Adenoviral gene transfer of nitric oxide synthase increases cerebral blood flow in rats.
OBJECTIVE
Depletion of nitric oxide may play a role in the development of vasospasm after aneurysmal subarachnoid hemorrhage. Replenishment of nitric oxide might be a useful treatment for vasospasm. Using rats, we performed intracisternal injections of replication-defective adenovirus containing the endothelial nitric oxide synthase (eNOS) gene and determined the localization of and effect on cerebral blood flow of transgene expression.


METHODS
Rats underwent baseline measurement of cortical cerebral blood flow using laser Doppler flowmetry. Replication-defective adenovirus containing the Escherichia coli LacZ gene (Ad327beta-Gal, n = 2/time point) or the bovine eNOS gene (AdCD8-NOS, n = 4/time point) or physiological saline solution was injected into the cisterna magna. Cerebral blood flow was measured 1, 2, 4, 7, or 14 days later, and the animals were killed. Expression of beta-galactosidase activity from the LacZ gene was examined by histochemical staining and that of eNOS was examined by polymerase chain reaction assays of messenger ribonucleic acid. Brains were histopathologically examined for inflammation.


RESULTS
Beta-galactosidase activity was observed throughout the leptomeninges and in some cells in the adventitia of small subarachnoid blood vessels in the Ad327beta-Gal-injected rats. Messenger ribonucleic acid for eNOS was detected in the leptomeninges and brainstem 1 and 2 days after injection of AdCD8-NOS. Rats injected with Ad327beta-Gal or physiological saline solution exhibited decreased cerebral blood flow beginning 2 days after virus injection and lasting up to 14 days after injection. Rats injected with AdCD8-NOS developed significant transient increases in cerebral blood flow 2 days after virus injection, followed by slight decreases in blood flow. There was inflammation in the subarachnoid space of all animals; the inflammation was qualitatively worse in animals injected with Ad327beta-Gal, compared with rats injected with AdCD8-NOS or saline solution.


CONCLUSIONS
Intracisternal injection of replication-defective adenovirus containing the eNOS gene can transiently increase cerebral blood flow.
Lüders
J C
JC
Section of Neurosurgery, Pritzker School of Medicine, University of Chicago Medical Center, Illinois 60637, USA.
Weihl
C C
CC
Lin
G
G
Ghadge
G
G
Stoodley
M
M
Roos
R P
RP
Macdonald
R L
RL
D000818:Animals; D001921:Brain / Q000098:blood supply / Q000201:enzymology / Q000821:virology; D002560:Cerebrovascular Circulation / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D018014:Gene Transfer Techniques; D005814:Genes, Viral / Q000235:genetics*; D007763:Lac Operon / Q000235:genetics*; D017078:Laser-Doppler Flowmetry / Q000379:methods; D017921:Mastadenovirus / Q000201:enzymology* / Q000235:genetics*; D019001:Nitric Oxide Synthase / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013346:Subarachnoid Space; D013997:Time Factors; D019076:Transgenes / Q000235:genetics; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D012333:RNA, Messenger; D012367:RNA, Viral; D019001:Nitric Oxide Synthase; D001616:beta-Galactosidase