Analysis of upstream elements in the HuC promoter leads to the establishment of transgenic Zebrafish with fluorescent neurons

Hae Chul Park, Cheol Hee Kim, Young Ki Bae, Sang Yeob Yeo, Seok Hyung Kim, Sung Kook Hong, Jimann Shin, Kyeong Won Yoo, Masahiko Hibi, Toshio Hirano, Naomasa Miki, Ajay B. Chitnis, Tae Lin Huh

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260 Citations (Scopus)

Abstract

HuC encodes an RNA binding protein homologous to Drosophila elav that serves as an excellent early marker for differentiating neurons. We have characterized the promoter of the zebrafish HuC gene by examining the ability of 5'-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos. We determined that 2.8 kb of the 5'-flanking sequence is sufficient to restrict GFP gene expression to neurons. The core promoter spans 251 base pairs and contains a CCAAT box and one SP1 sequence but no TATA box is present near the transcription start site. A putative MyT1 binding site and at least 17 E-box sequences are necessary to maintain the neuronal specificity of HuC expression. Interestingly, sequential removal of the putative MyT1 binding site and 14 distal E boxes does not appear to abolish neuronal expression; rather, it leads to a progressive expansion of GFP expression into muscle Cells. Further removal of the three proximal E boxes eliminates neuronal and muscle specificity of GFP expression and leads to ubiquitous expression of GFP in the whole body. Identification of key components of the HuC promoter has led to the establishment of a stable zebrafish transgenic line (HuC-GFP) in which GFP is expressed specifically in neurons. We crossed mind bomb (mib) fish with this line to visualize their neurogenic phenotype in live mib-/- mutant embryos. This cross illustrates how HuC-GFP fish could be used in the future to identify and analyze zebrafish mutants with an aberrant pattern of early neurons. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)279-293
Number of pages15
JournalDevelopmental Biology
Volume227
Issue number2
DOIs
Publication statusPublished - 2000 Nov 15
Externally publishedYes

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Keywords

  • E box
  • Elav
  • HuC
  • Mind bomb
  • Myt1
  • Neuron
  • Promoter
  • Transgenic
  • Zebrafish

ASJC Scopus subject areas

  • Developmental Biology

Cite this

Park, H. C., Kim, C. H., Bae, Y. K., Yeo, S. Y., Kim, S. H., Hong, S. K., Shin, J., Yoo, K. W., Hibi, M., Hirano, T., Miki, N., Chitnis, A. B., & Huh, T. L. (2000). Analysis of upstream elements in the HuC promoter leads to the establishment of transgenic Zebrafish with fluorescent neurons. Developmental Biology, 227(2), 279-293. https://doi.org/10.1006/dbio.2000.9898