Double electroporation in two adjacent tissues in chicken embryos

Area of Science:

• Developmental Biology
• Molecular Biology
• Genetics

Background:

• In ovo electroporation is a standard technique for introducing transgenes into chicken embryos.
• It is used for studying cell lineage, morphology, and molecular pathways via fluorescent reporters.
• Gain- and loss-of-function studies are possible with transgene or gene-silencing constructs.

Purpose of the Study:

• To develop and demonstrate a novel double-electroporation technique.
• To enable simultaneous introduction of different plasmids into opposing tissues in chicken embryos.
• To facilitate the study of cellular and molecular interactions between adjacent embryonic structures.

Main Methods:

• Development of a specialized electroporation chamber.
• Simultaneous electroporation of dorsal surface ectoderm with mCherry reporter construct.
• Simultaneous electroporation of subjacent somites with EGFP reporter construct.

Main Results:

• Successful co-expression of two different fluorescent reporter proteins (mCherry and EGFP) in adjacent tissues.
• Demonstration of the technique's feasibility in chicken embryos.
• Validation of the method for investigating protein localization in neighboring structures.

Conclusions:

• The developed double-electroporation technique is effective for studying interactions between adjacent tissues.
• This method enhances the ability to investigate cellular and molecular crosstalk during embryonic development.
• It provides a valuable tool for future developmental biology research.