Fast optical recordings of membrane potential changes from dendrites of pyramidal neurons
Fast optical recordings of membrane potential changes from dendrites of pyramidal neurons
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DOI：
10.1152/jn.1999.82.3.1615
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发表时间：
1999-09-01
影响因子：
2.5
通讯作者：
Zecevic, D
中科院分区：
文献类型：
--
作者：
Antic, S;Major, G;Zecevic, D
了解单个神经元的生物物理特性以及它们如何处理信息是了解大脑如何工作的基础。一种技术，将允许记录的时间和空间动态的电活动在神经过程中具有足够的分辨率将有助于进一步的研究。在这里，我们报告的应用程序的光学记录膜电位瞬变在许多网站上的脊椎动物神经元在脑切片的神经元过程中使用细胞内电压敏感染料。我们获得的证据表明：1）使用贴片电极用电压敏感染料加载神经元是可能的，而不会污染细胞外环境; 2）脑切片在所使用的激发/发射波长下不显示任何自发荧光; 3）染料的药理学作用是完全可逆的; 4）光动力损伤的水平已经允许有意义的测量，并且可以进一步降低; 5）染料的敏感性与无脊椎动物神经元的报道相当; 6）在2小时的孵育期内，染料扩散到远端突起中约500 μ m。该距离应随着孵育时间的延长而增加; 7）来自基底外侧树突（难以或不可能通过贴片电极接近）和顶端树突的光学记录的动作电位信号显示，直接索马刺激和突触刺激都触发了起源于索马附近的动作电位。尖峰backpropagated到两个基底外侧树突和顶端的过程，传播是有点快，在顶端树突。
Understanding the biophysical properties of single neurons and how they process information is fundamental to understanding how the brain works. A technique that would allow recording of temporal and spatial dynamics of electrical activity in neuronal processes with adequate resolution would facilitate further research. Here, we report on the application of optical recording of membrane potential transients at many sites on neuronal processes of vertebrate neurons in brain slices using intracellular voltage-sensitive dyes. We obtained evidence that 1) loading the neurons with voltage-sensitive dye using patch electrodes is possible without contamination of the extracellular environment; 2) brain slices do not show any autofluorescence at the excitation/emission wavelengths used; 3) pharmacological effects of the dye were completely reversible; 4) the level of photodynamic damage already allows meaningful measurements and could be reduced further; 5) the sensitivity of the dye was comparable to that reported for invertebrate neurons; 6) the dye spread similar to 500 mu m into distal processes within 2 h incubation period. This distance should increase with longer incubation; 7) the optically recorded action potential signals from basolateral dendrites (that are difficult or impossible to approach by patch electrodes) and apical dendrites show that both direct soma stimulation and synaptic stimulation triggered action potentials that originated near the soma. The spikes backpropagated into both basolateral dendrites and apical processes; the propagation was somewhat faster in the apical dendrites.