大鼠三叉神经脊束核尾侧亚核浅层内囊泡膜谷氨酸转运体样和P物质样阳性终末与Sindibis病毒转染表达GFP神经元之间的联系

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

ASSOCIATION OF VESICULAR GLUTAMATE TRANSPORTERS- AND SP-LIKE IMMUNOREACTIVE TERMINALS WITH NEURONS REVEALED BY INJECTION OF RECOMBINANT GFP-SINDIBIS VIRUS IN THE CAUDAL SPINAL TRIGEMINAL NUCLEUS OF THE RAT

In order to provide further morphological evidence for the excitatory neurotransmitters and substance P involved in transmission and modulation of orofacial noxious information and to explore a new method that more research will be made by using recombinant Sindibis virus injection and GFP (green fluorescent protein) expression fluorescence staining method in future, in the present study, the immunofluorescence histochemical staining method and recombinant Sindibis virus injection and GFP expression fluorescence staining technique were used to observe the association of two kinds of vesicular glutamate transporters (VGluT1 LI and DNPI LI) and SP LI terminals with GFP labeled neurons in the caudal spinal trigeminal nucleus (Vc) of the rat under a confocal laser scanning microscopy. The results showed: (1) 5～9 GFP labeled neurons were found ipsilaterally in the area near injection site of laminae Ⅰ～Ⅲ of Vc after recombinant Sindibis virus was injected unilaterally into the superficial laminae of Vc, and the bodies of these neurons were small(≤15 μm), circular, multipolar or in irregular shape; (2) A considerable number of VGluT1 LI, DNPI LI and SP LI terminals and varicosities were widely distributed in the superficial layer of Vc (laminae Ⅰ～Ⅲ). Within Vc, VGluT1 LI terminals and varicosities were observed most frequently in the inner part of lamina Ⅱ(Ⅱi)and lamina Ⅲ, although lower density of distribution was also seen in lamina Ⅰ and the outer part of laminaⅡ(Ⅱo).On the other hand, the highest density of DNPI LI terminals and varicosities was found in laminaeⅠand Ⅱ, whereas relatively lower density was observed in lamina Ⅲ. The dense terminals and varicosities with SP LI were seen almost in laminaeⅠandⅡ; (3) In the superficial laminae of Vc, axonal varicosities with dense VGluT1 LI or DNPI LI and SP LI frequently surrounded the somata and dendrites of Vc neuronal profiles labeled with GFP respectively and formed close contact with GFP labeled somata and dendrites of neuronal profiles. The present results suggested that the two kinds of vesicular glutamate transporter LI and SP LI axonal terminals are involved in the nociceptive information transmission and modulation. (Figure 1 on plate 44)