• Title/Summary/Keyword: ecdysteroid

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Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Shen, Xing-Jia;Yi, Yong-Zhu;Tang, Shun-Ming;Li, Yi-Ren;He, Jia-Lu;Wu, Xiang-Fu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.169-174
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    • 2004
  • Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (lue) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 - 2.0 $\mu\textrm{g}$/$m\ell$ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5-2.0 ${\mu}g$/$m\ell$ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Molecular characterization of juvenile hormone signaling pathway-related genes in the brackish water flea Diaphanosoma celebensis (기수산 물벼룩의 유충 호르몬(Juvenile hormone) 신호전달경로 관련 유전자의 특성 분석)

  • Hayoung Cho;Jewon Yoo;Young-Mi Lee
    • Korean Journal of Environmental Biology
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    • v.40 no.3
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    • pp.255-266
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    • 2022
  • In crustaceans, molting is regulated by interactions between ecdysteroid and juvenile hormone (JH) signaling pathway-related genes. Unlike the ecdysteroid signaling pathway, little information on the role of JH signaling pathway-related genes in molting is available in zooplanktonic crustaceans. In this study, three genes (juvenile hormone acid O-methyltransferase (JHAMT), methoprene-tolerant (Met), and juvenile hormone epoxide hydrolase (JHEH)) which are involved in the synthesis, receptor-binding, and degradation of JH were identified using sequence and phylogenetic analysis in the brackish water flea, Diaphanosoma celebensis. Transcriptional changes in these genes during the molting cycle in D. celebensis were analyzed. Sequence and phylogenetic analysis revealed that these putative proteins may be functionally conserved along with those of insects and other crustaceans. In addition, the expression of the three genes was correlated with the molting cycle of D. celebensis, indicating that these genes may be involved in the synthesis and degradation of JH, resulting in normal molting. This study will provide information for a better understanding of the role of JH signaling pathway-related genes during the molting process in Cladocera.

Ecdysteroids from Melandrii Herba (왕불류행의 Ecdysteroid성분)

  • 김호경;전원경;고병섭
    • YAKHAK HOEJI
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    • v.45 no.5
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    • pp.455-459
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    • 2001
  • Melandrii Herba has been used for tympanitis, sore throat, dysentery and dehumidifying diuretic. From the BuOH fraction of methanol extracts, three ecdysteroids (ecdysterone, ecdysterone 22-acetate and inokosterone) were isolated by column chromatography using Amberlite XAD-4, ODS and Sephadex LH-20 gel and by HPLC method. The structures of these compounds were identified on the basis of spectroscopic methods.

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Molecular Cloning and Sequencing of the Ecdysteroid UDP-Glucosyl-transferase Gene, EGT, from Bombyx mori Nuclear Polyhedrosis Virus K1

  • Park, Hye-Jin;Chung, Eun-Hwa;Lee, Kwang-Sik;Han, Ji-Hee;Lee, Seong-Jin;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.37-41
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    • 2001
  • The ecdysteroid UDP-glucosyltransferase (egt) gene isolated from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain was compared to its homologue from Autographa californica NPV (AcNPV) and Bm NPV T3. The egt gene of BmNPV-K1 encoded 506 amino acid open reading frame, and was 99.6% identical at the amino acid level and 99.2% identical at the nucleotide level to BmNPV T3. The BmNPV-K1 egt gene showed highly identity to AcNPV and BmNPV T3 strain. The BmNPV-K1 egt gene was different from amino acid sequence at 2 positions, 19 and 72, in BmNPV T3. The genomic location of egt gene in the BmNPV-K1 was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Northern hybridization analysis. Transcripts of the egt of Bm NPV-K1 peaked around 12 hrs postinfection (p.i.) and reduced at 24 hrs p.i.

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Effects of Phytoecdysteroid on the Proliferation and Activity of Bone Cells (Phytoecdysteroid가 조골세포와 파골세포의 성장과 활성에 미치는 영향)

  • Ko, Seon-Yle
    • Journal of Oral Medicine and Pain
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    • v.32 no.2
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    • pp.129-135
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    • 2007
  • Ecdysteroids are known as insect molting hormone. At the same time, ecdysteroids and plant ecdysteroids (phytoecdysteorids) reveal beneficial effects on mammal. The present study was undertaken to determine the possible cellular mechanism of action of phytoecdysteroids in bone metabolism. The effects on the osteoblasts were determined by measuring cell proliferation, alkaline phosphatase (ALP) activity, and gelatinase activity. The effects on the osteoclasts were investigated by measuring tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation after culturing osteoclast precursors. Phytoecdysteroid treatment showed a increase in ALP activity of osteoblasts. Phytoecdysteroid increased the activity of gelatinase. In addition, phytoecdysteroid decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL) in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, phytoecdysteroid may be a regulatory protein within the bone marrow microenvironment.

Cytotoxicological Effect of Tebufenozide, an Insect Growth Regulator (IGR): Stimulation of Filamentous Actin Reorganization and Enhancement of Hsp27 Expression in Drosophila Kc Cells

  • Hwang, Jee-Na;Jung, Hwa-Jin;Seo, Young-Rok
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.6
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    • pp.351-354
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    • 2004
  • The cytotoxicological responses to insect growth regulator (IGR), using tebufenozide as ecdysteroid mimic, were investigated in Drosophila Kc cells. Treatment of Kc cells with tebufenozide showed significant growth inhibition and striking morphological changes including aggregation and elongation of the cells. In order to understand the cellular mechanism underlying the response of Drosophila cells to tebufenozide, immunofluorescence microscopy was performed. We found that treatment of Kc cells with tebufenozide enhanced the reorganization of f-actin and stimulated the expression of hsp27. These data suggest a possible association of filamentous actin (f-actin) and hsp27 in the cytotoxicological mechanisms of growth regulators in Drosophila cells.

곤충호르몬의 작용기작과 그 이용방안

  • 문재유
    • Journal of Sericultural and Entomological Science
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    • v.38 no.2
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    • pp.186-188
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    • 1996
  • 곤충애 있어서 호르몬의 작용기작과 그 이용에 관하여 요야하면 다음과 같다. 곤충호르몬에는중추신경계의 신경분비세포에서 합성, 분비되는 peptide의 neuropeptide hormone (PTTH, bombyxin, diapause hormone등)이 있고, 상피계의 내분비선에서 합성, 분비되는 sesquiterpene의 유약호르몬 ( juvenile hormone)과 steroid의 탈피호르몬 (ecdysone)이 있다. 곤충호르몬은 특정한 표족세포에 있는 수용체와 높은 특이성과 높은 친화성을 가지고 결합해서 세포의 작용을 조절한다. 일반적으로 peptide hormone은 표적세포의의 세포막을 통과할 수 없으므로 표적세포의 막표면에 있는 수용체와 결합하는 것에 의해 세포내 대사제를 활성화시킴으로서 peptide hormone의 특이적인 발현이되는 것으로 알려지고 있다. 한편, ecdysone과 같은 steriod hormone이나 juvenlie hormone은 표적세포의 세포막을 용이하게 통과할수 있으므로 세포내의 세포막을 용이하게 통과할 수 있으므로 세포내로 들어가 수용체와 결합해서 hormone-receptor comlpex는 핵내로 들어가 DNA의 특이적인 영역에 결합하므로서 이들 호르몬 특이적인 기능이 발현되는 것으로 알려져 있다. Ecodysone의 활성이 있는 ecdysteroid가 여러 식물에서 발견되고 있어, 금후 양잠의 상족에 이용이 기대되고 있다. 또한, 향유약호르몬(AJH) 활성물지인 imidazole계 화합물은 양잠에 있어서 세섬도고치 생산에 그 이용이 기대되고 있다.

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Genetic Engineering for Detection of Endocrine Disruption using I-18 C Gene Expression in Chironomus riparius

  • Kwak Inn-Sil
    • Korean Journal of Environmental Biology
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    • v.23 no.3 s.59
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    • pp.269-274
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    • 2005
  • The 2D/E gel analysis for polypeptide expression reflecting I-18 C gene (early-ecdysterone inducible gene) has conducted the emerged C. riparius adults from larval phase exposure to tebufenozide acting as an ecdysteroidal molting hormone. Control group, the amount of ORE II of the I-18 C gene was larger than that of ORE I of this gene. After treatments, ORE I of the I-18 C gene was overexpressed as the polypeptide, whereas ORF II of this gene was expressed as the polypeptide and was clearly reduced expression. Accordingly, we consider that tebufenozide exhibited endocrine disruptions related processing of ecdysteroid receptor protein reflecting ORF II of I-18 C gene. Also, earlier emergence day was related overexpressed polypeptide reflecting ORE I of I-18 C gene. In this study result, tebufenozide induced changing of physiological condition, and then polypeptide expression reflecting early-ecdysterone inducible I-18 C gene was different between control group and exposure group.

Expression Changes of Chironomus riparius Proteome reflecting Ecdysterone Inducible Genes and Mitogenome exposure to Bisphenol-A

  • Kwak Inn-Sil
    • Korean Journal of Environmental Biology
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    • v.23 no.3 s.59
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    • pp.275-280
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    • 2005
  • Development of the fourth-instar larvae of Chironomus riparius has a sensitive to ecdysteroid hormones. The 2D/E gel analysis for polypeptide expression reflecting early-ecdysterone inducible gene has conducted the emerged female from larval phase exposure to bisphenol A (BPA). In the 2D/E gel 1108 protein spots were identified. The visualized protein spots allowed extraction of 17 protein spots differed more than 3 fold in BPA treated animals, which was approximately $1.6\%$ of the total protein spots. However, polypeptide expression reflecting early-ecdysterone inducible gene didn't change after treatments. In addition, detection for the damages or changes in mitogenome level was observed. The conserved cytochrome oxidase I in DNA level affected exposure to BPA $(1{\mu}gL^{-1})$ in this preliminary study.