Premium
Live imaging of apoptotic cells in zebrafish
Author(s) -
Ham Tjakko J.,
Mapes James,
Kokel David,
Peterson Randall T.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.10-161018
Subject(s) - zebrafish , apoptosis , phosphatidylserine , annexin , microbiology and biotechnology , biology , programmed cell death , annexin a5 , live cell imaging , in vitro , in vivo , green fluorescent protein , phospholipid scramblase , cell , phospholipid , membrane , biochemistry , gene , genetics
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 .—Van Ham, T. J., Mapes, J., Kokel, D., Peterson, R. T. Live imaging of apoptotic cells in zebrafish. FASEB J . 24, 4336–4342 (2010). www.fasebj.org