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D G Warnock

Publications and source records attributed to D G Warnock.

107 records · Page 6Linked to original sources
PubMed1981

Chloride uptake by brush border membrane vesicles isolated from rabbit renal cortex. Coupling to proton gradients and K+ diffusion potentials.

Brush border membrane vesicles were isolated from rabbit renal cortex by Mg(++)-precipitation and differential centrifugation. (36)Cl(-) and [(3)H]glucose uptakes were simultaneously determined by a rapid filtration technique. Lysis of the vesicles with distilled water abolished 90-95% of the radioactivity on the filters, suggesting that nearly all of the (36)Cl(-) and [(3)H]glucose counts represented uptake into an osmotically reactive intravesicular space. Inwardly directed K(+) gradients plus valinomycin stimulated (36)Cl(-) uptake, demonstrating a conductive pathway for chloride uptake into brush-border membrane vesicles. (36)Cl(-) uptake could also be stimulated by inwardly directed proton gradients (pH(outside) < pH(inside)). This effect was seen in the absence of sodium, as well as in the presence of valinomycin when the vesicles had equal K(+) concentrations inside and out. An "overshoot" phenomenon was observed when external (36)Cl(-) was 2 mM and the external pH was lowered from 7.5 to 6.0 or to 4.5. The effect of the proton gradient was presumed to be different from the conductive mechanism because (a) the stimulation of (36)Cl(-) uptake by inwardly directed K(+) diffusion potentials was additive to the proton gradient effect, and (b) competition studies revealed statistically significant effects of thiocyanate on the conductive pathway, but not on the proton-driven pathway.HCl cotransport or anion exchange are electrically neutral mechanisms which could couple (36)Cl(-) uptake to inwardly-directed proton gradients in a brush border membrane vesicle. If both electrically neutral and conductive path ways for chloride transport are present in the luminal membrane of the proximal tubule, then the mechanism as well as the direction of net chloride transport will be influenced by the nature of the accompanying cation transport process.

Animals↗
PubMed1979

Carbonic anhydrase-dependent bicarbonate reabsorption in the rat proximal tubule.

The extent to which bicarbonate reabsorption in the rat proximal convoluted tubule depends on carbonic anhydrase has been examined by in vivo microperfusion and the measurement of total CO2 concentration by microcalorimetry. Tubules were perfused with an ultrafiltrate-like solution at 13 nl/min, and volume reabsorptive rate (JV) was measured using [14C]inulin. Addition of either 800 or 100 microM acetazolamide to the perfusion solution completely inhibited the reabsorption of total CO2. The control total CO2 reabsorptive rate (JtCO2) was 147 +/- 23 pmol/mm.min, and acetazolamide reduced JtCO2 to -3 +/- 5 pmol/mm.min. Acetazolamide reduced JV by 65% from a control of 2.3 +/- 0.4 to 0.8 +/- 0.1 nl/mm.min. The dose-response curve for acetazolamide showed that the I50 for inhibition of JtCO2 was 4 microM. The inactive congener of acetazolamide, t-butyl acetazolamide, did not reduce JV or inhibit bicarbonate reabsorption, indicating that the effect of acetazolamide on JtCO2 was specific for carbonic anhydrase inhibition. Since bicarbonate reabsorption was completely blocked by carbonic anhydrase inhibition, there is no need to postulate either carbonic acid recycling or carbonic anhydrase-independent bicarbonate reabsorption.

Acetazolamide↗
PubMed1979

Effects of anion-transport inhibitors on NaCl reabsorption in the rat superficial proximal convoluted tubule.

The effects of anion-transport inhibitors on volume reabsorption, and total CO(2) concentrations were examined by in vivo microperfusion of superficial proximal convoluted tubules of rats. The luminal perfusion solution was a high-chloride, low-bicarbonate solution like that in the in vivo late proximal tubule. The anion-transport inhibitors were only added to the luminal perfusion solutions. In tubules perfused with the control high-chloride solution, the rate of volume reabsorption (J(v)) was 2.3+/-0.2 nl/mm.min (n = 18), and the collected total CO(2) concentration was 4.0+/-0.3 mM. Furosemide (3 mM) caused a marked reduction in volume reabsorption to 0.8+/-0.3 nl/mm.min (n = 20) and only a slight increase in the total CO(2) concentration of collected samples of perfusate (7.8+/-0.5 mM). 0.8 mM acetazolamide caused a more pronounced rise in the collected total CO(2) concentrations to 10.7+/-0.5 mM but only a slight fall in J(v) to 1.7+/-0.3 nl/mm.min (n = 19). Hence, we inferred that inhibition of carbonic anhydrase only partially accounted for the inhibition of J(v) by furosemide. 4-acetamido-4'-iso-thiocyanato-stilbene-2,2'-disulphonic acid (0.1 mM), a well-characterized inhibitor of erythrocyte anion exchange mechanisms, also reduced J(v) to 1.6+/-0.3 nl/mm.min (n = 15) without changing the total CO(2) concentrations of the collected perfusates (3.6+/-0.4 mM). The effect of 4-acetamido-4'-iso-thiocyanato-stilbene-2,2'-disulphonic acid on volume reabsorption could not be explained by carbonic anhydrase inhibition because there was no increase in the total CO(2) concentration of the collected fluids. Furosemide did not significantly inhibit the rate of tracer glucose efflux out of the tubules, which suggests that the effect of furosemide on volume reabsorption was not a result of some nonspecific depression of active sodium transport. These results are discussed with respect to the possible effects of anion-transport inhibitors on the paracellular shunt pathway, active sodium reabsorption, and neutral sodium chloride transport.

Absorption↗
PubMed1978

Occupancy of aldosterone binding sites in rat kidney cytosol.

The binding sites for aldosterone and a potent aldosterone antagonist (SC-26304) were studied in kidney cytosol from adrenalectomized rats. Preformed cytosol and kidney slices were incubated with 3H-labeled steroids in a wide range of concentrations. The recovery and characteristics of the binding sites were affected by the incubation and homogenization conditions. High-affinity, Type I mineralocorticoid binding was reduced by more than 95% when cytosol was incubated at 25 degrees C in the presence of calcium. Tissue dilution also affected the binding sites. SC-26304 was bound to high- and low-affinity receptors, similar to the binding of aldosterone. The physiologic response to aldosterone could result from binding to either or both sets of sites. Some of the physiologic responses to spirolactones could represent antagonism of the binding of aldosterone to either or both sites. A convenient method is presented for describing the relative occupancy of several different sites by any particular steroid.

Adrenalectomy↗
PubMed1978

Contribution of leaked load to solute transport by renal tubules.

Renal tubules reabsorb solutes from the glomerular filtrate. The relationship between "filtered load" and reabsorption has been previously discussed and analyzed in detail. One aspect which has not been emphasized, however, is that, when reabsorption of a solute causes its concentration (or activity) in the tubule lumen to decrease below the level in the blood, solute may enter the tubule down this concentration gradient adding a "leaked load" to the filtered load. The leaked load should be taken into account when quantifying tubular transport. In the present study we derived equations for estimating the leaked load and its contribution to transport. The importance of the leaked load of glucose in the rabbit proximal convoluted tubules is evaluated with parameters derived from in vitro perfusion and by solving the equations numerically. It is shown that, depending on the conditions, the leaked load of glucose may account for a substantial part of the glucose present in the tubule lumen and reabsorbed from the tubule. Also, the leaked load could conceivably be an important factor in the transport of other solutes such as lactase and bicarbonate in proximal tubules.

Animals↗
PubMed1978

Ion selectivity and proximal salt reabsorption.

Electrophysiological techniques were used in isolated perfused superficial (S) and juxtamedullary (JM) rabbit proximal convoluted tubules (PCT) to examine the relative sodium-to-chloride (PNa/PCl) and bicarbonate-to-chloride (PHCO3/PCl) permeability ratios. We found that the great majority of PCT are sodium selective and that PHCO3/PCl depends on the experimental conditions. In the presence of active sodium transport, PHCO3/PCl is high and increases with PNa/PCl. When PHCO3/PCl is determined after inhibition of active sodium transport or at 25 degrees C, PHCO3/PCl approximates the free solution anion mobility ratio of 0.5 and is independent of PNa/PCl. The difference between PHCO3/PCl determined in the presence of and in the absence of active transport suggests that the lowering of bath bicarbonate concentration in the presence of active transport changes both paracellular and transcellular current flow. In addition, we found that during luminal perfusion with high chloride, low bicarbonate, organic solute-free solutions, the transepithelial electrical potential depends on PNa/PCl and PHCO3/PCl. This potential is approximately 4.0 mV in S PCT with low PNa/PCl and falls progressively to zero in JM PCT with high PNa/PCl. From these data we conclude that anion concentration gradients drive an important diffusive flux of sodium chloride through the paracellular pathway only in PCT with low PNa/PCl ratios.

Animals↗
PubMed1977

Urinary acidification: CO2 transport by the rabbit proximal straight tubule.

Proximal straight tubules from rabbit kidneys were perfused in vitro in order to study transport of bicarbonate. Total CO2 content was measured in perfused and collected tubule fluid, using microcalorimetry. When the initial perfusate and bath contained 25 mM bicarbonate, the concentration of total CO2 decreased in the collected tubule fluid, indicating net reabsorption of bicarbonate. When the initial perfusate contained no bicarbonate and the bath contained 25 mM bicarbonate, total CO2 appeared in the collected tubule fluid. The rate at which total CO2 appeared in the tubule fluid was rapid, indicating a high permeability. Proximal straight tubules from superficial and juxtamedullary nephrons were compared and found to differ in permeability to CO2 and in transport rate. This functional heterogeneity may affect urinary acidification when there is redistribution of renal blood flow.

Animals↗
PubMedDate not supplied

The epithelial sodium channel in hypertension.

Our understanding of Na(+) transport defects has exploded in the past several years, and has provided unique insights into epithelial transport processes, and unusual clinical syndromes resulting from mutations of specific ion transporters. These genetic disorders affect Na(+) balance, with both Na(+) retaining and Na(+) wasting conditions being the consequence. A major focus of these studies has been the epithelial sodium channel (ENaC), which can be directly affected by mutations (eg, Liddle syndrome, autosomal recessive pseudohypoaldosteronism, type I) or by changes in the response to (autosomal recessive pseudohypoaldosteronism, type I), or production of mineralocorticoids (apparent mineralocorticoid excess syndrome, glucocorticoid-remediable aldosteronism). As a result, we now have clearly defined syndromes in which ENaC activity is dysregulated with subsequent development of disorders of systemic blood pressure that can be attributed to a primary renal mechanism.

Epithelial Sodium Channels↗
PubMedDate not supplied

Inverse relationship of urinary cyclic GMP to blood pressure reactivity in the CARDIA study: vasodilatory regulation of sympathetic vasoconstriction. Coronary Artery Risk Development in Young Adults.

OBJECTIVE: To determine whether urinary cyclic GMP (cGMP), which mediates the actions of the vasodilators nitric oxide and atrial natriuretic factor, is inversely related to blood pressure (BP) reactivity. In previous work, we found that urinary cGMP was inversely related to diastolic BP, but cGMP levels were higher among individuals presumed to have increased adrenergic activity, increased reactivity, and increased risk of hypertension (blacks, individuals with a family history of hypertension). METHOD: We measured 24-hour urinary cGMP levels in a substudy of 529 individuals in the Coronary Artery Risk Development in Young Adults (CARDIA) study; the sample was 23 to 35 years of age and approximately balanced for race (black/white) and gender. BP reactivity to stressors (video game, star-tracing, cold pressor) was tested 3 years earlier. Baseline BP was included as a covariate in all analyses. RESULTS: Diastolic BP reactivity to cold pressor was inversely related to cGMP excretion (p < .05); the relationship was strongest among black women with a family history of hypertension (partial r = -.33, p < .01). Systolic BP reactivity to star-tracing was also inversely related to cGMP (p < .01); the relationship for both star-tracing and video game stressors was strongest among black men (partial r values = -.25 and -.24, respectively; p values < .01). CONCLUSIONS: The results indicate that vasodilatory activity may impact the BP response to stress through modulation of adrenergic activation, particularly among blacks.

Adolescent↗
PubMedDate not supplied

Disorders of the epithelial Na(+) channel in Liddle's syndrome and autosomal recessive pseudohypoaldosteronism type 1.

The epithelial Na(+) channel (ENaC) is the key step in many Na(+)-absorptive epithelia, such as kidney and distal colon, that controls the overall rate of transepithelial Na(+) transport. ENaC is composed of three homologous subunits, alpha, beta, and gamma. The alpha subunit is the key subunit for the formation of a functional ion channel, while the beta and gamma subunits can greatly potentiate the level of expressed Na(+) currents. ENaCs belong to the recently identified DEG/ENaC supergene family, sharing the same basic structure with cytoplasmic amino and carboxy termini, two transmembrane regions, and a large extracellular loop. The human ENaC genes have been cloned, and using genetic linkage analysis the involvement of ENaC gene mutations in two distinct human diseases, Liddle's syndrome and autosomal recessive pseudohypoaldosteronism type 1 (PHA-1), has been demonstrated. In Liddle's syndrome, gain-of-function mutations in the beta or gamma ENaC subunits have been found; all identified mutations so far reside in the carboxy terminus of the protein, either deleting or modifying the functionally important PY motif. In PHA-1, loss-of-function mutations in the alpha, beta, or gamma subunits have been found; these mutations either truncate a significant portion of the structure or modify an amino acid that plays an important role in channel function. In this review, our current understanding about ENaC and the pathophysiology of Liddle's syndrome and PHA-1 caused by ENaC mutations will be discussed.

Animals↗