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1 Stroke and Neurovascular Regulation Laboratory, Departments of Neurosurgery and Neurology and 2 Cardiovascular Research Center, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129
We evaluated the effects of superfusing
1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one
(ODQ), eNOS null (B)an inhibitor of
soluble guanylyl cyclase, and 7-nitroindazole sodium (7-NI), a
selective neuronal nitric oxide synthase (nNOS) inhibitor, on the
acetylcholine (ACh) response in endothelial NOS (eNOS) null
mice. Pial arteriolar diameter was measured by intravital
microscopy through a closed cranial window under 
-chloralose
anesthesia. NOS activity was measured by
[3H]arginine-to-[3H]citrulline
conversion in subjacent cortex in vitro. The density and distribution
of muscarinic receptors in the brain were determined by quantitative
[3H]quinuclidinyl
benzilate autoradiography and did not differ between the eNOS mutants
and wild-type mice. ACh superfusion (1 and 10 µM) dose dependently
dilated pial arterioles in eNOS null and wild-type mice. ODQ (10 µM)
attenuated ACh-induced dilation in both eNOS mutants (41% decrease at
10 µM ACh, P < 0.01, n = 6) and wild-type strains
(n = 5 per group). By contrast,
topical superfusion of 7-NI (100 µM) attenuated the ACh response in
eNOS mutants only (66%, P < 0.05, and 25% decrease, P < 0.05, at 1 and 10 µM ACh, respectively). Our findings suggest that
nNOS-guanosine 3',5'-cyclic monophosphate (cGMP)-dependent
pathways dilate pial arterioles by compensatory mechanisms after eNOS
gene disruption.
endothelial nitric oxide synthase; neuronal nitric oxide synthase; soluble guanylyl cyclase; mutant mice; acetylcholine; cerebral circulation; closed cranial window
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