Expression and function of recombinant S1179D endothelial NO synthase in human pial arteries.
OBJECTIVE
Mutation of serine 1179 to aspartate on the endothelial NO synthase (eNOS) increases NO production in the absence of stimulation by agonists. The present study was designed to determine the effect of recombinant S1179DeNOS gene expression on the vasomotor function of human pial arteries.


METHODS
Pial arteries were isolated from 28 patients undergoing temporal lobectomy for intractable seizures. Adenoviral vectors (10(10) pfu/mL) encoding beta-galactosidase (AdCMVLacZ) or S1179DeNOS (AdCMVS1179DeNOS) were used for ex vivo gene transfer, and vasomotor function was evaluated in control and transduced arteries.


RESULTS
Contractions to cumulative additions of U46619 were not affected by expression of LacZ or S1179DeNOS. Endothelium-dependent relaxations to bradykinin or endothelium-independent relaxations to Diethylaminodiazen-1-ium-1,2-dioate were significantly reduced in arteries expressing S1179DeNOS. A superoxide dismutase mimetic, manganese (III) tetrakis (4-benzoic acid) porphyrin chloride, failed to improve the reduced relaxations to bradykinin. The levels of cGMP were significantly elevated in arteries expressing S1179DeNOS.


CONCLUSIONS
Our results support the concept that high local production of NO in pial arterial wall causes adaptive reduction of vasodilator reactivity to NO.
Sorenson
Joseph
J
Department of Anesthesiology, Mayo Clinic,Rochester, Minn 5590, USA.
Santhanam
Anantha Vijay R
AV
Smith
Leslie A
LA
Akiyama
Masahiko
M
Sessa
William C
WC
Katusic
Zvonimir S
ZS
D000328:Adult; D001224:Aspartic Acid / Q000235:genetics; D002536:Cerebral Arteries / Q000201:enzymology* / Q000503:physiopathology; D006152:Cyclic GMP / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000378:metabolism / Q000503:physiopathology; D004827:Epilepsy / Q000201:enzymology / Q000503:physiopathology; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D019001:Nitric Oxide Synthase / Q000235:genetics* / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D017354:Point Mutation; D011994:Recombinant Proteins / Q000378:metabolism; D012694:Serine / Q000235:genetics; D014664:Vasodilation
D011994:Recombinant Proteins; D001224:Aspartic Acid; D012694:Serine; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D006152:Cyclic GMP