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1 Department of Physiology, College of Medicine, University of Arizona, Tucson, Arizona 85724-5051; and 2 Department of Anatomy, University of California, San Francisco, California 94143-0452
Inhibition of nitric oxide (NO) synthesis
using NG-nitro-L-arginine methyl
ester (L-NAME) or
NG-monomethyl-L-arginine
(L-NMMA) increases venular
permeability in the rat mesentery (I. Kurose, R. Wolf, M. B. Grisham,
T. Y. Aw, R. D. Specian, and D. N. Granger. Circ.
Res. 76: 30-39, 1995), but the cellular mechanisms
of this response are not known. This study was performed to determine
whether such venular leaks are associated with changes in the
endothelial actin cytoskeleton. In anesthetized Sprague-Dawley rats,
the microvasculature of a mesenteric window was perfused with buffered
saline, with or without 10
5
M L-NAME,
L-NMMA, or the inactive
enantiomer
NG-nitro-D-arginine
methyl ester for 3 or 30 min. FITC-albumin was added to the perfusate
for the last 3 min. The vasculature was perfusion fixed, stained for
filamentous actin and for mast cells, and viewed microscopically. In
control preparations, venules showed few FITC-albumin leaks and the
endothelial actin cytoskeleton consisted of a peripheral rim along the
cell-cell junctions. Preparations treated with
L-NAME or
L-NMMA showed significantly more
leakage, the actin rims in leaky venules were discontinuous, and short, randomly oriented fibers appeared within the cells. In nonleaky venules, the peripheral actin rims sometimes contained small, equally
spaced discontinuities not seen in control preparations. Although a
mast cell stabilizer was used, 27-70% of the mast cells were
degranulated in the presence of
L-NMMA. Thus inhibition of NO
synthesis alters the endothelial cytoskeleton and increases albumin
leakage from mesenteric venules, either directly or indirectly via the
involvement of mast cells.
rat mesentery; confocal microscopy; endothelium; mast cells
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