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

Research output: Contribution to journalArticle

31 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1793-1801
Number of pages9
JournalJournal of Neurochemistry
Volume66
Issue number5
Publication statusPublished - 1996 May 1

Keywords

  • Gene expression
  • Inositol 1,4,5-trisphosphate receptor
  • Promoter
  • Purkinje cell
  • Transgenic mice
  • lacZ

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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    Furutama, D., Shimoda, K., Yoshikawa, S., Miyawaki, A., Furuichi, T., & Mikoshiba, K. (1996). Functional expression of the type 1 inositol 1,4,5-trisphosphate receptor promoter-lacZ fusion genes in transgenic mice. Journal of Neurochemistry, 66(5), 1793-1801.