‘Shocking’ developments in chick embryology: electroporation and in ovo gene expression

Abstract

Efficient gene transfer by electroporation of chick embryos in ovo has allowed the development of new approaches to the analysis of gene regulation, function and expression, creating an exciting opportunity to build upon the classical manipulative advantages of the chick embryonic system. This method is applicable to other vertebrate embryos and is an important tool with which to address cell and developmental biology questions. Here we describe the technical aspects of in ovo electroporation, its different applications and future perspectives.

The emergence of ectomesenchyme

“…Electroporation of chick embryos at Hamburger and Hamilton (HH) stages 6 -10 was performed following the wellestablished procedure (Itasaki et al, 1999). DNA was injected in the midbrain/rostral hindbrain at the concentration of 2 g/ l. Platinum/iridium electrodes (0.25 mm) were placed on either side of the embryo, with the anode at the right side.…”

Optimized cationic lipid‐based gene delivery reagents for use in developing vertebrate embryos

“…Therefore, the advantages and disadvantages of each method must be weighed when choosing the best transfection method for each particular experiment. To date, viral-mediated gene delivery (Sato Leber et al, 1996;Mor-gan and Fekete, 1996;Bell and Brickell, 1997;Sato et al, 2002) and electroporation (Itasaki et al, 1999;Swartz et al, 2001;Colas and Schoenwolf, 2003) have been the most common and effective techniques for ectopic gene expression within the developing embryo. Viral-mediated gene delivery has the advantage of highly efficient, long-term expression of the gene of interest, an important attribute for many misexpression studies.…”

Distinct susceptibility of developing neurons to death following Bax overexpression in the chicken embryo

“…Accordingly, to introduce Bax specifically into MNs during the period of early PCD, we have used in ovo electroporation 26 coupled with the tet regulatory system. 23 To target gene expression to spinal MNs, the reverse tetracycline-controlled transactivator is expressed under the control of the MN-specific HB9 promoter.…”

“…Electroporation was performed using a square wave electroporator (CUY21EDIT) as described elsewhere. 26 Doxycycline (10 mg; Sigma) was added at 24 and again at 36 h after electroporation, and embryos were harvested at 48 h (E4.5) after electroporation.…”