Real-Time Endocytosis Measurements by Membrane Capacitance Recording at Central Nerve Terminals.
Real-Time Endocytosis Measurements by Membrane Capacitance Recording at Central Nerve Terminals.
复制标题
通过中枢神经末梢膜电容记录进行实时内吞测量。
DOI：
10.1007/978-1-4939-8719-1_8
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Lou,Xuelin
中科院分区：
文献类型：
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作者：
Lou,Xuelin
内吞作用是细胞功能的基础。它可以通过膜片钳记录下的电容测量来监测。膜电容记录技术以高的时间分辨率和灵敏度测量细胞膜表面积及其变化，是胞吐和胞吞研究领域的一种强有力的生物物理手段。一个流行的方法是频域方法，需要处理被动正弦膜电流感应的正弦电压。这种技术需要在膜片钳记录期间以硬件或软件实现的相敏检测器或“锁定放大器”。它已被广泛应用于许多分泌细胞，但它的应用直接在中央突触前末梢是技术上的挑战。我们已经应用这种技术来研究突触内吞在杯的Held，一个大的突触末梢，以及小鼠胰腺β细胞。突触前电容的测量提供了一个独特的替代测量发射释放和突触前内吞。在这里，我们描述了这种方法在急性脑切片的杯举行，并提供了一个实用的指南，以获得高品质的电容测量突触前终端。
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.