Real-Time Endocytosis Measurements by Membrane Capacitance Recording at Central Nerve Terminals. Methods Mol Biol 2018;1847:95-108
Date
08/22/2018Pubmed ID
30129012DOI
10.1007/978-1-4939-8719-1_8Scopus ID
2-s2.0-85052248659 (requires institutional sign-in at Scopus site)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.
Author List
Lou XAuthor
Xuelin Lou PhD Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsData Interpretation, Statistical
Electric Capacitance
Endocytosis
Membrane Potentials
Mice
Patch-Clamp Techniques
Presynaptic Terminals
Synaptic Vesicles
