Tracing retinal fiber trajectory with a method of transposon-mediated genomic integration in chick embryo.
We report convenient retinal fiber tracing by transfecting the tracer cDNA by in ovo electroporation. Long-term and stable expression of tracer proteins such as green fluorescent protein is achieved by transposon-mediated genome integration of the tracer protein expression cassette. We carried out coelectroporation of a plasmid containing CAGGS-tracer cDNA flanked by the Tol2 transposable element along with a transposase expression vector to the optic vesicle of chick embryos at stage 11. By selecting electrodes, we can label a large group of retinal ganglion cells, or a small group of retinal ganglion cells; parallel electrodes assure transfection of large areas of the retina, and needle type electrodes label small areas of the retina. The retinal fiber trajectory and terminal zone (TZ) could be detected in the precise retinotopic manner on the contra-lateral side of the optic tectum. The method has advantage in that we can show the retinal fiber trajectory in relation to the molecules that are responsible for pathfinding for the retinal fibers in the same specimen.
Harada
Hidekiyo
H
Department of Molecular Neurobiology, Graduate School of Life Sciences and Institute of Development, Aging and Cancer, Tohoku University, Seiryo-machi 4-1, Aoba-ku, Sendai 980-8575, Japan.
Takahashi
Yoshiko
Y
Kawakami
Koich
K
Ogura
Toshihiko
T
Nakamura
Harukazu
H
D000818:Animals; D001369:Axons / Q000378:metabolism; D002642:Chick Embryo; D018274:Electroporation / Q000379:methods; D018014:Gene Transfer Techniques; D016678:Genome; D009412:Nerve Fibers / Q000378:metabolism*; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D018626:Retroelements / Q000235:genetics*; D013194:Staining and Labeling / Q000379:methods*
D018626:Retroelements