Skip to main content
. Author manuscript; available in PMC: 2014 Aug 19.
Published in final edited form as: Neuron. 2014 Feb 19;81(4):779–786. doi: 10.1016/j.neuron.2014.01.036

Figure 4. Cas Adaptor Proteins are Required Rosette-autonomously for GCL Cell Positioning and Organization.

Figure 4

(A–H) Developmental progression of ILM formation and maintenance in Control (A, C, E, G) and Six3Cre; TcKO (B, D, F, H) retinas, here immunolabeled with anti-laminin (Red), and TOPRO3 (Blue). (A and B) The ILM forms normally in Control and Six3Cre; TcKO retinas at e14.5. Ectopic cell aggregates begin to appear in Six3Cre; TcKO retinas before any disruption of the ILM is evident (e17.5, D; n=3 for each stage and genotype). (I and J) Retrograde labeling of WT (I) and Six3Cre; TcKO (J) RGCs by injection of CTB-Alexa546 into the LGN. Ganglion cells in ectopic rosettes that form in Six3Cre; TcKO retinas (J) are labeled by CTB. (K and L) Histological analysis of Control (K) and Pax6αCre;TcKO (L) P14 retinas stained with Topro3 (T3, Blue) and antibodies against EGFP (green) and laminin (Lam, red). Cells that form ectopic aggregates in Pax6αCre;TcKO (L, inset) animals are Cre/EGFP+.

(M and N) Phospho-Cas (PY-Cas, green) imunohistochemistry in P4 Control (M) and Six3Cre; TcKO (N) retinas. Though some PY-Cas staining remains in the Six3Cre; TcKO retinas (white asterisk, N), cells that form aggregates show no PY-Cas expression compared to Control (M). (O and P) Cryo-sections of WT (O) and Cas TcKO (P) e18.5 retinas electroporated in utero with pCAGGs-Cre-IRES-EGFP at e13.5. Sections were immunolabelled with anti-GFP (Green), TOPRO3 (T3, Blue) and anti-Laminin (Red). In utero electroporation of Cre-EGFP into Cas TcKO retinas results in formation of ectopic aggregates that contain EGFP+ cells (5/5 aggregates, n=3 independent animals); no aggregates form in WT retinas (n=3). White arrows: ectopic aggregates; yellow arrowheads: ILM. Scale bar, 50 μm.