High resolution electrophysiological techniques for the study of calcium-activated exocytosis.
High resolution electrophysiological techniques for the study of calcium-activated exocytosis.
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DOI：
10.1016/j.bbagen.2011.12.011
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发表时间：
2012-08
影响因子：
3
通讯作者：
Lindau, Manfred
中科院分区：
文献类型：
--
作者：
Lindau, Manfred
神经末梢和神经内分泌细胞的分泌囊泡通过钙激活的胞吐作用释放神经递质、神经肽和激素。在这个过程中的一个关键步骤是囊泡膜和质膜之间的融合孔的形成。胞吐融合导致质膜面积的增加，其可以测量为质膜电容的成比例增加。随着膜片钳技术的发展，单细胞、神经末梢和小膜片的高分辨率电容测量成为可能。本文综述了全细胞膜片钳电容测量方法及其与电压钳脉冲刺激和笼状钙光释放刺激的结合使用。它还讨论了在神经内分泌细胞和神经末梢的单个胞吐事件和融合孔特性的研究的补丁电容测量。电容测量提供了高分辨率的信息融合的程度和时间过程的表征囊泡池和胞吐的动力学。它们允许融合模式的表征，包括区分单个囊泡完全融合、瞬时吻跑融合或多囊泡复合胞吐。此外，融合孔电导及其动态行为的测量使得能够以类似于通过单通道记录表征离子通道功能的方式表征融合孔性质。膜片钳电容测量与药理学和分子操纵的胞吐作用相结合，是一种强有力的方法来研究钙激活胞吐融合孔形成的分子机制。
Neurotransmitters, neuropeptides and hormones are released from secretory vesicles of nerve terminals and neuroendocrine cells by calcium-activated exocytosis. A key step in this process is the formation of a fusion pore between the vesicle membrane and the plasma membrane. Exocytotic fusion leads to an increase in plasma membrane area that can be measured as a proportional increase in plasma membrane capacitance. High resolution capacitance measurements in single cells, nerve terminals and small membrane patches have become possible with the development of the patch clamp technique. This review discusses the methods of whole cell patch clamp capacitance measurements and their use in conjunction with voltage clamp pulse stimulation and with stimulation by photorelease of caged calcium. It also discusses patch capacitance measurements for the study of single exocytotic events and fusion pore properties in neuroendocrine cells and nerve terminals. Capacitance measurements provide high-resolution information on the extent and time course of fusion for the characterization of vesicle pools and the kinetics of exocytosis. They allow the characterization of the mode of fusion including distinction of single vesicle full fusion, transient kiss-and-run fusion or multivesicular compound exocytosis. Furthermore, measurement of fusion pore conductances and their dynamic behavior has enabled the characterization fusion pore properties in a way that resembles the characterization of ion channel function through single channel recordings. The combination of patch clamp capacitance measurements with pharmacological and molecular manipulations of exocytosis is emerging as a powerful approach to investigate the molecular mechanisms of calcium-activated exocytotic fusion pore formation.
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DOI：
10.1083/jcb.1.1.47
发表时间：
1955-01
期刊：
The Journal of biophysical and biochemical cytology
影响因子：
--
作者：
DE ROBERTIS, E D;BENNETT, H S
通讯作者：
BENNETT, H S
影响因子：
21.3
作者：
Gong, Liang-Wei;Alvarez de Toledo, Guillermo;Lindau, Manfred
通讯作者：
Lindau, Manfred
影响因子：
64.8
作者：
DETOLEDO, GA;FERNANDEZCHACON, R;FERNANDEZ, JM
通讯作者：
FERNANDEZ, JM
影响因子：
5.3
作者：
Elhamdani, A;Azizi, F;Artalejo, CR
通讯作者：
Artalejo, CR
影响因子：
64.8
作者：
CHOW, RH;VONRUDEN, L;NEHER, E
通讯作者：
NEHER, E