Immune Netw.  2016 Dec;16(6):317-321. 10.4110/in.2016.16.6.317.

Neutrophil Extravasation Cascade: What Can We Learn from Two-photon Intravital Imaging?

Affiliations
  • 1Department of Anatomy, Yonsei University College of Medicine, Seoul 03722, Korea. ymhyun@yuhs.ac

Abstract

Immune cells (leukocytes or white blood cells) move actively and sensitively based on body conditions. Despite their important role as protectors inside the body, it is difficult to directly observe the spatiotemporal momentum of leukocytes. With advances in imaging technology, the introduction of two-photon microscopy has enabled researchers to look deeper inside tissues in a three-dimensional manner. In observations of immune cell movement along the blood vessel, vascular permeability and innate immune cell movements remain unclear. Here, we describe the neutrophil extravasation cascade, which were observed using a two-photon intravital imaging technique. We also provide evidence for novel mechanisms such as neutrophil body extension and microparticle formation as well as their biological roles during migration.

Keyword

Neutrophil extravasation; Microparticle formation; Two-photon microscopy; Intravital imaging

MeSH Terms

Blood Vessels
Capillary Permeability
Cell Movement
Leukocytes
Microscopy
Neutrophils*

Figure

  • Figure 1 Schema of neutrophil extravasation cascade. In the process of intravascular migration, neutrophil tethers itself to the endothelial cells via (1) rolling, (2) adhesion and (3) crawling. Thereafter neutrophil is (4) firmly adhered to the luminal surface of the vessel. After approaching to the proper site of extravasation, (5) leukocyte transmigrate through the endothelial cells, pericyte and basement membrane. In this process of extravasation, neutrophil undergoes (a) intrusion, (b) perivascular embedment & crawling, (c) protrusion, and then finally (d) uropod elongation & microparticle formation. Microparticles (red dot) are formed in this stage, and usually embedded between endothelial cells and pericytes. When extravasation is over, (6) leukocyte starts interstitial migration.

  • Figure 2 Uropod elongation and microparticle formation during neutrophil extravasation. The cremaster muscle of WT mouse was stimulated by CXCL2 post to intravenous injection of Alexa Fluor 488-anti-Gr1 antibody and Texas Red-dextran. The representative image shows uropod elongation of an extravasating neutrophil (dotted line with arrow) and deposited microparticles at the subendothelium (arrows) from neutrophils.


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