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Studies on the establishment of the analytical method for, and the distribution and change of the total D-amino acids in mammalian body

Research Project

Project/Area Number 08457592
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionThe University of Tokyo

Principal Investigator

IMAI Kazuhiro  The Univ.of Tokyo, Grad.School of Pharm.Sci., Professor, 薬学系研究科, 教授 (50012620)

Co-Investigator(Kenkyū-buntansha) FUKUSHIMA Takashi  The Univ.of Tokyo, Grad.School of Pharm.Sci., Research Associate, 薬学系研究科, 助手 (00272485)
SANTA Tomofumi  The Univ.of Tokyo, Grad.School of Pharm.Sci., Research Associate, 薬学系研究科, 助手 (30187306)
HOMMA Hiroshi  The Univ.of Tokyo, Grad.School of Pharm.Sci., Associate Professor, 薬学系研究科, 助教授 (50190278)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥5,200,000 (Direct Cost: ¥5,200,000)
KeywordsD-aspartic acid / pineal gland / NBD-F / HPLC / chiral stationary phase / anti D-aspartate antibody / マイクロダイアリシス / HPLCキラル固定相
Research Abstract

During the course of our studies on the determination of D-amino acids in mammals, we found that a large amount (mMorder) of D-aspartic acid (D-Asp) is present in the rat pineal gland. The method we usedisa chiral stationary phase-HPLC separation adn fluorometoric detection of NBD-labelled amino acids. In the present study, to increase the efficiency of the enantioneric separation, we developed 3 new chiral stationary phase containing the same chiral moieties as the previous phases with an extended long alkyl chain from the aminopropylsilica support. One of them, APS-11-OA-2500S was the best for the enantiomeric separation of NBD-amino acids, amino acids ester and beta-amino butyricacid. The interaction between the analyte and the chiral moiety was also studied with NMR,showing some interaction of proton of amino moiety of amino acids and carbonyl moiety of the chiral stationary phase.
The whole body autoradiography of rat injected with ^<14>C labelled D-Asp showed the incorpration of radioactivity to the tissues where the endogenous D-Asp existed.
The polyclonal antibody to D-Asp revealed the localization of the endogenous D-Asp in the cytoplasm of pinealocyte which locate in the distal region of the gland. The specifid location of D-Asp was also observed in the pituitary gland, suggesting the exsitence of some important physiological roles of D-Asp in the mammaliam body.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] M.Tsunoda, et al.: "Enantiomeric Separation of NBD-Amino Acids by Capillary Zone Electrophoresis Using Cyclodextrins as Chiral Selectors." Chromatogr.18. 21-25 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.A.Lee, et al.: "Immunohistochemical Localization of D-Aspartate in the Rat Pineal Gland." Biochem.Biophys.Res.Comm.231. 505-508 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Imai, et al.: "Accumulation of Radioactivity in Rat Brain and Peripheral Tissues Including Salivary Gland after Intravenous Administration of ^<14>C-D-Aspartic Acid." Proc.Japan Acad.73,Ser.B. 48-52 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Hamase, et al.: "Regional distribution and postnatal changes of D-amino acids in rat brain." Biochim.Biophys.Acta. 1334. 214-222 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 今井 一洋, 他: "D-アミノ酸研究の最近の進歩" 薬学雑誌. 117. 637-646 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tsunode, M.Kato, T.Fukushima, T.Santa, H.Homma, K.Imai: "Enantiomeric Separation of NBD-Amino Acids by Capillary Zone Electrophoresis Using Cyclodextrins as Chiral Selectors." Chromatogr.18. 21-25 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.A.Lee, H.Homma, K.Sakai, T.Fukushima, T.Santa, K.Tashiro, T.Iwatsubo, M.Yoshikawa, K.Imai: "Immunohistochemical Localization of D-Aspartate in the Rat Pineal Gland." Biochem.Biophys.Res.Commun.231. 505-508 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Imai, T.Fukushima, T.Santa, H.Hommma, J.Sugihara, H.Kodama, M.Yoshikawa: "Accumulation of Radioactivity in Rat Brain and Peripheral Tissues Including Salivary Gland after Intravenous Administration of ^<14>C-D-Aspartic Acid." Proc.Japan Acad.73, Ser.B. 48-52 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Hamase, H.Homma, Y.Takigawa, T.Fukushima, T.Santa, K.Imai: "Regional distribution and postnatal changes of D-amiono acids in rat brain." Biochim.Biophys.Acta.1334. 214-222 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tsunoda, et al.: "Enantiomeric Separation of NBD-Amino Acids by Capillary Zone Electrophoresis Using Cyclodextrins as Chiral Selectors." Chromatogr.18. 21-25 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] J.A.Lee, et al.: "Immunohistochemical Localization of D-Aspartate in the Rat Pineal Gland." Biochem.Biophys.Res.Comm.231. 505-508 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Imai, et al.: "Accumulation of Radioactivity in Rat Brain and Peripheral Tissues Including Salivary Gland after Intravenous Administration of ^<14>C-D-Aspartic Acid." Proc.Japan Acad.73,Ser.B. 48-52 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Hamase, et al.: "Regional distribution and postnatal changes of D-amino acids in rat brain." Biochim.Biophys.Acta. 1334. 214-222 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 今井一洋、他: "D-アミノ酸研究の最近の進歩" 薬学雑誌. 117. 637-646 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Matsunaga,H.et al.: "Boron Trifluoride-Etherate(Lewis Acid)as an Efficient Acid at Cyclization/Cleavage Reaction of D/L-Amino Acid Affording the Retention of their Original Configuration" Biomed.Chromatogr.10. 95-96 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Matsunaga,H.et al.: "A Major Factor for the Racemization and the Dehydration at the Cyclization/Cleavage Stage in the Edman Sequencing Method." Anal.Chem.68. 2850-2856 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nakashima,K.& Imai,K.: "LC-CHEMILUMINESCENCE DETECTION" Methods in Chromatography. 1. 99-122 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Imai,K.et al.: "Analytical Chemistry and Biochemistry of D-Amino Acids" Biomed.Chromatogr.10. 303-312 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 今井一洋: "D-アミノ酸と疾患" ファルマシア. 32. 1214-1218 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hamase,K.et al: "Regional distribution and postnatal changes of D-amino acids in rat brain" Biochim.Biophys.Acta. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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