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Expression of exogenous genes in regenerating newt retina

Research Project

Project/Area Number 12640661
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 動物生理・代謝
Research InstitutionOsaka University

Principal Investigator

HISATOMI Osamu  Osaka Univ., Grad. Sch. of Sci., Assoc. Prof., 大学院・理学研究科, 助教授 (60231544)

Co-Investigator(Kenkyū-buntansha) TOKUNAGA Fumio  Osaka Univ., Grad. Sch. of Sci., Prof., 大学院・理学研究科, 教授 (80025452)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordsretina / regeneration / expression / newt / EST / visual pigment / GFP / 視物質
Research Abstract

In this project, we attempted to express exogeneous genes and knock down endogeneous genes with antisence oligonucleotides in newt retinas. Our results can be summarized as follows.
An expression vector of EGFP gene under the control of a CMV promotor was introduced with cation liposome in viteras of adult newt eyes. After three days from the operation, retinal sections were examined with a fluorescent microscope, but no fluorescence from EGFP was detected. Then we attempted to detect the EGFP by immunohistochemical investigation using anti-GFP antibodies. Antibodies were recognized ganglyon cell layer, inner plexiform layer and inner nuclear layer of the retinas suggesting that EGFP genes were expressed in the newt retinas. However; the amount of EGFP expressed in the retina is likely limited. So, we are now investigating the best condition for gene expression such as expression vectors, lipofection reagents, and the amount of introduced vectors.
In order to knock down a gene expression … More with anti-sense oligonucleotides, accurate nucleotide sequence of each newt gene should be clarified. We isolated mRNAs from adult normal newt retinas and regenerating newt retinas after 18-19 days from the surgical removal of normal retinas. Two kinds of cDNA libraries, normal and regenerating retinal cDNA libraries, were constructed. Several hundred clones were randomly isolated from these libraries, and nucleotide sequences were determined. Homology of these sequences to the sequences deposited in the databases, and function of each gene was elucidated. We found cDNAs encoding four kinds of opsins (protein moieties of visual pigments), OTX2 (a homeodomain protein responcible retinal development), and stathmin (a protein responcible for axonal regeneration). Expression of these genes were investigated by in situ hybridization. It is clarified that rod cells begin to express opsin before outer segments were formed, and that otx2 and stathmin genes are strongly expressed in regenerating newt retinas. Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (42 results)

All Other

All Publications (42 results)

  • [Publications] Hisatomi, O.: "Molecular evolution of proteins involved in vertebrate"Comparative Biochemistry Physiology. 133. 509-522 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakakibara, S.: "Opsin expression in adult, developing, and regenerating newt retinas"Molecular Brain Research. 103. 28-35 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kobayashi, Y.: "Distribution of rod-and cone-specific phosducins in retinas of non-mammalian vertebrates"Comparative Biochemistry Physiology. 133B. 77-83 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Konkawa, H.: "Distribution of blue-sensitive photoreceptors in amphibian retinas"Interdisciplinary Information Science. 8. 25-32 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O.: "Evolution of Visual Pigments and Related Molecules"Journal of Photoscience. 9. 41-43 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O.: "EST analysis of regenerating newt retina"Journal of Photoscience. 9. 267-268 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 久富 修: "視細胞分化における視物質の転写調節"Molecular Medicine. 39巻別冊. 19-27 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Taniguchi, Y.: "Pinopsin expressed in retinal photoreceptors of a diurnal gecko"FEBS Letters. 496. 69-74 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshikawa, T.: "Immunohistochemical localization of opsins and alpha-subunit of transducins in the pineal complex and deep brain of the Japanese grass lizard, Takydromus tachydromoides"Zoological Science. 18. 325-330 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takahashi, Y.: "Distribution of blue-sensitive photoreceptors in amphibian retinas"FEBS Letters. 501. 151-155 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Imamoto, Y.: "Roles of Amino Acid Residues near the Chromophore of Photoactive Yellow Protein"Biochemistry. 40. 4679-4685 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kobayashi, Y.: "Identification of phosducins expressed in teleost retinas"FEBS Letters. 502. 117-121 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakakibara, S., Hiramatsu, H., Takahashi, Y., Hisatomi, O., Kobayashi, Y., Sakami, S., Saito, T. and Tokunaga, F.: "Opsin expression in adult, developing, and regenerating newt retinas"Mol. Brain Res.. 103. 28-35 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Honkawa, H., Hisatomi, O., Kishida, Y. and Tokunaga, F.: "Two kinds of guanylate cyclase activating proteins in medaka retina"Interdisciplinary Info. Sci.. 8. 25-32 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kobayashi, Y., Hisatomi, O., Yamamoto, S. and Tokunaga, F.: "Distribution of rod- and cone-specific phosducins in retinas of non-mammalian vertebrates"Comp. Biochem. Physiol.. 133B. 77-83 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O. and Tokunaga, F.: "Molecular evolution of proteins involved in vertebrate phototransduction"Comp. Biochem. Physiol.. 133. 509-522 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O., Yamamoto, S., Kobayashi, Y., Honkawa, H., Takahashi, Y. and Tokunaga, F.: "Evolution of Visual Pigments and Related Molecules"J. Photosci.. 9. 41-43 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O., Hasegawa, A., Goto, T., Yamamoto, S., Sakami, S., Kobayashi, Y. and Tokunaga, F.: "EST analysis of regenerating newt retina"J. Photosci.. 9. 267-268 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O. and Tokunaga, F.: "Functional differentiation of vertebrate photoreceptors and regulation of opsin expression (in Japanese)"Molecular Medicine. 39. 19-27 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kobayashi, Y., Hisatomi, O., Satoh, T. and Tokunaga, F.: "Identification of phosducins expressed in teleost retinas"FEBS Lett.. 502. 117-121 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takahashi, Y., Hisatomi, O., Sakakibara, S., Tokunaga, F. and Tsukahara, Y.: "Distribution of blue-sensitive photoreceptors in amphibian retinas"FEBS Lett.. 501. 151-155 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Taniguchi, Y., Hisatomi, O., Yoshida, M. and Tokunaga, F.: "Pinopsin expressed in retinal photoreceptors of a diurnal gecko"FEBS Lett.. 496. 69-74 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Imamoto, Y., Koshimizu, H., Mihara, K., Hisatomi O., Mizukami, T., Tsujimoto, K., Kataoka, M., Tokunaga, F.: "Roles bf Amino Acid Residues near the Chromophore of Photoactive Yellow Protein"Biochemistry. 40. 4679-4685 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshikawa, T., Okano, T., Kokame, K., Hisatomi, O., Tokunaga, F., Oishi, T. and Fukada, Y.: "Immunohistochemical localization of opsins and alpha-subunit of transducins in the pineal complex and deep brain of the Japanese grass lizard, Takydromus taclrydromoides"Zool. Sci.. 18. 325-330 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O. and Tokunaga, F.: "Molecular basis of teleost vision (in Japanese)"Tanpakushitsu Kakusan Koso. 45. 2924-2930 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hisatomi, O.: "Molecular evolution of proteins involved in vertebrate"Comparative Biochemistry Physiology. 133・4. 509-522 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakakibara, S.: "Opsin expression in adult, developing, and regenerating newt retinas"Molecular Brain Research. 103・1-2. 28-35 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kobayashi, Y.: "Distribution of rod-and cone-specific phosducins in retinas of non-mammalian vertebrates"Comparative Biochemistry Physiology. 133B. 77-83 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Honkawa, H.: "Distribution of blue-sensitive photoreceptors in amphibian retinas"Interdisciplinary Information Science. 8・1. 25-32 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hisatomi, O.: "Evolution of Visual Pigments and Related Molecules"Journal of Photoscience. 9・2. 41-43 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hisatomi, O.: "EST analysis of regenerating newt retina"Journal of Photoscience. 9・2. 267-268 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Taniguchi, Y.: "Pinopsin expressed in retinal photoreceptors of a diurnal gecko"FEBS Letters. 496・2/3. 69-74 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yoshikawa, T.: "Immunohistochemical localization of opsins and alpha-subunit of transducins in the pineal complex and deep brain of the Japanese grass lizard, Takydromus tachydromoides"Zoological Science. 18・3. 325-330 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Imamoto, Y.: "Roles of Amino Acid Residues near the Chromophore of Photoactive Yellow Protein"Biochemistry. 40・15. 4679-4685 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takahashi, Y.: "Distribution of blue-sensitive photoreceptors in amphibian retinas"FEBS Letters. 501・2/3. 151-155 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kobayashi, Y.: "Identification of phosducins expressed in teleost retinas"FEBS Letters. 502・3. 117-121 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 久冨 修: "視細胞分化における視物質の転写調節"Molecular Medicine. 39巻別冊. 19-27 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Honkawa, H.: "Two kinds of guanylate cyclase activating proteins in medaka retina"Interdisciplinary Information Science. (印刷中).

    • Related Report
      2001 Annual Research Report
  • [Publications] Sakakibara, S.: "Opsin expression in intact and regenerating newt retinas"Molecular Brain Research. (印刷中).

    • Related Report
      2001 Annual Research Report
  • [Publications] 久冨修: "魚類の視物質の分子進化"タンパク質核酸酵素(共立出版). 45・17. 2924-2930 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yoshikawa,T.: "Immunohistochemical localization of opsins and alpha-subunit of transducins in the pineal complex and deep brain of the Japanese grass lizard, Takydromus tachydromoides"Zoological Science. (印刷中).

    • Related Report
      2000 Annual Research Report
  • [Publications] Imamoto,Y.,: "Roles of Amino Acid Residues near the Chromophore of Photoactive Yellow Protein."Biochemistry. (印刷中).

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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