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Developmental shift to a mechanism of synaptic vesicle endocytosis requiring nanodomain Ca2+

Nature Neuroscience volume 13, pages 838–844 (2010)Cite this article

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Abstract

Ca2+ is thought to be essential for the exocytosis and endocytosis of synaptic vesicles. However, the manner in which Ca2+ coordinates these processes remains unclear, particularly at mature synapses. Using membrane capacitance measurements from calyx of Held nerve terminals in rats, we found that vesicle endocytosis is initiated primarily in Ca2+ nanodomains around Ca2+ channels, where exocytosis is triggered. Bulk Ca2+ outside of the domain could also be involved in endocytosis at immature synapses, although only after extensive exocytosis at more mature synapses. This bulk Ca2+-dependent endocytosis required calmodulin and calcineurin activation at immature synapses, but not at more mature synapses. Similarly, GTP-independent endocytosis, which occurred after extensive exocytosis at immature synapses, became negligible after maturation. We propose that nanodomain Ca2+ simultaneously triggers exocytosis and endocytosis of synaptic vesicles and that the molecular mechanisms underlying Ca2+-dependent endocytosis undergo major developmental changes at this fast central synapse.

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Figure 1: Different effects of EGTA and BAPTA on vesicle endocytosis at the developing calyces of Held.
Figure 2: Ca2+ dependence of fast and slow endocytosis elicited by repetitive stimulation at the calyx of Held.
Figure 3: Effects of inhibitors for CaM and CaN in fast and slow endocytosis at P7–9 calyces.
Figure 4: No effect of inhibitors for CaM and CaN in fast and slow endocytosis at P13–14 calyces.
Figure 5: Developmental decline of CaN expression at the calyx of Held presynaptic terminals.
Figure 6: Developmental decline of GTP-independent endocytic component.

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Acknowledgements

We thank E. Johnson and S. Takamori for helpful comments. This work was supported by the Core Research for Evolutional Science and Technology of Japan Science and Technology Agency (T.T.), US National Institutes of Health grant EY014043 (H.v.G.) and Grant-in-Aid for Young Scientists from the Japanese Ministry of Education, Culture, Sports, Science and Technology #20700357 (T.Y.).

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Authors and Affiliations

  1. Cellular & Molecular Synaptic Function Unit, Initial Research Project, Okinawa Institute of Science and Technology Promotion Corporation, Okinawa, Japan

    Takayuki Yamashita, Kohgaku Eguchi & Tomoyuki Takahashi

  2. Department of Neurophysiology, Doshisha University Faculty of Life and Medical Sciences, Kyoto, Japan

    Naoto Saitoh & Tomoyuki Takahashi

  3. The Vollum Institute, Oregon Health and Science University, Portland, Oregon, USA

    Henrique von Gersdorff

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Contributions

T.Y. and T.T. designed the experiments. H.v.G. designed the experiments on nonhydrolysable GTP analogues. T.Y., K.E. and N.S. performed the experiments and analyzed the data. T.Y., H.v.G. and T.T. wrote the manuscript. All of authors revised and approved the final manuscript.

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Correspondence to Takayuki Yamashita or Tomoyuki Takahashi.

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Yamashita, T., Eguchi, K., Saitoh, N. et al. Developmental shift to a mechanism of synaptic vesicle endocytosis requiring nanodomain Ca2+. Nat Neurosci 13, 838–844 (2010). https://doi.org/10.1038/nn.2576

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