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本文阐明了利用细胞内神经生物素（Ｎｅｕｒｏｂｉｏｔｉｎ）染色与ＨＲＰ逆行追踪相结合的方法，研究大鼠三叉神经本体感觉的传导通路．首先将ＨＲＰ分别注射于或微电泳亚对侧丘脑腹后内侧核和后核群，同侧颈髓Ｃ２～Ｃ４平面；同侧闭口肌或同侧三叉神经运动核。２４～４８小时后，用电生理的方法鉴定１～５根闭口肌肌梭传入轴突，分别于细胞内注射神经生物素。ＨＲＰ用ＴＭＢ成色并以钨酸钠作为稳定剂，神经生物素用免疫组化的方法显示．待ＨＲＰ和神经生物素呈色后光镜下发现；在三叉神经运动核，三叉神经感觉核簇和邻近网状结构的ＨＲＰ逆行标记细胞被神经生物素细胞内染色的肌梭传入终扣紧密相贴（Ａｐｐｏｓｉｔｉｏｎ）．这些紧密相贴的结构在电镜下证实为细胞内染色的终扣和ＨＲＰ逆行标记细胞的胞体和的突之间的突触联系．因此，这些ＨＲＰ标记的神经元分别中继来自闭口肌肌梭的三叉神经本体感觉信息至丘脑、颈髓和三叉神经运动核等，分别形成三叉－丘脑和三叉－颈髓的本体感觉传导通路以及与三叉神经运动核之间的直接联系．细胞内神经生物素注射与ＨＲＰ逆行追踪相结合的方法提供了一种追踪中枢神经系统神经环路和神经通路的可靠手段．本方法优越于亲神经病毒追除、原癌基因表达以及神经元溃?
This article elucidated the use of intracellular neurobiotin (Neurobiotin) staining and HRP retrograde tracing method to study the sensory pathway of the trigeminal nerve in rats. Firstly, HRP was injected into the ventromedial nucleus and posterior nucleus of the subtropicalated rat on the microvillage, C2 ~ C4 plane of the ipsilateral cervical spinal cord, the ipsilateral closed muscle or ipsilateral trigeminal motor nucleus. After 24 to 48 hours, electrophysiological methods were used to identify the passage of 1 to 5 muscular spindle into the axon, and neurobiotin was injected intracellularly respectively. HRP TMB color with sodium tungstate as a stabilizer, neurobiotin immunohistochemical method was shown. After HRP and neurobiotin staining, the retrograde labeling of HRP retrogradely labeled cells in the trigeminal motor nucleus, the trigeminal sensory nucleus cluster and the adjacent reticular network was carried into the terminal sprocket tightly by the muscle spindle stained by neurobiotin Apposition. These closely-attached structures demonstrated electron microscopy to be the synaptic connection between the terminal end of intracellular staining and the soma of the HRP retrogradely labeled cells. Therefore, these HRP-labeled neurons relay the sensory information of the trigeminal nerve from the muscle spindles to the thalamus, cervical spinal cord and trigeminal motor nucleus, respectively, to form ontological sensory pathways of trigeminal-thalamus and trigeminal-cervical spinal cord and Direct contact with the trigeminal motor nucleus. The combination of intracellular neurobiotin injection and HRP retrograde tracing provides a reliable means of tracing the neural and neural pathways of the CNS. This method is superior to the pro-neuroviral recovery, proto-oncogene expression and neuronal collapse?