谷氨酸脱羧酶67-绿色荧光蛋白基因敲入小鼠三叉神经尾侧亚核内GFP阳性神经元的分布及与小白蛋白的共存  

DISTRIBUTION OF GFP-POSITIVE NEURONS AND THE COLOCALIZATION WITH PARVALBUMIN IN THE CAUDAL SPINAL TRIGEMINAL NUCLEUS IN THE GAD67-GFP KNOCK-IN MOUSE

作　　者：李金莲[1] 董玉琳[1] 熊抗辉[1] 金子武嗣[2] 

机构地区：[1]第四军医大学解剖学教研室暨梁琚脑研究中心,西安710032 [2]日本京都大学医学部高级脑形态学研究室

出　　处：《解剖学报》2006年第1期1-5,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30470907);第四军医大学骨干人才资助计划资助

摘　　要：目的观察谷氨酸脱羧酶67-绿色荧光蛋白（GAD67-GFP）基因敲入小鼠三叉神经尾侧亚核（Vc）浅层内，表达GFP的GABA能神经元的分布及其与小白蛋白（PV）的共存。方法分别运用原位分子杂交与免疫组织化学相结合；GFP与神经元标记物——神经元核蛋白（NeuN）或PV免疫荧光染色相结合的双重标记方法，在光学显微镜和激光共聚焦显微镜下进行观察。结果1．Vc浅层内90％以上的GFP阳性神经元同时表达GAD67 mRNA，而几乎所有表达GAD67 mRNA的阳性神经元都呈GFP阳性；2．GFP阳性神经元主要分布于Ｖc的Ⅰ－Ⅱ层内，细胞较小，尤其在Ⅱ层内可见大量密集分布的GFP阳性细胞和突起。GFP阳性神经元分别占Ⅰ、Ⅱ层内NeuN阳性神经元总数的19．4％和24．3％；3．GFP／PV双标神经元主要分布于Ｖc的Ⅰ－Ⅱ层，这些双标神经元大约占PV阳性神经元的62．4％，占GFP阳性神经元的12．8％。结论在Ｖc表达GFP的GABA能神经元主要密集分布于与外周伤害性信息传递关系密切的板层内，且大部分PV样阳性神经元属于GABA能神经元。Objective To observe the distribution of GABA-containing neurons revealed by GFP expression and the colocalization of the GFP with parvalbumin（PV） in the caudal spinal trigeminal nucleus（Vc）, the glutamate decarboxylase 67- green fluorescence protein（GAI）67-GFP） knock-in mice were used in the present study. Methods Double-labeled techniques were used by in situ hybridization combined with immunohistochemistry for GFP and double immunofluorescence histochemistry for GFP and NeuN（ neuronal nuclei protein, a neuronal marker） or PV. The stained sections were observed under light microscope and a confocal laser-scanning microscope. Results 1. Over 90% of GFP-positive neuronal cell bodies in the laminae Ⅰ and Ⅱ of the Vc showed hybridization signals for GAD67 mRNA, and that almost all neuronal cell bodies with GAD67 mRNA signals were GFP-positive; 2. GFP-positive neurons were mainly distributed in the laminae Ⅰ and Ⅱ of the Vc and the vast majority of them were small neurons. The considerable number of GFP-positive processes and somata were most densely observed in the lamina Ⅱ of the Vc. The proportion of GFP-positive neurons in the NeuN-labeled neurons of the Vc was about 19.4% and 24.3% in laminae Ⅰ and Ⅱ, respectively; 3. Double-labeled neurons for GFP/PV are mainly found in laminae Ⅰ and Ⅱ of the Vc. The proportion of GFP/PV double-labeled neurons was about 62.4% and 12.8% of total population of PV-and GFP-positive neurons in laminae Ⅰ - Ⅱ of the Vc, respectively. Conclusion GABAergic neurons are mainly distributed in the laminae Ⅰ and Ⅱ of the Vc which related closely to transmission of the nociceptive primary afferent information, and the majority of PV-positive neurons are GABAergic neurons.

关 键 词：三叉神经尾侧亚核 γ-氨基丁酸 绿色荧光蛋白 小白蛋白 原位分子杂交 免疫荧光组织化学 GAD 67-GFP基因敲入小鼠 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]