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Hybridohistochemical analysis of latently infected virus

Research Project

Project/Area Number 04454111
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 基礎獣医学
Research InstitutionHokkaido University

Principal Investigator

SATO Fumiaki  Hokkaido University, Faculty of Veterinary Medicine, Professor, 獣医学部, 教授 (10162471)

Co-Investigator(Kenkyū-buntansha) ENDOH Daiji  Hokkaido University, Faculty of Veterinary Medicine, Instructor, 獣医学部, 助手 (40168828)
KON Yasuhiro  Hokkaido University, Faculty of Veterinary Medicine, Assistant Professor, 獣医学部, 助教授 (10178402)
HAYASHI Masanobu  Rakuno Gakuen University, Department of Veterinary Radiology, Professor, 酪農学部, 教授 (10130337)
KUWABARA Mikinori  Hokkaido University, Faculty of Veterinary Medicine, Assistant Professor, 獣医学部, 助教授 (10002081)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1992: ¥3,000,000 (Direct Cost: ¥3,000,000)
KeywordsHybridohistochemistry / Latent infection / Herpesvirus / Marek's disease virus / ICP4 / Renin / Methylation / Mouse hepatitis virus / in situ ハイブリダイゼーション / LEC / マレック病ウイルス / ICP4ホモローグ / 電子顕微鏡 / インサイチュウハイブリダイゼーション / ジゴキシゲニン / 潜在感染 / 燐酸化蛋白質 / テロメア
Research Abstract

In this project, we focused on the latent infection of a gallid herpesvirus, Marek's disease virus (MDV). It was reported that some latently infected viruses may cause tumors and monsters. We observed the gene expression of the latently infected MDV using hybridohistochemistry. Prior to observation of the expression of a virus gene, we estimated the lower limit of the detection using a digoxigenin-labeled cRNA probe. We used mouse renin as a model. The lower limit of the detection was estimated to be the same range of abundance of beta-actin. Using the same method, we observed transcription of sense and antisense strands of Marek's disease virus (MDV) ICP4 homologue gene in MDV-infected chickens (3-4 weeks postinfection). A cRNA probe of MDV ICP4 was hybridized on paraformaldehyde-fixed and paraffin-embedded sections of feather follicle epithelium (FFE) and some lymphocytes in kidney, liver, nerve and spleen. Sense-strand transcripts of the MDV ICP4 gene were abundantly detected in the FFE compared with lymphocytes. Signals for antisense transcripts were seen in some kidney, liver, nerve and spleen lymphoid cells. Northern blot analysis confirmed the existence of transcripts in these tissues. Abundance and the sense/antisense ratio were also confirmed by Northern blot analysis. Immunoblot analysis also confirmed the expression of sense transcripts in FFE,kidney and liver. Thus, the results from lymphocytes suggested that sense and anti-sense transcripts of MDV ICP4 were expressed in some lymphocytes in the kidney, liver and spleen during the latent period. DNA of MDV genome was not detected because of its low copy umber. These data confirmed the reports on MD tumor cells that MDV genome was integrated in host genome and had a nucleosomal structure.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] Kon,Y.,Endoh,D.et al.: "A simple method of hybridohistochemistry for detection of renin mRNA in mouse" J.Vet.Med.Sci. 55. 461-463 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] N.Sasaki,Hayash,M.et al.: "Transgenic mice with antisense RNA and ribozyme" J.Vet.Med.Sci.55. 549-554 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Endoh,D.Niikura,M.et al.: "Expression and purification of recombinant Marek's disease virus serotype 1 specific phosphorylated protein pp38 in E.cdi" J.Vet.Med.Sci.56. 823-826 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hayashi,M.,Furuichi,T.et al.: "Enhancement of mRNA synthesis from Marek's disease virus genome in the lymphoblastoid cell line MDCL-MSB1 by 5-azac." J.Vet.Med.Sci.56. 287-291 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okui,T.,Hayashi,M.et al.: "A high frequency of induction of chromosome aberations in bone marrow cells of LEC strain rats by X-irradiation" Mutat.Res.324. 165-169 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kon,Y.,Endoh,D.et al.: "Expression of renin in coaglating glands is regulated by tests sterone" Anatomical Rec.241. (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Hayashi, D,Endoh, Y.Kon, T.Yamashita, N.Hashimoto, et al.: "Higher sensitivity of LEC strain rat in radiation-induced acute intestinal death" J Vet Med Sci.54. 269-273 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, D.Endoh, A.Fukamizu, K.Murakami, Y.Hashimoto, et al.: "Comparative study of renin expression in the coagulating glands of C57BL/6 and Balb/c mice" J Anat.181. 335-343 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, Y.Hashimoto, K.Murakami, M.Sugimura: "Whole-mount preparation of renin-containing cells in hydronephrotic mice" J Vet Med Sci.54. 589-590 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, Y.Hashimoto, K.Murakami, M.Sugimura: "An immunoelectron-microscopical observation of mouse juxtaglomerular cells in the case of experimental hydronephrosis" Acta Anat Basel.144 (4). 354-62 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, S.Takahashi, A.Fukamizu, k.Murakami, Y.Hashimoto, et al.: "Morphological and northern blot analysis of juxtaglomerular cells in experimental hydronephrotic mice" Anat Rec.232 (3). 393-400 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Mizutani, M.Hayashi, A.Maeda, T.Yamashita, H.Isogai, et al.: "Inhibition of mouse hepatitis virus multiplication by an oligonucleotide complementary to the leader RNA" J Vet Med Sci.54 (3). 465-72 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Niikura, Y.Matsuura, D.Endoh, M.Onuma, T.Mikami: "Expression of the Marek's disease virus (MDV) homolog of glycoprotein B of herpes simplex virus by a recombinant baculovirus and its identification as the B antigen (gp100, gp60, gp49) of MDV" J Virol.66 (5). 2631-8 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Hayashi, D,Endoh, Y.Kon, T.Yamashita, F.Sato, et al.: "Hypersensitivity of LEC strain rats in radiation-induced acute bone-marrow death" J Vet Med Sci.55 (1). 13-8 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, D.Endoh, A.Fukamizu, K.Murakami, Y.Hashimoto, et al.: "A simple method of hybridohistochemistry for detection of renin mRNA in the mouse kidney" J Vet Med Sci.55 (3). 461-3 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, Y.Hashimoto, M.Sugimura, K.Murakami: "Adrenal renin is localized in lysosomal granules" Anat Histol Embryol.22 (4). 324-7 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Mizutani, M.Hayashi, A.Maeda, N.Sasaki, T.Yamashita, et al.: "Inhibition of mouse hepatitis virus multiplication by antisense oligonucleotide, antisense RNA,sense RNA and ribozyme" Adv Exp Med Biol.342. 129-35 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] N.Sasaki, M.Hayashi, S.Aoyama, T.Yamashita, I.Miyoshi, et al.: "Transgenic mice with antisense RNA against the nucleocapsid protein mRNA of mouse hepatitis virus" J Vet Med Sci. 55 (4). 549-54 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] D.Endoh, M.Niikura, K.Hirai, N.Inagaki, M.Hayashi, et al.: "Expression and purification of recombinant Marek's disease virus serotype 1 specific phosphorylated protein pp38 in E.coli" J.Vet.Med.Sci.56 (5). 823-826 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] N.Hashimoto, D.Endoh, M.Kuwabara, H.Satoh, F.Sato: "Induction of lung tumors in C3H strain mice after single or fractionated irradiation with X-rays" J Vet Med Sci.56 (3). 493-8 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Hayashi, T.Furuichi, S.Ren, E.Isogai, M.Nonoyama, et al.: "Enhancement of mRNA synthesis from Marek's disease virus genome in the lymphoblastoid cell line, MDCC-MSB1, by 5-azacytidine" J Vet Med Sci.56 (2). 287-91 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Hayashi, T.Okui, D.Endoh, F.Sato, N.kasai, et al.: "Radiation hypersensitivity of LEC strain rats controlled by a single autosomal recessive gene" Mutat Res.314 (2). 135-42 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, D.Alcorn, K.Murakami, M.Sugimura, G.B.Ryan: "Immunohistochemical studies of renin-containing cells in the developing sheep kidney" Anat Rec.239 (2). 191-7 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Mizutani, M.hayashi, A.Maede, K.Ishida, T.Watanabe, et al.: "The inhibitory effects of MgSO4 on the multiplication and transcription of mouse hepatitis virus" Jpn J Vet Res.42 (2). 95-102 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Mizutani, A.Maeda, M.Hayashi, H.Isogai, S.Namioka: "Both antisense and sense RNAs against the nucleocapsid protein gene inhibit the multiplication of mouse hepatitis virus" J Vet Med Sci.56 (2). 211-5 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Okui, M.Hayashi, D.Endoh, F.Sato, N.Kasai, et al.: "A high frequency of induction of chromosome aberrations in the bone marrow cells of LEC strain rats by X-irradiation" Mutat Res.324 (4). 165-9 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kon, D.Endoh, K.Murakami, T.Yamashita, T.Watanabe, et al.: "Expression of renin in coagulating glands is regulated by testosterone" Anatomical Rec.241 (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Okai et at: "A high freguescy of induction of chromosome aberrations in bam massous alls" Mutation Res. 324. 165-169 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] D.Eudoh et al: "Expression and purification of recombinant MDU pp38in E.col" J.Vet.Med.Sci. 56. 823-826 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Mizutani et al.: "Inhibition of MHU multipli cation by digomueleotion" J.Vet.Med.Sci. 56. 211-215 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Kon et al.: "Expression of Remiu in coaylating Glands is regulated by Iestosterons" The Anat.Rec.241(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kon,Y.: "A simple method of hybridohistochemistry for detection of renin mRNA in the mouse kidney." J.Vet.Med.Sci.55. 461-463 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hayashi,M.: "Higher sensitivity of LEC strain rats in radiation-induced acute bone-marrow death." J.Vet.Med.Sci.55. 13-18 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kon-Ogura,Y.: "Distribution of T cell subsets in the chicken lymphoid tissues." J.Vet.Med.Sci.55. 59-66 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Sasaki,N.: "Transgenic mouse with antisense RNA against nucleocapsid protein mRNA of mouse hepatitis virus." J.Vet.Med.Sci.55. 549-554 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kon,Y.: "Adrenal renin is localized in lysosomal granules." Anat.Histol.Embryol.22. 324-327 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Mizutani,T.: "Both antisense and sense RNA against nucleocapsid protein gene ingibit the multiplication of mouse hepatitis virus." J.Vet.Med.Sci.56(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kon,Y.: "Immunohistochemical studies of renin-containing cells in the developing sheep kidney." Anat.Rec.(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hayashi,M.: "Enhancement of mRNA synthesis from Marek's disease virus genome in lymphoblastoid cell line,MDCC-MSB1,by 5-aza-cytidine." J.Vet.Med.Sci.56(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yasuhiro Kon: "A Simple Method of Hybridohistochemistry for detection of renin mRNA in the mouse kidney" J.Uet.Med.Sci. (1993)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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