PUBLICATION
Activator effect of coinjected enhancers on the muscle-specific expression of promoters in zebrafish embryos
- Authors
- Müller, F., Williams, D.W., Kobolak, J., Gauvry, L., Goldspink, G., Orbán, L., and Maclean, N.
- ID
- ZDB-PUB-970812-10
- Date
- 1997
- Source
- Molecular reproduction and development 47(4): 404-412 (Journal)
- Registered Authors
- Kobolak, Julianna, Maclean, Norman, Müller, Ferenc, Orban, Laszlo, Williams, Darren W.
- Keywords
- myosin heavy chain; myosin light chain; skeletal muscle; transgenic; coinjection
- MeSH Terms
-
- Actins/genetics
- Animals
- Embryo, Nonmammalian/metabolism*
- Enhancer Elements, Genetic/genetics*
- Gene Expression Regulation, Developmental*
- Genes, Reporter
- Histocytochemistry
- Lac Operon
- Microinjections
- Muscle, Skeletal/embryology
- Muscle, Skeletal/metabolism*
- Myosin Heavy Chains/genetics
- Myosin Light Chains/genetics
- Promoter Regions, Genetic/genetics*
- Rats
- Zebrafish/embryology
- Zebrafish/genetics
- beta-Galactosidase/genetics
- beta-Galactosidase/metabolism
- PubMed
- 9211424 Full text @ Mol. Reprod. Dev.
Citation
Müller, F., Williams, D.W., Kobolak, J., Gauvry, L., Goldspink, G., Orbán, L., and Maclean, N. (1997) Activator effect of coinjected enhancers on the muscle-specific expression of promoters in zebrafish embryos. Molecular reproduction and development. 47(4):404-412.
Abstract
The transient expression of reporter gene constructs in embryos provides a powerful tool to characterise cis-acting transcriptional elements of the genes involved in development. In the present study, we have analysed the expression pattern of several muscle-specific and ubiquitous regulatory sequences in microinjected zebrafish embryos. By using a fast and reproducible coinjection strategy, the mosaic expression of lacZ reporter gene was monitored in wholemount embryos injected with sequences containing putative enhancer elements and a carp myosin heavy chain promoter/lacZ reporter construct. We have found that a 0.9-kb myosin heavy chain (MyHC) proximal promoter containing several putative myogenic regulatory factors (MRF) binding sites is sufficient to restrict lacZ expression to the skeletal muscle fibres of prim-6 stage zebrafish embryos. Expression of a rat-derived foetal myosin light chain enhancer (MyLC) and different fragments of a carp beta-actin regulatory region together with the MyHC promoter were compared by accumulating the type, number and spatial distribution of beta-galactosidase-expressing cells on an expression map. beta- galactosidase activity increased similarly whether the MyLC enhancer was ligated to the promoter/ reporter construct directly or when coinjected as a separate fragment whilst skeletal muscle specificity was retained. The coinjection of two different forms of the beta-actin regulatory elements also showed a marked effect on the MyHC promoter activity. The coinjection of putative enhancers with minimal promoter constructs and subsequent analysis of the transient expression pattern in the developing embryos provides a rapid and simple technique to identify cis acting activator elements of genes expressed in the vertebrate embryo.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping