File:A-dynamin-1--dynamin-3--and-clathrin-independent-pathway-of-synaptic-vesicle-recycling-mediated-by-elife01621f006.jpg

English: (A) Neuronal cultures were stimulated at 80 Hz for 10 s and then allowed to recovery in resting buffer for 30 min. CTX-HRP was added either during both stimulation and recovery, or during recovery only, as indicated. The endocytic intermediates observed under these conditions, as well as their labeling pattern, were similar to those observed upon high K+ stimulation. (B) quantification of the results shown in A. (C) Pie charts illustrating SVs observed under the various conditions, based on the results shown in panel B, with sectors coded as indicated in the legend of Figure 2I. As in the case of high K+ stimulation (Figure 2), SV reformation during the stimulus was virtually abolished in Dyn1/3 DKO neurons (black sectors). Concerning the recovery of SVs after the stimulus (striped sectors), the major defect was observed in the reformation of labeled SVs from bulk endosomes (black stripes), this value was extrapolated (as for Figure 2I) by subtracting from the total striped area the fraction of SVs that become labeled when CTX-HRP was added only after the stimulus. Scale bar = 100 nm. ****, ** indicate p-values of <0.0001 and <0.01, respectively. n.s., ‘not significant’. Black asterisks refer to comparisons between total vesicles, colored asterisks to comparisons between HRP-labeled vesicles. Bars: standard error of the mean.