title: Live imaging of apoptotic cells in zebrafish.
abstract: Many debilitating diseases, including neurodegenerative diseases, involve apoptosis. Several methods have been developed for visualizing apoptotic cells in vitro or in fixed tissues, but few tools are available for visualizing apoptotic cells in live animals. Here we describe a genetically encoded fluorescent reporter protein that labels apoptotic cells in live zebrafish embryos. During apoptosis, the phospholipid phosphatidylserine (PS) is exposed on the outer leaflet of the plasma membrane. The calcium-dependent protein Annexin V (A5) binds PS with high affinity, and biochemically purified, fluorescently labeled A5 probes have been widely used to detect apoptosis in vitro. Here we show that secreted A5 fused to yellow fluorescent protein specifically labels apoptotic cells in living zebrafish. We use this fluorescent probe to characterize patterns of apoptosis in living zebrafish larvae and to visualize neuronal cell death at single-cell resolution in vivo.
authors.affiliation: Cardiovascular Research Center, Division of Cardiology, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA.
authors.forename: Tjakko J
authors.initials: TJ
authors.lastname: van Ham
authors.forename: James
authors.initials: J
authors.lastname: Mapes
authors.forename: David
authors.initials: D
authors.lastname: Kokel
authors.forename: Randall T
authors.initials: RT
authors.lastname: Peterson
mesh_terms: D000818:Animals; D017209:Apoptosis; D005456:Fluorescent Dyes / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D008853:Microscopy / Q000379:methods*; D015335:Molecular Probes / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology*
chemical_list: D005456:Fluorescent Dyes; D015335:Molecular Probes