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首页> 外文期刊>Journal of Neuroscience Methods >Voltage-sensitive dye recording using retrogradely transported dye in the chicken spinal cord: staining and signal characteristics.
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Voltage-sensitive dye recording using retrogradely transported dye in the chicken spinal cord: staining and signal characteristics.

机译：在鸡脊髓中使用逆行运输的染料进行电压敏感染料记录：染色和信号特征。

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摘要

We describe a novel method for retrogradely labeling specific neuronal populations using voltage-sensitive dyes. Styryl dyes were injected into the ventral roots of the isolated embryonic chick spinal cord. After waiting several hours, the dye labeled motoneurons and autonomic preganglionic neurons. Neuronal cell bodies, dendrites and axons were labeled; we presume that the dye traveled either by retrograde transport or by diffusion within the membrane of the axon to which the dyes were initially applied. Using either a photodiode array or a photomultiplier, fluorescence changes could be recorded from motoneurons following antidromic or synaptic activation. Several characteristics of the fluorescence changes were measured indicating that the signals did indeed reflect changes in the motoneuron membrane potential. The best labeling and optical signals were obtained using the relatively hydrophobic dyes di-8-ANEPPQ and di-12-ANEPEQ. In the great majority of cases these dyes responded with an increase in fluorescence of 1-3% (delta F/F) in response to synaptic or antidromic depolarization of the motoneurons. We anticipate that these techniques should be useful in the mapping of activity patterns and connectivity in neural networks within a defined population of neurons.
机译：我们描述了一种新的方法，用于使用电压敏感染料逆向标记特定的神经元种群。将苯乙烯基染料注射到分离的胚胎雏鸡脊髓的腹侧根中。等待数小时后，该染料标记了运动神经元和自主神经节前神经元。对神经元细胞体，树突和轴突进行标记;我们假设染料是通过逆行运输或通过最初在其上施加染料的轴突膜内扩散而行进的。使用光电二极管阵列或光电倍增管，可以在反线或突触激活后从运动神经元记录荧光变化。测量了荧光变化的几个特征，表明信号确实反映了运动神经元膜电位的变化。使用相对疏水的染料di-8-ANEPPQ和di-12-ANEPEQ获得最佳的标记和光学信号。在大多数情况下，这些染料响应于运动神经元的突触或反显性去极化，荧光增强了1-3％（δF / F）。我们期望这些技术在定义的神经元群体内的神经网络中的活动模式和连通性的映射中应该有用。

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