TY - JOUR
T1 - pHMA, a pH-sensitive GFP reporter for cell engulfment, in Drosophila embryos, tissues, and cells
AU - Fishilevich, Elane
AU - Fitzpatrick, James A.J.
AU - Minden, Jonathan S.
PY - 2010/2
Y1 - 2010/2
N2 - 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.
AB - 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.
KW - Cell engulfment
KW - Drosophila
KW - pHMA
UR - http://www.scopus.com/inward/record.url?scp=75149113698&partnerID=8YFLogxK
U2 - 10.1002/dvdy.22180
DO - 10.1002/dvdy.22180
M3 - Article
C2 - 20014104
AN - SCOPUS:75149113698
SN - 1058-8388
VL - 239
SP - 559
EP - 573
JO - Developmental Dynamics
JF - Developmental Dynamics
IS - 2
ER -