Double electroporation in two adjacent tissues in chicken embryos

Abstract

In ovo electroporation is a well established method to introduce transgenes into a number of tissues in chicken embryos, e.g., neural tissue, limb mesenchyme, and somites. This method has been widely used to investigate cell lineage, cell morphology, and molecular pathways by localized expression of fluorescent reporter constructs. Furthermore gain- and loss-of-function experiments can be performed by electroporating transgenes or gene-silencing constructs. We have developed a new technique to electroporate tissues positioned opposite to each other with different plasmids using an electroporation chamber. As proof of principle, we electroporated the dorsal surface ectoderm with a reporter construct expressing mCherry and the subjacent somites with a reporter construct expressing EGFP. This double-electroporation technique allows investigation of the localization of two different proteins of interest in two adjacent tissues and will be useful to examine the cellular and molecular interaction of neighboring structures during embryonic development. Developmental Dynamics 247:1211–1216, 2018. © 2018 Wiley Periodicals, Inc.