Abstract-The renin-angiotensin system is a major regulator of body sodium, predominantly through the actions of intrarenal angiotensin II of unclear origin. We show that polarized epithelium of the proximal tubule synthesizes and secretes angiotensinogen at its apical side and that the protein can be detected in urine as a function of dietary sodium. Furthermore, we demonstrate that renin is expressed and secreted in a restricted nephron segment, the connecting tubule, also in a sodium-dependent fashion. A paracrine renin-angiotensin system operating along the entire nephron may contribute to long-term arterial pressure regulation by integrating distant tubular sodium-reabsorbing functions.(Hypertension. 1999;34:1265-1274.)

Abstract-Impaired insulin signaling via phosphatidylinositol 3-kinase/Akt to endothelial nitric oxide synthase (eNOS) in the vasculature has been postulated to lead to arterial dysfunction and hypertension in obesity and other insulin resistant states. To investigate this, we compared insulin signaling in the vasculature, endothelial function, and systemic blood pressure in mice fed a high-fat (HF) diet to mice with genetic ablation of insulin receptors in all vascular tissues (TTr-IR Ϫ/Ϫ ) or mice with genetic ablation of Akt1 (Akt1Ϫ/Ϫ). HF mice developed obesity, impaired glucose tolerance, and elevated free fatty acids that was associated with endothelial dysfunction and hypertension. Basal and insulin-mediated phosphorylation of extracellular signal-regulated kinase 1/2 and Akt in the vasculature was preserved, but basal and insulin-stimulated eNOS phosphorylation was abolished in vessels from HF versus lean mice. In contrast, basal vascular eNOS phosphorylation, endothelial function, and blood pressure were normal despite absent insulinmediated eNOS phosphorylation in TTr-IR Ϫ/Ϫ mice and absent insulin-mediated eNOS phosphorylation via Akt1 in Akt1Ϫ/Ϫ mice. In cultured endothelial cells, 6 hours of incubation with palmitate attenuated basal and insulinstimulated eNOS phosphorylation and NO production despite normal activation of extracellular signal-regulated kinase 1/2 and Akt. Moreover, incubation of isolated arteries with palmitate impaired endothelium-dependent but not vascular smooth muscle function. Collectively, these results indicate that lower arterial eNOS phosphorylation, hypertension, and vascular dysfunction following HF feeding do not result from defective upstream signaling via Akt, but from free fatty acid-mediated impairment of eNOS phosphorylation. Key Words: arterial insulin signaling Ⅲ hypertension Ⅲ endothelial dysfunction Ⅲ mice Ⅲ diabetes S timulation of insulin receptors in the vasculature leads to increased activation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway and the mitogen-activated protein (MAP) kinase (eg, extracellular signaling-regulated kinase [ERK]1/2) pathway. [1][2][3][4] Insulin receptor (IR)-mediated stimulation of PI3K/Akt leads to endothelial nitric oxide (NO) synthase (eNOS) phosphorylation, NO production, and vasorelaxation. 4 -7 Insulin-mediated activation of ERK1/2 leads to endothelin (ET)-1 production, inhibition of eNOS phosphorylation, and subsequent vasocontraction. 1,4,8 Evidence from several experimental models of insulin resistance reveals impaired insulin-mediated PI3K/Aktdependent signaling in the vasculature, whereas ERK1/2 pathways are preserved or even augmented. 2,8,9 Collectively, these observations have led to the hypothesis that an imbalance in vascular insulin-mediated signaling can precipitate cardiovascular complications including endothelial dysfunction and hypertension. 1,4 Recently, it was shown that insulinmediated Akt phosphorylation was preserved, but NOmediated vasorelaxation was blunted, in arteries from obese, glucose ...

Abstract-Elements of a renin-angiotensin system expressed along the entire nephron, including angiotensinogen secreted by proximal tubule and renin expressed in connecting tubule, may participate in the regulation of sodium reabsorption at multiple sites of the nephron. The response of this tubular renin-angiotensin system to stepwise changes in dietary sodium was investigated in 2 mouse strains, the sodium-sensitive inbred C57BL/6 and the sodium-resistant CD1 outbred. Plasma angiotensinogen was not affected by sodium regimen, whereas plasma renin increased 2-fold under low sodium. In both strains, the variation in urinary parameters did not parallel the changes observed in plasma. Angiotensinogen and renin excretion were significantly higher under high sodium than under low sodium. Water deprivation, by contrast, induced significant activation in the tubular expression of angiotensinogen and renin. C57BL/6 exhibited significantly higher urinary excretion of angiotensinogen than did CD1 animals under both conditions of sodium intake. The extent to which these urinary parameters reflect systemic or tubular responses to challenges of sodium homeostasis may depend on the relative contribution of sodium restriction and volume depletion. Key Words: angiotensinogen Ⅲ renin Ⅲ sodium Ⅲ mouse Ⅲ genetics Ⅲ urine W e have advanced the hypothesis that a paracrine tubular renin-angiotensin system operates along the entire nephron. 1 Although angiotensinogen (AGT) is not filtered across the glomerular membrane, the protein 2 and its mRNA 3,4 have been detected in proximal tubule (PT), the protein is secreted to the apical side of PT cell monolayers, 1 has been detected in final urine under normal physiological conditions, 5 and was detected in luminal fluid of PT epithelium collected by micropuncture. 6 Systemic renin is filtered and reabsorbed in the PT. 7 Although not detected in situ, it may be expressed at low level in the PT. 8,9 We have found that renin was also synthesized and secreted in connecting tubule (CNT). 1 ACE and angiotensin (Ang) II receptors are expressed along the nephron. 10,11 High luminal Ang II has been observed in the PT, 12,13 where it stimulates sodium reabsorption. 14 Some observations support a similar role in terminal segments of the nephron. 15 The potential significance of this tubular renin-angiotensin system in blood pressure regulation is underlined by the observation that double transgenic animals overexpressing human renin systemically and human AGT in the PT develop hypertension. 16 The impact of dietary sodium on the expression of renin and tubular AGT and the significance of their urinary excretion as indicators of the activity of this tissue system were tested in the mouse. Two strains were investigated, C57BL/6 and CD1. The C57BL/6 inbred differs from other inbred lines in its response to dietary sodium 17 ; its sodium sensitivity has been demonstrated 18,19 and exploited in an attempt to map genetic determinants of the arterial pressure response to dietary sodium. 19 We have verified...