A novel method for monitoring surface membrane trafficking on hippocampal acute slice preparation
A novel method for monitoring surface membrane trafficking on hippocampal acute slice preparation
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DOI：
10.1016/s0165-0270(03)00050-5
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发表时间：
2003-05-30
影响因子：
3
通讯作者：
Rivera, C
中科院分区：
文献类型：
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作者：
Thomas-Crusells, J;Vieira, A;Rivera, C
蛋白质运输作为神经元可塑性的潜在调节因子引起了相当大的关注。因此，在这方面常用的实验范式中研究蛋白质运输的机制是有意义的。在这里，我们提出了一种在急性海马切片中进行细胞表面蛋白生物素化的方法，这是电生理记录最常用的准备。我们用两个先前表征的细胞表面受体，谷氨酸受体亚基A （GluR A）和转铁蛋白受体（TfR）验证了这一过程。我们观察到谷氨酸依赖于表面GluR a降解的增加，而TfR在使用的时间窗内没有显示出显著的降解。此外，所提出的方法提供了研究内化和再循环等过程的机会，也可以应用于检查正常和病理活动模式对常用电生理记录制剂中膜蛋白运输的影响。(C) 2003 Elsevier Science B.V.版权所有
Protein trafficking has attracted considerable attention as a potential regulator of neuronal plasticity. Therefore, it is of interest to study the mechanism involved in protein trafficking in experimental paradigms commonly used in this context. Here, we present a method for cell surface protein biotinylation in the acute hippocampal slice, the most commonly used preparation for electrophysiological recordings. We validated this procedure with two previously characterized cell surface receptors, glutamate receptor subunit A (GluR A) and the transferrin receptor (TfR). We observed a glutamate-dependent increase in the degradation of surface GluR A, whereas the TfR did not show significant degradation in the time window used. In addition, the presented method offers the opportunity to study processes such as internalisation and recycling, and can also be applied to examine the effect of normal and pathological patterns of activity on membrane protein trafficking in commonly used preparations for electrophysiological recordings. (C) 2003 Elsevier Science B.V. All rights reserved.