The Journal of Neuroscience, December 14, 2005, ():
Distinct Mechanisms for Neurotrophin-3-Induced Acute and Long-Term Synaptic Potentiation
J. Neurosci. Je et al.
25: 11719
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1.
Representative SSCs (downward deflections of varying amplitudes) of a triplet. (-) NT-3, a control synapse; (+) NT-3, a NT-3-expressing synapse.
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Supplemental Figure 2.
A, Sample recordings showing SSCs from four synapses, one expressing DN-Dynamin presynaptically (2 lower traces) while the other not (2 upper traces), in the same culture treated with or without NT-3 for 48 hours.
B, Expression of DN-dynamin for two days could not block the acute effect of NT-3.
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Supplemental Figure 3.
Inhibition of NT-3-induced long-term, but not acute, synaptic modulation by rapamycin at low concentration (20nM). Rapamycin was directly applied to nerve-muscle co-cultures. A and B, Rapamycin successfully blocked the change in SSC frequency induced by long-term treatment with NT-3. Rapamycin did not have significant effect on SSC amplitude (p=0.5). C, Blockade of NT-3 induced increase in the amplitude of ESC by rapamycin. There was no statistically significant difference between the “NT-3” and “rapamycin + NT-3” groups (p=0.3). D, Attenuation of NT-3-induced changes in synaptic varicosity by rapamycin. Cultures were treated together with rapamycin and NT-3, and synaptic varicosity was analyzed as described in Fig. 1B. E, Two-day pretreatment of rapamycin could not block the NT-3 mediated acute synaptic modulation (p=0.01). F, Rapamycin treatment at low (20 nM, right) or high (200 nM, middle) did not affect neuronal morphology. M: myocytes, N: neurons. Scale: 20 μm.
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Supplemental Figure 4.
Expression of dominant negative Akt blocks NT-3 induced S6 phosphorylation. DN-Akt and EGFP was co-expressed in Xenopus spinal neurons. White arrowheads indicate the location of neuron. Upon application of NT-3 more than 80% of neurons showed positive staining of phosphorylated ribosomal S6 protein (See Figure 7B). However, expression of Myc-DN-Akt in Xenopus spinal neurons inhibited the NT-3 induced increase in pS6 (A), whereas expression of EGFP or non-expressing control neurons still showed NT-3 induced S6 phosphorylation (B and C). More than 20 neurons per group were examined and pS6 results are summarized in a table.
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Supplemental video
