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Hyo-Jin Park,Jin-Woo Kim,Seul-Gi Yang1,Min-Ji Kim,Ho-Guen Jegal,Joung Jun Park,Deog-Bon Koo 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06
Bisphenol-A (BPA) as an organic synthetic compound of exhibiting estrogen-mimicking and hormone-like properties, which is commonly used to induce cellular stress or female reproductive toxicity. In addition, BPA induces the increasing of mitochondrial derived reactive oxygen species (ROS) such as superoxide, and production of these ROS affects to the meiotic maturation and cumulus cells expansion on in vitro maturation (IVM) of porcine cumulus-oocyte complexes (COCs). However, anti-oxidative effect of melatonin for reduction of BPA-induced superoxide on porcine oocyte maturation has not been reported. Therefore, in present study, we confirmed that the reduction of BPA-derived superoxide by melatonin related to the reducing of mitochondria mediated apoptosis on meiotic maturation and cumulus cells expansion of porcine COCs. Then, to investigate the effects of superoxide specific scavenger, Mito-TEMPO, during porcine oocyte maturation progression, COCs cultured in maturation medium with Mito-TEMPO (0.1 μM) after pre-treatment of BPA (75 μM) for 22 h. Reduced meiotic maturation rate and cumulus cells expansion of COCs in the BPA (75 μM) treated group were recovered (p<0.01) by Mito-TEMPO treatment. Also, increasing of mitochondria derived apoptotic factors (AIF, Cleaved Caspase 3 and Cleaved PARP 1) protein levels by BPA treatment were reduced by Mito-TEMPO treatment in porcine COCs maturation. Positive effects of Mito-TEMPO for superoxide reduction on oocyte maturation and reducing mitochondrial apoptosis showed the same pattern in melatonin (0.1 μM) treated COCs. In case of supplemented with BPA and melatonin, superoxide production in COCs was not changed compared to control or melatonin treated groups. Based on these results, we concluded that melatonin as a regulator of superoxide such as Mito-TEMPO improves oocyte maturation through reduction of mitochondria derived apoptosis during IVM of porcine COCs.
Effect of Ovarian Extract on Oocyte Maturation and Early Embryonic Development in Pigs
Seul-Gi Yang(Seul-Gi Yang),Jae-Hun Choi(Jae-Hun Choi),Young-Seo Jo(Young-Seo Jo),Ye-Won Kim(Ye-Won Kim),Dong-Mok Lee(Dong-Mok Lee),Hyo-Jin Park(Hyo-Jin Park),Deog-Bon Koo(Deog-Bon Koo) 한국동물보건학회 2022 한국동물보건학회지 Vol.1 No.2
Various factors in the ovary are known to regulate oocyte maturation and hormone secretory functions; however, the effect of ovarian extract (OE) on oocyte maturation and embryonic development in pigs remains unknown. In this study, we first evaluated whether OE supplementation in the in vitro maturation (IVM) medium alters the oocyte maturation capacity by affecting glucose/amino acid metabolites, meiotic maturation, cumulus cell (CC) expansion, and antioxidants. Various OE concentrations (50, 100, 200, 500, and 5000 μg/mL) were included in the IVM medium. Only the oocytes treated with 100 μg/mL OE exhibited an improved meiotic maturation rate when compared with that of the other groups (non-treated group, 78.6 ± 3.0% vs. 100 μg/mL OE-treated group, 81.6 ± 4.3%); however, the difference was not significant. To observe the changes in glucose and amino acid metabolism in the OE-treated oocytes, we measured the amounts of diverse constituents (glucose, lactate, glutamine, and ammonia) in the IVM medium containing OE. Lactate and ammonia levels in the OE-treated group after 44 h of IVM were higher (p < 0.01) than those in the non-treated group. In addition, the expression of the CC expansion factors (Has2 and Tnfaip6) significantly increased (p < 0.05), whereas the mRNA expression levels of antioxidative enzymes (Sod1, Cat, and Gpx1) significantly diminished (p < 0.05) in the OE-treated group. Moreover, mature oocytes treated with 100 μg/mL OE demonstrated increased subsequent embryonic development rates after 144 h of IVM. Thus, the addition of OE in IVM mediums may improve oocyte maturation capacity which could enhance antioxidative enzyme activation, energy metabolism, and expression of the CC expansion factors in porcine oocytes.
Seul-Gi Yang,Hyo-Jin Park,Jin-Woo Kim,Jae-Min Jung,Min-Ji Kim,Ho-Guen Jegal,In-Su Kim,Deog-Bon Koo 한국동물생명공학회(구 한국동물번식학회) 2017 Reproductive & Developmental Biology(Supplement) Vol.41 No.2
Mitochondrial functions are required for early embryonic development in pigs. Reactive oxygen species (ROS) can produce from the maintenance of mitochondrial functions during in vitro culture (IVC) of embryos. However, the role of mitochondria specific ROS (mito-ROS) in porcine early embryonic development remains unknown. Therefore, the objective of present study was to confirm the changes in functions, aggregation and dynamics of mitochondria by the regulation of mito-ROS in porcine embryo development. Here, we performed the IVC from classified two groups (G1; high lipid and G2; low lipid contents) at the zygote stage. Blastocyst developmental rate and total nuclei numbers in G2 embryos were lower (p<0.05) than that of G1 embryos. To investigate the superoxide as mitochondrial-target ROS on preimplantation development of G1 and G2 embryos, we performed the MitoSOX staining. Superoxide levels significantly increased (p<0.05) in G2 embryos. And, changes of mitochondrial membrane potential (MMP), ATP levels and expression of mitochondria fission protein were investigated by using JC-1 kit, ATP determination kit and Western blot analysis. MMP and ATP levels were significantly decreased (p<0.05) in G2 embryos compared with G1 embryos. Protein level of mitochondrial fission protein Drp1 was increased (p<0.05) in the cleavage stage of G2 embryos. To confirm the effects of reducing mito-ROS on the porcine embryo development, we treated with the Mito-TEMPO, mitochondria specific ROS scavenger, for G2 embryos. Mito-TEMPO improved preimplantation development to the blastocyst stage and reduced mito-ROS level in the G2 embryos (p<0.05). These results demonstrated that regulation of mito-ROS improves the blastocysts development and their qualities through the control of mitochondrial functions and dynamics of porcine embryos.
Records of the Marsh Grassbird Locustella pryeri sinensis in Korea
Seul-Gi Seo,Se-Young Park,Chang-uk Park,Sook-Young Cho,Seung-Yeon Lee,Gi-Chang Bing,Yang-Mo Kim,Jong-Hyoun Park,Young-Soo Kwon 한국조류학회II 2015 한국조류학회지 Vol.22 No.1
On 16 May 2014, a Marsh Grassbird (1<SUP>st</SUP> summer plumage) was observed at baenanggimi marsh (34°41"21.03"N, 125°25"32.37"E), Heuksan-myeon, Shinan-gun, Jeollanam-do in South Korea. This is the second record of Marsh Glassbird (L. pryeri) in Korea since it has been collected in 1962.
Reduction of mitochondria derived superoxide by Mito-TEMPO improves the porcine oocyte maturation
Seul-Gi Yang,Hyo-Jin Park,Jin-Woo Kim,Min-Ji Kim,Ho-Guen Jegal,In-Su Kim,Min-Young Guk,Sun-Mi Park,Ji-Eun Lee,Deog-Bon Koo,Joung Jun Park 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11
In general, the shape of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage is important roles on meiotic maturation of porcine oocyte during in vitro maturation (IVM). Then, mitochondria produce reactive oxygen species (ROS) such as superoxide from electron transport system during oocyte maturation. ROS levels on oocytes are regulated by various antioxidant enzymes in cumulus cells (CCs). However, the effect of mitochondria derived superoxide production from CCs during IVM of porcine oocyte has not been reported. Firstly, we divided groups according to large number of CCs (Grade 1: G1) and small number of CCs (Grade 2: G2). Then, we counted cumulus cells of G1 and G2 oocyte by using haemocytometer. The oocyte maturation rate was significant decreased (p < 0.05) in G2 oocytes than that of G1 oocytes. We measured mitochondria derived superoxide in G1 and G2 COCs by using Mito-SOX staining. Mitochondrial superoxide was higher in G2 COCs than G1 COCs. Then, the mRNA expression levels of antioxidant enzymes (SOD1, SOD2 and PRDX3) in G2 COCs were decreased compared to G1 COCs. To reduce mitochondria derived superoxide, we used Mito-TEMPO as mitochondrial superoxide scavenger. Oocyte maturation rates in both G1 and G2 groups treated with Mito-TEMPO were increased than that of non-treated groups. Mitochondrial superoxide was lower in G1 and G2 groups treated with Mito-TEMPO than that of non-treatment groups. The mRNA expression levels of antioxidant enzymes in G1 and G2 COCs treated with Mito-TEMPO were increased compared to non-treated groups. Based on these findings, we suggest that reduction of mitochondria derived superoxide by Mito-TEMPO assists maturation competence in porcine oocytes.
Seul-Gi Yang,Hyo-Jin Park,Jin-Woo Kim,Jae-Young Park,Jae-Min Jung,Min-Ji Kim,Deog-Bon Koo 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
Mitochondria are well known to regulate the mammalian embryo development. Recent studies showed that the mitochondrial dynamics responses are mainly generated through mitochondrial membrane potential (MMP) and cellular ATP production, which is dependent on mitochondrial reactive oxygen species (ROS). However, these mechanisms are unclear on development process of preimplantation porcine embryos. The aim of this study was to evaluate that difference of the mitochondrial dynamics-derived various functions on the embryo development according to lipid composition of zygote. First, zygote was classify two groups (high lipid, grade 1: G1 and low lipid, grade 2: G2) by lipid composition of cytoplasm. And, we performed the in vitro culture (IVC) using zygote of divided groups. The nuclei numbers and developmental rates of blastocysts were lower in G2 than those of G1 embryos. Next, we investigated the intracellular ROS and mitochondrial derived superoxide production in porcine embryos by using DCF-DA and Mito-SOX staining. As expected, both intracellular ROS and mitochondrial derived superoxide were significantly increased (p<0.05) in the preimplantation stage embryos of G2 group compared with G1 group. In addition, to observe difference of the mitochondrial functions, we investigated the mitochondrial membrane potential (MMP, ΔΨ) and contents of ATP in the preimplantation stage embryos by using JC-1 kit and ATP determination kit. These functions of mitochondria were dramatically reduced in cleavage stage embryos or blastocysts of G2 group. Finally, to verify the difference of the mitochondrial dynamics-derived various functions, we investigated the expressions of mitochondrial fission (Drp1, pDrp1-616) and fusion (Mfn1, Mfn2) factors by Western blotting analysis. Interestingly, the protein levels of pDrp1-616 in embryos of G1 group were continuously increased until blastocyst stage. Whereas, the expression patterns of Mfn1/2 in embryos of G2 group were significantly reduced during IVC progression. The expression patterns of mitochondria dynamic between the two groups were shown opposite. These results demonstrated that the lipid contents of zygote were related the positive-correlation with mitochondrial dynamics-derived functions in porcine embryos. Moreover, we suggest that lipid of zygote is play a important role on mitochondrial functions and dynamics during preimplantation embryos development in pigs.
단보 : 배추뿌리혹병균(Plasmodiophora brassicae)의 인공접종을 위한 효율적인 저장조건
양슬기 ( Seul Gi Yang ),박주영 ( Ju Young Park ),서문원 ( Mun Won Seo ),김홍기 ( Hong Gi Kim ) 한국균학회 2015 韓國菌學會誌 Vol.43 No.4
Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is a severe soilborne disease of Brassicaceae. Storage of clubroot gall is important for studies on pathogenicity and race identification. As the current storage method has been used for more than 100 years, a new storage method should be developed and the most efficient way maintaining pathogenicity should be determined. Effects of storage conditions with different storage periods on pathogenicity in galls of kimchi cabbage were examined in a greenhouse. The experiments were performed under six conditions and four temperatures in order to determine the most effective storage conditions for maintenance of pathogenicity. The most effective conditions for clubroot gall storage was the storage of whole gall at -70oC or storage of filtrate at the same temperature through eight layers of gauze after homogenization of the galls.