Critical hypertensive roles for angiotensin II (Ang II) have been postulate for its effects on the major resistance vessels of the renal cortex (afferent arteriole) and medulla (outer medullary descending vasa recta, OMDVR). Prolonged exposure to Ang II leads to slow pressor response to enhancement of renal vascular resistance. This proposal will examine the concept that Ang II-induced oxidative stress underlies these renal hypertensive mechanisms. This proposal will examine the concept that Ang II-induced oxidative stress underlies these renal hypertensive mechanisms. Ang II-induced oxidative stress implies either an enhanced generation, or decreased metabolism of reactive oxygen species (ROS), notably superoxide anion (P2), hydrogen peroxide (H2O2)and hydroxyl radical (OH). This concept will be studied in selected knockout models to assess the effects of deletion of ROS generated via p47/phox NAD(P)H oxidase, or NO generated via eNOS and of oxidant defense from the effects of deletions of extracellular superoxide dismutase, (EC-SOD) or dopamine 5 receptor (D5-R). Subproject 1 will utilize novel methods for intra-tubular and arteriolar measurements of pO2 combined with micropuncture and microperfusion in vivo. It will study the hypothesis that Ang II stimulates NAD(P)H oxidase-dependent ROS. This causes functional NO deficiency, a fall in renal O2 delivery and inefficient O2 utilization. The resulting fall in renal pO2 restrains ongoing ROS generation. Subproject 2 will contrast responses in isolated renal afferent and mesenteric resistance vessels during changes in Ang II combining measurements of contractility with [NO] and [ROS] developed in subprojects #3 and #4. It will test the concept that selective renal cortical vasoconstrictor actions of Ang II are due to ROS-dependent reduction in [NO], thereby promoting vasoconstriction of the afferent arteriole, whereas ROS actually relax mesenteric vessels via endothelium- dependent hyperpolarization. Subproject 3 will use novel fluorescence microscopy studies of vascular [ROS] and [NO], combined with direct measures of NO releases from single OMDVR to study the regulation of ROS and NO by Ang II and pO2 (defined in subproject #1) in isolated perfused OMDVR. Subproject 4 will use the novel model for hypertension and oxidative stress in the dopamine 5 receptor (D5-R) knockout mouse. It will investigate the interaction of the constitutively active D5-R with Ang II in the renal regulation of NAD(P)H oxidase, and other cellular oxidant or defense pathways. The functional effects of D5- R knockout will be explored in subprojects #1, #2, and #3. This is a proposal for an integrated functional genomics approach to the study of hypertensive mechanisms mediated by Ang II-stimulated ROS within the kidneys.
Showing the most recent 10 out of 207 publications
