Pineal-specific expression of green fluorescent protein under the control of the serotonin-N-acetyltransferase gene regulatory regions in transgenic zebrafish

Gothilf, Y., Toyama, R., Coon, S.L., Du, S.-J., Dawid, I.B., and Klein, D.C.
Developmental Dynamics : an official publication of the American Association of Anatomists   225(3): 241-249 (Journal)
Registered Authors
Dawid, Igor B., Du, Shao Jun (Jim), Gothilf, Yoav, Toyama, Reiko
pineal gland; zebrafish embryo; serotonin-N-acetyltransferase; transgenic fish; green fluorescent protein; mind-bomb; floating head
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Arylamine N-Acetyltransferase/genetics*
  • Base Sequence
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter/genetics
  • Green Fluorescent Proteins
  • Indicators and Reagents/metabolism
  • Luminescent Proteins/genetics
  • Molecular Sequence Data
  • Mutation/genetics
  • Pineal Gland/embryology*
  • Pineal Gland/physiology*
  • Promoter Regions, Genetic/genetics
  • Zebrafish/genetics*
12412006 Full text @ Dev. Dyn.
Zebrafish serotonin-N-acetyltransferase-2 (zfAANAT-2) mRNA is exclusively expressed in the pineal gland (epiphysis) at the embryonic stage. Here, we have initiated an effort to study the mechanisms underlying tissue-specific expression of this gene. DNA constructs were prepared in which green fluorescent protein (GFP) is driven by regulatory regions of the zfAANAT-2 gene. In vivo transient expression analysis in zebrafish embryos indicated that in addition to the 5'- flanking region, a regulatory sequence in the 3'-flanking region is required for pineal-specific expression. This finding led to an effort to produce transgenic lines expressing GFP under the control of the 5' and 3' regulatory regions of the zfAANAT-2 gene. Embryos transiently expressing GFP were raised to maturity and tested for germ cell transmission of the transgene. Three transgenic lines were produced in which GFP fluorescence in the pineal was detected starting 1 to 2 days after fertilization. One line was crossed with mindbomb and floating head mutants that cause abnormal development of the pineal and an elevation or reduction of zfAANAT-2 mRNA levels, respectively. Homozygous mutant transgenic embryos exhibited similar effects on GFP expression in the pineal gland. These observations indicate that the transgenic lines described here will be useful in studying the development of the pineal gland and the mechanisms that determine pineal -specific gene expression in the zebrafish.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes