• Title/Summary/Keyword: Gene regulation

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Expression of Coat Color Associated Genes in Korean Brindle Cattle by Microarray Analysis

  • Lee, Hae-Lee;Park, Jae-Hee;Kim, Jong Gug
    • Journal of Embryo Transfer
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    • v.30 no.2
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    • pp.99-107
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    • 2015
  • The aim of the present study was to identify coat color associated genes that are differentially expressed in mature Korean brindle cattle (KBC) with different coat colors and in Hanwoo cows. KBC calves, before and after coat color appearance, were included. Total cellular RNA was isolated from the tail hair cells and used for microarray. The number of expressed coat color associated genes/probes was 5813 in mature KBC and Hanwoo cows. Among the expressed coat color associated genes/probes, 167 genes were the coat color associated genes listed in the Gene card database and 125 genes were the pigment and melanocyte genes listed in the Gene ontology_bovine database. There were 23 genes/probes commonly listed in both databases and their expressions were further studied. Out of the 23 genes/probes, MLPH, PMEL, TYR and TYRP1 genes were expressed at least two fold higher (p<0.01) levels in KBC with brindle color than either Hanwoo or KBC with brown color. TYRP1 expression was 22.96 or 19.89 fold higher (p<0.01) in KBC with brindle color than either Hanwoo or KBC with brown color, respectively, which was the biggest fold difference. The hierarchical clustering analysis indicated that MLPH, PMEL, TYR and TYRP1 were the highly expressed genes in mature cattle. There were only a few genes differentially expressed after coat color appearance in KBC calves. Studies on the regulation and mechanism of gene expression of highly expressed genes would be next steps to better understand coat color determination and to improve brindle coat color appearance in KBC.

Toxicogenomic Analysis and Identification of Estrogen Responsive Genes of Di (n-ethylhexyl) Phthalate in MCF-7 Cells

  • Kim, Youn-Jung;Yun, Hye-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.149-156
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    • 2005
  • Di (n-ethylhexyl) phthalate (DEHP) is thought to mimic estrogens in their action, and are called endocrine disrupting chemicals. DEHP is used in numerous consumer products, especially those made of flexible polyvinyl chloride and have been reported to be weakly estrogenic. In this study, DEHP were tested for estrogenic properties in vitro models and with microarray analysis. First, the E-screen assay was used to measure the proliferation of DEHP in MCF-7 cells, a human breast cancer cell line. DEHP induced an increase in MCF-7 cell proliferation at concentration of $10^{-4}M$. Second, we carried out a microarray analysis of MCF-7 cells treated with DEHP using human c-DNA microarray including 401 endocrine system related genes. Of the genes analyzed, 60 genes were identified showing significant changes in gene expression resulting from DEHP. Especially, 4 genes were repressed and 4 genes were induced by DEHP compared to $17{\beta}-estradiol$. Among these genes, trefoil factor 3 (intestinal), breast cancer 1, early onset and CYP1B1 are involved in estrogen metabolism and regulation. Therefore it suggests that these genes may be associated with estrogenic effect of the DEHP on transcriptional level. The rationale is that, as gene expression is a sensitive endpoint, alterations of these genes may act as useful biomarkers to define more precisely the nature and level of exposure to kinds of phthalates.

Molecular Characterization and Expression Analysis of Creatine Kinase Muscle (CK-M) Gene in Horse

  • Do, Kyong-Tak;Cho, Hyun-Woo;Badrinath, Narayanasamy;Park, Jeong-Woong;Choi, Jae-Young;Chung, Young-Hwa;Lee, Hak-Kyo;Song, Ki-Duk;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.12
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    • pp.1680-1685
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    • 2015
  • Since ancient days, domestic horses have been closely associated with human civilization. Today, horse racing is an important industry. Various genes involved in energy production and muscle contraction are differentially regulated during a race. Among them, creatine kinase (CK) is well known for its regulation of energy preservation in animal cells. CK is an iso-enzyme, encoded by different genes and expressed in skeletal muscle, heart, brain and leucocytes. We confirmed that the expression of CK-M significantly increased in the blood after a 30 minute exercise period, while no considerable change was observed in skeletal muscle. Analysis of various tissues showed an ubiquitous expression of the CK-M gene in the horse; CK-M mRNA expression was predominant in the skeletal muscle and the cardiac muscle compared to other tissues. An evolutionary study by synonymous and non-synonymous single nucleotide polymorphism ratio of CK-M gene revealed a positive selection that was conserved in the horse. More studies are warranted in order to develop the expression of CK-M gene as a biomarker in blood of thoroughbred horses.

Comparison of Gene Expression between Cumulus Oocyte Complexes and Naked Oocytes by Suppression Subtractive Hybridization in Swine

  • Xiang, Zhi Feng;Zhang, Jin Zhou;Li, Xue Bin;Xie, Hong Bin;Wang, Qing Hua
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.1
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    • pp.17-24
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    • 2010
  • In the antral follicle phase, several layers of cumulus cells surround the oocyte and play an important support and regulation role in oocyte development and maturation via intercellular communications and interactions between oocytes and cumulus cells. However, information on stage specific gene expression in swine during the phase is not well understood. To investigate the function of cumulus cells during in vitro maturation of porcine oocytes and gene expression, suppression subtractive hybridization (SSH) was performed to screen genes that were differentially expressed between cumulus-oocyte complexes (COCs) and naked oocytes (NOs). Utilizing mRNAs from in vitro maturation oocytes, a SSH cDNA library from COCs as the tester and NOs as the driver was constructed. The SSH cDNA library was then screened using dot blot analysis. Results showed that a total of 70 clones randomly selected from the library were differentially expressed. Among these, 41 exhibited high homology to known genes and 11 were novel expressed sequences tags (ESTs). Four differentially expressed genes, including bfgf, sprouty 2, egr and btc, were further studied by real time quantitative PCR; results confirmed an increased expression of respective mRNA in COCs compared with NOs, which suggests that these factors may play an important role in oocyte development and maturation.

Modulation of Aqueous Extracted Angelicae sinensis Radix on Nitric Oxide Production and Pro-inflammatory Cytokine Gene Expressions in RAW 264.7 Macrophage Cells

  • Lee Young Sun;Han Ok Kyung;Shin Sang Woo;Park Jong Hyun;Kwon Young Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.6
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    • pp.1514-1518
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    • 2003
  • Angelica sinensis radix, Danggui, is a traditional oriental medication, which has been used to modulate immune response. We report here that aqueous extract of Angelica sinensis radix (ASR) can induces NO production, and inhibit LPS-induced NO production in dose-dependent manner in RAW 264.7 macrophage cells. ASR also induces iNOS mRNA and iNOS protein expression, and exhibit inhibitory effect on iNOS mRNA and protein expression in a dose-dependent manner in LPS-stimulated RAW 264.7 macrophage cells. Cytokines involved in the regulation of inflammatory reaction and immune response may play a role in the pathogenesis. ASR induces. pro-inflammatory cytokine gene expression (IL-1α, IL-1β and IL-6 gene) in a dose-dependent manner, and inhibits the expressions of these cytokines in LPS-stimulated RAW 264.7 macrophage cells. These data indicate that (1) ASR may be a potential therapeutic modulator of NO synthesis in various pathological conditions, and (2) the immunomodulatory effects of ASR may be, in part, associated with the inducing or suppression of pro-inflammatory cytokine gene expressions.

Inhibition of the Expression of Matrix Metalloproteinases in Articular Chondrocytes by Resveratrol through Affecting Nuclear Factor-Kappa B Signaling Pathway

  • Kang, Dong-Geun;Lee, Hyun Jae;Lee, Choong Jae;Park, Jin Sung
    • Biomolecules & Therapeutics
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    • v.26 no.6
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    • pp.560-567
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    • 2018
  • In the present study, we tried to examine whether resveratrol regulates the expression of matrix metalloproteinases (MMPs) through affecting nuclear factor-kappa B ($NF-{\kappa}B$) in articular chondrocytes. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-${\beta}$ ($IL-1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. Effect of resveratrol on $IL-1{\beta}$-induced secretion of MMP-3 was investigated in rabbit articular chondrocytes using western blot analysis. To elucidate the action mechanism of resveratrol, effect of resveratrol on $IL-1{\beta}$-induced $NF-{\kappa}B$ signaling pathway was investigated in SW1353, a human chondrosarcoma cell line, by western blot analysis. The results were as follows: (1) resveratrol inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) resveratrol reduced the secretion of MMP-3; (3) resveratrol inhibited $IL-1{\beta}$induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa $B{\alpha}$ ($I{\kappa}B{\alpha}$); (4) resveratrol inhibited $IL-1{\beta}$-induced phosphorylation and nuclear translocation of $NF-{\kappa}B$ p65. This, in turn, led to the down-regulation of gene expression of MMPs in SW1353 cells. These results suggest that resveratrol can regulate the expression of MMPs through affecting $NF-{\kappa}B$ by directly acting on articular chondrocytes.

Translational Control of The Heat Shock Gene Expression in Carrot (당근의 열충격 유전자 발현의 번역과정에서의 조절)

  • Cheol Ho HWANG
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.1
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    • pp.19-23
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    • 1995
  • Carrot cultured cells are able to respond to a temperature increase by inducing a set of new proteins, heat shock proteins (HSP). Such an induction of the HS gene was known to be achieved mainly at the level of transcription. However there has been an increasing number of evidences showing that a translational control was involved in the regulation of the HS gene expression. A comparison of HSP synthesized in vivo to in vivo (represent for mRNA level since the amount of the proteins produced by in vivo translation system will be proportional to an amount of the corresponding mRNA)showed no correlation between the amount of HS mRNA and the amount of the corresponding HSP at $30^{\circ}C$, It appears that a translational control may exert a major role in the expression of HS gene in carrot callus cells at $30^{\circ}C$.

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The Role of Adenylyl Sulfate Reductase to Abiotic Stress in Tomato

  • Seong, Eun-Soo;Lee, Ji-Yeon;Yu, Chang-Yeon;Yang, Deok-Chun;Eom, Seok-Hyun;Cho, Dong-Ha
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.173-180
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    • 2007
  • The full-length cDNA of LeAPR1 encoded a protein of 461 amino acid residues, which contained homology with phosphoadenosine phosphosulphate reductase (PAPS reductase) in N-terminal and an adenylylsulfate reductase in N-term and C-terminal. Analysis of the deduced amino acid sequence of LeAPR1 revealed that it shares high sequence identity with potato StAPR (96% identity)(Gene bank accession no. CDC44841). We found that multiple copies of LeAPR1 gene are present in the tomato genome through southern blot using genomic DNA was digested with 3 different restriction enzymes. The expression of LeAPR1 was also examined in various organs and its expression was also detected at high levels in roots and stems. Only high amounts of LeAPR1 transcripts were detected at high transcripts in the leaves at time 0, and then reduced as the plant stressed by the NaCl and abscisic acid (ABA). After 24h treatment of NaCl and ABA were showed increasing patterns of LeAPR1 gene. Time course of LeAPR1 gene expression was examined under oxidative stresses from metyl viologen (MV) and hydrogen peroxide ($H_2O_2$). In the presence of 10 mM $H_2O_2$ and $50\;{\mu}M$ MV, the levels of LeAPR1 transcript in leaves decreased after 1 h, and then increased strongly, peaked at 24 h. Our results indicated that LeAPR1 may play a role function of circadian regulation involved in abiotic stresses signaling pathways.

Methylation Status of H19 Gene in Embryos Produced by Nuclear Transfer of Spermatogonial Stem Cells in Pig

  • Lee, Hyun-Seung;Lee, Sung-Ho;Gupta, Mukesh Kumar;Uhm, Sang-Jun;Lee, Hoon-Taek
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.67-75
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    • 2011
  • The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.

Effect of Astragali radix Extract on Acetaminophen-induced Hepatotoxicity in Mice (황기 추출액이 Acetaminophen으로 유발된 마우스의 간 손상에 미치는 영향)

  • Lee Young Sun;Han Ok Kyung;Jean Tae Won;Lee Eun Sil;Kim Kwang Joong;Park Chan Woo;Kim Hyo Jung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.4
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    • pp.707-713
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    • 2002
  • Astragali radix (AR) is one of the oldest and mast frequently used crude drug for traditional medicine in many Asian countries. This study designed to investigate the hepatoprotective effects of the aqueous extracted AR (ARE) against acetaminophen (APAP)-induced hepatic damage in ICR mice. APAP at the dose of 450 mg/kg i.p produced liver damage in ICR mice. Serum enzyme activities of alanine aminotransferase, aspartate aminotransferase and sorbitol dehydrogenese was dramatically decreased up to control level by pretreatment of ARE. However, hepatic glutathione level did not show a significant change between the tested groups. We also investigated TNF α mRNA gene expression on APAP-induced liver damage by RT-PCR. APAP dramatically induced TNF α mRNA gene expression in ICR mice. Pretreatment of mice with ARE led to a marked decrease of TNF α mRNA gene expression. These data indicate that 1) ARE has clearly revealed a hepatoprotective effect against APAP-induced hepatic damage in ICR mice, and 2) the protective effect of ARE may be, in part, associated with the regulation of TNF α mRNA gene expression.