kinetochore-microtubule (kMT) attachments. Intriguingly, chromosome misalignment mediated by SAC inactivation was repaired, when anaphase onset was delayed by treating oocytes with proteasome inhibitor MG132. However, surprisingly, chromosomal defects following Zwint-1 knockdown were not restored by delaying anaphase onset. This result suggests that chromosomal defects induced by Zwint-1 knockdown are less likely associated with the failure of SAC activation. In addition, we observed that Aurora B/C kinase activity was not affected by Zwint-1 knockdown. Nevertheless, the meiotic defects induced by Zwint-1 knockdown were similar to those observed in Aurora B/C inhibition, suggesting that Zwint-1 is a downstream effector of Aurora B/C kinase during meiosis. Consistent with this, in Zwint-1 knockdown oocytes chromosomal defects following Aurora B/C inhibition were not restored when Aurora B/C inhibitor was removed, whereas the defects were well rescued in control oocytes after removing Aurora B/C inhibitor. This result suggests that the role of Aurora B/C kinases that correct erroneous kMT attachment is primarily regulated by Zwint-1. Collectively, our results demonstrated for the first time that Zwint-1 is an essential downstream effector of Aurora B/C kinase that corrects erroneous kMT attachment and regulates SAC activity, which ensures accurate homologous chromosome segregation during oocyte meiosis.

포스터 발표

19.
2015.09 서비스 종료(열람 제한)
Early growth response 1 (Egr1) is an inducible zinc finger transcription factor. Egr1 binds specific GC-rich region. Egr1 mRNA is rapidly and transiently expressed in pre-ovulatory follicles by LH and expressed in decidual cell by estrogen. Progesterone receptor (PR) is a nuclear transcription factor that is induced in granulosa cells of pre-ovulatory follicles following the LH surge. PR regulates ADAMTS1, which downstream gene of PR. In previous study, we observed ADAMTS1 mRNA expression pattern changed in Egr1 KO mice. Therefore, we expected that progesterone receptor gene expression is directly regulated by early growth response 1 in mouse ovarian granulosa cell. We could found the ER binding domain, Egr1 binding domain and CAAT box in PR promoter using the web tool AliBaba 2.1. We construct the PR promoter vectors truncated ER binding domain, Egr1 binding domain, CAAT box, respectively. We also construct the Egr1 expression vector using pcDNA 3.1 (+) vector. Granulosa cells are isolated from female ICR mice after 12h PMSG injection. To confirm the Egr1 overexpression, we performed western blot. For reporter assay, we used Dual-Luciferase reporter assay system. In conclusion, Egr1 may regulate PR expression in granulosa cell.
20.
2015.09 서비스 종료(열람 제한)
During implantation period uterus undergoes functional and histological changes including cell proliferation, differentiation, apoptosis. This delicate condition is regulated by various factors such as steroid hormones, cytokines, and growth factors. From previous our studies, we showed the spatio-temporal expression of Müllerian inhibiting substance (MIS) and MIS receptor Ⅱ (MISR II) in mouse uteri during pregnancy. In present study investigated the role of MISRⅡ in proliferation and differentiation in the uterus. The decidualization markers were dramatically decreased during treated rhMIS and MISRⅡ viral particles in in vitro decidualization model. The intensity of PAS staining and Oil-Red-O staining were dramatically decreased by treatment of rhMIS and overexpression of MISRⅡ. MIS/MISRⅡ also suppressed stromal proliferation regulate the differentiation of stroma cell during decidualization. MIS suppress the stromal proliferation and decidual differentiation along with its receptor. Put together it is suggested that MIS works as paracrine and autocrine factors for cell proliferation regulator and differentiation regulator during implantation.
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