• Development and Reproduction
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• v.14 no.2
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• pp.115-121
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• 2010

The in vitro effects of nonylphenol (NP) and 2,2',4,6,6'-pentachlorobiphenyl (PCB104) on ovarian steroidogenesis of the ribbed gunnel, Dictyosoma burgeri were investigated. Oocytes taken during maturation stage were incubated with 100 ng/$m{\ell}$ of NP and PCB104 in the presence of exogenous precursor, $[^3H]-17{\alpha}$-hydroxyprogesterone ($[^3H]-17{\alpha}OHP$). Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography. The identities of the major metabolites were testosterone (T) and estradiol-$17{\beta}$ (E2). NP treatment inhibited production of E2 metabolite in the oocytes of 1.2, 1.3 and 1.4 mm although NP inhibited production of T metabolite at the oocytes of 1.1, 1.3 and 1.4 mm. PCB104 treatment inhibited production of T metabolite in the oocytes of all groups and E2 metabolite in the oocytes of 1.2, 1.3 and 1.4 mm. In conclusion, these results suggested that NP and PCB104 had an inhibitory effects on conversion of $[^3H]-17{\alpha}OHP$ to T and E2 during the oocyte maturation process of ribbed gunnel.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.91-98
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• 2001

We evaluated the effects of the substrate and inhibitors related to phosphatidylinositol metabolism on in vitro maturation and fertilization of porcine oocytes. Cumulus-oocyte complexes were cultured in mTLP-PVA medium supplemented with or without inositol (250 mM) fur 46h. Subsequently, these oocytes were inseminated with fresh boar semen in mTALP-PVA medium for 6h. At 6h after insemination, oocytes were cultured for further 12 h in TCM-199 supplemented with 10% FBS (fetal bovine serum). The higher percentage of oocytes in inositol-supplemented medium reached metaphase of the second meiotic division compared to those in control (81.4% vs. 67.3%; P<0.()5). following 18 h of insemination, more number of male pronuclei were formed in the oocytes matured in inositol-supplemented medium than in those of control experiment (42.0% vs. 27.3%; P<0.05). When oocytes were cultured in medium with 10mM LiCl (chloride lithium) or 0.5mM dbcAMP (dibutyryl cyclic adenosine monophosphate) to determine the role of inositol on the maturation of oocytes, these two drugs inhibited the meiotic division of oocytes (P<0.05). However, addition of inositol to the culture medium did overcome the inhibitory effect of these drugs on the oocyte maturation. DbcAMP and verapamil supplemented synergistically arrested the meiotic division of oocytes. Addition of verapamil did not inhibit germinal vesicle breakdown, but it severly inhibited the second meiotic division of oocytes. These results suggest that inositol exert its improving effects on maturation, by activating the PI (phosphatidylinositol) cycle and causing beneficial changes in both cytoplasm and membrane of oocytes.

• Proceedings of the Korean Nutrition Society Conference
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• 2000.11a
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• pp.67-67
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• 2000

• Proceedings of the Korean Nutrition Society Conference
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• 2000.11a
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• pp.73-73
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• 2000

• Proceedings of the Korean Nutrition Society Conference
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• 2000.11a
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• pp.74-74
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• 2000

• The Korean Journal of Zoology
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• v.36 no.1
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• pp.77-83
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• 1993

난모세포성숙기 (peripheral germinal vesicle stage)의 무지개송어(Oncorhynchus mykiss)를 대상으로 방사성 표지 전구체(3H-pregnenolone, 3H-l7$\alpha$-hydroxyprogesterone 그리고 3H-androstenedione)를 이용하여 유 nitro에서 관찰한 협막세포층의 스테로이드 주 대사 경로는 다음과 같으며, pregnenolone -) 17a-hydroxypregnenolone$\longrightarrow$ 17u-hydroxyprogesterone $\longrightarrow$ androstenedione $\longrightarrow$ testosterone 이는 $\Delta$5-스테로이드 경로의 존재가능성을 지적하고 있다.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.95-95
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• 2003

Inositol은 세포의증식 및 정보전달과정에 관여하는 phosphatidylinositol (PI)의 구성성분으로서 중요한 세포내 기능을 수행한다. P1는 세포내에서 특이적 인산화효소에 의하여 Pl4-phosphate(PIP), Pl4,5-phosphate($PIP_2$)로 변환되며 PIP$_2$는 phospholipase C(PLC)에 의하여 세포내 second messengers인 1,2-diacylglycerol (DAG)와 inositol 1,4,5-triphosphate ($IP_3$)로 변환된다. 이렇게 생산된 DAG와 IP3는 각각 protein kinase C의 활성과 $Ca^{2++}$의 동원에 관여하여 다양한 세포내 신호전달에 관여하는 것으로 보고되고 있다. 또한 mouse에서 $IP_3$의 작용에 의한 $Ca^{2++}$의 상승은 난모세포의 성숙분열이 촉진되고 돼지 난포세포에 있어서도 PI대사가 일어나고 있는 것으로 보고되고 있다. 본 연구에서는 돼지 미성숙 난모세포의 성숙과 단위발생에 미치는 inositol의 영향을 확인하기 위하여 실시하였다. inositol의 농도가 난포란의 성숙에 미치는 영향을 검토하기 위하여 난포란을 각각 $150 \mu mole$, $250 \mu mole$, $350 \mu mole$, inositol을 포함하는 Whitten's 배양액에서 44시간 성숙시킨 결과 $93.57 \pm 4.21, 93.91 \pm 2.71, 92.96 \pm 3.58%$가 성숙되어 대조구의 $87.10 \pm 4.21$보다 유의적(P<0.05)으로 차이를 나타내었다. 난포란의 등급에 따른 inositol의 영향을 확인하기 위하여 형태적으로 난구세포가 치밀한 난포란과 난구세포가 치밀하지 않은 난포란을 $250 \mu mole$ inositol을 포함하는 Whitten's 배양액에서 성숙을 유도한 결과 난구세포가 치밀한 난포란과 난구세포가 치밀하지 않은 난포란에서 inositol을 첨가하였을 때 성숙율은 각각 $95.35 \pm 2.22와 63.55 \pm 8.12$로 inositol을 첨가하지 않은 대조구보다($89.21 \pm 3.69와 48.56 \pm 8.99$) 유의적인 차이를 보였다. 난구세포가 inositol에 의한 성숙에 미치는 영향을 확인하기 위하여 난구세포를 제거한 난포란을 inositol을 포함하는 배지에서 성숙을 유도한 결과 inositol을 첨가하지 않은 처리구보다 양호한 성숙율을 보였다.

• Development and Reproduction
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• v.12 no.1
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• pp.107-112
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• 2008

To investigate the $C_{21}$-steroids produced from maturating oocytes in the longchin goby, Chasmichthys dolichognathus, the oocytes ($0.74{\sim}0.97\;mm$) were incubated with radiolabeled $17{\alpha}$-hydroxyprogesterone ($^3H-17{\alpha}OHP$) for 24 hours. The resulting metabolites were analyzed by thin layer chromatography and identified by gas chromatography-mass spectrometry. Two $C_{21}$-steroids, $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$) and $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$), were converted from $^3H-17{\alpha}OHP$ in the maturing oocytes. These two main metabolites were detected at 0.80 mm diameter oocytes or greater. In addition, the effects of these metabolites on in vitro germinal vesicle breakdown (GVBD) were tested. The sensitivity of oocytes to the induction of GVBD was greater at $17{\alpha}20{\beta}P$ than $17{\alpha}20{\alpha}P$. This result showed that $17{\alpha}20{\beta}P$ is a major maturation inducing steroid (MIS) in longchin goby, suggesting $17{\alpha}20{\alpha}P$ may play a role in regulating the oocyte maturation process.

• Development and Reproduction
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• v.11 no.3
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• pp.263-272
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• 2007

Contrast to mouse where its in vitro maturation rates are high without specific supplements or presence of the cumulus cells, there are some species, such as porcine, where its in vitro oocyte maturation rates are still very low. This comparative study was conducted to investigate the role of malate dehydrogenase(Mor2) during oocyte maturation by RNAi in the mouse and porcine. The Mor2 double-stranded RNA(dsRNA) was prepared speciesspecifically and microinjected into the cytoplasm of denuded germinal vesicle(GV) oocytes. Oocytes were cultured for 48 h(porcine) and 16 h(mouse) in M199 with 10% porcine follicular fluid, pyruvate, p-FSH, EGF, cystein, and estradiol-$17{\beta}$. We measured changes in oocyte morphology, maturation rates and mRNA levels after Mor2 RNAi. We confirmed gene sequence-specific knock down of Mor2 mRNA in both species after Mor2 RNAi. In contrast to our previous finding that mMor2 RNAi resulted in GV arrest in the mouse, we found that pMor2 RNAi resulted in MI arrest in denuded porcine oocytes(58%), but developed to MII(84.4%) in COCs. To determine whether this difference between mouse and porcine RNAi is due to differences in culture media, we cultured mouse oocytes in the M199 media for 16 h after mMor2 RNAi. Mouse oocytes were developed to MII stage(62%) and there was no statistical difference compared to that of non-injected(76.8%) and buffer-injected(73.3%) control groups. Therefore, we concluded that the mouse and porcine oocytes are having different metabolic systems in relation to malate dehydrogenase for oocyte maturation. This could be a basis for differences in maturation rates in vitro in two species. Further scrutinized studies on the metabolic pathways would led us in finding better culture system to improve oocyte maturation rates in vitro, especially in more challenging species like the porcine.

• Journal of Bio-Environment Control
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• v.25 no.1
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• pp.9-15
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• 2016

High temperature is one of the important environmental factors limiting cultivation of apple (Malus domestica Borkh). The expression of genes related with anthocyanin synthesis and sugar accumulation in response to high temperature was studied in the 'Hongro' apple fruits at different developmental stages in different temperature conditions through real-time PCR. Expression of ${\hat{a}}$-amylase (BMY) and polygalacturonase (PG) genes related with sugar synthesis was higher in late ripening stages than in initial ripening stages. Expression of four genes such as phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), and malate dehydrogenase (MDH), which were related with fruit skin coloration, increased gradually in apple fruits of the middle and late ripening stages. Interestingly, the expressions of all genes were highly inhibited expressed at $30-35^{\circ}C$ compared to $25^{\circ}C$ in all ripening stages. In the further work, investigation of expression levels of various genes could be conducted in the level of transcriptomics in fruits at the middle ripening stages to get meaningful information of ripening metabolism in apple in high temperatures.