Calcium triggers calcineurin-dependent synaptic vesicle recycling in mammalian nerve terminals

BACKGROUND Following exocytosis at the synapse, synaptic vesicle components are recovered by endocytosis. Morphological analysis has suggested that this occurs by a clathrin-mediated pathway, and the GTPase dynamin is thought to be involved in 'pinching off' endocytosing vesicles. The finding that the calcium-dependent phosphatase calcineurin can dephosphorylate dynamin and two other proteins implicated in endocytosis (amphiphysin and synaptojanin) has suggested a potential role for calcium and dephosphorylation in regulating synaptic vesicle endocytosis. RESULTS We tested this hypothesis with an endocytosis assay in isolated nerve terminals (synaptosomes) that relies on the use of the fluorescent dye FM2-10. In synaptosomes, vesicle recycling occurs predominantly via a pathway dependent on both dynamin and amphiphysin. We found that endocytosis could be stimulated maximally at calcium concentrations that yielded only low levels of exocytosis, suggesting that the two processes had different calcium sensitivities cyclosporin A and Fk506, we identified calcineurin as a calcium sensor for endocytosis and showed that its activity is essential for synaptic vesicle endocytosis in synaptosomes. CONCLUSIONS Our results suggest that dynamin-dependent synaptic vesicle endocytosis is triggered by calcium influx occurring upon nerve-terminal depolarisation. An essential mediator of calcium's effect is calcineurin, the activation of which leads to dephosphorylation of at least four proteins implicated in endocytosis-dynamin, amphiphysin 1, amphiphysin 2 and synaptojanin. Our findings also imply that endocytosis and exocytosis may occur in tandem in vivo simply because they share a responsiveness to calcium influx, rather than because they are mechanistically coupled.