Ecological characteristics of a brown alga, Scytosiphon lomentaria, were investigated from January 2021 to December 2021 in its natural habitat off Sodol, Jumunjin, eastern coast of Korea. The S. lomentaria population at the site formed widespread patches on mid shore. During the investigation, environmental conditions including seawater temperature, salinity, and dissolved oxygen were monitored at the site. Growth and maturation of the S. lomentaria population were identified through qualitative and quantitative investigations. An estimation of the effective cumulative temperature for maturation of the alga was obtained based on growth data and a biological zero temperature of 8°C. Sporangia were observed from February to May when seawater temperatures ranged from 7.7°C to 16.4°C. A maturation peak was detected in April when seawater temperature was 12.1°C. After zoospore release, the alga became bleached and only the crust remained after June. Developmental initiation of the thallus occurred at temperatures above 8°C. Its maturation required approximately 162 degree-days.

큰열매모자반은 제주도 연안 생태계에서 수관층을 형성하는 중요한 다년생 갈조류이다. 이 연구는 2018년 5월부터 2019년 6월까지 제주연안에서 큰열매모자반 개체군의 생장 및 생식패턴을 밝히기 위하여 수행되었다. 큰열매 모자반 개체군의 정량조사를 통해 월별 형질분석, 밀도 및 현존량 분석을 실시하였다. 또한 큰열매모자반의 생식자원 배분 특성을 조사하기 위하여 전체 엽체길이 및 엽중량에 대한 생식엽길이와 중량비율을 측정하였다. 조사지역에서 큰열매모자반의 최대 엽장은 6월에 135.3±20.0 cm, 최대 엽중량은 5월에 평균 3.6±2.1 kg·wet-wt., 평균밀도는 4.5 individuals·m-2 및 평균현존량은 4.6 kg·wet-wt.·m-2였다. 생식기탁의 형성은 4~8월 (수온 16.1~25.0°C)까지 관찰되었으며, 난방출 시기는 6~7월 (수온 19.3~22.9°C)이 었다. 엽체의 발달은 수온 14.1°C 이상의 조건에서 이루어졌으며, 성숙에 요구되는 유효 적산온도는 726.3 degreedays였다. 큰열매모자반의 생식배분은 6월에 최대 69.3% 로 나타났다. 큰열매모자반의 생장과 성숙패턴은 생장기 (10~1월), 생식시작기 (2~4월), 성숙기 (5~6월), 난 방출기 (6~7월) 및 휴지기 (8~9월)로 구분되었다.

The objective of this study was to establish an in vitro culture system for ovarian preantral follicles of B6D2F1. First, we optimized the in vitro preantral-follicle culture by culture duration, follicle stimulating hormone (FSH) type, and activin A concentration. Duration of in vitro culture for 9, 11, and 13 days was sufficient for the normal development of preantral follicles to antral follicles. Formation of cumulus cell–oocyte complex (COC) was induced by treatment with human chorionic gonadotropin (hCG; 2.5 IU/mL) and epidermal growth factor (EGF; 5 ng/mL). In addition, metaphase II (MII) oocytes formed during this in vitro culture of preantral follicles. In vitro preantral-follicle culture for 9 days showed higher rates of growth and maturation, thus yielding a greater number of antral follicles, and there were significant differences (p < 0.05) in the number of MII oocytes (that formed from these preantral follicles via differentiation) between the 9-day culture and 11-day or 13-day culture. The follicles cultured for 9 days contained a tightly packed well-defined COC, whereas in follicles cultured for 11 days, the COC was not well defined (spreading was observed in the culture dish); the follicles cultured for 13 days disintegrated and released the oocyte. Second, we compared the growth of the preantral follicles in vitro in the presence of various FSH types. There were no significant differences in the growth and maturation rates and in differentiation into MII oocytes during in vitro culture between preantral follicles supplemented with FSH from Merck and those supplemented with FSH from Sigma. To increase the efficiency of MII oocyte formation, the preantral follicles were cultured at different activin A concentrations (0 to 200 ng/mL). The control follicles, which were not treated with activin A, showed the highest rate of differentiation into antral follicles and into MII oocytes among all the groups (0 to 200 ng/mL). Therefore, activin A (50 to 200 ng/mL) had a negative effect on oocyte maturation. Thus, in this study, we propose an in vitro system of preantral-follicle culture that can serve as a therapeutic strategy for fertility preservation of human oocytes for assisted reproductive medicine, for conservation of endangered species, and for creation of superior breeds.

This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.

The expression of MMPs in the development of the fertilized egg has a very important role in cell configuration. Objective To evaluate the clinical, the effect of differentially expressed MMPs on serum and serum - free medium on the maturation of blastocysts. The expression patterns of MMPs in serum and serum-free medium were compared at 6 h, 18 h and at the blastocyst stage using real-time PCR, ELISA and immunofluorescence. The results showed that the expression of MMPs was increased in the embryos of the serum medium, as a result of analysis of MMPs and TIMPs, MMP-2 was expressed in the cytoplasm of embryos in the serum-free medium, And it was found to be higher in expression than MMP-9. The serum medium was different from the bloodless badge: overall, TIMPs showed a higher expression in the ovarian cells than cyanosis, and TIMP-3 was more pronounced. Development rate of blastocyst according to in vitro culture method was higher than that of serum - free medium (61.22% 60/98) and serum - free medium (48.28% 28/58). Analysis of the protein release locations of MMPs and TIMPs showed that MMPs and TIMPs are highly expressed in serum mediums, focusing on the inner cell mass. However, very low expression appeared in the tropoblast. On the other hand, serum - free medium showed different expression from serum medium and TIMPs expression was generally low. Therefore, in the case of serum media, the expression of MMPs is highly expressed in the cytoplasm of the fertilized egg, increasing the reconstruction of cells.

The aim of this study was to investigate the role of Src homology 2-containing phosphotyrosine phosphatase SHP2 in intricate signaling network invoked by oocyte to achieve cytoplasmic maturation and also blastocyst development. Activation of SHP2 regulates multicellular differentiation, proliferation and survival through numerous signal pathways. The most prominent pathway is RAS/PI3K and p-AKT signaling cascade, as a result mitogenic effect become enhanced. Oocytes were cultured in cisplatin an anticancer drug, but selective activator of SHP2 and our grouping were SOF medium alone, SOF + EGF, SOF + CISPLATIN 0.3 μM, and SOF + EGF + CISPLATIN 0.3 μM. We evaluated that EGF neutralizes the apoptotic effect of cisplatin as well as maintain the high expression of SHP2, as a result blastocyst development become boosted up. We also found that inhibition of SHP2 with its specific inhibitor PHPS1 5 μM decreases the blastocyst development and neutralizes growth factors effect. The developmental ability and quality of bovine embryos were determined by assessing their cell number, gene expression, immunofluorescence, and immunoblot. The differences in embryo development between experimental groups were analyzed by one-way ANOVA. Our results show that SHP2 have significant effect on MAP kinase pathways which expand the cumulus cells during oocyte maturation and blastocyst development as compare to inhibition of SHP2 with PHPS1. SHP2 not only transduce the signaling of epidermal growth factor but it also has a role in signal transduction of FGF and IGF. The expression of ERK, PI3K/p-AKT and mTOR was increased with EGF, but with the treatment of SHP2 inhibitor the expression of these genes become drop done. So we can conclude from these results that SHP2 is important for oocyte maturation as well as for blastocyst development.

본 연구는 방사무늬김(Pyropia yezoensis) 사상체의 생장에 대한 단색광의 영향을 연구하기 위해 blue light(480 ㎚), green light(550 ㎚), yellow light(600 ㎚) and red light(730 ㎚)에서 방사무늬김(P. yezonesis)의 유리 사상체를 배양한 후 colony 직경, chlorphyll a 함량, 각포자낭 형성률을 측정하여 생장특성을 비교 분석하였다. 그 결과, colony 직경과 chlorophyll a 함량이 480(blue light) ㎚에서 각각 2,472.6±27.0 ㎛ and 1.55±0.03 mg g.dw-1로 최대값이 나타났고 각포자낭 형성률 역시 37.87±1.08 %로 blue light에서 최대값으로 확인되었다. 따라서 blue light는 다른 단색광(green, yellow, red light)에 비해 김 사상체의 성장 및 성숙을 촉진하는 데에 효과적으로 작용하며, 본 연구결과 는 향후 김의 실내 양식 기술 개발에 활용될 수 있을 것이라 사료된다

The oocyte undergoes various events during maturation and requires many substances for the maturation process. Various intracellular organelles are also involved in maturation of the oocyte. During the process glucose is essential for nuclear and cytoplasmic maturation, and adenosine triphosphate is needed for reorganization of the organelles and cytoskeleton. If mitochondrial function is lost, several developmental defects in meiotic chromosome segregation and maturation cause fertilization failure. The endoplasmic reticulum, a store for Ca2+, releases Ca2+ into the cytoplasm in response to various cellular signaling molecules. This event stimulates secretion of hormones, growth factors and antioxidants in oocyte during maturation. Also, oocyte nuclear maturation is stimulated by growth factors such as epidermal growth factor. This review summarizes roles of organelles with focus on the Golgi apparatus during maturation in oocyte.

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=

The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from 2~6mm follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, 0.5㎍/ml FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at 39℃ in a humidified atmosphere of 5% CO2 in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with 1% orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm (1 x10 5 cells/ml) in mTBM with 0.3% BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with 0.4% BSA. At 6hr of culture, the embryos were fixed in 3.7% formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and ≥3 PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group (67%), but it did not differ among the all added groups (86%, 85%, 79% and 81%, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups (25%, 30%, 33%, 29% and 29%, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased (66% vs 58%, 54%, 52% and 55%, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

우리 나라의 서해와 남해 연안을 분포하는 꽃새우는 북쪽으로는 강화도에서 남쪽으로 사랑도까지 널리 분포한다. 본 연구에서 대상으로 한 거문도 인근해역 꽃새우의 암ㆍ수 성비는 암컷이 48.6%로 나타났고, 이 개체군은 7월과 8월에 성숙하고, 년 1회 신생 개체군이 발생되었다. 교미한 개체는 7월과 8월에 출현하기 시작하고, 갑각장 19 ㎜ 이상의 암컷에서는 반 이상이 교미한 것으로 나타났으며, 숙도지수는 7-8월에 최고치를 나타내었다. 암컷의 최소 성숙개체는 갑각장 18 ㎜이고, CL<SUB>50</SUB> 성 성숙체장과 교미체장은 각각 갑각장 18.89 ㎜와 19.19 ㎜로 나타났다. 체장 빈도 변화에 의해 추정된 거문도 개체군의 수명은 암컷 14-15개월이고, 수컷은 13-147개월 나났다. 변형된 von Bertalanffy 성장식에 추정된 성장계수(K)는 암ㆍ수 각각 1.40/년과 2.00/년이고, 최대 갑각장(L<SUB>∞</SUB>)은 암ㆍ수 각각 29.54 ㎜와 18.95 ㎜이었으며, 암컷이 수컷 보다 성장이 빠르고, 큰 것으로 나타났다.

The stimulatory effect of EGF and FSH on oocyte maturation have been reported in various mammalian species. And some reports presented FSH enhanced the effect of EGF on oocyte maturation. But, the interaction between EGF and FSH on nuclear maturation of mammalian oocytes is not fully understood. We observed the effect of EGF and FSH on nuclear maturation during in vitro maturation of mouse oocytes. Also, we examined the interaction between EGF and FSH on nuclear maturation of mouse oocytes using the EGFR inhibitor or FSH inhibitor. Germinal vesicle (GV) stage oocytes were obtained from 3-4weeks PMSG primed BCFI hybrid mice and cultured in TCM-199 medium with 0.4%PVP supplemented with/without EGF (1ng/ml), FSH (1ug/ml), EGFR specific tyrosine kinase inhibitors: Tyrphostin AG 1478 (500nM), MAP kinase kinase inhibitor : U0126 (20uM) or PD 98059 (100uM) for 14-l5hr. Rapid staining method were used for the assessment of nuclear maturation. Nuclear maturation rates of EGF indjor FSH-treated group were significantly higher than those of control group. Treatment of EGFR inhibitor significantly block the nuclear maturation of GV oocyte in EGF-treated group, but it did not block those of GV oocyte in FSH-treated or FSH and EGF-treated group. Treatment of FSH inhibitor(U0126, PD98059) significantly block the nuclear maturation of EGF-treated group, FSH-treated and FSH and EGF-treated group. These results show that EGF has a stimulatory effect as well as different action pathway with FSH on in-vitro maturation of mouse oocyte in vitro. Therefore, further studies will be needed to find the signaling pathway of EGF associated with nuclear maturation.

본 연구는 성장인자 EGF, IGF-1, EGF+IGF-1 이 소 난포란의 체외성숙에 미치는 영향을 규명하기 위하여 실시하였다. 소 난포란의 체외성숙 시 EGF를 농도별 (10, 50 및 100ng/m1)로 첨가하여 실험한 결과 통계학적 유의성은 인정되지 않았으나 24시간 배양 후 성숙율은 대조군에 비하여 높은 경향을 나타내었다. 소난포란의 체외성숙 시 EGF와 IGF-1을 병용 처리하여 실험한 결과, EGF 또는 IGF-1 단독처리 군에 비하여 병용처