Whole Retinal Explants from Chicken Embryos for Electroporation and Chemical Reagent Treatments.
The retina is a good model for the developing central nervous system. The large size of the eye and most importantly the accessibility for experimental manipulations in ovo/in vivo makes the chicken embryonic retina a versatile and very efficient experimental model. Although the chicken retina is easy to target in ovo by intraocular injections or electroporation, the effective and exact concentration of the reagents within the retina may be difficult to fully control. This may be due to variations of the exact injection site, leakage from the eye or uneven diffusion of the substances. Furthermore, the frequency of malformations and mortality after invasive manipulations such as electroporation is rather high. This protocol describes an ex ovo technique for culturing whole retinal explants from chicken embryos and provides a method for controlled exposure of the retina to reagents. The protocol describes how to dissect, experimentally manipulate, and culture whole retinal explants from chicken embryos. The explants can be cultured for approximately 24 hr and be subjected to different manipulations such as electroporation. The major advantages are that the experiment is not dependent on the survival of the embryo and that the concentration of the introduced reagent can be varied and controlled in order to determine and optimize the effective concentration. Furthermore, the technique is rapid, cheap and together with its high experimental success rate, it ensures reproducible results. It should be emphasized that it serves as an excellent complement to experiments performed in ovo.
Department of Neuroscience, Biomedical Center, Uppsala University.
Shahrzad
S
Shirazi Fard
Department of Neuroscience, Biomedical Center, Uppsala University.
Maria
M
Blixt
Department of Neuroscience, Biomedical Center, Uppsala University; finn.hallbook@neuro.uu.se.
Finn
F
Hallböök
D000818:Animals; D002642:Chick Embryo / Q000502:physiology* / Q000601:surgery*; D018274:Electroporation / Q000379:methods*; D009924:Organ Culture Techniques / Q000379:methods*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D012160:Retina / Q000196:embryology / Q000502:physiology* / Q000601:surgery*