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Real-Time Endocytosis Measurements by Membrane Capacitance Recording at Central Nerve Terminals

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Clathrin-Mediated Endocytosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1847))

Abstract

Endocytosis is fundamental to cell function. It can be monitored by capacitance measurements under patch-clamp recordings. Membrane capacitance recording measures the cell membrane surface area and its changes at high temporal-resolution and sensitivity, and it is a powerful biophysical approach in the field of exocytosis and endocytosis. A popular one is the frequency domain method that entails processing passive sinusoidal membrane currents induced by a sinusoidal voltage. This technique requires a phase-sensitive detector or “lock-in amplifier” implemented in hardware or software during patch-clamp recordings. It has been widely used in many secretory cells, but its application directly at central presynaptic terminals is technically challenging. We have applied this technique to study synaptic endocytosis in the calyx of Held, a large glutamatergic synaptic terminal, as well as mouse pancreatic β-cells. The presynaptic capacitance measurements provide a unique alternative to measuring transmitter release and presynaptic endocytosis. Here, we describe this method at the calyx of Held in acute brain slices and provide a practical guide to obtaining high quality capacitance measurements at presynaptic terminals.

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 is partially supported by the National Institutes of Health (NIH) grants R01DK093953 (X.L.), 1R21NS101584-01 (X.L.), and the grant AAB1425-135-A5362 (X.L.). I thank Meyer Jackson for his comments on the manuscript.

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Authors and Affiliations

  1. Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI, USA

    Xuelin Lou

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  1. Xuelin Lou
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Correspondence to Xuelin Lou .

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Editors and Affiliations

  1. Department of Cellular & Molecular Physiology, University of Liverpool, Institute of Translational Medicine, Liverpool, UK

    Laura E. Swan

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Lou, X. (2018). Real-Time Endocytosis Measurements by Membrane Capacitance Recording at Central 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_8

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  • DOI: https://doi.org/10.1007/978-1-4939-8719-1_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8717-7

  • Online ISBN: 978-1-4939-8719-1

  • eBook Packages: Springer Protocols

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