© 1995 American Heart Association, Inc.
Articles |
Simultaneous Measurements of Ca2+ and Nitric Oxide in Bradykinin-Stimulated Vascular Endothelial Cells
From the Department of Physiology (L.A.B.), Loyola University Chicago, Maywood, Ill; the Department of Chemistry and Institute of Biotechnology (Z.T., S.M., T.M.), Oakland University, Rochester, Mich; and the Department of Physiology (P.S.S., W.G.W.), University of Maryland, Baltimore, Md.
Correspondence to Tadeusz Malinski, PhD, Department of Chemistry, Institute of Biotechnology, Oakland University, Rochester, MI 48309-4401.
Abstract The production of
endothelium-derived relaxing factor (EDRF), known to be
nitric oxide (NO), is triggered by a rise in the cytoplasmic calcium
concentration ([Ca2+]i) subsequent to
receptor binding of vasoactive agonists. In vascular endothelial cells,
NO is synthesized from L-arginine by the
Ca2+/calmodulin–dependent NO synthase. In
this study, we report the first simultaneous measurements of
[Ca2+]i and [NO] at the level of
single endothelial cells. In cultured bovine aortic endothelial cells,
extracellular application of bradykinin (BK, 10 to 20 µmol/L) caused
transient (sometimes oscillatory) increase in
[Ca2+]i, which was measured
with the fluorescent Ca2+ indicator fura 2 and
fluorescence imaging microscopy. BK caused an increase in
[Ca2+]i, primarily through
release from intracellular stores. Under identical experimental
conditions, BK caused a transient increase in [NO], which was
measured by application of a porphyrinic NO microsensor. [NO] peaked
at 
0.5 µmol/L. Simultaneous measurements of
[Ca2+]i and [NO] in BK-stimulated
endothelial cells revealed that a transient increase in
[Ca2+]i was rapidly followed by an
increase in [NO] that outlasted the
[Ca2+]i transient.
Key Words: endothelium-derived relaxation factor/nitric oxide • bradykinin • cytoplasmic calcium concentration • porphyrinic NO microsensor • fura 2
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