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J Biol Chem, Vol. 274, Issue 1, 175-182, January 1, 1999
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From the We present evidence in astrocytes that
5,6-epoxyeicosatrienoic acid, a cytochrome P450 epoxygenase metabolite
of arachidonic acid, may be a component of calcium influx factor, the
elusive link between release of Ca2+ from
intracellular stores and capacitative Ca2+ influx.
Capacitative influx of extracellular Ca2+ was inhibited by
blockade of the two critical steps in epoxyeicosatrienoic acid
synthesis: release of arachidonic acid from phospholipid stores by
cytosolic phospholipase A2 and cytochrome P450 metabolism of arachidonic acid. AAOCF3, which inhibits cytosolic
phospholipase A2, blocked thapsigargin-stimulated release
of arachidonic acid as well as thapsigargin-stimulated elevation of
intracellular free calcium. Inhibition of P450 arachidonic acid
metabolism with SKF525A, econazole, or
N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide, a
substrate inhibitor of P450 arachidonic acid metabolism, also blocked
thapsigargin-stimulated Ca2+ influx. Nano- to picomolar
5,6-epoxyeicosatrienoic acid induced [Ca2+]i
elevation consistent with capacitative Ca2+ influx. We have
previously shown that 5,6-epoxyeicosatrienoic acid is synthesized and
released by astrocytes. When 5,6-epoxyeicosatrienoic acid was applied
to the rat brain surface, it induced vasodilation, suggesting that
calcium influx factor may also serve a paracrine function. In summary,
our results suggest that 5,6-epoxyeicosatrienoic acid may be a
component of calcium influx factor and may participate in regulation of
cerebral vascular tone.
Department of Pharmacology and Toxicology,
Medical College of Virginia, Virginia Commonwealth University,
Richmond, Virginia 23298-0613 and the ¶ Department of Biochemistry
and Pharmacology, University of Texas, Southwestern Medical Center,
Dallas, Texas 75235
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