Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels.
Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels.
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DOI：
10.1016/j.neuron.2013.07.012
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发表时间：
2013-09-18
期刊：
影响因子：
16.2
通讯作者：
Korchev YE
中科院分区：
文献类型：
--
作者：
Novak P;Gorelik J;Vivekananda U;Shevchuk AI;Ermolyuk YS;Bailey RJ;Bushby AJ;Moss GW;Rusakov DA;Klenerman D;Kullmann DM;Volynski KE;Korchev YE
到目前为止，对突触前离子通道的直接电接触仅限于大型专门化终末，如HOLD或海马苔藓状纤维束的花冠。更丰富的小突触终末(≤1和μm)的电生理和离子通道补充仍然知之甚少。在这里，我们报道了一种基于培养中小突触的超分辨率扫描离子电导成像的方法，该方法以大约100-150 nm的3D分辨率，允许以所有四种形式(细胞连接、内向外、外向外和全细胞)进行突触前膜片钳记录。使用这项技术，我们报道了突触前K+，Na+，Cl，−和钙通道的记录。这种半自动的方法可以直接研究小型中央突触的突触前离子通道的分布和性质。细胞贴附式膜片钳记录小型中央突触中离子通道的地形成像Novak和他的同事已经开发出一种使用超分辨率扫描离子电导显微镜识别的亚微米中央突触中纳米级靶向膜片钳突触前记录的方法。这种半自动的方法为小型突触前终末的生理学打开了一扇新的窗口。
Direct electrical access to presynaptic ion channels has hitherto been limited to large specialized terminals such as the calyx of Held or hippocampal mossy fiber bouton. The electrophysiology and ion-channel complement of far more abundant small synaptic terminals (≤1 μm) remain poorly understood. Here we report a method based on superresolution scanning ion conductance imaging of small synapses in culture at approximately 100–150 nm 3D resolution, which allows presynaptic patch-clamp recordings in all four configurations (cell-attached, inside-out, outside-out, and whole-cell). Using this technique, we report presynaptic recordings of K+, Na+, Cl−, and Ca2+ channels. This semiautomated approach allows direct investigation of the distribution and properties of presynaptic ion channels at small central synapses. Topographic imaging of live synaptic boutons at nanoscale resolution Cell-attached patch-clamp recordings of ion channels in small central synapses Whole-cell small presynaptic bouton recordings Novak and colleagues have developed a method for nanoscale-targeted patch-clamp presynaptic recordings in submicrometer central synapses identified using superresolution scanning ion conductance microscopy. This semiautomated approach opens a new window on the physiology of small presynaptic terminals.
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影响因子：
48
作者：
Novak, Pavel;Li, Chao;Shevchuk, Andrew I.;Stepanyan, Ruben;Caldwell, Matthew;Hughes, Simon;Smart, Trevor G.;Gorelik, Julia;Ostanin, Victor P.;Lab, Max J.;Moss, Guy W. J.;Frolenkov, Gregory I.;Klenerman, David;Korchev, Yuri E.
通讯作者：
Korchev, Yuri E.
影响因子：
5.3
作者：
Hu, H;Shao, LR;Storm, JF
通讯作者：
Storm, JF
影响因子：
4.5
作者：
BOHLE, T;BENNDORF, K
通讯作者：
BENNDORF, K
影响因子：
16.2
作者：
Han Y;Kaeser PS;Südhof TC;Schneggenburger R
通讯作者：
Schneggenburger R
影响因子：
3.4
作者：
Gorelik, J;Gu, YC;Korchev, YE
通讯作者：
Korchev, YE