Transducin-like enhancer protein 1(TLE-1) is protein associated with cell proliferation. This study analyzed change of TLE-1 mRNA expression during in vivo and in vitro maturation in porcine oocytes. Oocytes and granulose cells were collected from follicles of <2 mm, 2～6 mm and >6 mm in diameter in slaughtered pig’s ovaries. Oocytes collected from follicles of 2～6 mm in diameter were used after in vitro maturation for 0, 10, 20 and 44 h. Cumulus cells and granulose cells were collected after treatment with hyaluronidase. In results, TLE-1 mRNA expression in oocytes collected from follicle >6 mm in diameter is increased, TLE-1 RNA expression in cumulus cells and granulosa cells from follicles <2 mm, 2 mm 6 mm and >6 mm in diameter. However, there is no significant difference. On the other hand, TLE-1 mRNA expression from oocytes cultured for 10 h and 44 h is increased, TLE-1 mRNA in cumulus cells cultured for 10 h is significant increased(p<0.05) than other culture periods. In conclusion, these results show that TLE-1 is expressed in all cell types of oocytes, cumulus cells and granulose cells, and associated with oocyte maturation.

The objective of this study was to evaluate the effects of co-culture of bovine oocytes with cumulus cells on in vitro maturation and development following in vitro fertilization in bovine oocytes. Bovine cumulus-oocyte complexes (COCs) and denuded oocytes (DO) were co-cultured with the cumulus cells in TCM199 for 20~22 hr, and evaluated the nuclear type of oocyte. After in vitro maturation, oocytes were coincubated for in vitro fertilization with frozen-thawed spermatozoa selected by 65% percoll in DM-Heparin and DM-Caffeine for 15~18 hr. Presumptive zygotes were cultured for 48 hr in CR1aa in vitro culture medium with 10% FBS, and evaluated the cleavage rates. The results confirmed that the highest percentage of metaphase II (M-II) stage was observed in COCs (30.1±3.5%, 24.2±1.8%) as compared to DO (7.1±1.3%, 17.4±13.9%) (p<0.05). In addition, the increased cleavage rates were obtained from COCs (69.6±2.1%, 75.6±2.9%) when compared to DO (21.6±7.5%, 29.5±12.6%) (p<0.05). In conclusion, this study suggested that cumulus cells secreted positive factors during in vitro maturation of oocytes and early embryonic development after in vitro fertilization of bovine oocytes.

Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, OC and COCs. Activity of MMP-2 in the OC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the COCs 36 hr. Expression of TIMP-3 protein in the COCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

멸종위기에 처한 꼬치동자개(Pseudobagrus brevicorpus)의 보전과 복원을 위한 기초 자료를 제공하기 위하여 인공사육 환경에서 사육된 종묘에서 친어까지의 성장과 성성숙 특징들을 조사하였다. 부화 후 698일의 인공사육 기간 동안 전장과 체중은 각각 89.22±10.29mm와 70.93±7.68g로 성장하였고, 전장 (TL)과 체중(BW)의 관계는 BW=5×10-5 TL2.678 (R2=

조류가 식물 열매의 성숙도와 크기에 따라 어떻게 선택하는지를 알아보기 위하여 사육 중인 3종의 새에게 성숙도와 크기가 다른 식물 3종의 열매를 주어 그 먹는 양을 계수하고, 다변량통계로 분석하였다. 실험에 사용한 새는 금계(Chrysolophus pictus, 꿩과), 양비둘기(Columba rupestris, 비둘기과)와 자바공작 (Pavo muticus, 꿩과)이고, 열매는 왕벚나무(Prunus yedoensis, 장미과), 뜰보리수나무 (Elaea

억새의 최적 조직배양조건을 확립하기 위하여 성숙종자로부터 배발생 캘러스 유도 및 캘러스로부터 식물체 재분화에 미치는 식물생장 조절물질의 영향을 조사하였다. 배발생 캘러스 유도시 첨가되는 auxin으로는 2,4-D가 가장 효율적이었으며, 3 mg/L 2,4-D가 첨가된 배지에서 배발생 캘러스가 가장 높은 빈도로 유도되었다. 식물체 재분화는 배발생 캘러스를 1 mg/L 2,4-D와 2 mg/L BA가 첨가된 재분화 배지에서 배양했을 때 가장 높은 재분화효율

페레니얼 라이그라스의 5가지 품종을 이용하여 성숙종자로부터 배발생 캘러스 유도 및 캘러스로부터의 식물체 재분화 효율을 조사하였다. 성숙종자로부터 4주 동안 배양한 캘러스 유도율은 'Accent', 'Topgun', 'Renenge GLX', 'Bison', 'Tetrellite' 순으로 높은 캘러스 유도율을 보였고 암상태에서 배양한 캘러스가 광상태에서 배양한 것보다 7~9% 정도 높은 효율을 나타내었다. 식물체로의 재분화에 적합한 배지는 MS 배지보다

This study examined the maturation and main spawning season of Japanese spear lobsters, Linuparus tirgonus(Von Siebold) captured around Jeju island from January to September, 2008. Carapace length(CL), body weight(BW) and gonad weight(GW) were measured. Gonadosomatic index(GSI) and sex ratio were calculated. In female group, CL showed the highest value in June and decreased after August. GW showed a peak in July and decreased rapidly after August. The mean gonadosomatic Index(GSI) reached a maximum value between June and August. Number of egg ranged from 143,360 to 189,504.

Maturation and spawning of the jack mackerel, Trachurus japonicus was investigated based on the samples captured in the coastal waters around Jeju Island from January 2007 to December 2008. Gonadosomatic index began to increase in March, and reached a maximum between April to June. After spawning it began to decrease from August. Reproductive season was estimated to March-July, with peak in April. Fecundity was proportional to the size of the female, with the clutch size varying from 33,493 eggs in the smallest female(FL=27.0cm) to 627,061 eggs in the largest(FL=40.6cm). Size at 50% sexual maturity(FL50), determined from mature females, was 26.6cm FL. Annual reproductive cycles of this species could be divided into six successive stages; immature stage(September-December), nucleolus stage(January-February), yolk vesicle stage(February-March), vitellogenic stage(March-April), ripe stage(April-July) and spent stage(July-September).

The objective of this study was to examine the effect of eCG and various concentrations (20, 40, and 80 ) of porcine FSH on nuclear maturation and intracellular glutathione (GSH) level of oocytes, and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (10 IU/ml hCG and 10 IU/ml eCG or FSH) for the first 22 h and then further cultured in hormone-tree medium for an additional 22 h. Nuclear maturation of oocytes () was not influencem foreCG and various concentrations FSH. Embryonic development to the cleavage stage () and mean number of cells in blastocyst ( cells) after PA were not altered but blastocyst formation e-treignificaddlor(p<0.05) improvem forthe supplementation eith 80 FSHr(64%) compared to 47%, io8%, iand 47% in oocytes that were treated with eCG, 20,i and 40 FSH,i numectivelo. In SCNT, fusion () of cell-cytoplast couplets and siosequent embryo cleavage () were not influencem fordifferent gonadotropins but blastocyst formation tended to increase forthe supplementation eith 80 FSHr(25% vs. ). Our nuults demonstrated that oocyte maturation and embryonic development after PA and SCNT e-frinfluencem fortype of gcem fortype of gits concentration. In this study, supplementation of maturation medium eith 80 FSHrimproved preimplantation development of PA and SCNT pig embryos, probably by increasing intracellular GSH concentration of matured oocytes.

This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.

본 실험은 지베렐린이 ‘신고’배 숙기와 비대에 미치는 영향을 조사하기 위하여 실시되었다. 과실중 조사에서 모든 Gibberellic acid A4+7 처리구는 대조구인 지베렐린 도포제와 비등한 약효가 나타났으며, 무처리구에 비해서는 증가를 나타내었다. 횡경은 GA4+7을 만개후 35일째에 도포한 것이, 종경은 만개후 40일째 도포한 것이 가장 효과적인 것으로 나타났으며, 과형지수는 모든 처리구에서 비슷한 수치로 나타났다. ‘신고’배 발달 초기에는 GA4+7 2.4% 처리구가 GA3+GA4+7 2.7% 처리구나 무처리구보다 현저하게 큰 차이로 비대가 되는 것을 볼 수가 있으나, 성숙기에 들어서면 GA4+7 2.4% 처리구와 GA3+GA4+7 2.7% 처리구의 크기와 가시적인 착색도가 거의 비슷해지는 것을 볼 수 있었다. GA3+GA4+7의 성숙촉진 효과를 검토하기 위하여 에테폰 액제와 비교하여 실험한 결과, GA3+GA4+7를 만개후 35일째에 어린 과실에 도포한 것이 에테폰과 비슷한 정도로 착색이 되었으며, 무처리구와 비교했을 때에는 모든 GA3+GA4+7 처리구와 에테폰 처리 모두 착색이 6일정도 빨라 착색속도에 현저한 차이를 보여주었다. 모든 GA3+GA4+7 처리구는 무처리구에 비하여 당도는 증가하고 산도는 감소하였으며, 과실의 경도는 무처리구보다 낮은 것으로 나타났으나 유의한 차이는 없었다. 상기의 결과로 GA3+GA4+7 처리구 모두가 에테폰이나 무처리구보다 과실의 비대에 있어 효과가 큰 것으로 나타났다. 특히 GA3+GA4+7를 만개후 35일째에 처리한 것이 횡경 및 종경, 과형지수 모두 가장 높게 나타났으며 착색 또한 깔끔하게 되어 상품성이 높게 평가되었다.

티모시의 colt 품종을 이용하여 최적 조직배양조건을 확립하기 위하여 성숙종자를 배양함에 있어 첨가되어지는 식물 생장조절 물질의종류와 농도가 배발생 캘러스 유도 및 식물체재분화에 미치는 영향을 조사하였다. 배발생 캘러스 유도시 첨가되는 2,4-D는 3 mg/L 첨가 구에서 78%로 캘러스 유도율이 가장 높았으며, 3 mg/L 2,4-D와 0.1 mg/L의 BA를 첨가하여 배양했을 때 형성된 캘러스는 조직적으로 치밀하며 유백색의 배발생 캘러스가 가장 많이