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St. Luke’s-Roosevelt Hospital Center, Departments of
* Physiology and Cellular Biophysics, and
Medicine, College of Physicians and Surgeons, Columbia University, New York, NY 10019;
Departments of Pediatrics, Microbiology-Immunology and Pathology, Dalhousie University, Halifax, Nova Scotia, Canada
Cytokine-induced lung expression of the endothelial cell (EC)
leukocyte receptor P-selectin initiates leukocyte rolling. To
understand the early EC signaling that induces the expression, we
conducted real-time digital imaging studies in lung venular
capillaries. To compare receptor- vs nonreceptor-mediated effects, we
infused capillaries with respectively, TNF-
and arachidonate. At
concentrations adjusted to give equipotent increases in the cytosolic
Ca2+, both agents increased reactive oxygen species (ROS)
production and EC P-selectin expression. Blocking the cytosolic
Ca2+ increases abolished ROS production; blocking ROS
production abrogated P-selectin expression. TNF-, but not
arachidonate, released Ca2+ from endoplasmic stores and
increased mitochondrial Ca2+. Furthermore, Ca2+
depletion abrogated TNF- responses partially, but arachidonate
responses completely. These differences in Ca2+
mobilization by TNF- and arachidonate were reflected in spatial
patterning in the capillary in that the TNF- effects were localized
at branch points, while the arachidonate effects were nonlocalized and
extensive. Furthermore, mitochondrial blockers inhibited the TNF--
but not the arachidonate-induced responses. These findings indicate
that the different modes of Ca2+ mobilization determined
the spatial patterning of the proinflammatory response in lung
capillaries. Responses to TNF- revealed that EC mitochondria
regulate the proinflammatory process by generating ROS that activate
P-selectin expression.
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