• Title/Summary/Keyword: Early embryo

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Lmbr1 Expression in Early Embryo Development Stages in White Leghorn and Chinese Silky

  • Zhang, Ying;Xu, Weizhuo;Li, Ning
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.7-12
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    • 2009
  • Lmbr1 is regarded as a key gene that controls the digital model formation in early developmental stages of the chicken. However, there are few reports of lmbr1 expression levels and tendencies in 4-toe and 5-toe chicken species. Therefore, the objective of this study was to compare the lmbr1 expression in White Leghorn (4-toe) and Chinese Silky (5-toe). Firstly, total RNA was extracted from 14 different embryonic development stages (HH3 to HH31) in White Leghorn and Chinese Silky. Secondly, dramatic gene expression changes of lmbr1 were monitored by RT-PCR, which indicated a general up-down-up tendency with subtle differences between these two species. Moreover, Q-PCR reactions were performed to quantitate the expression level of lmbr1 in the 14 selected developmental stages. These data demonstrated a first lmbr1 expression peak of 18.68 and 15.32, a lmbr1 expression trough of 6.61 and 1.80, and a second lmbr1 expression peak of 22.33 and 12.48 in White Leghorn and Chinese Silky, respectively. Finally, embryonic in situ hybridization analysis identified that lmbr1 expressed in the ectoderm in HH21, HH23 and HH24 developmental stages in both species.

Alteration of DNA Methylation in Oct-4 Gene in Mouse Preimplantation Embryos by the Interference RNA

  • Kim, Jong-Mu;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Chung, Hak-Jae;Chang, Won-Kyong;Kim, Nam-Hyung
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.21-28
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    • 2007
  • During early embryo development, Oct-4 is an important transcription factor for the early differentiation the present study was first examined methylation status in distal enhancer and promoter region of Oct-4 during mouse pre-implantation embryo development. In oocyte and sperm, high methylation was observed in both distal and proximal of promoter in Oct-4. Following fertilization relatively high methylation level remained until 8-cell stage embryos, but decreased at the morula and blastocyst stage. Specific gene knock down of Oct-4 by siRNA injection into zygote induced higher methylation rates of both distal and proximal region of promoter of Oct-4. These results suggest a functional link between the DNA methylation status of distal and promoter resign in the Oct-4 gene and the gene sequence-specific transcriptional silencing by exogenous siRNA injection during mouse preimplantation embryos.

Cloning and Characterization of Bovine 5-Cytosine DNA Methyltransferase I cDNA

  • Lee, Poongyeon;Min, Kwan-Sik;Lee, Hyun-Gi;Kim, Soon-Jeung;Chung, Hee-Kyoung;Seo, Myung-Kyu;Lee, Yun-Keun;Kim, Sung-Woo;Park, Jin-Ki
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.39-39
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    • 2003
  • Methylation of DNA 5-cytosine in mammalian early embryo affects great deal in nuclear reprogramming and chromatin remodeling of developing embryo. Current efforts to clone and produce cloned animals including transgenic animals face various problems including low birth rate, irregular development, and so on. In this report, cDNA for the one of house keeping methyltransfcrase, Dnmt1 was cloned from bovine somatic tissues and was analyzed for its nucleotide sequences to investigate the structure and function of the gene in bovine early development. Nucleotide sequence of bovine Dnmt1 homologue showed 76.8% identity with that of human Dnmtl and 66.4% with mouse Dnmt1. Translated amino acid sequence showed 88.4% homology with human homologue and 75.8% homology with mouse counterpart. Three types of Dnmt1 are reported in mouse and human, and are likely present in bovine tissues. Understanding of role of Dnmt1 in bovine development may shed a light in the field of animal, especially bovine cloning.

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Effect of Nanomaterials on the Early Development of Fish Embryos: (2) Metallic Nanomaterials (어류수정란 발달에 미치는 나노독성 연구동향: (2) 금속계 나노물질)

  • Shin, Yu-Jin;An, Youn-Joo
    • Journal of Korean Society on Water Environment
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    • v.28 no.6
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    • pp.943-953
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    • 2012
  • Because of their unique properties, nano-sized metallic nanomaterials (NMs) have been used in extensive applications of biomedicine, electronics, optics, engineering, and personal care products. Accordingly, with the increasing release of NMs into the environment, numerous studies of nanoecotoxicity have been conducted. Fish embryo toxicity test (FET) has many benefits in evaluating toxicity of NMs as an alternative to a whole-body test in fish. In this study, we collected and analyzed the toxicity studies of metallic NMs on freshwater fish embryos. Most studies have demonstrated that metallic NMs are highly toxic during the early development of fish embryos. However, it should be noted that the results for the same NMs on the same test species show variation due to differences in the size or surface properties of the test NMs and exposure conditions. For the safe use of metallic NMs, we need to analyze their effects based on their properties, test species, environmental media, and diverse conditions.

Comparison of Mechanical and Enzymatic Methods for the Isolation of Bovine Ovarian Follicles

  • Lim, Hyun-Joo;Kim, Dong-Hoon;Im, Gi-Sun;Lim, Jeong-Mook
    • Journal of Embryo Transfer
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    • v.25 no.2
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    • pp.93-96
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    • 2010
  • The isolation of preantral follicles from the ovaries of bovine was performed under mechanical and enzymatic methods. A significant increase in the total number of follicles retrieved was detected when tissue chopper was used. Micro-dissection could supply good quality, larger sized follicles (400 to $700{\mu}m$) but with the lowest yield ($9.0{\pm}1.0$). The isolated preantral and early antral follicles were cultured for 14 days. Follicles isolated by the mechanical method had a greater growth during a culture period than follicles collected enzymatically. Morphologically normal bovine oocytes from early antral follicles after 14 days culture were 59.6% after culture and after 24 h of maturation culture, 12.9% of in vitro-grown oocytes reached the second metaphase. In conclusion, this study showed that mechanical method can be used effectively to isolate intact preantral follicles from bovine ovaries.

Role of dipeptidyl peptidase-4 as a potentiator of activin/nodal signaling pathway

  • Park, Dong-Seok;Kim, Kyuhee;Jang, Minjoo;Choi, Sun-Cheol
    • BMB Reports
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    • v.51 no.12
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    • pp.636-641
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    • 2018
  • DPP4 (dipeptidyl peptidase-4), a highly conserved transmembrane glycoprotein with an exo-peptidase activity, has been shown to contribute to glucose metabolism, immune regulation, signal transduction, and cell differentiation. Here, we show that DPP4 is involved in control of activin/nodal signaling in Xenopus early development. In support of this, gain of function of DPP4 augmented Smad2 phosphorylation as well as expression of target genes induced by activin or nodal signal. In addition, Dpp4 and Xnr1 showed synergistic effect on induction of ectopic dorsal body axis, when co-injected at suboptimal doses in early embryos. Conversely, saxagliptin, a DPP4 inhibitor repressed activin induction of Smad2 phosphorylation. Notably, overexpression of Dpp4 disrupted specification of dorsal body axis of embryo, leading to malformed phenotypes such as spina bifida and a shortened and dorsally bent axis. Together, these results suggest that DPP4 functions as a potentiator of activin/nodal signaling pathway.

A Maternal Transcription Factor, Junction Mediating and Regulatory Protein is Required for Preimplantation Development in the Mouse

  • Lin, Zi-Li;Li, Ying-Hua;Jin, Yong- Xun;Kim, Nam-Hyung
    • Development and Reproduction
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    • v.23 no.3
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    • pp.285-295
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    • 2019
  • Junction-mediating and regulatory protein (JMY) is a regulator of both transcription and actin filament assembly. The actin-regulatory activity of JMY is based on a cluster of three actin-binding Wiskott-Aldrich syndrome protein homology 2 (WH2) domains that nucleate actin filaments directly and promote nucleation of the Arp2/3 complex. In addition to these activities, we examined the activity of JMY generation in early embryo of mice carrying mutations in the JMY gene by CRISPR/Cas9 mediated genome engineering. We demonstrated that JMY protein shuttled expression between the cytoplasm and the nucleus. Knockout of exon 2, CA (central domain and Arp2/3-binding acidic domain) and NLS-2 (nuclear localization signal domain) on the JMY gene by CRISPR/Cas9 system was effective and markedly impeded embryonic development. Additionally, it impaired transcription and zygotic genome activation (ZGA)-related genes. These results suggest that JMY acts as a transcription factor, which is essential for the early embryonic development in mice.

Evaluation of Offshore Seawater Qualify using Gametes and Embryos of Starfishes (Asterina pectinifera) (불가사리 알을 이용한 연안해수의 수질 평가)

  • Yu Chun Man;Lee Jong Bin;Park Jong Cheon;Joo Hyun Soo
    • Korean Journal of Environmental Biology
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    • v.22 no.3
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    • pp.376-380
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    • 2004
  • In August 2003, the water quality of offshore waters along the Incheon coast of Korea was evaluated by biological evaluation using gametes, embryos and early development systems of a starfish species (Asterina pectinifera). As the result of performing biological evaluations on seawater samples from a total thirteen sites, the formation rate of normal larva was 16-68%. At seawater sample from site 5 and 13, formation rate of normal larva averaged 16%, the most abnormal rate hindering the early embryo development of the experimental animal, while that of site 3 averaged 68%, the highest formation rate of normal larva. At seawater sample from site 2, 4, 7, 9, 10, 11, 12, formation rate of normal larva averaged 33-54%, those which damage the development of early embryos slightly. At seawater sample from site 1, 5, 6, 8, 13, formation rate of normal larva averaged 16-28%, those which damage the development of early embryos strongly.

Screening of Green Fluorescent Protein Gene and Sexing by PCR in Bovine Embryos (소 수정란에서 Green Fluorescent Protein 유전자 검색 및 PCR에 의한 성감별)

  • Lee, H. J.;Kang, T. Y.;Rho, G. J.;Chae, Y. J.;Lee, H.;Choe, S. Y.
    • Journal of Embryo Transfer
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    • v.15 no.2
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    • pp.157-165
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    • 2000
  • The efficiency of transgenic livestock production could be improved by early screening of transgene-integration and sexing of embryos at preimplantational stages before trasferring them into recipients. We examined the effciency of multiplex PCR analysis for the simultaneous confirmation of the trasgene and sex during the preimplantational development of bovine embryos and the possibility of green fluorescent protein(GFP) gene as a non-invasive marker for the early screening of transgenic embryos. The GFP gene was microinjected into the male pronuclei of bovine zygotes produced in vitro. The injected zygotes were co-cultured in TCM-199 containing 10% FCS with boving oviductal epithelial cells in a 5% CO2 incubator. Seventeen(13.0%) out of 136 gene-injected bovine zygotes developed by multiplex PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Eight(67%) of 12 embryos at 2-cell to blastocyst stage were positive in the PCR analysis, but only two(11.8%) of 17 blastocysts expressed the GFP gene. Their sex was determined as 7 female and 5 male embryos by the PCR analysis. The results indicate that the screening of GFP gene and sex in bovine embryos by PCR analysis and fluorescence detection could be a promisible method for the preselection of transgenic embryos.

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Detection of genetic abnormalities in human sperm, oocytes, and preimplantation embryos using fluorescence in situ hybridization (FISH) (Fluorescence in situ hybridization(FISH) 기법을 이용한 인간 생식세포 및 착상전 배아의 유전이상 검색)

  • 방명걸
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.12-18
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    • 1998
  • Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

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