脑区特异性单分子荧光成像
Brain Region Specific Single-Molecule Fluorescence Imaging.
机构信息
Department of Chemistry , University of Kentucky , Lexington , Kentucky 40506 , United States.
Department of Physiology , University of Kentucky , Lexington , Kentucky 40536 , United States.
出版信息
Anal Chem. 2019 Aug 6;91(15):10125-10131. doi: 10.1021/acs.analchem.9b02133. Epub 2019 Jul 24.
Abstract
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 α4β2 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.
摘要
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中蛋白质分布的动态。该方法用于确定尼古丁对不同小鼠大脑区域中受体数量分布的影响。尼古丁诱导的α4β2 烟碱型乙酰胆碱受体 (nAChR) 的上调与它们的表达、运输和数量分布的变化有关。使用单分子技术，已经在基于细胞培养的系统中对 nAChRs 的结构组装进行了定量。然而，这些方法无法定量测量发生在活体动物中的生物分子组装。尼古丁诱导的上调和 nAChR 数量分布的变化在不同的大脑区域都有所不同。我们的单分子方法表明，尼古丁在不同的大脑区域以不同的方式作用，以改变组装来响应暴露和戒断。
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