The Journal of Neuroscience, August 17, 2005, ():
Src-Dependent Tyrosine Phosphorylation at the Tips of Growth Cone Filopodia Promotes Extension
J. Neurosci. Robles et al.
25: 7669
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. Model for Src-dependent regulation of filopodial motility. Positive guidance cues activate Cdc42. Active Cdc42 binds PAK, which disrupts its autoinhibitory dimerization, thereby allowing phosphorylation by Src. Tyrosine-phosphorylated PAK is fully active and targeted to the tips of filopodia, where it may regulate filopodial extension by promoting actin monomer addition or preventing severing/depolymerization of growing actin filaments. Src catalytic activity may also be directly modulated by guidance cue receptors.
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Movie 1. Fluorescence and DICimage sequences of a GFP-dSH2-expressing Xenopus spinal neuron growth cone extending on LN. Note PY enrichment within stable, oblong structures in the central domain of the growth cone and proximal regions of filopodia. In contrast, PY puncta at the tips of filopodia exhibit rapid movement during periods of filopodial extension. Time lapse, 60x.
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Movie 2. Fluorescence image sequences of Xenopus spinal neuron growth cones on PDL coexpressing GFP and either KD-Src (left) or CA-Src (right). Note that although the KD-Src expressing growth cone contains multiple filopodia these are very stable and no new filopodia are generated during the image sequence. In contrast, the CA-Src expressing growth cone contains many highly motile filopodia and generates many new filopodia during the image sequence. Time lapse, 60x.
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Movie 3. Fluorescence and DIC image sequences of a GFP-dSH2-expressing Xenopus spinal neuron growth cone on PDL during acute BDNF stimulation. Two min after BDNF addition there is a rapid increase in filopodial number along the axon shaft. Five min after BDNF stimulation (t = 500 s) a majority of new and pre-existing filopodia are motile and contain PY-positive tips. Note that filopodia retain PY even during periods of substratum detachment and lateral motion. Time lapse, 60x.
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Movie 4. Dual-colored image sequence of a growth cone on PDL expressing DsRed-PAK1 (red) and GFP-dSH2 (green). Note the high degree of colocalization between DsRed-PAK1 and GFP-dSH2 at the tips of filopodia during extension. Time lapse, 60x.
