Single-cell imaging of retinal ganglion cell apoptosis with a cell-penetrating, activatable peptide probe in an in vivo glaucoma model

被引:100
作者
Barnett, Edward M. [3 ]
Zhang, Xu [3 ]
Maxwell, Dustin [1 ,2 ]
Chang, Qing [3 ]
Piwnica-Worms, David [1 ,2 ]
机构
[1] Washington Univ, Sch Med, Mol Imaging Ctr, Mallinckrodt Inst Radiol, St Louis, MO 63110 USA
[2] Washington Univ, Sch Med, Dept Dev Biol, St Louis, MO 63110 USA
[3] Washington Univ, Sch Med, Dept Ophthalmol & Visual Sci, St Louis, MO 63110 USA
基金
美国国家卫生研究院;
关键词
caspase; molecular imaging; near-infrared fluorescence; RED FLUORESCENT PEPTIDE; RAT RETINA; PROTEIN TRANSDUCTION; MEMBRANE-PERMEANT; DEATH; INHIBITION; MATRIX-METALLOPROTEINASE-9; NEURODEGENERATION; DEGENERATION; TRANSPORTERS;
D O I
10.1073/pnas.0812884106
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Molecular imaging probes have potential for in vivo identification of apoptosis and other intracellular processes. TcapQ, a cell-penetrating, near-infrared fluorescent peptide probe designed to be optically silent through intramolecular fluorescence quenching and activated by effector caspases, has been previously described and validated in vitro. Herein, using NMDA-induced apoptosis of retinal ganglion cells (RGCs), representing an in vivo rat model of glaucoma, we assessed the ability of TcapQ to image single-cell apoptosis through effector caspase activity. Following intravitreal injection, intracellular TcapQ activation occurred specifically in RGCs, identified individual apoptotic cells, showed a clear dose-response relationship with NMDA, and colocalized with TUNEL labeling in the retina. There was a significant diminution of probe activation following pretreatment with a specific inhibitor of caspase-3. Stereospecificity was also exhibited by the lack of intracellular fluorescence upon administration of the noncleavable isomer, dTcapQ. TcapQ has potential utility in detecting and monitoring single-cell apoptosis in glaucoma in vivo.
引用
收藏
页码:9391 / 9396
页数:6
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