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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
来源:
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.
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页码:9391 / 9396
页数:6
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