Adenovirus-mediated gene transfer into the NTS in conscious rats. A new approach to examining the central control of cardiovascular regulation.
The nucleus tractus solitarii (NTS) is an important site for the regulation of sympathetic nerve activity. It receives the signals through afferent fibers from arterial baroreceptors, chemoreceptors, cardiopulmonary receptors, and other visceral receptors. Many studies have examined the role of nitric oxide (NO) in the NTS in cardiovascular regulation. However, most of these studies were conducted in an acute state with anesthesia. We have developed a novel technique of endothelial nitric oxide synthase (eNOS) gene transfer into the NTS in vivo. Adenovirus vectors encoding either the beta-galactosidase gene (Ad beta gal) or the endothelial nitric oxide synthase gene (AdeNOS) gene were transfected into the NTS. In the Ad beta gal-treated rats, the local expression of beta-galactosidase was confirmed by X-Gal staining, and beta-galactosidase activity was quantified using a colorimetric assay. In the AdeNOS-treated rats, the local expression of eNOS protein was confirmed by immunohistochemistry, and eNOS production was measured by in vivo microdialysis. Blood pressure and heart rate were monitored by a radiotelemetry system in a conscious state. The expression of each gene was observed from day 5 to day 10 after the gene transfer. In the AdeNOS-treated rats, blood pressure and heart rate significantly decreased from day 5 to day 10, and then thereafter gradually recovered over time. Our method may be useful in examining the local effect of a particular substance produced by a specific gene in the brain on cardiovascular function.
Hirooka
Y
Y
Department of Cardiovascular Medicine, Cardiovascular Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. hyoshi@cardiol.med.kyushu-u.ac.jp
Sakai
K
K
Kishi
T
T
Takeshita
A
A
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001794:Blood Pressure / Q000502:physiology; D002417:Cattle; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006339:Heart Rate / Q000502:physiology; D008297:Male; D009566:Nitrates / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000235:genetics; D052250:Nitric Oxide Synthase Type III; D009573:Nitrites / Q000378:metabolism; D051381:Rats; D011921:Rats, Inbred WKY; D017552:Solitary Nucleus / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D009566:Nitrates; D009573:Nitrites; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D001616:beta-Galactosidase