The Isolation and Identification of Spinal Neurons That Control Movement in the Xenopus Embryo
The Isolation and Identification of Spinal Neurons That Control Movement in the Xenopus Embryo
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非洲爪蟾胚胎中控制运动的脊髓神经元的分离和鉴定
DOI：
10.1111/j.1460-9568.1991.tb00039.x
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发表时间：
1991
影响因子：
3.4
通讯作者：
N. Dale
中科院分区：
文献类型：
--
作者：
N. Dale
设计了一种分离爪蟾胚胎脊髓神经元的方法。使用多种方法，可以在体外区分前面描述的八种神经元类别中的四种。首先，由于神经元的许多解剖特征在分离后仍然存在，因此可以根据解剖标准分离出感觉神经元和甘氨酸能抑制性中间神经元。其次，在分离前用荧光素结合的葡聚糖胺对完整脊髓中的神经元进行选择性标记，可以鉴定出三种分离的神经元：运动神经元、假定的兴奋性中间神经元和抑制性中间神经元。利用甘氨酸免疫细胞化学证实了抑制性中间神经元的身份。分离神经元的生理特性与体内神经元相似。解离的神经元表现出较强的膜调节和向外整流，这两种作用均可被注射Cs+离子阻断。神经元还保留了激动剂N -甲基- d -天冬氨酸、海钠酸盐、半qualate、甘氨酸和GABA的受体。因此，分离的神经元的膜特性和药理学敏感性似乎不受分离过程的影响。因此，从非洲爪蟾胚胎神经系统中分离出来的四类神经元，可以为详细研究单个神经元的特性如何促进脊椎动物运动模式产生的电路功能的有效模型奠定基础。
A procedure for isolating spinal neurons of the Xenopus embryo has been devised. Using a variety of methods four of the eight previously described categories of neuron can be distinguished in vitro. Firstly, since many anatomical features of the neurons survived dissociation, sensory neurons and glycinergic inhibitory interneurons could be identified after isolation using anatomical criteria. Secondly, by selective labelling of the neurons in the intact spinal cord with fluorescein‐conjugated dextran amines prior to dissociation, three classes of isolated neuron could be identified: motoneurons, putative excitatory interneurons and, once again, inhibitory interneurons. The identity of the inhibitory interneurons has been confirmed using glycine immunocytochemistry. The physiological properties of the isolated neurons were similar to those of their in vivo counterparts. The dissociated neurons exhibited strong membrane accommodation and outward rectification, both of which could be blocked by the injection of Cs+ ions. The neurons also retained their receptors for the agonists N‐methyl‐d‐aspartate, kainate, quisqualate, glycine and GABA. Both the membrane properties and pharmacological sensitivity of the isolated neurons therefore appeared to be unaltered by the dissociation procedure. Thus the four classes of neuron from the Xenopus embryonic nervous system that can be identified after isolation constitute the basis for a valid model for detailed study of how the properties of individual neurons contribute to the functioning of the circuitry that underlies locomotor pattern generation in vertebrates.
改进的荧光化合物用于追踪细胞谱系。
影响因子：
2.7
作者：
Gimlich,RL;Braun,J
通讯作者：
Braun,J