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. 2002 Dec 12;99(26):17143–17148. doi: 10.1073/pnas.222657399

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Copyright © 2002, The National Academy of Sciences

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Fig 4. — Electron microscopy of synaptojanin 1-deficient synapses shows a persistent, stimulus-dependent increase in clathrin-coated vesicles. (a–e) Electron micrographs show tracer-labeled (HRP and HRP-WGA as indicated) recycling vesicles in control (a and c) and knockout (b, d, and e) nerve terminals after a 900-AP train (10 Hz) and 10-min recovery period. In control nerve terminals, labeled vesicles are randomly intermixed in the cluster of synaptic vesicles. In knockout nerve terminals, there is an accumulation of labeled and unlabeled clathrin-coated vesicles (circles in b) that are often segregated at the periphery of the synaptic vesicle cluster. Note the row of clathrin-coated vesicles in e. (f) Morphometry of total clathrin-coated vesicles (labeled plus unlabeled) showing the persistent accumulation of clathrin-coated vesicles after stimulation. (g) HRP-WGA-labeled vesicles make up a large fraction of the accumulated clathrin-coated vesicles in knockout neurons, confirming that they are truly endocytic. (h) Fluid-phase HRP-labeled clathrin-coated vesicles decrease after a 10-min recovery period, showing that at least some of the vesicles progress to downstream recycling steps.