• Title/Summary/Keyword: Oct-4

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Genetic Approach to Identify Critical Factors for Mouse Early Embryogenesis

  • Park, Joon-Hyun;Kim, Ji-Soo;Sonn, Sung-Keun;Rhee, Kun-Soo
    • Animal cells and systems
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    • v.10 no.1
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    • pp.41-47
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    • 2006
  • Development of the mammalian pre-implantation embryos has unique features, such as a slow and unsynchronized cell division, compaction, and eventual formation of blastocysts with inner cell mass and trophectoderm. In order to have a clue on molecular mechanisms that reside in mouse early development, we suppressed expression of early embryo-specific genes with RNAi and observed their development in vitro. We observed developmental defects in embryos microinjected with dsRNAs for Oct4 or Nanog among the tested genes. Careful examinations revealed that development of the most of the Oct4- or Nanog-suppressed embryos were arrested at the morula stage. These results suggest that the Oct4 and Nanog activities are also required for embryogenesis earlier than the blastocyst stage.

Effects of Seeding Date and Rate on Growth and Yield of Barley Cultivar 'Duwonchapssalbori' in Southern Region

  • Chun, Jong-Un;Park, Hae-Sik;Nam, Jung-Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.1
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    • pp.14-19
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    • 2000
  • In order to develop an appropriate cultural practice for a newly bred 2-rowed, waxy, naked barley cultivar 'Duwonchapssalbori' in the southern region, 2 varieties, 3 seeding dates, 4 seeding rates were designed with 2 replications. The heading and maturity dates of 'Duwonchapssalbori' were' earlier than 'Saessalbori by 2-5 and 4 days, respectively. Culm length of 'Duwonchapssalbori' was 60 to 66cm with lodging resistance. Grain yields of both varieties were 4.28-4.33 ton/ha in the plot of Oct. 20 seeding, 4.11-4.18 ton in the plot of Oct. 28 seeding, and 3.7-13.89 ton/ha in the Nov. 5 seeding. Grain yield with different seeding rates highly significantly fitted with secondary polynomial equations in the plots of Oct. 20 and Oct. 28 seedings ($R^2$=0.84-0.85), showing the greatest yield with seeding rate of 140kg per ha. However, the grain yield with various seeding rates in the plot of Nov. 5 seeding showed simple linear regression ($R^2$=0.71), showing the seeding rate over 160kg in the case of later seeding to be optimal. Number of spikes per $m^2$ significantly contributed to grain yield, and fitted to simple linear regression ($R^2$=0.881-0.891), suggesting that plenty of early stands should be established at early seedling stage for high grain yield. Young spikes of 'Duwonchapssalbori' in the plot of Oct. 20 seeding elongated with linear increments since Feb. 10 (6.7mm), and showed rapid increment since March 7 (15.1mm). However, elongation of young spikes of 'Saessalbori' began from March 15 with slow increments. From these studies, the optimum seeding date and seeding rate in 'Duwonchapssalbori' were Oct. 20-28 and 140kg per ha for high grain yield in the southern regions.

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Effect of cell-penetrating peptide-conjugated estrogen-related receptor ${\beta}$ on the development of mouse embryos cultured in vitro

  • Yang, Ning Jie;Seol, Dong-Won;Jo, Junghyun;Jang, Hyun Mee;Yoon, Sook-Young;Lee, Dong Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.1
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    • pp.1-8
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    • 2014
  • Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

Very Small Putative Stem Cells Detected in Human Endometrium (인간 자궁내막에서 발견되는 극소형 추정줄기세포)

  • Choi, Jong-Ryeol;Joo, Jong-Kil;Jun, Eun-Sook;Ko, Kyoung-Rae;Lee, Hong-Gu;Lee, Kyu-Sup;Kim, Won-Gyu
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.2
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    • pp.99-113
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    • 2010
  • Objective: It has been recently reported that very small stem cells with pluripotency are detected in murine and human. The purposes of this study are to confirm whether very small putative stem cells (VSPSCs), which have the proper characteristics of stem cells as well as the expression of stem cell markers, are detected in human endometrium. Methods: The endometrial cells of 5 women, which were obtained by endometrial biopsy, were cultured for 2 weeks and were confirmed for the expressions of alkaline phosphatase, OCT-4 and CXCR4 by immunochemistry. Subsequently VSPSCs were separated by percoll density gradient method and were cultured. Also VSPSCs and their derived cells were confirmed for the expressions of OCT-4 and CXCR4. Results: The colonies, which is composed with VSPSCs less than 3 ${\mu}m$ and the 5~15 ${\mu}m$ sized hyperchromatic round cells, were detected in the endometrium of all of women and showed the strong expressions of alkaline phosphatase, OCT-4 and CXCR4. In culture after the separation of VSPSCs by percoll, these cells showed the morphological and functional characteristics of stem cells; self-renewal, colony formation, embryoid body-like formation and differential plasticity. VSPSCs formed gradually the 5~15 ${\mu}m$ sized hyperchromatic round cells and the 10~20 ${\mu}m$ sized sphere-shaped cells by cell-to-cell aggregation or cell fusion. Then these cells differentiated the various cells including fibroblast-like cells, nerve-like cells and endothelium-like cells. VSPSCs and their derived cells often showed the strong expressions of OCT-4 and CXCR4. Conclusion: VSPSCs less than 3 ${\mu}m$ and their derived cells are detected in human endometrium and these cells have the proper characteristics of stem cells and the expressions of stem cell markers as alkaline phosphatase, OCT-4 and CXCR4.

Haplotype Analysis and Single Nucleotide Polymorphism Frequency of Organic Cation Transporter Gene (OCT1 and 2) in Korean Subjects

  • Kim, Se-Mi;Lee, Sang-No;Yoon, Hwa;Kang, Hyun-Ah;Cho, Hea-Young;Lee, Il-Kwon;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.345-351
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    • 2009
  • Organic cation transporters (OCTs) are important for absorption, elimination of many endogenous small organic cations as well as a wide array of drugs and environmental toxins. This gene is located in a cluster on chromosome 6 and OCTs are in major organs such as intestine, liver, kidney, brain and placenta. Therefore, expression levels and function of OCTs directly affect plasma levels and intracellular concentrations of drugs and thereby determine therapeutic response. The aim of this study was to investigate the frequency of the SNPs on OCT1 (C181T and C1022T) and OCT2 (G808T) to analyze haplotype frequency in healthy Korean population. Human subjects have been genotyped for OCT1 (C181T for 195 subjects and C1022T for 825 subjects), using polymerase chain reaction-based diagnostic tests (RFLP). And for OCT2 (G808T), a total of 861 subjects have been genotyped, using pyrosequencing method. Haplotype was statistically inferred using an algorithm based on the expectation-maximization (EM). OCT1 C181T genotyping showed 100% homozygous wild-type (C/C). OCT1 C1022T genotyping showed wild-type (C/C), heterozygous (C/T) and homozygous mutant-type (T/T) and each accounted for 72.1, 24.5 and 3.4%, respectively. OCT2 G808T genotyping results also showed homozygous wild-type (G/G), heterozygous (G/T) and homozygous mutant-type (T/T) and each took 81.8, 17.9 and 0.3%, respectively. Based on these genotype data, haplotype analysis between OCT1 C181T and OCT1 C1022T has proceeded. The result has revealed that linkage disequilibrium between alleles is not obvious (P=0.0122).

Presence of Transcription Factor OCT4 Limits Interferon-tau Expression during the Pre-attachment Period in Sheep

  • Kim, Min-Su;Sakurai, Toshihiro;Bai, Hanako;Bai, Rulan;Sato, Daisuke;Nagaoka, Kentaro;Chang, Kyu-Tae;Godkin, James D.;Min, Kwan-Sik;Imakawa, Kazuhiko
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.5
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    • pp.638-645
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    • 2013
  • Interferon-tau (IFNT) is thought to be the conceptus protein that signals maternal recognition of pregnancy in ruminants. We and others have observed that OCT4 expression persists in the trophectoderm of ruminants; thus, both CDX2 and OCT4 coexist during the early stages of conceptus development. The aim of this study was to examine the effect of CDX2 and OCT4 on IFNT gene transcription when evaluated with other transcription factors. Human choriocarcinoma JEG-3 cells were cotransfected with an ovine IFNT (-654-bp)-luciferase reporter (-654-IFNT-Luc) construct and several transcription factor expression plasmids. Cotransfection of the reporter construct with Cdx2, Ets2 and Jun increased transcription of -654-IFNT-Luc by about 12-fold compared with transfection of the construct alone. When cells were initially transfected with Oct4 (0 h) followed by transfection with Cdx2, Ets2 and/or Jun 24 h later, the expression of -654-IFNT-Luc was reduced to control levels. OCT4 also inhibited the stimulatory activity of CDX2 alone, but not when CDX2 was combined with JUN and/or ETS2. Thus, when combined with the other transcription factors, OCT4 exhibited little inhibitory activity towards CDX2. An inhibitor of the transcriptional coactivator CREB binding protein (CREBBP), 12S E1A, reduced CDX2/ETS2/JUN stimulated -654-IFNT-Luc expression by about 40%, indicating that the formation of an appropriate transcription factor complex is required for maximum expression. In conclusion, the presence of OCT4 may initially minimize IFNT expression; however, as elongation proceeds, the increasing expression of CDX2 and formation of the transcription complex leads to greatly increased IFNT expression, resulting in pregnancy establishment in ruminants.

Crystal Structure of N,N'-di-tert-butoxycabonyl-2, 7-diazabicyclo[3.3.0]oct-4-ene. (N,N'-di-tert-butoxycabonyl-2,7-diazabicyclo[3.3.0]oct-4-ene의 결정구조)

  • 김문집;이재혁
    • Korean Journal of Crystallography
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    • v.8 no.2
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    • pp.132-137
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    • 1997
  • The crystal structure of N,N'-di-tert-butoxycabonyl-2,7-diazabicyclo[3.3.0]oct-4-ene has been determined from single crystal x-ray diffraction study; C16H26N2O4, Triclinic, P1, a=11.119(1) Å, b=13.638(1) Å, c=6.214(1) Å, α=92.14(1)°, β=103.49(1)°, γ=73.35(1)°, V=877.4(2)Å3, T=293(2)K, Z=2, CuKα(λ=1.5418Å). The structure was solved by direct method and refined by full-matrix least squares to a final R=5.38% for 2389 unique observed F0>4σ(F0) reflections and 225 parameters.

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n-Alkane Utilizing Capability and Location of the Genes for Alkane Hydroxylases in Pseudomonas maltophilia N246

  • Choi, Soon-Young;Lee, Myung-Hye;Hwang, Moon-Ok
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.252-255
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    • 1993
  • Pseudomonas maltophilia N246 carrying on OCT plasmid grew on n-alkanes of 6 to 14 carbon atoms, but not on n-alkanes of more carbon atoms. P. maltophilia strains with and without OCT plasmid could utilize primary alcohols. aldehydes and fatty acids derived from n-alkane. The N246 strain could also utilize monocarboxylic and dicarboxylic acids, and terminal branched dimethyloctane. Unlike the genes of alcohol dehydrogenase and aldehyde dehydrogenase which were located on both the chromosome and the OCT plasmid, genes for the alkane hydroxylase components were located only on the OCT plasmid in P. maltophilia N246.

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Genes for the Catabolism of Deoxyfructosyl Glutamine in pAtC58 Are Attributed to Utilization of Octopine in Agrobacterium tumefaciens Strain NT1

  • Baek, Chang-Ho;Park, Dae-Kyun;Lee, Ko-Eun;Hwang, Won;Kim, In-Hwang;Maeng, Jue-Son;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.822-828
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    • 2004
  • Nopaline-type Agrobacterium tumefaciens strain C58 cannot utilize octopine (Oct) as the sole carbon and nitrogen sources. This strain harbors two plasmids; a virulent plasmid, pTiC58, and a megaplasmid, pAtC58. From strain NT1, which is a derivative of C58 harboring only pAtC58, we isolated spontaneous mutants that utilize Oct as the sole nitrogen source. These Oct-catabolizing mutants, however, could not utilize the opine as the sole carbon source. In contrast, strain UIA5, a plasmid-free derivative of C58, could not give rise to such mutants. The mutations isolated from NT1 were mapped to socR in pAtC58, which is a negative regulator of the soc operon responsible for the uptake and catabolism of an Amadori opine, deoxyfructosyl glutamine (Dfg). A derivative of UIA5 carrying a clone of the soc operon with a transposon inserted in socR also utilizes Oct as the sole nitrogen source. However, UIA5 harboring the operon with mutations in each of the structural genes in the soc operon, socA, B, C, and D, lost the ability to generate spontaneous Oct-utilizing mutants, suggesting that soc genes in pAtC58 are required for the utilization of Oct as a nitrogen source, and that derepressed expression of these genes allows cells to utilize Oct. In contrast, Oct-catabolizing mutants derived from C58, which grew using Oct as the sole nitrogen source, could also utilize the opine as the sole carbon source. These mutants did not carry any detectable mutations in socR or the region upstream to the gene in pAtC58, suggesting that mutations occurring elsewhere in the genome, most likely in pTiC58, allow the uptake and catabolism of the opine.