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Blood, 1 December 2000, Vol. 96, No. 12, pp. 3816-3822
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Endothelial [Ca2+]i signaling during
transmigration of polymorphonuclear leukocytes
Wen-Hong Su,
Hsiun-ing Chen,
Ji-ping Huang, and
Chauying J. Jen
From the Department of Physiology, College of Medicine,
National Cheng-Kung University, Tainan, Taiwan, Republic of China.
Vascular endothelium plays an important role in regulating
the transendothelial migration of polymorphonuclear leukocytes (PMNs).
In this study, the intracellular calcium ion
([Ca2+]i) signaling of endothelial cells
(ECs) during PMN transmigration was examined at the single-cell level.
Human umbilical vein ECs were cultured on a thin layer of collagen gel.
The ECs were labeled with fura-2, immersed in formyl-Met-Leu-Phe, and
subsequently perfused with fresh buffer to establish a gradient of
chemoattractant across the EC monolayer. The entire process of PMN
rolling on, adhering to, and transmigrating across the EC monolayer was
recorded under both phase-contrast and fluorescence optics. The data
showed the following: (1) At high concentration (approximately
3 × 106/mL), both PMN suspension and its supernatant
stimulated frequent EC [Ca2+]i elevations
across the monolayer; (2) when used at lower concentration (approximately 5 × 105/mL) to avoid the interference of
soluble factors, PMN transmigration, but not rolling or adhesion, was
accompanied by EC [Ca2+]i elevation; (3) the
latter EC [Ca2+]i elevation occurred
simultaneously in ECs adjacent to the transmigration site, but not in
those that were not in direct contact with the transmigrating PMNs; (4)
this EC [Ca2+]i elevation was an
initial and required event for PMN transmigration; and (5) PMNs
pretreated with 5,5'-dimethyl-1,2-bis(2-aminophenoxy)ethane-N, N, N',
N'-tetraacetic acid transmigrated with the accompanying EC
[Ca2+]i elevation, but they became elongated
in the collagen gel. In conclusion, PMNs induce adjacent EC
[Ca2+]i signaling, which apparently mediates
the "gating" step for their subsequent transmigration.
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