(A, A′) In vivo electroporation. (A) Plasmid DNA is injected into the eye on P0, receives square waves pulses. (A′) P8 brains are examined for labeled RGC axons.
(B) Example of pCMV-tdTomato electroporated RGC. Arrowhead and inset: RGC axon expressing tdTomato. (C) High magnification of the RGC shown in (B) the RGC soma, dendrites and axon (arrowhead) express high levels of tdTomato. Scale = 125μm.
(D–G) Example of NOT-projecting tdTomato+ axons in wildtype (D, E) and CNTN4−/− mice (F, G); arrowhead: parent axon. (E) Wildtype, NOT-projecting RGC axon reconstruction from boxed region in D. (G) CNTN4−/−, NOT-projecting axon reconstruction from boxed region in F. Scale in D, F= 250μm., Scale in E, G = 50μm.
(H) Quantification of the average number (±SEM) of branches from branch order 1–8 for wildtype and CNTN4−/−, NOT-projecting axon (n=4 axons/mice wildtype, n=2 axons/mice CNTN4−/−).
(I) Quantification of the average arbor surface area for wildtype and CNTN4−/− mice (±SEM), p=0.040.
(J) Quantification of the average arbor volume for wildtype and CNTN4−/− mice (±SEM), p=0.062.
(K) The percentage of electroporated axons arborizing in the NOT after electroporation in wildtype (8/58) and CNTN4−/− (2/25) mice.