In response to atherogenic stimuli, blood monocytes transmigrate across the vascular endothelium not only through endothelial cell–cell junctions (para-cellular) but also through endothelial cells themselves (trans-cellular). The molecular mechanism of the latter is mostly unknown, because it rarely happens, especially in vitro. Although many reports have recognized trans-cellular migration from snapshot images of leukocytes halfway across the endothelium at non-junctional locations, it often produces a false-positive result, because some leukocytes that initiate trans-cellular migration withdraw and return to the apical endothelial surface. Thus, analyzing the entire process is essential. In this study, complete monocyte trans-cellular migration was successfully captured for live cells, with simultaneous visualization of endothelial PECAM-1. We suggest the possible existence of both PECAM-1-related migration at peri-junctional sites and PECAM-1-unrelated migration at sites remote from junctions. This is the first report to describe the entire process of monocyte trans-cellular migration for live cells and its relationship with endothelial PECAM-1.
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This work was supported in part by Grant-in-Aid for Scientific Research in Japan (18300156 and 19700391), and grants from Kawasaki Medical School (Project-Research 18-306T and 19-314Y). Ken Hashimoto is also supported by grants from Takeda Science Foundation (Japan) for research on monocyte trans-endothelial migration.
Conflict of interest
The authors declare that they have no conflict of interest.
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Supplemental movie 1. PECAM-1-unrelated trans-cellular migration at sites remote from junctions. Monocyte M1 (red) underwent trans-cellular migration at a site relatively remote (>20 μm) from the nearest junction (arrows at 38–47 min), but related endothelial PECAM-1 (green, GFP-tagged) was not observed. Beneath the endothelium, M1 seemed to be in contact with another monocyte M2 that had previously transmigrated (arrowheads at 43–47 min), and the two monocytes seemed tightly stuck each other at 47 min (arrow and arrowhead). The movie clip was integrated from sequential images in Fig. 2, together with other images from the same sample. Time after monocyte addition in minutes is shown in the lower-right corner. Maximum-projected xy-images toward the z-axis are shown, with xz/yz orthogonal images cut with the specified planes that are indicated as white lines on xy-images. Bar, 20 μm.
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Hashimoto, K., Kataoka, N., Nakamura, E. et al. Live-cell visualization of the trans-cellular mode of monocyte transmigration across the vascular endothelium, and its relationship with endothelial PECAM-1. J Physiol Sci 62, 63–69 (2012). https://doi.org/10.1007/s12576-011-0181-8
- Trans-cellular migration