Direct In Vivo Manipulation and Imaging of Calcium Transients in Neutrophils Identify a Critical Role for Leading-Edge Calcium Flux

被引:40
作者
Beerman, Rebecca W. [1 ]
Matty, Molly A. [1 ,2 ]
Au, Gina G. [1 ]
Looger, Loren L. [3 ]
Choudhury, Kingshuk Roy [4 ]
Keller, Philipp J. [3 ]
Tobin, David M. [1 ,5 ]
机构
[1] Duke Univ, Dept Mol Genet & Microbiol, Durham, NC 27710 USA
[2] Duke Univ, Univ Program Genet & Genom, Durham, NC 27710 USA
[3] Howard Hughes Med Inst, Ashburn, VA 20147 USA
[4] Duke Univ, Dept Biostat & Bioinformat, Durham, NC 27710 USA
[5] Duke Univ, Dept Immunol, Durham, NC 27710 USA
基金
美国国家科学基金会;
关键词
PSEUDOMONAS-AERUGINOSA INFECTION; CA2+ INFLUX; CELL-MIGRATION; ZEBRAFISH; PHAGOCYTOSIS; MOTILITY; RECEPTOR; POLARITY; CHANNEL; MODEL;
D O I
10.1016/j.celrep.2015.11.010
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Calcium signaling has long been associated with key events of immunity, including chemotaxis, phagocytosis, and activation. However, imaging and manipulation of calcium flux in motile immune cells in live animals remain challenging. Using light-sheet microscopy for in vivo calcium imaging in zebrafish, we observe characteristic patterns of calcium flux triggered by distinct events, including phagocytosis of pathogenic bacteria and migration of neutrophils toward inflammatory stimuli. In contrast to findings from ex vivo studies, we observe enriched calcium influx at the leading edge of migrating neutrophils. To directly manipulate calcium dynamics in vivo, we have developed transgenic lines with cell-specific expression of the mammalian TRPV1 channel, enabling ligand-gated, reversible, and spatiotemporal control of calcium influx. We find that controlled calcium influx can function to help define the neutrophil's leading edge. Cell-specific TRPV1 expression may have broad utility for precise control of calcium dynamics in other immune cell types and organisms.
引用
收藏
页码:2107 / 2117
页数:11
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