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pHMA, a pH‐sensitive GFP reporter for cell engulfment, in Drosophila embryos, tissues, and cells
Author(s) -
Fishilevich Elane,
Fitzpatrick James A. J.,
Minden Jonathan S.
Publication year - 2010
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.22180
Subject(s) - microbiology and biotechnology , biology , green fluorescent protein , cell , phagocytosis , reporter gene , apoptosis , embryo , hek 293 cells , cell culture , receptor , biochemistry , gene , genetics , gene expression
Engulfment of apoptotic cells by phagocytosis ensures the removal of unwanted and defective cells. We developed a genetically encoded marker for cell engulfment, pHMA, which consists of the pH‐Sensitive derivative of GFP, pHluorin, fused to the actin‐binding domain of Moesin. In healthy cells of Drosophila embryos and cultured cells, pHMA resides at the cell cortex. In dying cells, pHMA loses its cortical localization and reports a modest decrease in pH. In embryos, the dying cells lose their apical contacts, then move basally and are ultimately engulfed by neighboring cells or macrophages. The cell corpse material is strongly acidified soon after engulfment and persists in the phagocytic cell for several hours. Changes in the pHMA signal correlate well with increases or decreases in apoptosis. These data show that pHMA is a useful reporter for cell engulfment and can be used in screening for mutations that affect cell engulfment. Developmental Dynamics 239:559–573, 2010. © 2009 Wiley‐Liss, Inc.