Abstract
Acute perturbations of clathrin and associated proteins at synapses have provided a wealth of knowledge on the molecular mechanisms underlying clathrin-mediated endocytosis (CME). The basic approach entails presynaptic microinjection of an inhibitory reagent targeted to the CME pathway, followed by a detailed ultrastructural analysis to identify how the perturbation affects the number and distribution of synaptic vesicles, plasma membrane, clathrin-coated pits, and clathrin-coated vesicles. This chapter describes the methodology for acutely perturbing CME at the lamprey giant reticulospinal synapse, a model vertebrate synapse that has been instrumental for identifying key protein–protein interactions that regulate CME in presynaptic nerve terminals with broader extension to nonneuronal cell types.
The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-8719-1_19
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Change history
24 October 2018
This book was inadvertently published with the incorrect title as Clathrin-Mediated Endoytosis: Methods and Protocols. This has now been corrected throughout the book to Clathrin-Mediated Endocytosis: Methods and Protocols.
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Acknowledgments
This work was supported by NIH grants: NINDS/NIA R01 NS078165 (to J.R.M.) and NIGMS R01 GM118933 (to E.M.L.). The authors would like to thank Dr. Eileen M. Lafer (University of TX Health Sciences Center, San Antonio) for providing the AP-2 peptide, as well as Paul Oliphint and Kara Marshall for technical assistance.
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Walsh, R.B., Bloom, O.E., Morgan, J.R. (2018). Acute Manipulations of Clathrin-Mediated Endocytosis at Presynaptic Nerve Terminals. In: Swan, L. (eds) Clathrin-Mediated Endocytosis. Methods in Molecular Biology, vol 1847. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8719-1_6
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DOI: https://doi.org/10.1007/978-1-4939-8719-1_6
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