Storey E, Beyreuther K, Masters C L
Department of Pathology, University of Melbourne, Parkville, Victoria, Australia.
Brain Res. 1996 Oct 7;735(2):217-31. doi: 10.1016/0006-8993(96)00608-7.
Immunofluorescence on primary dissociated rat neuronal cultures (cortical, hippocampal, and cerebellar) and organotypic hippocampal cultures was used to investigate the pattern of distribution of cell-surface amyloid protein (APP). Antibodies directed against the extracellular (N-terminal) portion of APP or against the entire molecule, but not against the C-terminal portion, revealed a striking segmental pattern of immunoreactivity along both axons and dendrites of all neuronal types tested. The pattern first developed between 24 and 48 h in culture. The segments showed co-localization with beta 1-integrin and talin immunoreactivities, but not with GAP-43 or clathrin, indicating that they may mark adhesion patches. Confocal laser microscopy supported a surface location for the APP responsible for the segmented pattern on neurites, as did the reduction of segmental immunoreactivity after exposure to mu-calpain or trypsin. It is conjectured that APP may have a role in cell-substratum interactions in the medium term, during such events as synaptic plasticity and neurite stability during extension.
利用原代解离大鼠神经元培养物(皮质、海马和小脑)以及海马器官型培养物上的免疫荧光来研究细胞表面淀粉样蛋白(APP)的分布模式。针对APP细胞外(N端)部分或整个分子而非C端部分的抗体,在所有测试的神经元类型的轴突和树突上均显示出明显的节段性免疫反应模式。这种模式在培养24至48小时之间首次出现。这些节段与β1整合素和踝蛋白的免疫反应共定位,但与GAP-43或网格蛋白不共定位,表明它们可能标记黏附斑。共聚焦激光显微镜支持负责神经突节段模式的APP位于表面,暴露于μ-钙蛋白酶或胰蛋白酶后节段性免疫反应性降低也支持这一点。据推测,在诸如突触可塑性和神经突延伸过程中的稳定性等事件期间,APP可能在中期细胞与基质的相互作用中发挥作用。