Effects of Endoplasmic Reticulum Stress Inhibitor Treatment during the Micromanipulation of Somatic Cell Nuclear Transfer in Porcine Oocytes

Park, Yeo-Reum,Park, Hye-Bin,Kim, Mi-Jeong,Jung, Bae-Dong,Lee, Seunghyung,Park, Choon-Keun,Cheong, Hee-Tae The Korean Society of Developmental Biology 2019 발생과 생식 Vol.23 No.1

We examined the effects of endoplasmic reticulum (ER) stress inhibitor treatment during the micromanipulation of porcine somatic cell nuclear transfer (SCNT) on the in vitro development of SCNT embryos. ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxycholic acid (TUDCA; $100{\mu}M$) were added to the micromanipulation medium and holding medium. The expression of X-box binding protein 1 (Xbp1), ER-stress-associated genes, and apoptotic genes in SCNT embryos was confirmed at the one-cell and blastocyst stages. Levels of Xbp1 splicing and expression of ER-stress-associated genes in SCNT embryos at the one-cell stage decreased significantly with TUDCA treatment (p<0.05). The expression of ER-stress-associated genes also decreased slightly with the addition of both salubrinal and TUDCA (Sal+TUD). The expression levels of caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in the TUDCA and Sal+TUD treatments (p<0.05). At the blastocyst stage, there were no differences in levels of Xbp1 splicing, and transcription of ER-stress-associated genes and apoptosis genes between control and treatment groups. However, the blastocyst formation rate (20.2%) and mean blastocyst cell number ($63.0{\pm}7.2$) were significantly higher (p<0.05) for embryos in the TUDCA treatment compared with those for control (12.6% and $41.7{\pm}3.1$, respectively). These results indicate that the addition of ER-stress inhibitors, especially TUDCA, during micromanipulation can inhibit cellular damage and enhance in vitro development of SCNT embryos by reducing stress levels in the ER.

Endoplasmic Reticulum (ER) Stress and Apoptosis in Parthenogenetic Porcine Embryos Following Activation Methods

Yeo-Reum Park,Hye-Bin Park,Hwa-Yeon Lee,Hyo-Kyung Bae,Hui-Yeon Shin,Seunghyung Lee,Choon-Keun Park,Boo-Keun Yang,Hee-Tae Cheong 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10

This study was conducted to examine the effects of activation methods on the ER stress induction and subsequent apoptosis and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by four activation methods; 1) electric stimulus(ES) with two DC pulses of 1.25 kV/cm, for 30 ㎲ (E), 2) ES + 10 μM Ca-ionophore (A23187) treatment for 5 min (EC), 3) ES + 2 mM 6-dimethylaminopurine treatment for 3 h (ED), or 4) ES + A23187 + 6-DMAP (ECD). After activation, parthenogenetic embryos were in vitro cultured in PZM-3 medium and sampled to analyze the x-box binding protein 1 (Xbp1) mRNA, ER stress-associated genes and apoptotic genes at 3 h post ES and the 1-cell and blastocyst stages. The un-spliced and spliced x-box binding protein 1 (Xbp1) mRNA were confirmed by RT-PCR. Also ER stress-associated genes, such as the C/EBP homologous protein (CHOP), binding protein (BiP), activating transcription factor 4 (ATF4) and glucose-regulated protein 94 (GRP94), and apoptotic genes were analyzed by real-time quantitative RT-PCR (RT-qPCR). The band intensities of spliced Xbp1 (Xbp1s) mRNA was higher in the EC group than other three groups at 3 h and the 1-cell stage, while it was higher in the ED groups compared with E group at the blastocyst stage. Four ER stress-associated genes were showed the highest expression in the EC group and weakly expressed in the ED group at 3 h. However, most of those genes were highly expressed in EC and ECD groups at the 1-cell and blastocyst stages with some variation. The expressions of Bcl-2-associated X protein (Bax) and caspase-3 mRNAs were significantly higher in EC group than other three groups at all stages. The developmental rate to the blastocyst stage was higher (p<0.05) in ED and ECD groups (32.1±3.8 to 34.6±2.2%) than that of E group (26.1±3.9%). These results suggest that the intracellular ER stress of parthenogenetic porcine embryos is affected by activation method and subsequently lead to the apoptosis of embryos.

Endoplasmic Reticulum (ER) Stress and Apoptosis in Parthenogenetic Porcine Embryos following Different Combination of Activation Methods

Park, Hye-Bin,Park, Yeo-Reum,Lee, Hwa-Yeon,Bae, Hyo-Kyung,Lee, Seunghyung,Park, Choon-Keun,Yang, Boo-Keun,Cheong, Hee-Tae The Korean Society of Embryo Transfer 2017 한국동물생명공학회지 Vol.32 No.1

This study was conducted to investigate the effect of activation method on the endoplasmic reticulum (ER) stress induction, apoptosis and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by four activation methods; 1) electric stimulus (ES) (E), 2) $ES+10{\mu}M$ Ca-ionophore (A23187) treatment (EC), 3) ES+2 mM 6-dimethylaminopurine (6-DMAP) treatment (ED), or 4) ES+A23187 and 6-DMAP treatments (ECD). Parthenogenetic embryos were sampled to analyze x-box binding protein 1 (Xbp1) mRNA, ER stress-associated genes and apoptosis genes at 3 h after ES and the 1-cell and blastocyst stages. In the EC group, the band intensity of spliced Xbp1 (Xbp1s) mRNA was higher than those of the other groups at the 3 h and 1-cell stage, and higher than that of the E group at the blastocyst stage. Four ER stress-associated genes were expressed at the highest level in the EC group and weakly expressed in the ED group at 3 h after activation. However, most of the genes were highly expressed at the 1-cell and blastocyst stages with some variation in the EC and ECD groups. Expression of Bcl-2-associated X protein (Bax) and caspase-3 mRNA was significantly higher in the EC group than in the other groups at all development stages. The developmental rates to the blastocyst stage were higher in the ED and ECD groups than in the E and EC groups. These results suggest that the intracellular ER stress of parthenogenetic porcine embryos is affected by the activation method and subsequently lead to the apoptosis of embryos.

Endoplasmic Reticulum (ER) Stress and Apoptosis in Parthenogenetic Porcine Embryos Following Activation Methods

Yeo-Reum Park,Hye-Bin Park,Hwa-Yeon Lee,Hyo-Kyung Bae,Hui-Yeon Shin,Seunghyung Lee,Choon-Keun Park,Boo-Keun Yang,Hee-Tae Cheong 한국동물생명공학회(구 한국동물번식학회) 2016 발생공학 국제심포지엄 및 학술대회 Vol.2016 No.10

Cadmium-Substituted Concanavalin A and Its Trimeric Complexation

( Yeo Reum Park ),( Da Som Kim ),( Dong-heon Lee ),( Hyun Goo Kang ),( Jung Hee Park ),( Seung Jae Lee ) 한국미생물 · 생명공학회 2018 Journal of microbiology and biotechnology Vol.28 No.12

Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens. Structural studies have detailed the interaction between ConA and carbohydrates and the metal coordination environment with manganese and calcium ions (Mn-Ca-ConA). In this study, ConA was crystallized with a cadmium-containing precipitant, and the refined structure indicates that Mn²⁺ was replaced by Cd²⁺ (Cd-Ca-ConA). The structural comparison with ConA demonstrates that the metal-coordinated residues of Cd-Ca-ConA, that is Glu8, Asp10, Asn14, Asp19, and His24, do not have conformational shifts, but residues for sugar binding, including Arg228, Tyr100, and Leu99, reorient their side chains, slightly. Previous studies demonstrated that excess cadmium ions can coordinate with other residues, including Glu87 and Glu183, which were not coordinated with Cd²⁺ in this study. The trimeric ConA in this study coordinated Cd²⁺ with other residues, including Asp80 and Asp82, for complex generation. The monomer does not have specific interaction near interface regions with the other monomer, but secondary cadmium coordinated with two aspartates (Asp80 and Asp82) from monomer 1 and one aspartate (Asp16) from monomer 2. This study demonstrated that complex generation was induced via coordination with secondary Cd²⁺ and showed the application potential regarding the design of complex formation for specific interactions with target saccharides.

Endoplasmic Reticulum Stress Inhibitor Treatment during Somatic Cell Nuclear Transfer (SCNT) Process of Porcine Embryos

Yeo-Reum Park,Hye-Bin Par,Mi-Jeong Kim,Seunghyung Lee,Choon-Keun Park,Boo-Keun Yang,Hee-Tae Cheong 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

We examined the effect of endoplasmic reticulum (ER) stress inhibitor treatment during somatic cell nuclear transfer (SCNT) process on the reprogramming efficiency of porcine embryos. ER stress inhibitors such as Salubrinal (200 nM) and tauroursodeoxychloic acid (TUDCA, 100 μM) were added to the manipulation medium and holding medium. Porcine SCNT embryos were sampled at the 1-cell stage for mRNA extraction. The expression of x-box binding protein 1 (Xbp1), ER stress-associated genes and apoptotic genes were confirmed by RT-PCR or RT-qPCR. The levels of X-box binding protein 1 (Xbp1) splicing and the expression of ER stress-associated genes were significantly decreased by TUDCA treatment at the 1-cell stage of SCNT embryos (p<0.05). The expression of ER stress-associated genes was also slightly decreased by a combination treatment of salubrinal and TUDCA (Sal+TUD). The expression levels of Caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in TUDCA and Sal+TUD groups (p<0.05). The blastocyst formation rate (20.2%, 41/203) and the mean cells number in blastocysts (63.0±7.2, p<0.05) were significantly increased in the TUDCA treatment group compared with the control group (12.6% and 41.7±3.1, p<0.05). These results show that the treatment of ER stress inhibitor, especially TUDCA, during SCNT process can inhibit cellular damages and enhance in vitro development of SCNT embryos by reducing ER stress.

문장부호에 대한 교육과정 지도상 문제점과 아동 사용 실태의 연구

박여름(Park Yeo-reum) 한국어문교육학회 2008 어문학교육 Vol.37 No.-

  There are two different typical ways to express oneself : speaking and writing. Speaking may be concurrent with appropriate gestures to ensure smooth communication, and a speaker can represent what"s on his or her mind in a different tone depending on the context or content. However in terms of writing, letters are the only vehicle to describe one"s own ideas or feelings. we can"t use appropriate gestures in writing. we need punctuation marks to get more information : speaker"s mind on the context, section of direct speech or indirect speech, emphasize, words, phrases, etc.. Punctuation marks is really important to understand the structure of sentences in grammatically and logically.<BR>  Despite the importance of punctuation marks, however, the elementary curriculum just dealt with a limited number of punctuation marks without considering the real lives of children. And elementary schoolers often use punctuation marks that don"t comply with the spelling system of Korean.<BR>  The purpose of this study was to examine problems with punctuation marks in the 7th national curriculum and the use of the marks by elementary schoolers in an attempt to redetermine the types of punctuation marks to be taught in elementary school and what to teach.<BR>  Chapter Ⅱ described the concept, necessity and system of punctuation marks. The 7th elementary Korean curriculum were compared with the revised curriculum in terms of punctuation marks, and the frequency of punctuation marks in the 7th korean textbooks was investigated to have a critical discussion about the 7th curriculum and textbooks.<BR>  First, punctuation marks refer to various signs by which speakers can make a clear description of what they want to say and by which readers can have the right understanding of writing.<BR>  Second, in the current punctuation mark regulations, 19 punctuation marks were largely categorized into seven, and their names and how to use the marks were specified.<BR>  Third, in the 7th curriculum, students learned about punctuation marks in the first and third grade only, and just six out of 19 punctuation marks were taught : period, question mark, exclamation mark, comma, double quotation marks and single quotation marks. Besides, just some typical functions of the marks were explained without handling all their functions. In the textbooks, a wider variety of punctuation marks were used in addition to the above-mentioned marks. The textbooks contained some quotation marks that weren"t included in the regulations, which was likely to throw children into confusion. And there wasn"t enough time to teach the location and shape of punctuation marks.<BR>  Chapter Ⅲ investigated the perception of elementary school teachers about the state of punctuation marks education and the use of the marks among second to sixth graders. Students" writing was analyzed to see whether punctuation marks that were and weren"t included in the 7th curricula were used by them. Besides, to what extent different graders used punctuation marks and what types of errors they made were investigated.<BR>  As a result, first, it"s found that most of the teachers investigated believed that students should learn about punctuation marks in different grades, and that there should be more education about the use and notation of the marks. Some argued that the 7th national curricula should deal with a larger number of punctuation marks than the types of the marks. Ellipsis was most widely pointed out as a punctuation mark that should be additionally taught, followed by colon and parenthesis.<BR>  Second, in the event of the lower graders, the most frequently used punctuation mark that was included in the 7th national curricula was period, followed by comma, exclamation mark, double quotation marks and single quotation marks. And the mos

Endoplasmic Reticulum (ER) Stress and Apoptosis in Parthenogenetic Porcine Embryos following Different Combination of Activation Methods

Hye-Bin Park,Yeo-Reum Park,Hwa-Yeon Lee,Hyo-Kyung Bae,Seunghyung Lee,Choon-Keun Park,Boo-Keun Yang,Hee-Tae Cheong 한국수정란이식학회 2017 한국동물생명공학회지 Vol.32 No.1

This study was conducted to investigate the effect of activation method on the endoplasmic reticulum (ER) stress induction, apoptosis and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by four activation methods; 1) electric stimulus (ES) (E), 2) ES+10 μM Ca-ionophore (A23187) treatment (EC), 3) ES+2 mM 6-dimethylaminopurine (6-DMAP) treatment (ED), or 4) ES+A23187 and 6-DMAP treatments (ECD). Parthenogenetic embryos were sampled to analyze x-box binding protein 1 (Xbp1) mRNA, ER stress-associated genes and apoptosis genes at 3 h after ES and the 1-cell and blastocyst stages. In the EC group, the band intensity of spliced Xbp1 (Xbp1s) mRNA was higher than those of the other groups at the 3 h and 1-cell stage, and higher than that of the E group at the blastocyst stage. Four ER stress-associated genes were expressed at the highest level in the EC group and weakly expressed in the ED group at 3 h after activation. However, most of the genes were highly expressed at the 1-cell and blastocyst stages with some variation in the EC and ECD groups. Expression of Bcl-2-associated X protein (Bax) and caspase-3 mRNA was significantly higher in the EC group than in the other groups at all development stages. The developmental rates to the blastocyst stage were higher in the ED and ECD groups than in the E and EC groups. These results suggest that the intracellular ER stress of parthenogenetic porcine embryos is affected by the activation method and subsequently lead to the apoptosis of embryos.

Anti-hyperlipidemic Effect of Polyphenol Extract (Seapolynol^TM) and Dieckol Isolated from Ecklonia cava in in vivo and in vitro Models

Yeo, A-Reum,Lee, Jung-Lim,Tae, In-Hwan,Park, Seok-Rae,Cho, Young-Ho,Lee, Bong-Ho,Shin, Hyeon-Cheol,Kim, Seong-Ho,Yoo, Yung-Choon The Korean Society of Food Science and Nutrition 2012 Preventive Nutrition and Food Science Vol.17 No.1

The inhibitory effect of polyphenol extracts (Seapolynol$^{TM}$, SPN) of the marine brown algae Ecklonia cava and dieckol, a major component of SPN, on hyperlipidemia was investigated in ICR mice fed a high-fat diet (HFD) for five weeks. For analysis of the anti-hyperlipidemic effects of SPN and dieckol, these two agents were given orally on a daily basis to HFD-fed mice for four weeks, starting one week after the beginning of HFD feeding. Groups administered with SPN as well as dieckol showed lower body weight gains than the HFD only group. Administration of SPN and dieckol also resulted in a significant reduction of the level of total cholesterol (TCHO), triglyceride (TG), and low-density lipoprotein (LDL) cholesterol in the serum of HFD-fed mice. In Oil Red O staining using 3T3-L1 preadipocytes, it was shown that both SPN and dieckol markedly inhibited lipid accumulation of 3T3-L1 cells. Furthermore, SPN and dieckol (50 ${\mu}g$/mL) significantly inhibited 3-hydroxyl-methyl glutaryl coenzyme A (HMGCoA) reductase activity in vitro. Taken together, these results suggest that polyphenols of Ecklonia cava (SPN) and dieckol reduce body weight gain and fat accumulation in HFD-induced obese mice, and that their hypolipidemic effect is related to the inhibition of adipogenesis of adipocytes and HMGCoA reductase activity.

Molecular characterization of a white rot disease-resistance gene, AMet, from apple

Yeo Jin Kang,A Reum Park,Seung Kyu Oh,Dae Il Kim,Jong Han Park,Hee Jae Lee,In Jung Kim 한국육종학회 2007 한국육종학회 학술발표회 발표요지 Vol.39 No.1