Figure 3

Upregulation of Rtn4b in zebrafish RGCs after ONS. RGCs in retina whole mounts showed weak immunostainings in the cytoplasm after exposure to Rtn4b AB (A). (B,E,H) Labeling with Phalloidin against F-actin shows all cells and their cytoplasm. (C,F,I) Merge of (A,B), (C,D), and (G,H) with DAPI stainings to visualize nuclei. (D,E,F) 5 days after ONS, the RGCs exhibit a significant increase in size and increase in Rtn4b labeling intensity (48% in comparison to control, P < 0.01) in the cytoplasm. (G,H,I) 10 days after optic nerve sections, the size on the RGCs and the intensity of Rtn4b staining is still highly increased (53% over controls, P < 0.01). Scale bar, 10 μm. (J) This apparent increase in the 100 kd Rtn4b protein is also seen in Western blots with retinae at 5 (P < 0.05) and 10 days (P < 0.0001, Student’s T-test) after ONS. Anti-alpha tubulin served as loading control.