Journal
JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM
Volume 31, Issue 5, Pages 1311-1320Publisher
SAGE PUBLICATIONS INC
DOI: 10.1038/jcbfm.2010.233
Keywords
annexin A5; cell death; fluorescence imaging; propidium iodide; stroke; TUNEL staining
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Funding
- TSB Technologiestiftung Berlin-Zukunftsfonds Berlin
- European Union [201024, 202213]
- Bundesministerium fur Bildung und Forschung (BMBF)
- German Research Foundation
- Herman and Lilly Schilling Foundation
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To monitor stroke-induced brain damage and assess neuroprotective therapies, specific imaging of cell death after cerebral ischemia in a noninvasive manner is highly desirable. Annexin A5 has been suggested as a marker for imaging cell death under various disease conditions including stroke. In this study, C57BL6/N mice received middle cerebral artery occlusion (MCAO) and were injected intravenously with either active or inactive Cy5.5-annexin A5 48 hours after reperfusion. Some mice also received propidium iodide (PI), a cell integrity marker. Only in mice receiving active Cy5.5-annexin A5 were fluorescence intensities significantly higher over the hemisphere ipsilateral to MCAO than on the contralateral side. This was detected noninvasively and ex vivo 4 and 8 hours after injection. The majority of cells positive for fluorescent annexin A5 were also positive for PI and fragmented DNA as detected by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate-biotin nick end labeling (TUNEL) staining. This study demonstrates the high specificity of annexin A5 for visualization of cell death in a mouse model of stroke. To our knowledge, this is the first study to compare the distribution of injected active and inactive annexin A5, PI, and TUNEL staining. It provides important information on the experimental and potential clinical applications of annexin A5-based imaging agents in stroke. Journal of Cerebral Blood Flow & Metabolism (2011) 31, 1311-1320; doi:10.1038/jcbfm.2010.233; published online 19 January 2011
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