Visualization of cell death in mice with focal cerebral ischemia using fluorescent annexin A5, propidium iodide, and TUNEL staining

Peyman Bahmani, Eyk Schellenberger, Jan Klohs, Jens Steinbrink, Ryan Cordell, Marietta Zille, Jochen Mueller, Denise Harhausen, Leo Hofstra, Chris P. M. Reutelingsperger, Tracy Deanne Farr, Ulrich Dirnagl, Andreas Wunder*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

33 Citations (Web of Science)

Abstract

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
Original languageEnglish
Pages (from-to)1311-1320
JournalJournal of Cerebral Blood Flow and Metabolism
Volume31
Issue number5
DOIs
Publication statusPublished - May 2011

Keywords

  • annexin A5
  • cell death
  • fluorescence imaging
  • propidium iodide
  • stroke
  • TUNEL staining

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