• Evaluate the importance a novel renal specific ADPR cyclase and its cyclic ADP ribose / ryanodine receptor Ca2+ signaling pathway in renal vascular reactivity in vivo. Student will perform surgery on an anesthetized rat or mouse and measure renal blood flow using an ultrasonic transducer. Renal vascular reactivity is assessed in response to vasoactive agents injected iv or directly into the renal artery to produce transient renal vasoconstriction.
• Determine the functional importance of a novel renal specific ADPR cyclase and cyclic ADP ribose / ryanodine receptor Ca2+ signaling pathway in vascular smooth muscle cells of afferent arterioles. Student will isolate preglomerular resistance arterioles (<100 mm in diameter) using an iron oxide/sieving method or microdissection and measure cytosolic Ca2+ concentration utilizing the fluorophore fura-2 in imaging studies. Arterioles are isolated from mice lacking the conventional ADPR cyclase, CD38, due to gene targeting.
• Assess the relative activities of a novel renal ADP ribosyl cyclase vs. conventional CD38. Student will use a biochemical assay to measure production of the metabolite cyclic ADP ribose in afferent arterioles isolated from CD38-/- and wild-type mice.
• Identify specific ryanodine receptor subtypes (RyR1-3) in vascular smooth muscle cells of afferent arterioles. Student will utilize real-time RT-PCR to quantify mRNA of each subtype.
