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Functional Expression of the Type 1 Inositol 1,4,5-Trisphosphate Receptor Promoter-lacZ Fusion Genes in Transgenic Mice

Authors

  • Daisuke Furutama,

    1. Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, and
    2. First Department of Internal Medicine, Osaka Medical College, Osaka; and
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  • Kouji Shimoda,

    1. Laboratory Animal Center, Keio University School of Medicine, Tokyo;
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  • Shingo Yoshikawa,

    1. Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, and
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    • The present address of Dr. S. Yoshikawa is Department of Molecular Neurobiology, Institute of Basic Medical Science, University of Tsukuba, Ibaraki, Japan.

  • Atsushi Miyawaki,

    1. Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, and
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  • Teiichi Furuichi,

    Corresponding author
    1. Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, and
      Address correspondence and reprint requests to Dr. T. Furuichi at Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-Ku, Tokyo 108, Japan.
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  • Katsuhiko Mikoshiba

    1. Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, and
    2. Department of Molecular Neurobiology, Institute of Physical and Chemical Research (RIKEN), Tsukuba Life Science Center, Ibaraki, Japan
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Address correspondence and reprint requests to Dr. T. Furuichi at Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-Ku, Tokyo 108, Japan.

Abstract

Abstract: Inositol 1,4,5-trisphosphate receptor (IP3R) is an inositol 1,4,5-trisphosphate (InsP3)-gated Ca2+ release channel. Type 1 IP3R (IP3R1) is the neuronal member of the IP3R family in the CNS and is predominantly expressed in cerebellar Purkinje cells. To elucidate the molecular mechanisms responsible for coupling gene expression to neuronal InsP3/Ca2+ signaling, we have studied the structure and function of the 5′-flanking region of the mouse IP3R1 gene. The cloned 5′-flanking region has several sequences sharing identity with motifs for known transcriptional regulation. We have fused 5′-flanking regions 1N from −528 to +169 and 4N from −4,187 to +169 to a β-galactosidase gene (lacZ) as a reporter marker and have characterized their in vivo gene expression. Both 1N and 4N fusion genes functioned as a strong promoter in a neuroblastoma-glioma hybrid cell line NG108-15. Moreover, both 1N and 4N transgenic mouse lines carrying these 1N and 4N fusion genes showed characteristic patterns of β-galactosidase activity in the CNS that are almost consistent with that of the endogenous IP3R1 protein, thereby suggesting that the 1N region from −528 to +169 contains sequence elements responsible for regulating gene expression in neurons and for specifying predominant expression in cerebellar Purkinje cells.

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