This study was performed to evaluate the effects of fibroblast co-culture on in vitro maturation (IVM) of prepubertal mouse preantral follicles. The intact preantral follicles were obtained from the ovaries of 12－14 day old mice and these were cultured individually in α-minimal essential medium (α-MEM) supplemented with 5% fetal bovine serum (FBS), 100 mIU/㎖ recombinant follicle stimulating hormone (rFSH), 1% insulin-transferrin-selenium, 100 μg/ml penicillin and 50 ㎍/㎖ streptomycin as base medium for 12 days. A total of 200 follicles were cultured in base medium co-cultured with mouse embryonic fibroblast (MEF) (MEF group) (n=100) or only base medium as control group (n=100). Survival rate of follicles on day 12 of culture were significantly higher in the MEF group of 90.0%, compared with 77.0% of the control group (p=0.021). Follicle diameters on day 6 and 8 of the culture period were significantly larger in the MEF group than those in the control group (p=0.021, p=0.007, respectively). Estradiol levels in culture media on day 4, 6, 8, 10 and 12 of the culture period were significantly higher in the MEF group (p=0.043, p=0.021, p=0.006, p<0.001 and p=0.008, retrospectively). Our data suggest that MEF cell co-culture on IVM of mouse preantral follicle increases survival rate and promotes follicular growth and steroid production.

High temperature impediment in developing stages of crops has been occurred due to the impact of global warming. Rice production is notable to be sensitive to increasing environmental temperature and grain filling temperatures are already approaching threatening levels in many countries with rice cultivation. Recent proteomic analyses exposed impulsive changes of metabolisms during rice grain development. Interestingly, proteins involved in glycolysis, citric acid cycle, lipid metabolism, and proteolysis were accumulated at higher levels in mature grain than those of developing stages. High temperature (HT) stress in rice ripening period enhances damaged (chalky) grains which have loosely compacted shape starch granules. We carried out two-dimensional gel electrophoresis to analyze protein profiles during grain filling and different developmental stages of rice seed maturation. Proteins were separated from the fertilized seeds (seeds from 7 days and 21 days after fertilization) and seed maturation stage using IEF in the first-dimension and SDS-PAGE in the second dimension along with MALDI-TOF mass spectrometry. More than 1,000 protein spots were detected on a two-dimensional gel electrophoresis. A total of 120 different protein spots out of 140 protein spots were identified by MALDI-TOF and nano LCQ-TOF mass spectrometer. The identified proteins were categorized into six (6) different groups according to their expression patterns during grain filling and seed maturation. Some proteins were confirmed during seed development stages such as cytoplasmic malate dehydrogenase, whereas others were appeared at a specific stage like putative subtilisin-like protease, germin-like, seed allergenic proteins. Furthermore, the chalking mechanism of rice grain under the HT stress could be discussed in terms of grain starch glycome, transcriptome, and proteome.

This study was conducted to investigate the food value of Cudrania tricuspidata at its various maturation stages. The pH, total acid and reducing sugar contents of its fruit juice were determined to have been 4.2～5.1, 1.4～2.0% and 5.4～8.6%, respectively. The general chemical components of its fruit were observed as 76～80% moisture, 2.2～3.5% crude protein, 1.7～2.9% crude fat, 0.8～1.2% ash and 14.5～16.4% carbohydrate. Its free sugar, glucose and fructose contents were determined. The fructose contents of both its ripened and over-ripened fruits were higher than their glucose contents. Organic acids such as oxalic acid, citric acid, tartaric acid, malic acid and succinic acid were detected, and the concentration of the malic acid and the succinic acid were found to have been most abundant. The K content was higher than the amounts of other minerals, such as Ca, Fe, K, Mg, Na and P. Its vitamin C and the total amount of its dietary fiber were 127.5～149.2 mg% and 22.7～38.7%, respectively. Its insoluble dietary fiber content was higher than its soluble dietary fiber content. Its total polyphenol and flavonoid content were 18.9～19.6 mg% and 40.9～48.2 mg%, respectively.

본 연구는 콩에 함유된 아이소플라본 합성에 관여하는 요인 중 평균기온이 미치는 영향을 구명하고자 수행하였다. 시험품종은 생태형이 다른 일품검정콩, 청자콩, 흑청콩 3품종을 사용하였다. 파종은 5월 15일, 5월 30일 6월 15일 3시기에 재식밀도 60×15cm 로 하였다. 재배관리는 경기도 콩 표준재배법에 준하여 하였다. 아이소플라본 함량 분석은 개화기후 30일부터 5일 간격으로 수확기까지 채취하여 얻은 시료를 UPLC로 분석하여 얻은 결과는 다음과 같다. 1. 아이소플라본 함량은 시험품종 모두 파종기가 늦을수록 높았다. 2. 아이소플라본 함량은 품종과 파종기에 관계없이 genistein이 가장 높았고, 다음은 daidzein, glycitein 순이었는데, glycitein은 개화기 후 45~55일경 이후에는 뚜렷한 증가를 보이지 않았고, 등숙기간중 기온변화에 안정적이었다. 3. 아이소플라본 중 genistein 함량이 daidzein 함량보다 높아진 시기는 일품검정콩은 개화기 후 50~55일, 청자콩은 개화기 후 40~55일, 흑청콩은 개화기 후 60~65일이었다. 4. 등숙기간중 개화기 후 30일부터 5일 간격으로 평균기온과 아이소플라본 함량과의 관계를 분석한 결과, 일품검정콩에서 genistein은 y=-15.28x+407.9 (R2=0.505* ), 청자콩에서 diadzein은 y=-6.188x-164.5(R2=0.454* )로 유의한 상관관계를, genistein은 y=-11.59x+297.6(R2=0.545**)으로 고도의 유의한 상과관계를 보였다. 5. 아이소플라본 함량이 높은 검정콩 생산이 적합한 지역은 등숙기간중 립비대기(R5 )~생리적 성숙기(R7 )의 평균기온이 상대적으로 낮은 경기북부지역과 강원도, 충청북도, 경상북도의 내륙지역과 고지대일 것으로 고찰되었다.

Recently, we reported growth arrest-specific gene 6 (Gas6) as a new maternal effect gene (MEG), that expressed in the oocytes but functioned principally during embryogenesis. Gas6 RNAi-treated oocytes developed to metaphase II (MII) stage but they have affected M-phase promoting factor (MPF) activity and incurred abnormal pronuclear (PN) formation during fertilization. Gas6 is a ligand of TAM family members (Tyro3, Axl and Mertk) of receptor tyrosine kinase (RTK). Purpose of the present study was to evaluate the expression of Tyro3, Axl and Mertk transcripts in oocytes and early embryos. Expression of Gas6 and Mertk mRNA was detectable in oocytes and follicular cells, while Tyro3 and Axl mRNA was expressed only in follicular cells. Expression of Mertk mRNA was relatively constant during oocytes maturation and embryogenesis, but the other receptors, Tyro3 and Axl, were not expressed in oocytes and PN stage of embryos at all. Knockdown of Mertk mRNA and protein by using sequence-specific Mertk double strand RNA (dsRNA) did not affect oocytes maturation. In this case, however, contrary to the ligand Gas6 RNA interference (RNAi), MPF activity had not been changed by Mertk RNAi. Therefore, we concluded that the Gas6-Mertk signaling is not directly related to the oocyte maturation. It is still required to study further regarding the function of Mertk as the receptor of Gas6 during preimplantational early embryogenesis.

To evaluate the effects of benzo[a]pyrene (B[a]P), one of polycyclic aromatic hydrocarbons (PAHs), on in vitro oocyte maturation (GVBD) and sex steroid hormone production, maturing oocytes (oocyte diameters=0.74, 0.88 and 0.93 mm) of the longchin goby, Chasmichthys dolichognathus were incubated with B[a]P (1, 10 and 100 ng/mL) for 24 hours. After incubation, the oocytes were fixed with clearing solution (ethanol:formalin:glacial acrtic acid=6:3:1). The location of the germinal vesicle was observed under low-power magnification using a dissecting microscope. Steroids in aliquots of the incubation media were extracted twice using five volumes of ethylacetate:cyclohexane (1:1). Then, the T, E2 and 17α20βP levels were measured by RIA. In oocytes 0.74 mm diameter (vitellogenic oocytes), B[a]P had no significant effect on GVBD at the concentrations tested. In oocytes 0.88 mm diameter (fully vitellogenic oocytes), B[a]P inhibited GVBD significantly at 1 and 100 ng/mL. T production was decreased and the ratio of E2/T was increased significantly at 1 and 10 ng/mL compared with control. In 0.93 mm diameter oocytes (germinal vesicle located near the center of oocytes), B[a]P induced GVBD significantly at 10 and 100 ng/mL and decreased the ratio of E2/T significantly at 1 and 10 ng/mL compared with control. These findings suggest that B[a]P has different sensitivity to the oocyte maturation according to the oocyte diameters.

박대 생식소중량지수(gonadosomatic index, GSI)의 월간 변화는 2월에서 7월까지는 아주 낮은 값을 보였으나, 8월부터 생식소가 급격히 발달하기 시작하여 9월에 최고값을 나타내었다. 이후 10월부터 점차 낮아져 11월에 다시 낮아지는 경향을 보였다. 비만도(condition factor, CF)의 월간 변화도 9월에 최대값을 보였다. 간중량지수(hepatosomatic index, HSI)의 월간 변화는 6월에 최대값을 보인 후 7월에

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants derived from incomplete combustion of carbons and crude oil. In this study, we investigated the effects of benzo[a]pyrene (B[a]P), a representative PAHs on in vitro sex steroid hormone production and germinal vesicle breakdown (GVBD) using isolated oocytes of longchin goby (Chasmichthys dolichognathus) and chameleon goby (Tridentiger trigonocephalus). Oocytes in diameters of 0.8-0.9 (end vitellogenic stage) and 0.9-1.0 mm (germinal vesicle migratory stage) from longchin goby and 0.5 mm (fully vitellogenic stage) from chameleon goby were used. In GVBD assay, B[a]P at 10 nM stimulated GVBD in the oocytes of 0.8-0.9 mm from longchin goby. B[a]P at 1 nM stimulated GVBD in the oocytes with diameter 0.5 mm from chameleon goby. In steroid production from oocytes of longchin goby, B[a]P at 100 nM decreased testosterone (T) production, B[a]P at 1,000 nM increased estraiol-17 (J (E2) production and 10 and 100 nM increased -dihydroxy-4-pregnen-3-one () production in the oocytes with diameter 0.8-0.9 mm. B[a]P at 1,000 nM increased E2 production, 100 and 1,000 nM increased production in the oocytes with diameter 0.9-1.0 mm. In steroid production of oocytes from chameleon goby, B[a]P at 1,000 nM increased production. B[a]P at 10 nM increased production. In the ratio of to T (/T), B[a]P at 100 and 1,000 nM increased /T in the oocytes of longchin goby. B[a]P at 100 nM also increased /T in the oocytes of chameleon goby. Taken together, these results suggest that B[a]P have not only weak estrogenic effects but progestogenic effects on oocyte maturation.

The Egr family of zinc finger transcription factors consisting of 4 members (Egr1 to -4) regulates critical genetic programs involved in cellular growth, differentiation, and function. Especially, the critical role for Egr1 in regulating luteinizing hormone responsiveness was demonstrated by using gene-targeted mouse models. Other members of Egr family were shown to be involved in other cellular and developmental processes. To understand if Egr3 is implicated in ovarian functions, we focused on identifying cell type-specific and subcellular localization of Egr3 in cycling mouse ovaries and oocytes. RT-PCR analyses show that Egr3 mRNA is expressed in the mouse ovary and oocytes. By immunofluorescence staining, we observed that Egr3 is weakly expressed in subsets of granulosa cells. Interestingly, Egr3 seems to be co-localized with meiotic spindle in some oocytes in the ovarian section. Therefore, we examined Egr3 localization in MI oocytes cultured in vitro. We confirmed co-localization of Egr3 and microtubule in the mouse oocyte during meiosis I. Egr3 localization is noted around condensing chromosomes during prometaphase I (PMI). At metaphase I (MI) and MII, Egr3 is localized on meiotic spindle and also around each cytosolic microtubule organizing centers (MTOCs) in a punctate pattern. To examine if microtubule is required for correct positioning of Egr3 on this structure, we observed the pattern of Egr3 in oocytes matured under taxol or nocodazole. In taxol-treated oocyte, Egr3 and gamma-tubulin complex are enlarged. In nocodazole-treated oocyte, Egr3 localization on spindle and MTOCs are abolished. Thus, Egr3 localization seems to require the presence of intact microtubule. Collectively, our result shows for the first time that Egr3, a transcription factor, is localized on meiotic spindle of maturing mouse oocytes. The work suggests a novel role for Egr3 as a factor involved in MTOC dynamics during meiosis.

본 연구는 한방 약용자원으로 사용되고 있는 대추의 효능과 생리활성에 대한 연구의 일환으로 대추 열매의 열수 및 에탄올 추출물의 폴리페놀 화합물과 각 추출물의 항산화활성을 측정하였다. 대추 추출물의 수율은 미숙 대추가 가장 낮은 11% 내외의 수율을 보였으며, 완숙 대추가 미숙 대추보다 2배 이상 높은 수율을 보였고, 건조 대추의 열수 및 에탄올 추출물은 각각 55.67, 65.95%이었다. 대추 추출물의 폴리페놀 함량은 열수와 에탄올 추출물 모두

‘조망’은 조숙, 다수성이며 바이오디젤 생산 및 식용으로 이용이 가능한 유채로 2006년에 웅성불임인 목포-CGMS와 임성회복환분친인 8634-B-1-3-1-8을 교배하여 생산력검정시험(2007, 2008)과, 지역적응시험(2008)을 거쳐, 2008년에 육성된 품종이다. ‘조망’의 엽형은 하부익상전렬, 엽색 및 줄기색은 녹색, 결각은 중간이며 꽃색은 황색, 종피색은 흑색이다. ‘조망’의 성숙일는 6월 3일로 ‘선망’보다 1일, ‘탐미유채’보다는 4일

대하, Fenneropenaeus chinensis의 난소 성숙에 대한 serotonin(5-hydroxytryptamine, 5-HT), ecdysone(20-hydroxyecdysone, 20-E) 그리고 HCG(human chorionic gonadotropin)의 효과를 조사하였다. 대하에 5-HT(체중 g당 ), 20-E(체중g당 ) 그리고 HCG(체중 g당 5 IU, 10 IU)를 5일 간격으로 3회 주사한 후 10일이 더 지난 시점에 채집하

Previously, we obtained the list of genes differentially expressed between GV and MII oocytes. Out of the list, we focused on functional analysis of Zap70 in the present study, because it has been known to be expressed only in immune cells. This is the first report about the expression and its function of Zap70 in the oocytes. Synthetic 475 bp Zap70 dsRNA was microinjected into the GV oocytes, and the oocytes were cultured in vitro. In addition to maturation rates, meiotic spindle and chromosome rearrangements, and changes in expression levels of transcripts of three kinases, Erk1/2, JNK, and p38, were determined. Zap70 is highly expressed in immature GV oocytes, and gradually decreased as oocyte matured. When dsRNA of Zap70 was injected into the GV oocytes, Zap70 mRNA specifically and completely decreased by 2 hr and its protein expression also decreased significantly. Absence of Zap70 resulted in maturation inhibition at meiosis I (57%) with abnormalities in meiotic spindle formation and chromosome rearrangement. Concurrently, mRNA expression of Erk2, JNK, and p38, were affected by Zap70 RNAi. Therefore, we concluded that Zap70 is involved in MI-MII transition by affecting expression of MAP kinases.

Single seeds of common buckwheat cultivar Suwon No. 1 when subjected to SDS-PAGE revealed very high polymorphism. High variation existed for protein or protein subunits with molecular weight 54-47kDa, 45-25kDa and 16-11kDa. The electrophoregram showed variation for globulin as well as other protein fractions. About 300 proteins were separated by two-dimensional electrophoresis in common buckwheat (Fagopyrum esculentum Moench.) seed. Seed maturation is a dynamic and temporally regulated phase of seed development that determines the composition of storage proteins reserves in mature seeds. Buckwheat seeds from 5, 10, 15, 20, and 25 days after pollination and matured stage were used for the analysis. This led to the establishment of high-resolution proteome reference maps, expression profiles of 48 spots. It was identified 48 proteins from MALDI-TOF/MS analysis of wild buckwheat seed storage proteins. The 48 proteins were found identical or similar to those of proteins reported in buckwheat and other plants; it is belonging to 9 major functional categories including seed storage proteins, stress/defense response, protein synthesis, photosynthesis, allergy proteins, amino acid, enzyme, metabolism, and miscellaneous. It appears that the major allergenic storage protein separated played the important role in buckwheat breeding and biochemical characterization.

웅어 Coilia nasus의 생식소중량지수(GSI), 비만도지수, 난소 발달, 군성숙도, 포란수를 2007년 1월부터 2007년 12월 까지 조직학적 관찰 및 형태 측정 분석에 의해 조사하였다. GSI와 비만도지수는 2월에 증가하기 시작하여 난소가 성숙해지는 6월에 최대에 이르며, 그 후 산란이 일어나는 7월에 급격히 감소하였다. 그러므로 GSI와 비만도지수의 월별 변화는 난소의 성숙 및 산란과 밀접한 관련을 갖는다. 암컷의 난소 발달 단계는 연속적인

겨울철 사육조건이 양식산 뱀장어 Anguilla japonica의 인위적인 성성숙 유도 및 번식에 미치는 영향을 조사하였다. 봄철 salmon pituitary extracts(SPE)을 이용하여 성성숙 유도 예정인 양식산 암컷 뱀장어 친어를 8주의 겨울철 사육기간 동안, 4개의 사육환경(해수저온, ; 해수고온, ; 담수저온, ; 담수고온, )에 각각 순치하여 사육하였다. 그 후 각각의 실험구의 뱀장어를 해수 로 이동하여 SPE를 매주 1회 8주 동안

연어과 어류인 열목어 재조합 생식선자극호르몬(r-mtFSH 또는 r-mtLH) 투여에 따른 암컷 뱀장어(Anguilla japonica)의 성성숙 유도 효과를 조사하였다. 양식산 암컷 뱀장어를 해수에 적응시킨 후 매주 복강에 재조합 호르몬을 농도별(0.1, 1, 10 ㎍/㎖/fish)로 10주 반복 주사하였다. 매주 증체중을 측정하였고, 생식소중량지수 [GSI;(생식소중량/체중량×100]와 혈중 성호르몬 농도 변화를 조사하였다. 그 결과, 모든 실험군에서 GSI는 서서히 감소하는 경향을 나타내었다. 또한, 대조구에서 혈중 testosterone (T)과 estradiol-17β(E2)는 유의적인 증가를 보이지 않았지만, 재조합 호르몬을 투여한 실험군에서 투여 2주와 4주 후에 T와 E2 농도가 증가하였다. 또한, mt-rFSH(1, 10 ㎍/㎖/fish) 또는 mt-rLH(0.1, 1, 10 ㎍/㎖/fish)을 투여한 실험군에서는 난경이 대조구에 비해 유의하게 증가하였다. 이러한 결과는 열목어 재조합 생식선자극호르몬이 암컷 뱀장어의 초기 난성숙 발달을 유도함을 시사한다.