• Reproductive and Developmental Biology
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• 제28권4호
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• pp.267-273
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• 2004

Maturation of oocytes is maintained by complex procedures along with follicular genesis and is a critical step for embryonic development. Purine known as an oocyte maturation regulator is present in follicular fluid. In this study, the roles of guanosine as a strong inhibitor of GVBD and a modulator of cyclic GMP concentration in ooyctes were revealed. Denuded immature oocytes were treated with guanosine, and the maturation rates and cGMP concentration of oocytes were measured. GVBD was blocked in a concentration dependent manner by guanosine, but this effect was reversible. However, GVBD was lagged yet not significant by adenosine. Both guanosine and adenosine modified cGMP concentration in oocytes. The characteristic of the guanosine-treated oocyte was significantly higher cGMP compared with the adenosine-treated oocyes at initial time of the maturation. Based these results, guanosine may be a strong and reversible GVBD inhibitor. Although the precise mechanism of guanosine presently is unclear, the results suggest that guanosine may lead the accumulation of cGMP in oocyte cytoplasm, which in turn suppresses GVBD.

• 한국수정란이식학회지
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• 제33권4호
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• pp.271-280
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• 2018

Here, we evaluated the mode of programmed cell death during porcine oocyte maturation by comparing the two major pathways associated with programmed cell death, apoptosis (type I), and autophagy (type II). We investigated the expression and localization of major genes involved in autophagy and apoptosis at mRNA and protein levels. Furthermore, the effect of hormonal stimulation on autophagy and apoptosis was analyzed. We found that the activity of autophagy-associated genes was increased in the cumulus-oocyte complexes (COCs) following follicle-stimulating hormone (FSH) treatment, while the addition of luteinizing hormone (LH) reversed this effect. The expression of proteins associated with autophagy was the highest in FSH-treated COCs. On the other hand, caspase-3 protein level was maximum in COCs cultured with LH. The treatment with rapamycin resulted in the effect similar to that observed with FSH treatment and increased autophagy activity. Thus, hormonal stimulation of pig oocytes resulted in distinct patterns of maturation. The high-quality oocytes majorly relied on the type II pathway (autophagy), while the type I pathway (apoptosis) was more prominent among poor-quality oocytes. Further investigation of this distinction may allow the development of techniques to produce high-quality oocytes in porcine in vitro fertilization.

• 한국수정란이식학회지
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• 제12권3호
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• pp.293-300
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• 1997

Supplementation of maturation medium with additional granulosa cells has beneficial effect on in vitro maturation of bovine follicular oocytes and their subsequent cleavage and development in vitro. However, maturation system using granulosa cells have some disadvantages that collection of granulosa cells is cumbersome and metabolic activity of the cells is variable according to ovarian cycle or follicular size. We hypothesized that bovine immsture oocytes matured without granulosa cell coculture can fertilize and develop normally if the medium volume per oocyte is reduced during in vitro maturation. Immature oocytes were matured for 24 hours in a TCM199 containing 10% fetal calf serum, anterior pitultary hormone (0.02 AU /ml Antrinⓡ) and estradiol with or without granulosa cells in vitro. In Group 1, 35 to 40 oocytes were matured in a well of 4-well plastic dish containing 500 $\mu$l of maturation medium and granulosa cells, and 9 to 10 oocytes were matured in a 50-$\mu$l drop of maturation medium without granulosa cells in Group 2. After maturation, oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Inseminated oocytes were cultured in a microdrop (30 $\mu$l) of a synthetic oviduct fluld medium (SOFM) containing BSA, Minimum Essential Medium essential and non-essential amino acids for 9 days. As a preliminary experiment, we investigated the beneficial effect of granulosa cells during maturation on subsequent cleavage and development using the same type of culturedishes (4-well dish). Granulosa cells could not increase embryo cleavage after fertilization but significantly improved (p<0.05) embryo development to expanding blastocyst (Table1 and 2). In Group 1, 68 and 80% of inseminated oocytes have cleaved at 30 hours and 2 days after IVF, respectively, which is similar (p>0.05) to the result of Group 2 (69% at 30 hours and 78% at 2 days after IVF). The oocytes in Group 2 showed 21 and 11% of developmental rates to expanding and hatching blastocysts, respectively, which was not significantly different (p>0.05) from those (20 and 10%, respectively) of oocytes in Group 1. In conclusion, it has been clarified that a microdrop culture system without granulosa cells for in vitro maturation can support bovine embryonic development to blastocyst in vitro as readily as a granulosa cell coculture system.

• 한국동물학회지
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• 제28권2호
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• pp.99-107
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• 1985

생쥐난자의 성숙을 억제하는 것으로 알려진 dbcAMP와 progesterone을 동시에 처리하였을 때의 영향을 알아보기 위하여 본 실험을 행한 결과는 다음과 같다. 1) 10 $\\muM$/ml의 dbcAMP 혹은 2 $\\muM$/ml의 progesterone을 단독처리하였을 때에는 난자의 성숙을 억제하지 못했다. 2) 그러나 dbcAMP(10 $\\mug$/ml)와 progesterone(2 \\mug$/ml)을 동시에 배양액에 처리하였을 때에는 난자의 성숙이 억제되었다. 3) 난자를 먼저 4시간동안 위의 두 억제제가 들어있는 배양액에서 배양한 뒤 다시 정상배양액에 옮겨 계속 배양하면 나자들은 대조군에서와 같은 비율로 성숙을 하였다. 위의 결과로 보아 dbcAMP와 progesterone이 동시에 배양액에 존재할 때에는 각각 단독 존재하였을 때 영향이 없는 농도의 dbcAMP와 progesterone으로도 능히 난자의 성숙을 억제 시킬 수 있었으며 이 같은 억제효과는 가역적임을 알게되었다.

• Reproductive and Developmental Biology
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• 제28권2호
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• pp.95-99
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• 2004

본 연구는 미성숙돼지 난모세포의 체외 성숙에 있어서 myo-inositol의 영향을 알아보기 위하여 실시하였다. 미성숙 돼지 난모세포을 myo-inositol을 포함하는 또는 포함하지 않은 체외 성숙배지에서 44시간 체외 성숙을 유도하였을 때 성숙율은 myo-inositol을 첨가한 성숙배지에서 성숙시킨 실험구에 유의하게 높았다(P<0.05). Myo-inositol 에 의한 성숙율 향상에 있어서 난구세포의 영향을 알아보기 위하여 난구세포의 치밀도에 따라 분류하여 미성숙 난모세포을 myo-inositol을 포함하는 체외 성숙배지에서 배양하였을 때 난구세포가 치밀한 미성숙 난모세포에서가 더 높은 성숙율을 나타내었다(P<0.05). 그러나 난구세포가 치밀하지 않은 미성숙 난모세포도 myo-inositol을 포함하는 성숙배지에서 배양하면 성숙율은 대조구에 비하여 유의하게 높았다(P<0.05). 이러한 결과는 돼지 난모세포의 체외 성숙배지에 myo-inositol의 첨가는 체외 성숙율을 향상시킬 수 있음을 나타내고 있다.

• Journal of Animal Science and Technology
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• 제57권8호
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• pp.27.1-27.6
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• 2015

This study was designed to investigate the effects of ${\alpha}$-tocopherol and granulosa cells monolayer on nuclear maturation and cleavage rates of ovine cumulus-oocyte complexes (COCs). The COCs (n = 2814) were matured in maturation medium supplemented with various concentration of ${\alpha}$-tocopherol (0, 5, 10, $15{\mu}g/ml$), oocytes were incubated at $39^{\circ}C$ with 5 % $CO_2$ for 24 h in three culture systems: (a) maturation medium (MM; n = 884), (b) co-cultured with granulosa cells (CG; n = 982) and (c) co-cultured with granulosa cells and cells were further cultured in MM for 12 h (CG + 12hMM; n = 948). Our results showed that ${\alpha}$-tocopherol had no effect on GVBD and MII as compared to control group, but when ${\alpha}$-tocopherol added to maturation medium the rate of cleavage decreased. This indicates interaction of above mentioned factors in any of the treatments showed no significant differences on the rate of maturation and cleavage stages (MII, GVBD and cleavage) (p > 0.05). The oocytes co-cultured with granulosa cells for 24 h had beneficial effects on cleavage rate. The maximum MII and cleavage rates were achieved when oocytes had extra 12 h culture in the maturation medium without granulosa cells. Results also showed our modified co-culture system (CG + 12hMM), improved rates of MII and the cleavage in comparison with other studied maturation systems.

• 대한본초학회지
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• 제22권3호
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• pp.127-132
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• 2007

Objectives : FLOS CHRYSANTHEMI (FC) has been reported to possess a variety of pharmacological activities. However, the effect of FC on the dendritic cells has not been determined. Methods : To examine the effect of FC on the immune response, we used several methods such as flow cytometric analyses, enzyme-linked immunosorbent assay. Results : 1. FC inhibited lipopolysacchride (LPS)-induced maturation of bone marrow-derived dendritic cells (BMDC) such as down-regulation of MHC class II and CD40. 2. FC also inhibited uptake of FITC-Dextran in BMDC stimulated with LPS. 3. Furthermore, FC inhibited several kinds of cytokine production such as TNF-a, IL-6 and IL-12 in BMDC. Conclusions : These results suggest that FC plays pivotal role m the development of inflammatory diseases.

• 한국식품저장유통학회지
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• 제11권4호
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• pp.461-467
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• 2004

장기간 자연숙성시킨 전통된장의 항산화 효과는 숙성기간이 길어질수록 대체로 감소되었으며, 숙성기간 1년 된장에서 methanol 분획물이 가장 우수하였으며, 다음으로 hexane과 물 분획물의 순이었다. 지용성 및 수용성 추출물은 숙성기간이 길어질수록 항산화 효과가 점진적으로 오히려 증가되었으나, 분획물에 비하여 낮은 수치를 나타내었다. 전통된장의 수소공여능은 수용성 색소 추출물과 methanol 및 butanol 분획물에서 대체적으로 높은 수치였으나, chloroform과 ethylacetate 분획물은 매우 낮았다. 전통된장의 항암효과에서 인체 폐암 세포주(A549)는 물 분획물에서 가장 높았고, 인체 유방암 세포주(MCF-7)는 methanol 분획물에서 가장 높은 수치를 나타내었으며, 특히 전통된장의 숙성기간이 길어질수록 암세포 성장억제효과는 높게 나타났다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.242-242
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• 2004

This study was conducted to investigate the effects of various supplements added into maturation medium of immature porcine oocytes on quantity of cytoplasmic lipid droplets(LD), subsequent fertilization and development to the blastocyst stage in vitro. The basic maturation medium was TCM 199 + 1 ㎍/㎖ FSH, 0.57 mM cystein, 10 ng/㎖ EGF and was supplemented various supplements(10% FBS, 10% pFF, 0.4% BSA, 1.0% BSA, 0.4% PVP, 1.0% PVP). (omitted)

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.72-72
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• 2002

Oocytes maturation, characterized by germinal vesicle (GV) breakdown, formation of the first meiotic spindle, expulsion of the first polar body and arrest in metaphase of second meiotic division (MII), occurs in preovulatory follicles in response to the surge of gonadotropin and leads to an ovulated oocyte in vivo. However, meiotic resumption in vitro occurs spontaneously following removal of cumulus-oocytes complexes (COCs) from the follicle. (omitted)