Porchet Raymonde, Probst Alphonse, Bouras Constantin, Dráberová Eduarda, Dráber Pavel, Riederer Beat M
Institut de Biologie Cellulaire et de Morphologie, Université de Lausanne, Rue du Bugnon 9, 1005 Lausanne, Switzerland.
Proteomics. 2003 Aug;3(8):1476-85. doi: 10.1002/pmic.200300456.
Glial fibrillary acidic protein, GFAP, is a major intermediate filament protein of glial cells and major cytoskeletal structure in astrocytes. The entorhinal cortex has a key role in memory function and is one of the first brain areas to reveal hallmark structures of Alzheimer's disease and therefore provides an ideal tissue to investigate incipient neurodegenerative changes. Here we have analyzed age- and disease-related occurrence and composition of GFAP in the human entorhinal cortex by using one- and two-dimensional electrophoresis, Western blots and immunocytochemistry combined with confocal microscopy. A novel monoclonal antibody, GF-02, was characterized that mainly reacted with intact GFAP molecules and indicated that more acidic and soluble GFAP forms were also more susceptible to degradation. GFAP and vimentin increased with aging and in Alzheimer's disease (AD). Two-dimensional electrophoresis and Western blots revealed a complex GFAP pattern, both in aging and AD with different modification and degradation forms. Immunohistochemistry indicated that reactive astrocytes mainly accumulated in relation to neurofibrillary tangles and senile plaques in deeper entorhinal cortex layers. GFAP may be used as an additional but not exclusive diagnostic tool in the evaluation of neurodegenerative diseases because its levels change with age and respond to senile plaque and tangle formation.
胶质纤维酸性蛋白（GFAP）是神经胶质细胞的一种主要中间丝蛋白，也是星形胶质细胞中的主要细胞骨架结构。内嗅皮质在记忆功能中起关键作用，是最早出现阿尔茨海默病标志性结构的脑区之一，因此是研究早期神经退行性变化的理想组织。在这里，我们通过一维及二维电泳、蛋白质免疫印迹和免疫细胞化学结合共聚焦显微镜，分析了人类内嗅皮质中GFAP与年龄和疾病相关的出现情况及组成。我们鉴定了一种新型单克隆抗体GF-02，它主要与完整的GFAP分子反应，并表明酸性更强且更易溶解的GFAP形式也更容易降解。GFAP和波形蛋白在衰老过程以及阿尔茨海默病（AD）中均增加。二维电泳和蛋白质免疫印迹显示，在衰老和AD中，GFAP呈现出复杂的模式，具有不同的修饰和降解形式。免疫组织化学表明，反应性星形胶质细胞主要聚集在内嗅皮质深层的神经原纤维缠结和老年斑周围。GFAP水平会随年龄变化，并对老年斑和缠结的形成产生反应，因此可作为评估神经退行性疾病的辅助而非唯一诊断工具。
