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AN APPLICATION OF IN SITU HYBRIDIZATION AND ANTISENSE OLIGONUCLEOTIDE METHOD TO VISUALIZE AND MANIPULATE GENE EXPRESSION OF STEROID HORMONE RECEPTOR PROTEINS IN THE NERVOUS SYSTEM

Research Project

Project/Area Number 05454674
Research Category

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

Allocation TypeSingle-year Grants
Research Field Neuroscience in general
Research InstitutionKYOTO PREFECTURAL UNIVERSITY OF MEDICINE

Principal Investigator

KAWATA Mitsuhiro  KYOTO PREFECTURAL UNIVERSITY OF MEDICINE DEPARTMENT OF ANATOMY,PROFESSOR, 医学部, 教授 (60112512)

Co-Investigator(Kenkyū-buntansha) MORITA Noriyuki  KYOTO PREFECTURAL UNIVERSITY OF MEDICINE DEPARTMENT OF ANATOMY,ASSISTANT, 医学部, 助手 (50239662)
YURI Kazunari  KYOTO PREFECTURAL UNIVERSITY OF MEDICINE DEPARTMENT OF ANATOMY,ASSISTANT PROFESS, 医学部, 講師 (10220534)
UEDA Shuichi  KYOTO PREFECTURAL UNIVERSITY OF MEDICINE DEPARTMENT OF ANATOMY,ASSOCIATE PROFESS, 医学部, 助教授 (60150570)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1994: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥4,500,000 (Direct Cost: ¥4,500,000)
KeywordsAntisence / Gene manipulation / In situ hybridization / Immunohistochemistry / Steroid hormones / Steroid hormone receptor / Sexual difference / Hippocampus / 免疫組織細胞化学法 / 脳分化 / 細胞培養 / 視床下部視索前野 / 副腎髄質細胞 / 突起伸長 / 受容体 / 組織化学
Research Abstract

Chromaffin cells were found to contain glucocorticoid receptor-immunoreactivity in their nucleus. When dexamethasone was added into the culture medium, the extension of the processes, intensity of immunoreactivity of MAP-2 were significantly diminished. These results indicated that the differentiation of chromaffin cells to neurons is determined, at least in part, by humoral factor of glucocorticoid through its receptor. The addition of antisense oligos for glucocorticoid receptor cleary caused the significant extension of processes of chromaffin cells, indicating that the protein synthesis of steroid receptor was blocked by the antisense inhibitory action to its mRNA.
In the sexually dimorphic area of the hypothalamus, immunocytochemical studies show that estrogen induces a significant changes of numbers of immunoreactive never fibers containing neurotensin, calcitonin-gene related peptide (CGRP), Met-enkephalin, cholecystokinin-8 (CCK-8) and dopamine, indicating that axonal sprouting … More of neurons containing these chemical messengers occur in response to estrogen. The relationship between estrogen-receptor and gene expression of these biochemical messengers in concomitant with the process formation or elongation has been discussed, particularly in the context of male and female differences. Estrogen-receptor was found to localize in the cell nucleus by immunocytochemistry in conjunction with the use of estrogen-receptor antibody. Colchicine treatment blocked immunpositive reaction in the cell nucleus, suggesting that estrogen-receptor protein was synthesized in the cytoplasm followed by the transport to the nucleus through nuclear pore in association with microtubules. Monolayr cultures from male and female rat hypothalami were developed and the differences of neurite extension was investigated. In this point, it should be noted that developmental and estrogen-induced changes in mRNAs coding specific proteins for the cytoskeleton, including tubulin-like molecules, have been indentified in the hypothalamic preoptic area. Less

Report

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

    (34 results)

All Other

All Publications (34 results)

  • [Publications] Kawata,M.,Yuri,K.,Morita,N.,and Morimoto,M.: "Steroid hormone effect on the neuronal configuration-in vivo and in vitro studies." Neuronal Cytoskeleton. 193-203 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kawata,M: "An application of in situ hybridization and antisense oligonucleotidele method to visualize and mainpulate gene expression of specific proteins in the nervous system." Chin.J.Histochem.Cytochem.2. 69-70 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yuri,K.,Kawata,M.: "Region-specific changes of tyrosine hydroxylase-immunoreactivity by estrogen treatment in female rat hypothalamus." Brain Res.645. 278-284 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Morimoto,M.,Morita,N.,Kawata,M.: "The effects of NGF and glucocorticoid on the cytological features of rat chromaffin cells in vitro." NeuroReport. 5. 945-956 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kawata,M.,Yuri,K.and Morimoto.M.: "Steroid hormone effects on gene expression,neuronal structure and differentiation." Hormones and Behavior. 28. 477-482 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nishi,M.,Ueda,S.,Morita,N.and Kawata,M.: "Effects of glucocorticoid on neurones of rat dentate gyrus in dissociated culture." NeuroReport. 5. 2446-2448 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hirakawa, M.and Kawata, M.: "Distribution pattern of c-fos expression induced by sciatic nerve sectioning in the rat central nervous system." J.Hirnforsch.34. 433-446 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ueda, S., Matsumoto, Y., Azmitia, E.C., and Kawata, M.: "Intraocular co-grafts of fetal dorsal raphe nucleus and suprachiasmatic nucleus." Brain Res.605. 181-186 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ison, A., Yuri, K., Ueta, Y., Leng, G., Koizumi, Ko., Yamashita, H., and Kawata, M.: "Vasopressin and oxytocin-immunoreactive hypothalamic neurons of inbred polydipsic mice." Brain Res.Bull.31. 405-414 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koizumi, K., Zeballos, M., Kawata, M., Kannan, H., and Yamashita, H.: "The hypothalamic vasopressinergic neurons of inbred polydipsic mouse." Ann.New York Acad.Sci.689. 612-615 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yuri, K.and Kawata, M.: "Time-course analysis of changes in calcitonin gene-related peptide-and methionine-enkephalin-immunoreactivity in the female rat preoptic area after estrogen treatment." Neuroscience. 55. 1067-1074 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kawata, M., Yuri, K., Morita, N., and Morimoto, M.: "Steroid hormone effect on the neuronal configuration-in vivo and in vitro studies." In Neuronal Cytoskeleton (ed.by Hirokawa, N.). JSSP,CRC Press, Tokyo. 193-203 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kawata, M.: "An application of in situ hybridization and antisense oligonucleotide method to visualize and mainpulate gene expression of specific proteins in the nervous system." Chin.J.Histochem.Cytochem.Vol 2 Suppl.69-70 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kawahito, Y., Sano, H., Kawata, M., Yuri, K., Mukai, S., Yamamura, Y., Kato, H.: "Chrousos, G.P., Wilder, R.L.and Kondo, M, : Local secretion of corticotropin-releasing hormone by enterochromaffin cells in human colon." Gastroenterology. 106. 859-865 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ueda, S.and Kawata, M.: "Regeneration of serotonergic immunoreactive fibers in the brain of 5,6-dihydroxytryptamine treated rat." J.Brain Res.35. 159-180 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yuri, K.and Kawata, M.: "Estrogen affects calcitonin gene-related peptide-and methionine-enkephaline-immunoreactive neuron in the female rat preoptic area." Neurosci.Lett.169. 5-8 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yuri, K., Kawata, M.: "Region-specific changes of tyrosine hydroxylase-immunoreactivity by estrogen treatment in female rat hypothalamus." Brain Res.645. 278-284 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Morimoto, M., Morita, N., Kawata, M.: "The effects of NGF and glucocorticoid on the cytological features of rat chromaffin cells in vitro." NeuroReport. 5. 945-956 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kawata, M., Yuri, K.and Morimoto, M.: "Steroid hormone effects on gene expression, neuronal structure and differentiation." Hormones and Behavior. 28. 477-482 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nishi, M., Ueda, S., Morita, N.and Kwata, M.: "Effects of glucocorticoid on neurones of rat dentate gyrus in dissociated culture." NeuroReport. 5. 2446-2448 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ueda, S., Gu, X.F., Whitaker-Azmitia, P.M., Naruse, I., and Azmitia, E.C.: "Neuoro-glial neurotrophic interaction in the S-100b retarded mutant mouse (Polydactyly Nagoya).I.Immunocytochemical and neurochemical studies." Brain Res.633. 275-283 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ueda, S., Hou, X.P., Whitaker-Azmitia, P.M., and Azmitia, E.C.: "Neuoro-glial neurotrophic interaction in the S-100b retarded mutant mouse (Polydactyly Nagoya).II.Co-cultures studies." Brain Res.633. 284-288 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 河田光博、上田陽一: "ラジオアイソ-プ(RI)を用いるin situハイブリダイゼーション(ISH)法" 組織細胞化学1994. 140-143 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yuri,K.and Kawata,M.: "Estrogen affects calcitonin gene-related peptide-and methionine-enkephaline-immunoreactive neuron in the female rat preoptic area." Neurosci.Lett.,. 169. 5-8 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yuri,K.,Kawata,M.: "Region-specific changes of tyrosine hydroxylase-immunoreactivity by estrogen treatment in female rat hypothalamus." Brain Res.645. 278-284 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Morimoto,M.,Morita,N.,Kawata,M.: "The effects of NGF and glucocorticid on the cytological features of rat chromaffin cells in vitro." NeuroReport. 5. 945-956 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kawata,M.,Yuri,K.and morimoto,M: "Steroid hormone effects on geneexpression,neuronal structure and differentiation." Hormones and behavior. 28. 477-482 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Nishi,M.,Ueda,S.,Morita,N and Kawata,M.: "Effects of glucocorticoid on neurones of rat dentate gyrus in dissociated culture." NeuroReport. 5. 2446-2448 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ison,A,.Yuri,K.,et al.: "Vasopressinandoxytocin-immunoreactivehypothalamic---" Brain Res.Bull.31. 405-414 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ueda,S.et al.: "Intraocular co-grafts of fetal dorsal raphenucleusand---" Brain Res.605. 181-186 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yuri,K.and Kawate,M.: "Time-courseanalysis of changes in calcitonin gene-related peptide---" Neuroscience. 55. 1067-1074 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hirakawa,M.and Kawata,M.: "Distribution pattern ofc-fosexpression in duced by sciatic---" J.Hirnforsch.34. 433-446 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 河田光博ら: "下垂体の分子細胞化学" 日本臨床. 51. 2530-2535 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 河田光博ら: "合成DNAによる細胞内mRNAの同定と発現阻止、" 組織細胞化学1993. 76-85 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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