PUBLICATION
Frontline Science: Dynamic cellular and subcellular features of migrating leukocytes revealed by in vivo lattice lightsheet microscopy
- Authors
- Manley, H.R., Potter, D.L., Heddleston, J.M., Chew, T.L., Keightley, M.C., Lieschke, G.J.
- ID
- ZDB-PUB-200424-4
- Date
- 2020
- Source
- Journal of Leukocyte Biology 108(2): 455-468 (Journal)
- Registered Authors
- Keightley, M. Cristina, Lieschke, Graham J., Manley, Harriet
- Keywords
- lattice lightsheet microscopy, leukocytes, migration, phagocytes, uropod, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Biomarkers
- Cell Adhesion
- Cell Death
- Chemotaxis, Leukocyte/physiology*
- Endothelium, Vascular/metabolism
- Fluorescent Antibody Technique
- Intravital Microscopy*/methods
- Leukocytes/cytology*
- Leukocytes/physiology*
- Macrophages/cytology
- Macrophages/physiology
- Models, Biological
- Neutrophils/cytology
- Neutrophils/physiology
- Phagocytosis
- Zebrafish
- PubMed
- 32323898 Full text @ J. Leukoc. Biol.
Citation
Manley, H.R., Potter, D.L., Heddleston, J.M., Chew, T.L., Keightley, M.C., Lieschke, G.J. (2020) Frontline Science: Dynamic cellular and subcellular features of migrating leukocytes revealed by in vivo lattice lightsheet microscopy. Journal of Leukocyte Biology. 108(2):455-468.
Abstract
Neutrophil and macrophage (Mϕ) migration underpin the inflammatory response. However, the fast velocity, multidirectional instantaneous movement, and plastic, ever-changing shape of phagocytes confound high-resolution intravital imaging. Lattice lightsheet microscopy (LLSM) captures highly dynamic cell morphology at exceptional spatiotemporal resolution. We demonstrate the first extensive application of LLSM to leukocytes in vivo, utilizing optically transparent zebrafish, leukocyte-specific reporter lines that highlighted subcellular structure, and a wounding assay for leukocyte migration. LLSM revealed details of migrating leukocyte morphology, and permitted intricate, volumetric interrogation of highly dynamic activities within their native physiological setting. Very thin, recurrent uropod extensions must now be considered a characteristic feature of migrating neutrophils. LLSM resolved trailing uropod extensions, demonstrating their surprising length, and permitting quantitative assessment of cytoskeletal contributions to their evanescent form. Imaging leukocytes in blood vessel microenvironments at LLSM's spatiotemporal resolution displayed blood-flow-induced neutrophil dynamics and demonstrated unexpected leukocyte-endothelial interactions such as leukocyte-induced endothelial deformation against the intravascular pressure. LLSM of phagocytosis and cell death provided subcellular insights and uncovered novel behaviors. Collectively, we provide high-resolution LLSM examples of leukocyte structures (filopodia lamellipodia, uropod extensions, vesicles), and activities (interstitial and intravascular migration, leukocyte rolling, phagocytosis, cell death, and cytoplasmic ballooning). Application of LLSM to intravital leukocyte imaging sets the stage for transformative studies into the cellular and subcellular complexities of phagocyte biology.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping