A Novel Method for Imaging Apoptosis Using a Caspase-1 Near-Infrared Fluorescent Probe
View/ Open
71600 neo0602_0095.pdf (1.169Mb)
Access Status
Full text of the requested work is not available in DASH at this time ("restricted access"). For more information on restricted deposits, see our FAQ.Author
Messerli, Shanta M.
Prabhakar, Shilpa
Tang, Yi
Shah, Khalid
Cortes, Maria L.
Murthy, Vidya
Weissleder, Ralph
Breakefield, Xandra O.
Tung, Ching-Hsuan
Published Version
https://doi.org/10.1593/neo.03214Metadata
Show full item recordCitation
Messerli, Shanta M., Shilpa Prabhakar, Yi Tang, Khalid Shah, Maria L. Cortes, Vidya Murthy, Ralph Weissleder, Xandra O. Breakefield, and Ching-Hsuan Tung. 2004. “A Novel Method for Imaging Apoptosis Using a Caspase-1 Near-Infrared Fluorescent Probe.” Neoplasia 6 (2): 95–105. https://doi.org/10.1593/neo.03214.Abstract
Here we describe a novel method for imaging apoptosis in cells using a near-infrared fluorescent (NRF) probe selective for caspase-1 (interleukin 1beta-converting enzyme, ICE). This biocompatible, optically quenched ICE-NIRF probe incorporates a peptide substrate, which can be selectively cleaved by caspase-1, resulting in the release of fluorescence signal. The specificity of this probe for caspase-1 is supported by various lines of evidence: 1) activation by purified caspase-1, but not another caspase in vitro; 2) activation of the probe by infection of cells with a herpes simplex virus amplicon vector (HGC-ICE-lacZ) expressing a catalytically active caspase-1-lacZ fusion protein; 3) inhibition of HGC-ICE-lacZ vector-induced activation of the probe by coincubation with the caspase-1 inhibitor YVAD-cmk, but not with a caspase-3 inhibitor; and 4) activation of the probe following standard methods of inducing apoptosis with staurosporine, ganciclovir, or ionizing radiation in culture. These results indicate that this novel ICE-NIRF probe can be used in monitoring endogenous and vector-expressed caspase-1 activity in cells. Furthermore, tumor implant experiments indicate that this ICE-NIRF probe can be used to detect caspase-1 activity in living animals. This novel ICE-NIRF probe should prove useful in monitoring endogenous and vector-expressed caspase-1 activity, and potentially apoptosis in cell culture and in vivo.Citable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:41384215
Collections
- HMS Scholarly Articles [17852]
Contact administrator regarding this item (to report mistakes or request changes)