Functional expression of the type 1 inositol 1,4,5-trisphosphate receptor promoter-lacZ fusion genes in transgenic mice

Daisuke Furutama, Kouji Shimoda, Shingo Yoshikawa, Atsushi Miyawaki, Teiichi Furuichi, Katsuhiko Mikoshiba

研究成果: Article査読

33 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)1793-1801
ページ数9
ジャーナルJournal of Neurochemistry
66
5
DOI
出版ステータスPublished - 1996 5

ASJC Scopus subject areas

  • 生化学
  • 細胞および分子神経科学

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