title: In ovo electroporation in embryonic chick retina.
abstract: Chicken embryonic retina is an excellent tool to study retinal development in higher vertebrates. Because of large size and external development, it is comparatively very easy to manipulate the chick embryonic retina using recombinant DNA/RNA technology. Electroporation of DNA/RNA constructs into the embryonic retina have a great advantage to study gene regulation in retinal stem/progenitor cells during retinal development. Different type of assays such as reporter gene assay, gene over-expression, gene knock down (shRNA) etc. can be performed using the electroporation technique. This video demonstrates targeted retinal injection and in ovo electroporation into the embryonic chick retina at the Hamburger and Hamilton stage 22-23, which is about embryonic day 4 (E4). Here we show a rapid and convenient in ovo electroporation technique whereby a plasmid DNA that expresses green fluorescent protein (GFP) as a marker is directly delivered into the chick embryonic subretinal space and followed by electric pulses to facilitate DNA uptake by retinal stem/progenitor cells. The new method of retinal injection and electroporation at E4 allows the visualization of all retinal cell types, including the late-born neurons(1), which has been difficult with the conventional method of injection and electroporation at E1.5(2).
authors.affiliation: Department of Pharmacology, University of Medicine and Dentistry, New Jersey-Robert Wood Johnson Medical School.
authors.forename: Mohammed M
authors.initials: MM
authors.lastname: Islam
authors.forename: Sung Tae
authors.initials: ST
authors.lastname: Doh
authors.forename: Li
authors.initials: L
authors.lastname: Cai
mesh_terms: D000818:Animals; D002642:Chick Embryo; D004247:DNA / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics; D018274:Electroporation / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D012160:Retina / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology
chemical_list: D049452:Green Fluorescent Proteins; D004247:DNA