Bozyczko D, Horwitz A F
J Neurosci. 1986 May;6(5):1241-51. doi: 10.1523/JNEUROSCI.06-05-01241.1986.
We have used the CSAT (cell substrate attachment) monoclonal antibody (Mab), which is directed against a putative laminin and fibronectin receptor, to examine its role in the adhesive phenomena of neurons. This antibody was previously found to disturb the adhesion of several classes of fibroblasts and muscle. Here we report its effects upon neuronal-substrate adhesion. Two sources of neurons were investigated--the dorsal root and ciliary ganglia. Both responded similarly. Neurons plated in the presence of the CSAT Mab did not adhere to the substratum and process formation was inhibited completely for at least 24-48 hr. In explant cultures, when neurons were first allowed to extend processes prior to addition of the CSAT Mab, the results depended on the particular substrate. With some substrates, the neurites bundled and detached from the substratum; with others, they retracted and regrew to form large fascicles or bundles of processes. In dissociated cultures that already had extended processes, neurites fasciculated and cell bodies aggregated in response to the presence of the CSAT Mab. The magnitude of this response varied, depending upon the substrate. The antigen was localized, using immunofluorescence, on neuronal cell bodies, axons, and growth cones. This distribution correlated with its biological effects on all parts of the neuron. The antigen was isolated from neuronal cultures by immunoaffinity purification. It migrated in the molecular weight range of 140 kDa on reducing SDS-PAGE. This antigen is very similar to that isolated from fibroblasts, which is an integral membrane glycoprotein complex. The data presented implicate the participation of the CSAT antigen in neurite extension and fasciculation.
我们使用了CSAT（细胞底物附着）单克隆抗体（Mab），它针对一种假定的层粘连蛋白和纤连蛋白受体，来研究其在神经元黏附现象中的作用。先前发现这种抗体可干扰几类成纤维细胞和肌肉的黏附。在此我们报告其对神经元 - 底物黏附的影响。研究了两种神经元来源——背根神经节和睫状神经节。两者反应相似。在CSAT Mab存在的情况下接种的神经元不黏附于底物，并且至少在24 - 48小时内突起形成完全受到抑制。在外植体培养中，当神经元在添加CSAT Mab之前先被允许伸出突起时，结果取决于特定的底物。对于一些底物，神经突会成束并从底物上脱离；对于其他底物，它们会缩回并重新生长形成大的束状或突起束。在已经伸出突起的解离培养物中，神经突会成束，并且细胞体会因CSAT Mab的存在而聚集。这种反应的程度有所不同，取决于底物。使用免疫荧光法将抗原定位在神经元细胞体、轴突和生长锥上。这种分布与其对神经元所有部分的生物学效应相关。通过免疫亲和纯化从神经元培养物中分离出该抗原。在还原SDS - PAGE上，它在分子量范围为140 kDa处迁移。这种抗原与从成纤维细胞中分离出的抗原非常相似，是一种整合膜糖蛋白复合物。所呈现的数据表明CSAT抗原参与了神经突的延伸和成束过程。
