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2015 ; 13
(10
): 2107-17
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Direct In Vivo Manipulation and Imaging of Calcium Transients in Neutrophils
Identify a Critical Role for Leading-Edge Calcium Flux
#MMPMID26673320
Beerman RW
; Matty MA
; Au GG
; Looger LL
; Choudhury KR
; Keller PJ
; Tobin DM
Cell Rep
2015[Dec]; 13
(10
): 2107-17
PMID26673320
show ga
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.