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

Park, H.-C., Kim, C.-H., Bae, Y.-K., Yee, S.-Y., Kim, S.-H., Hong, S.-K., Shin, J., Yoo, K.-W., Hibi, M., Hirano, T., Miki, N., Chitnis, A.B., and Huh, T.-L.
Developmental Biology   227(2): 279-293 (Journal)
Registered Authors
Bae, Young Ki, Chitnis, Ajay, Hibi, Masahiko, Hong, Sung-Kook, Huh, Tae-Lin, Kim, Cheol-Hee, Kim, Seok-Hyung, Miki, Naomasa, Yoo, Kyeong-Won
HuC; elav; zebrafish; neuron; promoter; Myt1; E box; transgenic; mind bomb
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Base Sequence
  • Binding Sites/genetics
  • Cell Differentiation
  • DNA/genetics
  • DNA Primers/genetics
  • ELAV Proteins
  • ELAV-Like Protein 3
  • Gene Expression Regulation, Developmental
  • Green Fluorescent Proteins
  • Luminescent Proteins/genetics
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Mutation
  • Nerve Tissue Proteins/genetics*
  • Neurons/cytology
  • Neurons/metabolism*
  • Promoter Regions, Genetic*
  • Protein Serine-Threonine Kinases/metabolism
  • Protein-Tyrosine Kinases/metabolism
  • RNA-Binding Proteins/genetics*
  • Ribonucleoproteins/genetics
  • Zebrafish/embryology*
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish Proteins*
11071755 Full text @ Dev. Biol.
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.
Genes / Markers
Show all Figures
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes