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A novel assessment technique of reproductive and genetic toxicity in domestic animals using a nuclear magnetic resonance instrument

Research Project

Project/Area Number 06556048
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field Applied animal science
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

MIYAMOTO Hajime  Kyoto University, Department of Animal Sciences, Professor, 農学部, 教授 (00026618)

Co-Investigator(Kenkyū-buntansha) AZUMA Yasuyoshi  Research Institute of Environmental Sciences, Environmantal Toxicology Unit, Res, 環境毒生部, 研究員
IRIE Masakazu  Osaka Agricultural Research Center, Livestock Divislon, Researcher, 畜産部, 主任研究員
ISHII Takashi  Kyoto University, Department of Animal Sciences, Assistant, 農学部, 助手 (70111945)
MANABE Noboru  Kyoto University, Department of Animal Sciences, Associate Professor, 農学部, 助教授 (80243070)
葛西 孫三郎  高地大学, 農学部, 教授 (60152617)
丹羽 晧二  岡山大学, 農学部, 教授 (40089115)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥9,400,000 (Direct Cost: ¥9,400,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1994: ¥4,600,000 (Direct Cost: ¥4,600,000)
Keywordsnuclear magnetic resonance / environmental toxicants / reproductive and genetic toxicity / farm animals / embryonic teratogeneit / teratogenic substance / scanning laser microscopy / retinoic acids / 核磁気共鳴装置 / 卵胞細胞の生理特性 / 核磁気イメージング / 胎児の奇形 / エネルギー代謝 / リン酸代謝機能
Research Abstract

In vivo and in vitro phosphorus-31 and proton nuclear magnetic resonance (NMR) spectra of the ovary, liver, skeletal muscle and embryo of experimental and farm animals were acquired using a house-made surface coil, conventional round coil, and/or imaging coils. Beta-ATP/inorganic phosphate ratio which represents the high energy state and intracellular pH etc.were observed in vivo. Many environmental toxicants were identified and quantified without any extraction processes from these organs and embryos using a NMR spectroscopy. Proton NMR experiments were done by a modification of conventional techniques, and 31P NMR experiments were performed as follows : During the NMR examination, the sample was maintained in a probe chamber at controlled air temperature of 25. C.In.vivo 31P NMR spectra of the steady-state organs within intact animals were obtained using a JNM-alpha-400 FT-NMR spectromenter (JEOL,Tokyo, Japan) equipped with 89-mm bore super-conducting magnet (9.20 Tesla ; Oxford Inst … More ruments, Bedford, MA,USA). The animal was mounted on a surface coil probe (two-turns, 2.0cm diameter) with was tuned to 31P at 161.7MHz and used in conjunction with a ferrite screen composed of strips of computer tape (Scotch 700 ; Three M Co., St.Louis, MO) to eliminate 31P NMR signals arising from superficial muscles (7). Field homogeneity was controlled by adjusting shim parameters using a water 1H signal derived from the organ sample. Acquisition conditions were set as follows to obtain fully relaxd signals : the pulse width was 20 micro-seconds, the pulse repetition time was 3.0s, and 200 scans were accumulated. Thus, it took 5-25 min for each measurement. After exponential multiplication, the accumulated free induction decays were subjected to Fourier transformation. Individual spectral peak areas were calculated by computer integration with JEOL software (JEOL), and the creatine phosphate (CP) peak was used as a positional reference (0 ppm). The relative amount of each compound was converted to absolute concentration by comparison with the data of an external reference (20mM methylene diphosphonate in water) in a spherical bulb. The present study showed that NMR spectroscopy is a novel assessment tools for assessment of reproductive and genetic toxicity in experimental and domestic animals. Less

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (34 results)

All Other

All Publications (34 results)

  • [Publications] 眞鍋昇ら: "Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in the pig ovaries" Experientia. 52. 647-651 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 杉本実紀ら: "Follicle survival in neonatal rat ovaries cryopreserved by vitrification." Cryo Letter. 17. 93-98 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 眞鍋昇ら: "Ca2+/Mg2+-Dependent endonuclease but not Ca2+-Dependent, Mg2+-Dependent or cation-independent endonuclease is involved in granulosa cell apoptosis of pig atretic follicles." Journal of Reproduction and Development. 42. 247-253 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 眞鍋昇ら: "Effect of the mycelial extract of cultured cordyceps sinensis on in vivo hepatic energy metabolism in the mouse." Journal of Pharmacology. 70. 85-88 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 森田眞紀ら: "Urtrastructural features pf secretion by murine oviductal epithelium." Journal of Mammalian Ova Research. 14. 1-7 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 奥田智彦ら: "Retinoic acid induces malformations related to cell death in the developing mouse embryo." Journal of Reproduction and Development. 43. 43-54 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 森田眞紀ら: "Histochemistry and Cytochemistry : Changes in cell proliferation and morphology in murine oviductal epithelial during the estrous cycle." The Japan Society of Histochemistry and Cytochemistry Nakanishi Pr. Co., Kyoto, Japan, 1122 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 眞鍋昇ら: "Histochemistry and Cytochemistry : Apoptosis occurs in granulosa cells but not cumulus cells in the atretic Graafian follicles in the porcine ovaries." The Japan Society of Histochemistry and Cytochemistry Nakanishi Pr. Co., Kyoto, Japan, 1122 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Noboru Manabe, Yuzuru Imai, Hiroko Ohno, Yhoichi Takahagi, Miki Sugimoto, and Hajime Miyamoto: "Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in the pig ovaries" Experientia. 52. 647-651 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Miki Sugimoto, Hajime Miyamoto, Masashi Kabasawa, and Noboru Manabe: "Follicle survival in neonatal rat ovaries cryopreserved by vitrification" Cryo Letter. 17. 93-98 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Noboru Manabe, Yuzuru Imai, Yoshihiro Kimura, Akira Myoumoto, Miki Sugimoto, Hajime Miyamoto, Yoshinori Okamura, and Manabu Fukumoto: "Ca2+/Mg2+-dependent endonuclease but not ca2+Dependent, mg2+-Dependent or cation-independent endonuclease is involved in granulosa cell apoptosis of pig atretic follicles" Journal of Reproduction and Development. 42. 247-253 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Noboru Manabe, Miki Sugimoto, Yasuyoshi Azuma, Naoki Taketomo, Akio Yamashita, Hiroshi Tsuboi, Akinobu Tsunoo, Noriko Kinjo, Huang Nian-Lai, and Hajime Miyamoto: "Effect of the mycelial extract of cultured cordyceps sinensis on in vivo hepatic energy metabolism in the mouse" Journal of Pharmacology. 70. 85-88 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Maki Morita, Miki Sugimoto, Hajime Miyamoto and Noboru Manabe: "Ultrastructural features of secretion by murine oviductal epithelium" Journal of Mammalian Ova Research. 14. 1-7 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tomohiko Okuda, Hiroaki Takakuwa, Makiko Mikami, Hajime Miyamoto, and Noboru Manabe: "Retinoic acid induces malformations related to cell deathin the developing mouse embryo" Journal of Reproduction and Development. 43. 43-54 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Maki Morita, Miki Sugimoto, Noboru Manabe and Hajime Miyamoto: Histochemistry and Cytochemistry : Changesin cell proliferation and morphology in murine oviductal epithelial during the estrous cycle. The Japan Society of Histochemistry and Cytochemistry.Nakanishi Pr.Co., Kyoto, Japan, 1122 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Noboru Manabe, Yuzuru Imai, Akira Myomoto, Yoshihiro Kimura and Hajime Miyamoto: Histochemistry and Cytochemistry : Apoptosis occurs in granulosa cells but not cumulus cells in the atretic Graafian follicles in the porcine ovaries. The Japan Society of Histochemistry and Cytochemistry Nakanishi Pr.Co., Kyoto, Japan, 1122 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 眞鍋昇ら: "Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in the pig ovaries" Experientia. 52. 647-651 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 杉本実紀ら: "Follicle survival in neonatal rat ovaries cryopreserved by vitrification." Cryo Letter. 17. 93-98 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 眞鍋昇ら: "Ca2+/Mg2+-Dependent endonuclease but not Ca2+-Dependent,Mg2+-Dependent or cation-independent endonuclease is involved in granulosa cell apoptosis of pig atretic follicles." Journal of Reproduction and Development.42. 247-253 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 眞鍋昇ら: "Effect of the mycelial extract of cultured cordyceps sinensis on in vivo hepatic energy metabolism in the mouse." Journal of Pharmacology. 70. 85-88 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 森田眞紀ら: "Ultrastructural features of secretion by murine oviductal epithelium." Journal of Mammalian Ova Research. 14. 1-7 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 奥田智彦ら: "Retinoic acid induces malformations related to cell deathin the developing mouse embryo." Journal of Reproduction and Development. 43. 43-54 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 森田眞紀ら: "Histochemistry and Cytochemistry : Changes in cell proliferation and morphology in murine oviductal epithelial during the estrous cycle." The Japan Society of Histochemistry and Cytochemistry Nakanishi Pr.Co.,Kyoto,Japan, 1122 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 眞鍋昇ら: "Histochemistry and Cytochemistry : Apoptosis occurs in granulosa cells but not cumulus cells in the atretic Graafian follicles in the porcine ovaries." The Japan Society of Histochemistry and Cytochemistry Nakanishi Pr.Co.,Kyoto,Japan, 1122 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Miyamoto H., et al.: "Female reproductive properties and prenatal development of a senescence accelerated mouse strain" The Journal of Experimental Zoology. 272. 116-122 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 宮本 元ら: "潅流培養したヤギ卵巣の組織化学的研究" 日本胚移植学会雑誌. 17. 81-87 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Manabe N. et al.: "Immunohistochemical quantitation for extracellular matrix proteins in rats with glomerulonephritis" Nephron. 71. 79-86 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Manabe N. et al.: "Immunohistochemical micro quantification of fast-myosin in sections of mammalian skeletal muscles" Journal of Animal Science. 73. 88-95 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 眞鍋 昇、宮本 元: "卵胞退行における顆粒層細胞のアポトーシスの役割とその制御" 家畜人工受精誌. 167. 1-10 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang W. H. et al.: "Functional analysis using chlortetracycline fluorescence and in vitro fertilization of frozen-thawed ejaculated bore spermatozoa incubated in a protein-free chemically defined medium" The Journal of Reproduction and Fertility. 17. 81-87 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Magosaburo kasai: "Cryopreservation of mammalian embryos, Vitrification In: Methods in Molecular Biology, Vol. 38, Chapter 21" Humana Press Inc., Totowa, NJ, USA, 769 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Zhu,S.E.: "Optimization of the procedures fro the vitrification of expended mouse blastocysts in glycerol-based solutions" Journal of Peproduction and Development. 40. 293-300 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Lin,J.M.: "Effects of fetal calf serum in culturemedium on development of bovine oocytes matured and fertilized in vitro." Theriogenology. 41. 1091-1098 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Manabe,N.: "Physiomorphological changes in mouse embryos induced by retinoic acid evaluate NMR microscopy" Journal of Veterinary Medical Sucience. 〔発表予定〕.

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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