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Ca2+ and calmodulin initiate all forms of endocytosis during depolarization at a nerve terminal

Nature Neuroscience volume 12, pages 1003–1010 (2009)Cite this article

A Corrigendum to this article was published on 01 May 2010

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Abstract

Although endocytosis maintains synaptic transmission, how endocytosis is initiated is unclear. We found that calcium influx initiated all forms of endocytosis at a single nerve terminal in rodents, including clathrin-dependent slow endocytosis, bulk endocytosis, rapid endocytosis and endocytosis overshoot (excess endocytosis), with each being evoked with a correspondingly higher calcium threshold. As calcium influx increased, endocytosis gradually switched from very slow endocytosis to slow endocytosis to bulk endocytosis to rapid endocytosis and to endocytosis overshoot. The calcium-induced endocytosis rate increase was a result of the speeding up of membrane invagination and fission. Pharmacological experiments suggested that the calcium sensor mediating these forms of endocytosis is calmodulin. In addition to its role in recycling vesicles, calcium/calmodulin-initiated endocytosis facilitated vesicle mobilization to the readily releasable pool, probably by clearing fused vesicle membrane at release sites. Our findings provide a unifying mechanism for the initiation of various forms of endocytosis that are critical in maintaining exocytosis.

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Figure 1: Calcium influx triggers endocytosis.
Figure 2: Very slow endocytosis at a [Ca2+]i of 0.5–0.75 μM.
Figure 3: Calcium influx increases rateendo and triggers rapid endocytosis and endocytosis overshoot.
Figure 4: Calcium influx initiates bulk endocytosis and speeds up the fission pore closure rate.
Figure 5: Calmodulin blockers inhibit endocytosis.
Figure 6: Syt2 is not critical in mediating endocytosis.
Figure 7: CBD, calmidazolium and dynasore slow down the RRP replenishment.
Figure 8: Endocytosis overshoot retrieves vesicles stranded at the plasma membrane.

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Change history

  • 03 September 2009

    In the version of this article initially published, the units for the calcium concentration labels in Figure 2a,d are incorrect. The correct unit should be µM. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

This work was supported by the National Institute of Neurological Disorders and Stroke Intramural Research Program of the US National Institutes of Health.

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  1. Xin-Sheng Wu and Benjamin D McNeil: These authors contributed equally to this work.

Authors and Affiliations

  1. National Institute of Neurological Disorders and Stroke, Bethesda, Maryland, USA

    Xin-Sheng Wu, Benjamin D McNeil, Jianhua Xu, Junmei Fan, Lei Xue, Li Bai & Ling-Gang Wu

  2. Department of Pulmonary Medicine, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA

    Ernestina Melicoff & Roberto Adachi

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Contributions

X.-S.W. conducted the double-patch experiments for Figure 2 and many of the capacitance recordings for other figures. B.D.M. initiated the project, designed and conducted capacitance experiments and helped write the paper. J.X. and J.F. conducted capacitance experiments. L.X. performed the simulations. E.M. and R.A. provided the Syt2−/− mice. L.B. maintained the animal colonies and L.-G.W. supervised the project, designed experiments and wrote the paper.

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Correspondence to Ling-Gang Wu.

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Supplementary Figures 1–6, Supplementary Data 1, 2 and Supplementary Discussion (PDF 569 kb)

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Wu, XS., McNeil, B., Xu, J. et al. Ca2+ and calmodulin initiate all forms of endocytosis during depolarization at a nerve terminal. Nat Neurosci 12, 1003–1010 (2009). https://doi.org/10.1038/nn.2355

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