Live-cell vibrational imaging of choline metabolites by stimulated Raman scattering coupled with isotope-based metabolic labeling.
Live-cell vibrational imaging of choline metabolites by stimulated Raman scattering coupled with isotope-based metabolic labeling.
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DOI：
10.1039/c3an02281a
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发表时间：
2014-05-21
期刊：
影响因子：
--
通讯作者：
Min W
中科院分区：
文献类型：
--
作者：
Hu F;Wei L;Zheng C;Shen Y;Min W
胆碱是一种小分子，在所有生物体的生物化学中占据关键地位。最近的研究表明胆碱代谢物与癌症、动脉粥样硬化和神经系统发育密切相关。为了检测胆碱及其代谢物，现有的物理方法（例如磁共振波谱法和正电子发射断层扫描）往往受到空间分辨率差和辐射剂量大的限制。荧光成像虽然具有亚微米分辨率，但需要引入大量荧光团，因此很难标记小胆碱分子。通过将新兴的键选择性受激拉曼散射显微镜与氘代胆碱的代谢掺入相结合，我们在活哺乳动物细胞系、原代海马神经元和多细胞生物秀丽隐杆线虫中实现了含胆碱代谢物的高分辨率成像。在癌细胞和非癌细胞之间观察到胆碱代谢物的不同亚细胞分布，这可能揭示胆碱代谢和脂质介导的信号事件的功能差异。在神经元中，胆碱掺入在胞体和神经突中可见，与蛋白质相比，胆碱代谢物在其中分布更均匀。此外，在线虫幼虫的咽部区域也观察到胆碱定位，与其器官发生机制一致。这些应用证明了基于同位素的受激拉曼散射显微镜在未来体内胆碱相关疾病检测和发展监测中的潜力。
Choline is a small molecule that occupies a key position in the biochemistry of all living organisms. Recent studies have strongly implicated choline metabolites in cancer, atherosclerosis and nervous system development. To detect choline and its metabolites, existing physical methods such as magnetic resonance spectroscopy and positron emission tomography, are often limited by the poor spatial resolution and substantial radiation dose. Fluorescence imaging, although with submicrometer resolution, requires introduction of bulky fluorophores and thus is difficult in labeling the small choline molecule. By combining the emerging bond-selective stimulated Raman scattering microscopy with metabolic incorporation of deuterated choline, herein we have achieved high resolution imaging of choline-containing metabolites in living mammalian cell lines, primary hippocampal neurons and multicellular organism C. elegans. Different subcellular distributions of choline metabolites are observed between cancer cells and non-cancer cells, which may reveal functional difference in the choline metabolism and lipid-mediated signaling events. In neurons, choline incorporation is visualized within both soma and neurites, where choline metabolites are more evenly distributed compared to the protein. Furthermore, choline localization is also observed in the pharynx region of C. elegans larvae, consistent with its organogenesis mechanism. These applications demonstrate the potential of isotope-based stimulated Raman scattering microscopy for future choline-related disease detection and development monitoring in vivo.
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影响因子：
48
作者：
Wang, Meng C.;Min, Wei;Freudiger, Christian W.;Ruvkun, Gary;Xie, X. Sunney
通讯作者：
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影响因子：
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作者：
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影响因子：
3.3
作者：
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通讯作者：
Volkmer, A.
DOI：
10.1073/pnas.1215689109
发表时间：
2012-12-26
影响因子：
11.1
作者：
Craciun, Smaranda;Balskus, Emily P.
通讯作者：
Balskus, Emily P.
磷脂酰胆碱的肠道菌群代谢促进心血管疾病。
影响因子：
64.8
作者：
Wang, Zeneng;Klipfell, Elizabeth;Bennett, Brian J.;Koeth, Robert;Levison, Bruce S.;Dugar, Brandon;Feldstein, Ariel E.;Britt, Earl B.;Fu, Xiaoming;Chung, Yoon-Mi;Wu, Yuping;Schauer, Phil;Smith, Jonathan D.;Allayee, Hooman;Tang, W. H. Wilson;DiDonato, Joseph A.;Lusis, Aldons J.;Hazen, Stanley L.
通讯作者：
Hazen, Stanley L.