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首页> 外文期刊>Life sciences >Detection of alpha7 nicotinic acetylcholine receptors with the aid of antibodies and toxins.
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Detection of alpha7 nicotinic acetylcholine receptors with the aid of antibodies and toxins.

机译：借助抗体和毒素检测α7烟碱型乙酰胆碱受体。

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摘要

Neuronal nicotinic acetylcholine receptors (nAChRs) containing alpha7 subunit are well represented in the brain and some non-neuronal tissues, and their malfunctioning is associated with diverse pathologies. Therefore, detection and quantification of alpha7 nAChR are important tasks. The affinity-purified antibodies were prepared against the 1-23 and 179-190 fragments of the human and rat alpha7 nAChR extracellular domain. The specificity and selectivity of these alpha7 (1-23) and alpha7 (179-190) antibodies was tested by ELISA in model systems: the E. coli-expressed alpha7 subunit extracellular domain and the pituitary cell line GH(4)C(1) stably expressing human alpha7 nAChR. On the rat brain slices two antibodies and biotinylated alpha-cobratoxin specifically stained the hippocampus region known to be rich in alpha7 nAChR. Western blot analysis revealed that in the human thalamus membranes and in rat brain membranes, antibodies alpha7 (1-23) stained a single band of 62 kDa, while the alpha7 (179-190)antibodies stained a doublet of 53-54 kDa. The results obtained show that utilization of model systems and a combination of several antibodies with appropriately labeled toxins may provide better ways for detection of alpha7 nAChR.
机译：含有alpha7亚基的神经元烟碱型乙酰胆碱受体（nAChRs）在大脑和一些非神经元组织中得到了很好的表现，其功能异常与多种病理相关。因此，α7nAChR的检测和定量是重要的任务。制备了针对人和大鼠alpha7 nAChR细胞外域的1-23和179-190片段的亲和纯化抗体。这些alpha7（1-23）和alpha7（179-190）抗体的特异性和选择性通过ELISA在以下模型系统中测试：大肠杆菌表达的alpha7亚基胞外域和垂体细胞系GH（4）C（1 ）稳定表达人alpha7 nAChR。在大鼠脑切片上，两种抗体和生物素化的α-眼镜蛇毒素特异性染色了已知富含α7nAChR的海马区。 Western印迹分析表明，在人丘脑膜和大鼠脑膜中，抗体alpha7（1-23）染色了一条62 kDa的单条带，而alpha7（179-190）抗体则染色了53-54 kDa的双峰。获得的结果表明，利用模型系统以及几种抗体与适当标记的毒素的组合可能为检测α7nAChR提供更好的方法。

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