Brain Region Specific Single-Molecule Fluorescence Imaging
Brain Region Specific Single-Molecule Fluorescence Imaging
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DOI：
10.1021/acs.analchem.9b02133
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发表时间：
2019-08-06
影响因子：
7.4
通讯作者：
Richards, Christopher I.
中科院分区：
文献类型：
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作者：
Fu, Xu;Moonschi, Faruk H.;Richards, Christopher I.
我们开发了一种方法，利用从大脑区域提取的纳米级囊泡结合单分子成像来监测动物的生理状况如何调节不同大脑区域蛋白质分布的动态。采用该方法测定了尼古丁对小鼠不同脑区受体化学计量分布的影响。尼古丁诱导的α 4 β 2烟碱乙酰胆碱受体（nAChRs）的上调与其表达、运输和化学计量的变化有关。nachr的结构组装已经在基于细胞培养的系统中使用单分子技术进行了量化。然而，这些方法不能量化发生在活体动物体内的生物分子组装。尼古丁诱导的nAChR化学计量的上调和变化在不同的脑区是不同的。我们的单分子方法揭示了尼古丁在不同大脑区域的不同行为，以改变暴露和戒断反应的组装。
We developed an approach utilizing nanoscale vesicles extracted from brain regions combined with single molecule imaging to monitor how an animal's physiological condition regulates the dynamics of protein distributions in different brain regions. This method was used to determine the effect of nicotine on the distribution of receptor stoichiometry in different mouse brain regions. Nicotine-induced upregulation of alpha 4 beta 2 nicotinic acetylcholine receptors (nAChRs) is associated with changes in their expression, trafficking, and stoichiometry. The structural assembly of nAChRs has been quantified in cell culture based systems using single molecule techniques. However, these methods are not capable of quantifying biomolecule assembly that takes place in a live animal. Both nicotine-induced upregulation and changes in nAChR stoichiometry differ across brain regions. Our single molecule approach revealed that nicotine acts differentially across brain regions to alter assembly in response to exposure and withdrawal.