New Fluorescent Probes Targeting the Mitochondrial-Located Translocator Protein 18 kDa (TSPO) as Activated Microglia Imaging Agents
New Fluorescent Probes Targeting the Mitochondrial-Located Translocator Protein 18 kDa (TSPO) as Activated Microglia Imaging Agents
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DOI：
10.1007/s11095-011-0552-0
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发表时间：
2011-11-01
影响因子：
3.7
通讯作者：
Trapani, Giuseppe
中科院分区：
文献类型：
--
作者：
Denora, Nunzio;Laquintana, Valentino;Trapani, Giuseppe
为了评价新型转位蛋白18 kDa(TSPO)靶向荧光探针在活体小胶质细胞分子成像中的应用，合成了化合物2-4，并测定了它们对TSPO的稳定性和亲和力。化合物2-4与RA2细胞在脂多糖(一种有效的小胶质细胞激活剂)存在下以及与正常和化学损伤的脑组织切片孵育。将化合物2-4注射到小鼠颈动脉或直接注射到纹状体。这些体外和体内研究的细胞和组织切片经组织化学处理后在荧光显微镜下观察，化合物2-4在缓冲液和生理介质中都很稳定，对TSPO具有很高的亲和力，并被发现能有效地染色活的Ra2小胶质细胞。Mito Tracker Red双重染色表明，化合物2和3的结合部位可能存在于线粒体上。体内研究表明，化合物2-4可以部分渗透到脑内，并且小鼠纹状体细胞被化合物2-4和小胶质细胞标记物CD11b染色，化合物2-4在体外和体内都能荧光标记激活的小胶质细胞。
To evaluate the utility of new Translocator protein 18 kDa (TSPO)-targeted fluorescent probes for in vivo molecular imaging of activated microglia.Compounds 2-4 were synthesized; their stability and affinity for TSPO were determined. Compounds 2-4 were incubated both with Ra2 cells in the presence of LPS, a potent activator of microglia, and with tissue sections of normal and chemically injured brains. Compounds 2-4 were injected into carotid artery or directly in striatum of mice. Cells and tissue sections from these in vitro and in vivo studies were observed by fluorescence microscopy after histochemical treatments.Compounds 2-4 are stable in both buffer and physiological medium and showed high affinity for TSPO and were found to stain live Ra2 microglial cells effectively. Double staining with Mito Tracker Red suggested that binding sites of compounds 2 and 3 may exist on mitochondria. In vivo studies showed that compounds 2-4 may penetrate in part into brain; moreover, cells in mouse striatum were stained with compounds 2-4 and microglial marker CD11b.Compounds 2-4 can fluorescently label activated microglia in vitro and in vivo.