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应用 GFP基因重组病毒标记技术和免疫荧光组织化学技术 ,在荧光显微镜和激光扫描共聚焦显微镜下观察了大鼠三叉神经脊束核尾侧亚核 (Vc)浅层内两种囊泡膜谷氨酸转运体 (VGlu T1样和 DNPI样 )以及 SP样阳性纤维和终末与 Sindibis病毒转染表达绿色荧光蛋白阳性神经元之间的联系。目的在于为探索兴奋性神经递质和 SP参与面口部伤害性信息在 Vc浅层内的传递和调控提供形态学依据 ;并为今后更好地开展绿色荧光蛋白基因重组病毒标记法的研究探索新路。结果显示 :(1)绿色荧光蛋白基因重组 Sindibis病毒注入一侧 Vc浅层后 ,仅在同侧 Vc注射部位附近的 ～ 层内观察到 5～ 9个绿色荧光蛋白标记神经元 ,这些神经元的胞体为小型 (≤ 15μm) ,呈圆形、多角形或不规则形 ;(2 ) Vc浅层内均可见大量的 VGlu T1样、DNPI样和 SP样阳性纤维和终末 ,其中 VGlu T1样阳性纤维和终末主要密集分布于 层的内侧部和 层 , 层和 层外侧部较为稀疏 ;而 DNPI则主要密集分布于 层和 层 , 层相对地较为稀疏 ;SP样阳性纤维和终末主要分布于 层和 层 ;(3 ) VGlu T1样、DNPI样和 SP样阳性终扣分别聚集于绿色荧光蛋白标记神经元的胞体或树突周围 ,并与之形成密切接触。以上结果提示 ,Vc浅层内两种囊泡膜谷氨酸转运体样和 SP样神经终
GFP gene recombinant virus labeling technology and immunofluorescence histochemistry were used to observe the expression of two types of vesicle and glutamine in the superficial layer of caudal subnucleus (Vc) of the rat trigeminal nucleus under fluorescent microscope and laser scanning confocal microscope Acid transporters (VGluT1-like and DNPI-like) as well as SP-like positive fibers and terminals and Sindibis virus transfected expression of green fluorescent protein-positive neurons. The purpose of this study is to provide morphological evidence for exploring the transmission and regulation of excitatory neurotransmitters and SP involved in facial and nociceptive information in the Vc superficial layer, and to explore a new method for the future development of recombinant GFP labeling methods new road. The results showed that: (1) After injection of recombinant green fluorescent protein gene Sindibis into the superficial layer of Vc, 5-9 green fluorescent protein labeled neurons were observed only in the ~ layer near the injection site of ipsilateral Vc. (2) All the VGlu T1-like, DNPI-like and SP-like positive fibers and terminals were visible in the superficial layers of Vc, of which VGlu T1-like positive The fiber and terminal are mainly densely distributed in the inner part and the inner layer of the layer, while the outer part of the layer and the outer layer are sparse; while the DNPI is mainly densely distributed in the layer and the layer, the layer is relatively sparse; the SP-like positive fiber and the terminal are mainly distributed in (3) VGluT1-like, DNPI-like, and SP-like positive terminators were clustered around and closely contacted with the green fluorescent protein-labeled neurons in the cell body or dendrites, respectively. The above results suggest that two vesicular membrane glutamate transporters and SP-like neurons in the superficial Vc