包埋前ABC法和纳米金-银加强法双重标记技术在免疫电镜中的应用  

Preembedding double labeling combined ABC method with nanogold- silver enhancement: an immunoelectron microscopic study

作　　者：刘津平[1] 贾轶[1] 刘莹莹[1] 鞠躬[1] 

机构地区：[1]第四军医大学神经科学研究所,陕西西安710032

出　　处：《中国神经科学杂志》2002年第4期728-731,共4页

基　　金：国家自然科学基金 (3 9870 2 64 );第四军医大学创新工程 (CX 99A0 0 2 )资助项目

摘　　要：应用包埋前ABC法和纳米金 银加强法免疫双重标记技术 ,电镜观察GABA能抑制性突触传递与呼吸节律产生中枢pre B tzinger复合体神经激肽 1受体 (neurokinin 1receptor,NK1R)免疫反应神经元的关系 ,目的在于介绍一种简单、实用的包埋前免疫电镜双重标记技术 ,及其在神经生物学领域的应用。高电子密度的DAB免疫反应产物标记谷氨酸脱羧酶 (glutamicaciddecarboxylase,GAD)免疫反应神经末梢 ,免疫金颗粒主要标记pre B tzinger复合体NK1R免疫反应神经元胞体或树突的膜表面 ,有时胞质内也可见到免疫金颗粒标记。GAD免疫反应神经末梢与NK1R免疫反应神经元胞体或树突形成典型的对称性突触。这种包埋前免疫电镜双重标记技术具有以下优点 :1.包埋前实现免疫组化双重标记有利于组织抗原性的保存。 2 .有利于抗体的组织内渗透。 3.免疫金颗粒抗原定位精确 ,并可用于定量分析。By observation of GABAergic synapses onto neurokinin 1 receptor (NK1R) immunoreactive (ir) neurons in the pre Btzinger complex, the kernel for respiratory rhythmogenesis, we proposed to demonstrate a preembedding double labeling approach combining ABC method with nanogold silver enhancement under electron microscopy in rats. High dense DAB react product indicated glutamic acid decarboxylase (GAD, a marker of GABAergic terminals) ir terminals, while NK1R immunoreactivity was revealed by immunogold particles. NK1R ir immunogold particles presented mainly on the inner surface of the plasma membrane of both somas and dendrites in the pre Btzinger complex. At times, cytoplasm was also labeled with NK1R immunoreactivity. Symmetric synapses were identified between GAD ir terminals and NK1R ir somas or dendrites. The present preembedding immunoelectron microscopic double labeling has advantages as follows: 1. All of the immunoreactive processes are performed before embedding, when tissues preserve high antigen activity. 2. Antibodies are much easy to penetrate tissues. 3. Antigens are precisely recognized by immunogold particles, which are easily quantified under electron microscopy. \[

关 键 词：纳米金-银加强法 ABC法 免疫电镜 pre-Poetzinger复合体 

分 类 号：R392-33[医药卫生—免疫学]