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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 SL, Du SJ, Dawid IB, Klein DC.
Department
of Zoology, George S Wise Faculty of Sciences, Tel Aviv University, Tel
Aviv, Israel.
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. Published 2002
Wiley-Liss, Inc.
PMID: 12412006 [PubMed - indexed for MEDLINE]
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