关键词: 细胞型朊蛋白;受体;免疫组织化学

    摘 要:目的 表达和纯化以朊蛋白(PrP)胞外部分与人IgG Fc段构建的融合蛋白PrPFc，并以此为配体蛋白，定位细胞型朊蛋白(PrPc)的受体在小鼠不同发育阶段脑内的分布. 方法 收集表达PrPFc的细胞培养液，通过蛋白A-琼脂糖柱亲和层析纯化，以PrPFc纯化蛋白与新鲜弱固定的组织切片反应，免疫组织化学法显示受体在脑内分布模式. 结果 在正常成年小鼠脑内，细胞型朊蛋白的受体分布以位于大脑皮层(包括新皮层和旧皮层)和纹状体为最多，其次为丘脑、下丘脑及脑干，再次为海马的CA区和齿状回及小脑的蒲肯野和颗粒细胞层，嗅球部位也有一些细胞呈阳性染色;出生后14d小鼠脑内朊蛋白受体与成年动物分布模式及染色强度皆类似，而出生当日及7d后受体分布与14d和成年动物相似，但染色强度明显较弱. 结论 朊蛋白受体(或结合蛋白)在脑组织有广泛分布;朊蛋白与其受体在脑组织中分布相似.

    Localization of the binding proteins for cellular pri┐on protein in mouse brains of different developmen┐tal stages

    YANG Ting，RAO Zhi-Ren，JU Gong

    Institute of Neurosciences of Chinese PLA，Fourth Military Medical University，Xi'an710033，China

    Keywords:cellular prion protein;receptor;immunohis-tochemistry

    Abstract:AIM To recombinant protein PrPFc containing the extracellular portion of prion protein and human IgG Fc fragment was expressed and purified.This fusion protein was used as a ligand to localize the binding proteins for cellular prion protein in adult mouse brains.Distribution pattern of the binding proteins from different developmental stages of mice were detected and compared.METHODS PrPFc was secreted into the culture medium and purified through protein A-Sepharose column.The concentration and purity of the fu-sion protein were checked.Purified PrPFc was used to incu-bate with short fixed fresh frozen cryostat cut sections ......