http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
- 中文 을 입력하시려면 zhongwen을 입력하시고 space를누르시면됩니다.
- 北京 을 입력하시려면 beijing을 입력하시고 space를 누르시면 됩니다.
RISS 인기검색어
- 검색키워드
- 저자 : Junchul David Yoon (검색결과 33 건)
- 무료
- 기관 내 무료
- 유료
-
GDF8 enhances SOX2 expression and blastocyst total cell number in porcine IVF embryo development
Yoon, Junchul David,Hwang, Seon-Ung,Kim, Mirae,Lee, Gabsang,Jeon, Yubyeol,Hyun, Sang-Hwan Elsevier 2019 Theriogenology Vol.129 No.-
<P><B>Abstract</B></P> <P>Growth differentiation factor 8 (GDF8) is a member of the transforming growth factor-β family and a physiological regulator. According to recent studies, GDF8 can be detected in follicular fluid and the uterus, suggesting that GDF8 may affect preimplantation embryonic development and act in a paracrine manner to improve the success of late-blastocyst implantation <I>in vivo</I>. We investigated the effect of GDF8 supplementation during <I>in vitro</I> culture (IVC) of porcine embryos derived from <I>in vitro</I> fertilization (IVF) and parthenogenetic activation (PA) on cleavage, blastocyst formation rate, and total cell number and analysed gene transcription levels and cell linage specification in the resulting blastocysts. First, the concentration of GDF8 in porcine oviductal fluid was determined to be 139.8 pg/mL. Then, 0, 0.2, 2, or 20 ng/mL GDF8 was added to embryos throughout the entire IVC period. Our results showed that supplementation with GDF8 during porcine preimplantation embryo IVC enhanced blastocyst formation and total cell number and altered the transcriptional patterns of genes that regulate pluripotency and cavitation. Furthermore, using differential immunostaining, we demonstrated that supplementation with GDF8 enhanced the expression of the genuine inner cell mass (ICM) marker SOX2 and the ICM/trophectoderm ratio, improving IVF blastocyst quality. In conclusion, for the first time, we demonstrated the presence of the <I>in vivo</I> oviductal factor GDF8 in oviductal fluid. Furthermore, we found that GDF8 supplementation at 0.2 ng/mL increased the blastocyst total cell number and ICM/trophectoderm ratio by inducing the transcription of genes involved in developmental competence and the expression of genuine ICM marker SOX2 during porcine IVF embryo development <I>in vitro</I>.</P>
-
Supplement of GDF8 during Porcine Oocyte in vitro Maturation changed transcriptional landscape
Junchul David Yoon,Eunsong Lee,Sang-Hwan Hyun 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05
Growth differentiation factor 8 (GDF8) is a member of the transforming growth factor-β that has been identified as a strong physiological regulator. The purpose of this study is to investigate the effects of GDF8 on porcine oocytes during in vitro maturation (IVM). We investigated a specific gene transcription levels in oocytes and cumulus cells (CC) after IVM by realtime PCR arry, and specific protein expression and activation levels in matured CCs by western blotting. Each concentration (0, 1, 10, and 100 ng/ml) of GDF8 was added in maturation medium (TCM199) during process of IVM. Data were analyzed by ANOVA followed by Duncan using SPSS (Statistical Package for Social Science). Data are presented as the mean and Differences were considered significant at P < 0.05. After 44 h of IVM, oocytes are mechanically denuded from CCs with 0.1% of hyaluronidase, and then the separated oocytes and CCs were sampled following each group. To assess the effect of GDF8 on specific gene transcription level changes as a dose response during IVM, the realtime PCR array was performed. In CCs the 1- and 10 ng/ml of GDF8 supplement group showed the transcription co-factors CBP and SP1, cell metabolic regulator MAPK1, and cumulus expansion related genes Has2, Cox-2, Ptx3 and Areg transcription levels were significantly distinguished with control when hierarchically clustered by Euclidean distance with average linkage method after IVM. In matured oocytes the 10- and 100 ng/ml of GDF8 supplement group showed the maternal factors JMJD3 and Zar1, transcriptional regulator FOXO1, Sirt1 and Sirt2, mitochondrial activity factor Sirt3, ACSL3 and ACADL, anti-apoptosis gene BCL-2, and oocyte secrete factor BMP15 mRNA transcription levels were significantly distinguished compared with control. To determine effect of GDF8 supplement during IVM, the GDF8 down steam canonical regulator SMAD2/3 protein phosphorylation levels analyzed in CCs by western blotting. The 10- and 100 ng/ml supplement groups showed significantly increase phosphorylated (P)-SMAD3 (1.56 and 1.34 times higher than control) protein levels (P < 0.05). In conclusion, supplement of GDF8 during IVM activates FOXO homolog transcription and induced cumulus cells expansion via activation of SMAD3 signaling in CCs. While process of IVM, the transcriptional landscape changes in CCs may consequently result maternal factors accumulation and mitochondrial activation in oocytes.
-
Junchul David Yoon,Eunsong Lee,Sang-Hwan Hyun 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10
Growth differentiation factor8 (GDF8) is a member of the transforming growth factor-β that has been identified as a robust physiological regulator. According to recent studies, the GDF8 is detected in oviduct fluid and uterus which led us to suggest that the GDF8 may effect on preimplantation embryonic development and act paracrine role to correlate with successful late-blastocyst implantation in in vivo. We investigated the effect of GDF8 supplement during in vitro culture (IVC) of porcine embryos derived from in vitro fertilization (IVF) on cleavage and blastocyst (BL) formation rate, and gene transcription level analysis in BL. Data were analyzed by one way ANOVA, followed by Tukey’s range test. Respectively 0.2, 2 and 20 ng/mL of GDF8 were added during IVC, and experimental groups were as follows; control (0), 0.2, 2, and 20 GDF8 supplement groups. At IVC 48hr, there was no significant difference on cleavage rate from the different concentration of GDF8 supplement groups (65.7%, 66.0%, 66.3%, and 65.8%, respectively). After additional 120hr of embryo culture, 0.2 group was shown significantly (p<0.05) higher than control in blastocyst formation rate and total cell number (32.5% and 88.0±7.3 VS 40.4% and 118.4±12.7, respectively). Using the achieved IVF BLs, the specific gene expression pattern were evaluated. The embryo development competence marker Pcna, Pou5f1 and Sox2 (1.81, 2.85 and 2.09 times, respectively), and cell junction assembly regulator Adam10, Adam17, Tjp1, Cdh1 (1.40, 1.98, 1.79 and 1.80 times, respectively) genes mRNA transcript levels in 0.2 group were significantly increased in 0.2 group compared with control. Moreover, pro-apoptotic factor Cas3 gene mRNA transcript levels was significantly decreased in 0.2 group than control (0.62 times). In conclusion, the supplementation of 0.2 ng/mL GDF8 during IVC significantly improved embryonic developmental potential via regulating embryo developmental competence markers and cell junction assembly regulator transcriptional levels.
-
Effect of GDF8 on Porcine Chimeric Embryo Development
Junchul David Yoon,Jongpil Kim,Eunsong Lee,Sang-Hwan Hyun 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10
Recent decade, the studies for generation of chimeric organs have been advanced remarkably with the production of chimeric animals using by pluripotent stem cells (PSCs). However, the distribution rate of injected cells into an embryo is still low and it is considered as one of the major reasons of low efficiency of chimera production. The purpose of this study is to investigate the effect of growth differentiation factor 8 (GDF8) on porcine chimeric embryo development. In first, we produced mCherry-marked porcine iPSCs (mCh-iPSCs). The mCh-iPSCs were injected into 8 cells or morula stage of in vitro fertilized (IVF) embryos at day 2 under a microscope with the micromanipulator. These chimeric embryos were then cultured porcine embryonic stem cell medium (DEME F10/low glucose with 15% FBS) for 18hr and then transferred to fresh porcine zygote medium 5 (PZM5 with or without 150 pg/mL of GDF8). We investigated the effect of supplemented GDF8 during in vitro culture (IVC) of these chimeric embryos on their cleavage patterns, blastocyst formation ratio, and injected cells’ distributions. Data were analyzed by ANOVA followed by Tukey’s range test using SPSS (Statistical Package for Social Science). After day 5 of in vitro chimeric embryo culture, the GDF8 supplement group was shown significantly higher Using the chimeric embryos of the blastocyst stage, we evaluated distributions of the injected cells by immune staining of SOX2 as porcine inner cell mass (ICM) marker. The GDF8 supplement group was shown significantly increased mCherry and SOX2 per SOX2 expressing cells ratio than control (49.2% VS 20.7, respectively). In conclusion, the supplementation of GDF8 during IVC of porcine chimeric embryos improved embryonic developmental competence and enhanced the distributions of injected cells into ICM of chimeric embryos at the pre-implantation stage.
-
-
-
Junchul David Yoon,Eunsong Lee,Sang-Hwan Hyun 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05
Growth differentiation factor8 (GDF8) is a member of the transforming growth factor-β that has been identified as a strong physiological regulator. Overall of the current studies, the GDF8 is detected in oviduct fluid and uterus which led us to suggest that the GDF8 may effect on preimplantation embryonic development and act paracrine role to correlate with successful late-blastocyst implantation in in vivo. The purpose of this study is the effects of GDF8 on porcine parthenogenesis (PA) embryo development during in vitro culture (IVC). We were investigated the effect of GDF8 supplement during PA embryo IVC by cleavage and blastocyst formation rate and patterning analysis. Data were analyzed by on way ANOVA, followed by Tukey’s range test. Respectively 0.2, 2 and 20 ng/mL of GDF8 were added during IVC followed experiment design as control, 0.2, 2, and 20 GDF8 supplement groups. After 48h of embryo culture time, no significant difference was observed on cleavage rate from the different concentration (0, 0.2, 2, and 20 ng/ml) of GDF8 supplement groups (65.7%, 66.0%, 66.3%, and 65.8%, respectively). After 120h of embryo culture time, the 0.2 and 2 group showed significantly (p<0.05) higher blastocyst formation rate than control (40.4% and 36.4% VS 40.4%, respectively). In embryo developmental pattern analysis, the 0.2 ng/ml GDF8 supplement groups showed significantly higher (p<0.05) 2-3 cell cleavage- and early blastocyst pattern compared with control (12.0% and 10.4% VS 6.6% and 6.2%, respectively). However there are no significantly different pattern was observed in other groups. In conclusion, the 0.2 ng/ml of GDF8 supplementation during porcine PA embryo IVC significantly changed embryonic developmental patterns. However there are further studies are required such as analysis of blastocyst total number, specific gene transcription pattern, and ICM/TE rate to make clarify and support the conclusion.
-
Yoon, Junchul David,Hwang, Seon-Ung,Kim, Mirae,Jeon, Yubyeol,Hyun, Sang-Hwan Society for the Study of Reproduction [etc.] 2019 BIOLOGY OF REPRODUCTION Vol.101 No.1
<P> Growth differentiation factor 8 (GDF8), also known as myostatin, is a member of the transforming growth factor-β (TGF-β) family and has been identified as a strong physiological regulator of muscle differentiation. Recently, the functional role of GDF8 in reproductive organs has received increased interest following its detection in the human placenta and uterus. To investigate the effects of GDF8 during porcine oocyte in vitro maturation (IVM), we assessed the quality of matured oocytes. Furthermore, we investigated the specific gene transcription and protein activation levels in oocytes and cumulus cells after IVM and subsequent embryonic development after in vitro fertilization and parthenogenetic activation. Prior to these experiments, the concentration of GDF8 in porcine follicular fluid was determined. During the entire IVM period, 1.3 ng/mL GDF8 and its signaling inhibitor SB431542 (SB) at 5 μM were added as control, SB, SB + GDF8, and GDF8 groups, respectively. Our results demonstrate that supplementation with GDF8 during porcine oocyte IVM enhanced both meiotic and cytoplasmic maturation, with altered transcriptional patterns, via activation of Sma- and Mad-related protein 2/3 (SMAD2/3). Using the pharmacological inhibitor SB431542, we demonstrated that inhibition of GDF8-induced Smad2/3 signaling reduces matured oocyte quality. In conclusion, for the first time, we demonstrated paracrine factor GDF8 in porcine follicular fluid in vivo. Furthermore, we showed that GDF8 supplementation improved mature oocyte quality by regulating p38 mitogen-activated protein kinase phosphorylation and intracellular glutathione and reactive oxygen species levels during porcine IVM. </P>
-
-
연관 검색어 추천
이 검색어로 많이 본 자료
활용도 높은 자료
