Figure 5

Both retinal projection neurons and their substrate pathway can be manipulated separately in the same embryo. a-d: Eye-targeted electroporation can be combined with brain electroporation. a: A dorsal view of an embryo doubly transfected. Retinal axons (red in b and c) navigate normally to the tectum, passing through a transfected region of the diencephalon (green in c) (dashed line indicates the OT boundary). Eye- and ventral-targeted electroporation can be combined (d). Frontal section showing axons from the transfected retina (red) that have crossed the transfected midline (GFP-transfected) and growing dorsally towards tectum (arrow). e-g: Electroporation can be performed on embryos lipofected in the eyes. e: High magnification of two GFP lipofected axons passing through a cluster of electroporated tectal cells. f and g: Frontal sections of an embryo lipofected in the eye and electroporated in the brain. Retinal axons in the dorsal brain (green: f, g) traversed the transfected cells (red: g). Outlines of brains in wholemounts (b, c, e) and sections (f, g) were drawn based on bright field images and DAPI counterstainings respectively. Epi., epiphysis; Di., diencephalon; OT, optic tectum; Tel, telencephalon. Scale bars: 400 μm in a; 100 μm in b-g.