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Figure 3


Fig. 3. Constitutively active RhoA-induced RhoA-kinase-dependent axonal F-actin bundles. (A,B) Chariot-peptide-mediated transfection of axons with constitutively active RhoA (L63RhoA) induced axon bundles in axons that (A) underwent retractions and also in axons that (B) did not retract but exhibited partially collapsed growth cones with distal accumulation of F-actin. Insets show magnified and color-inverted regions of the axons as denoted by the white lines. Bar, 10 µm. (C) Quantification (in percent) of axon segments exhibiting F-actin bundles (as shown in Fig. 2) revealed that treatment with y-27632 at the time of L63RhoA transfection completely inhibited the formation of F-actin bundles. (D) L63RhoA decreased the percentage of time that axon spent extending relative to BSA-treated controls. Data are normalized to BSA-transfected axons. (E) Inhibition of ROCK minimized L63RhoA-induced axon retraction. The percentage of axons simultaneously transfected with L63RhoA and treated with y-27632 that exhibit retraction during imaging was decreased by 69% relative to axons transfected with L63RhoA.





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