Li E. Yang

Mechanisms of Pressure Natriuresis

Abstract: An acute increase in blood pressure provokes a rapid decrease in proximal tubule salt and water reabsorption that is central to tubuloglomerular feedback regulation of renal blood flow and glomerular filtration rate and contributes to pressure natriuresis. The molecular mechanisms responsible for this critical homeostatic adjustment were studied. When blood pressure is acutely elevated, apical proximal tubule NHE3 are rapidly redistributed out of the microvilli to intermicrovillar clefts and then endosomal pools, and Na,K‐ATPase activity is suppressed. Depressing apical Na+ entry without hypertension is not sufficient to decrease Na,K‐ATPase activity, and depressing Na,K‐ATPase activity alone is not sufficient to decrease proximal tubule Na+ and water reabsorption; thus, it appears that coordinated decreases in both NHE3 surface distribution and Na,K‐ATPase activity may be important for the response to acute hypertension. Clamping plasma angiotensin II levels blunts the retraction of NHE3 from the cell surface to endosomal pools. The increased volume flow of salt and water to the loop of Henle stimulates Na,K‐ATPase activity in this region and provides evidence for a downstream shift in sodium transport during acute hypertension. These same responses in the proximal tubule and loop develop and persist in the spontaneously hypertensive rat. These studies demonstrate that sodium transporters along the nephron are very dynamic, responding quickly to normal fluctuations of blood pressure, and are key to generating the macula densa tubuloglomerular feedback signal and for accommodating increased volume flow through the loop of Henle.

Redistribution of Myosin VI from Top to Base of Proximal Tubule Microvilli during Acute Hypertension

During acute hypertension, Na(+)/H(+) exchangers (NHE3) retract from top to base of proximal tubule microvilli (MV) and Na(+) reabsorption decreases in proximal tubule. This study aimed to determine whether the actin-based motor myosin VI coordinately retracts with NHE3 in response to acute hypertension. BP was raised approximately 50 mmHg in rats for 20 to 30 min or sham treated, and kidneys were analyzed by subcellular fractionation or microscopy. During acute hypertension, myosin VI redistributed from low density apical MV-enriched membranes (from 23 +/- 2.4 to 11.4 +/- 2.2%) into higher density membranes (from 23.2 +/- 0.7 to 36.9 +/- 2.6%). By confocal microscopy, myosin VI was detected over the whole length of the MV in controls, then became completely focused at the base of MV during acute hypertension. For electron microscopic analysis using immunogold labeling, MV were divided into five zones from top (z1) to base (z5). In controls, myosin VI was evenly distributed through the five MV zones. In acute hypertension, myosin VI decreased in z1 (from 20.6 +/- 1.9 to 10.5 +/- 2.3%) and z2 (from 21.0 +/- 2.0 to 13.2 +/- 1.4%) and increased in z5 (from 21.1 +/- 3.3 to 38.6 +/- 3.0%). These results provide the first observation that acute hypertension causes myosin VI redistribution and support the idea that myosin VI may serve as the molecular motor for NHE3 retraction from top to base of MV during acute hypertension.

The proximal convoluted tubule is a target for the uroguanylin-regulated natriuretic response.

Objectives and Methods: Guanylin and uroguanylin are peptides synthesized in the intestine and kidney that are postulated to have both paracrine and endocrine functions, forming a potential enteric-renal link to coordinate salt ingestion with natriuresis. To explore the in vivo role of guanylin and uroguanylin in the regulation of sodium excretion, we used gene-targeted mice in which the uroguanylin, guanylin or the peptide receptor guanylate cyclase C gene expression had been ablated. Results: Metabolic balance studies demonstrated that there was impaired excretion of a sodium load in uroguanylin (but not in guanylin or guanylate cyclase C) knockout mice. Uroguanylin-dependent natriuresis occurred without an increase in circulating prouroguanylin. A distinct morphological phenotype was present in the proximal convoluted tubules of uroguanylin knockout animals after an enteral salt loading. Marked vacuolization of the proximal convoluted tubule epithelial cells was observed by using light and electron microscopy. There was also a change in the distribution of the sodium hydrogen exchanger 3 (NHE3) after an enteral salt loading. In wild-type animals, there was a partial redistribution of NHE3 from the villus fraction to the less accessible submicrovillus membrane compartment, but this effect was less apparent in uroguanylin knockout animals, presumably resulting in greater Na+/H+ exchange. Conclusions: Together, these findings further establish a role for uroguanylin in fluid homeostasis and support a role for uroguanylin as an integral component of a signaling mechanism that mediates changes in Na+ excretion in response to an enteral salt loading. Proximal tubular NHE3 activity is a possible target for uroguanylin-mediated changes in Na+ excretion.