Richard W. Osgood

The Effect of Bradykinin on Proximal Tubular Sodium Reabsorption in the Dog: Evidence for Functional Nephron Heterogeneity

In a previous study we have found that acetylcholine, a renal vasodilator, inhibits fractional and absolute reabsorption of sodium in the proximal tubule of the dog. To delineate whether this effect on proximal tubular sodium reabsorption was related to alterations in renal hemodynamics or to a direct tubular action of the drug, free-flow micropuncture studies were performed in the dog in which the tubular fluid to plasma inulin ratio and nephron filtration rate were determined before and during the administration of a structurally different renal vasodilator, bradykinin. This agent increased sodium excretion from 12 to 96 muEq/min and decreased total kidney filtration fraction from 0.35 to 0.25. However, sodium reabsorption in the proximal tubule of the superficial nephrons was unchanged during bradykinin administration. Since it has been shown that a decrease in filtration fraction and presumably peritubular capillary protein concentration will decrease proximal tubular sodium reabsorption, studies were performed to determine whether the fall in total kidney filtration fraction seen with both vasodilators is paralleled by a similar change in the circulation of superficial nephrons. The results of these studies indicate that neither agent altered superficial nephron capillary protein concentration, hematocrit, or filtration fraction. In contrast, a decrease in capillary protein concentration, hematocrit, and filtration fraction was consistently demonstrated during the intrarenal infusion of 7.5-15 ml/min of Ringers solution while an increase in these parameters occurred during the i.v. administration of norepinephrine, 60 mug/min. In the Ringers infusion studies, both fractional and absolute sodium reabsorption in the proximal tubule were decreased concomitant with the fall in capillary protein concentration and hematocrit. THIS DATA SUGGESTS THAT: (a) the hemodynamic effect of renal vasodilatation is not the same in the circulation of all nephrons; (b) the inhibitory effect of acetylcholine on proximal tubular sodium reabsorption is due to a direct tubular action; (c) a decrease in capillary protein concentration and/or hematocrit does decrease proximal tubular sodium reabsorption; (d) although proximal reabsorption of sodium is unchanged in the superficial nephrons during bradykinin administration, a decrease in reabsorption may be present in deeper nephrons in which filtration fraction is decreased.

Effect of renal vasodilatation on the distribution of cortical blood flow in the kidney of the dog

Studies were performed to evaluate the validity of using the radioactive microsphere technique to measure regional blood flow in the renal cortex. A technique was developed in which the renal cortex was divided into four equal zones, and the fractional and absolute distribution of blood flow in these zones was determined. It was consistently found that approximately 70% of the renal blood flow was distributed to the two outer cortical zones with the remaining 30% going to the two inner cortical zones. In addition, there was a reproducible pattern of distribution of blood flow in different areas of the same kidney after a single injection of microspheres and in the same area of the kidney after multiple injections of microspheres. Using this method, the distribution of renal blood flow was determined before and during the intrarenal administration of either acetylcholine (40 mug/min) or bradykinin (5 mug/min). Both agents decreased the per cent of blood flow to outer cortical zone 1, caused no change in zone 2, and increased the fractional blood flow in inner cortical zones 3 and 4. When this data was evaluated in terms of total blood flow, there was no change in zone 1, an increase in zone 2 commensurate with the change in total blood flow, and a marked increase in inner cortical zones 3 and 4 which accounted for 60 and 65% of the increase in total blood flow during acetylcholine and bradykinin administration, respectively.Therefore, the natriuresis of renal vasodilatation is associated with a redistribution to inner cortical nephrons.

Studies on the Mechanism of Oliguria in a Model of Unilateral Acute Renal Failure

To further evaluate the mechanism of the oliguria of acute renal failure, a model was utilized in which intense and prolonged vasoconstriction produced the unilateral cessation of urine flow. The radioactive microsphere method was used to measure total and regional blood flow before and after the intrarenal infusion of norepinephrine, 0.75 mug/kg/min, for 2 h in the dog. In the control kidney, renal blood flow increased 32% 48 h after norepinephrine in association with a fall in the fractional distribution of flow to the outer cortex. In the experimental kidney, total renal blood flow fell from 190 ml/min before norepinephrine to 116 ml/min at 48 h (P < 0.025) with a uniform reduction in cortical blood flow. After the administration of 10% body wt Ringers solution, there was a marked redistribution of flow to inner cortical nephrons in both the control and experimental kidney. In addition, there was a marked increase in total blood flow in both kidneys. On the experimental side, flow rose to 235 ml/min, a value greater than in either the control period (P < 0.05) or at 48 h after norepinephrine (P < 0.001). However, in spite of this marked increase in blood flow, there was essentially no urine flow from the experimental kidney. In separate studies, the animals were prepared for micropuncture. In all studies, the surface tubules were collapsed, and there was no evidence of tubular obstruction or leakage of filtrate. Over 99% of the 15-muM spheres were extracted in one pass through the experimental kidney. An analysis of the forces affecting filtration suggested that an alteration in the ultrafiltration coefficient may be responsible, at least in part, for the anuria in this model. In this regard, transmission and scanning electron microscopy revealed a marked abnormality in the epithelial structure of the glomerulus. It is suggested that a decrease in glomerular capillary permeability may be present in this model of acute renal failure.

Studies on the Mechanism of Reduced Urinary Osmolality after Exposure of the Renal Papilla

Studies were performed in Munich-Wistar rats to determine whether changes in papillary plasma flow might be responsible for the concentrating defect which occurs after exposure of the extrarenal papilla. Papillary plasma flow was measured by (125)I-albumin accumulation. Initial studies in hydropenic animals revealed that papillary plasma flow was 40% higher in the kidney with the exposured papilla, 41 vs. 29 ml/min per 100 g of papilla (P < 0.001). This increase in papillary plasma flow was detectable 15 or 45 min after removing the ureter. Because it was unclear whether the rise in papillary plasma flow was a cause or the result of the fall in urine osmolality, similar studies were performed in animals undergoing a water diuresis. In this setting, papillary plasma flow still increased on the exposed side compared to the control side, 81 vs. 60 ml/min per 100 g, despite similarly low urine osmolalities of 155 and 174 mosmol/kg, respectively. This finding is compatible with the possibility that papillary exposure per se causes an increase in papillary plasma flow and that this hemodynamic alteration may lead to a reduction in urinary osmolality secondary to washout of the medullary interstitium. A final group of hydropenic rats was given either indomethacin or meclofenamate before removing the ureter. In these studies, there was no difference in either the papillary plasma flow or the urine osmolality between control and exposed kidneys. It is therefore suggested that opening the ureter induces an increase in papillary plasma flow by some mechanism which may involve an alteration in prostaglandin synthesis.

A Comparison of the Segmental Analysis of Sodium Reabsorption during Ringer's and Hyperoncotic Albumin Infusion in the Rat

Studies were designed to compare the segmental analysis of sodium reabsorption along the nephron during volume expansion with either 10% body weight Ringers or 0.6% body weight hyperoncotic albumin. Total kidney and nephron glomerular filtration rate increased similarly with both, but urinary sodium excretion (12.7 vs. 4.0 mueq/min, P < 0.001) and fractional sodium excretion (5.0 vs. 1.6%, P < 0.001) increased to a greater extent with Ringers. Fractional reabsorption of sodium in the proximal tubule was diminished in both groups but to a significantly greater extent during Ringers (P < 0.005). Absolute reabsorption was inhibited only in the Ringers group. Delivery of filtrate out of the proximal tubule was greater in the Ringers studies, 45 vs. 37 nl/min (P < 0.001). However, both fractional and absolute sodium delivery to the early and late distal tubule were not significantly different in the two groups. Fractional reabsorption in the collecting duct decreased from 96% in hydropenia to 31% during Ringers but fell only slightly to 80% in the albumin studies. Absolute collecting duct reabsorption was also greater in the albumin studies, 0.55 vs. 0.21 neq/min (P < 0.001), which could totally account for the difference in urinary sodium excretion between the two groups. (22)Na recovery in the final urine after end distal microinjections was 71% during Ringers infusion and 34% during albumin (P < 0.001). From these data we conclude that: (a) Ringers solution has a greater inhibitory effect on proximal tubular sodium reabsorption, and (b) in spite of this effect, differences in mucosal to serosal collecting duct sodium transport are primarily responsible for the greater natriuresis during Ringers infusion.

Further Studies on Segmental Sodium Transport in the Rat Kidney during Expansion of the Extracellular Fluid Volume

The present studies were designed to further investigate the possibility of heterogeneity of nephron function during Ringer loading in the rat, and to determine the specific nephron segment responsible for this finding. As in previous studies from this laboratory with smaller rats (50-125 g), net addition of sodium between late distal tubule and papillary base (6.9 vs. 10.4% of the filtered load, respectively, P <0.005) was found in more mature rats (170-230 g). In contrast, there was net reabsorption of sodium between these two segments in nonvolume-expanded animals, 1.70 vs. 0.45% of the filtered sodium load, P <0.005. Because nephron heterogeneity of sodium transport during extracellular volume expansion is the most likely explanation for these findings, further studies were performed to determine the specific juxtamedullary nephron segment responsible for the net addition pattern between late distal tubule and papillary base in Ringer-loaded animals. First, a comparison was made of sodium delivery to the late proximal tubule of superficial nephrons vs. the delivery rate to the bend of Henles loop of juxtamedullary nephrons in both hydropenia and Ringer loading. Fractional sodium delivery was quite comparable between the superficial and juxtamedullary nephrons in both hydropenia and Ringer loading although the absolute level was much greater in both groups of nephrons in the Ringer studies. Chlorothiazide (15 mg/kg loading and 15 mg/kg per h) given during Ringer loading markedly increased late distal sodium delivery, 19% of the filtered load, but did not prevent net addition of sodium at the papillary base. In contrast, furosemide (5 mg/kg loading and 5/mg/kg per h) given during Ringer loading completely reversed the segmental pattern, 35.5 and 28.8% at late distal tubule and papillary base, respectively, P <0.005. These studies demonstrate that the net addition of sodium between late distal tubule and papillary base during Ringer loading is not limited to immature rats and that the segmental pattern does not occur in non-volume-expanded animals. Further, the reversal of the net addition pattern with furosemide, but not chlorothiazide, and the comparable proximal nephron delivery rates in Ringer loading suggest that the loop of Henle of juxtamedullary nephrons reabsorbs less sodium than the same portion of superficial nephrons in this setting. A model is proposed to explain this finding.

The measurement of nephron filtration rate and absolute reabsorption in the proximal tubule of the rabbit kidney.

Micropuncture studies were performed in the rabbit to determine nephron filtration rate and absolute fluid reabsorption in the proximal tubule in order to compare the latter value with data obtained with the in vitro microperfusion technique. New Zealand white rabbits, 2-2.8 kg, were studied. Nephron filtration rate was 21 nl/min (n equal to 48) and absolute reabsorption along the length of the accesible portion of the proximal convoluted tubule was 10.3 nl/min. Correcting this value for tubular length gives a fluid reabsorption of approximately 1.9 nl/mm per min. In view of the marked difference between the in vivo and in vitro techniques and the various sources of error with each, this is reasonably similiar to the value of 1.3 nl/mm per min obtained in the isolated proximal convoluted tubule.

The Effect of Beta Adrenergic Stimulation on Proximal Tubular Sodium Reabsorption

Summary Studies were designed to determine whether beta adrenergic stimulation has a direct effect on proximal tubular sodium transport. The recollection micropuncture technique was utilized before and during the intrarenal infusion of isoproterenol, 0.018 μg/kg/min, the highest dose of the drug which had no renal or systemic hemodynamic effects. In the first group of five studies, the tubular fluid to plasma inulin (TF/P inulin) ratio was 1.47 ± 0.04 SEM and 1.46 ± 0.08 before and during beta stimulation, respectively. In a second group of six experiments in which alpha blockade was maintained throughout with phenoxybenzamine, 0.09 μg/kg/min, the TF/P inulin ratio was again unchanged at 1.48 ± 0.09 in the control period and 1.52 ± 0.09 in the experimental period. Nephron filtration rate and absolute reabsorption were unchanged in both groups of studies. In addition, total GFR, renal plasma flow, sodium excretion and mean arterial pressure were unchanged during isoproterenol infusion. From these data, there is no indication that beta stimulation has an inhibitory effect on proximal tubular sodium reabsorption.

Absence of Aglomerular Blood Flow During Renal Vasodilatation and Hemorrhage in the Dog

Summary The radioactive microsphere method was used to quantitate aglomerular blood flow during intrarenal acetylcholine administration (6 studies) and hemorrhagic hypotension (5 studies). In each of the 11 studies, less than 1% of the injected radioactivity was recovered in renal venous effluent. These findings strongly suggest that a significant diversion of renal blood flow by a non-glomerular pathway does not occur in these two models.

Delineation of the site of action of guanabenz in the renal tubule.

Guanabenz, a centrally acting alpha 2-agonist, has been shown to lower blood pressure (BP) while maintaining glomerular filtration rate and increasing water and solute excretion. The site and mechanism of the increased solute excretion are still unclear. To evaluate the effect of guanabenz on water and solute reabsorption in the collecting duct, Munich Wistar rats were utilized in micropuncture experiments. Micropuncture samples were obtained from the most proximal portion (base) of the collecting duct and the most distal part (tip). In an initial group of hydropenic rats, collecting duct water and chloride (Cl) reabsorption were evaluated during a control and experimental period during which guanabenz was infused at 20-40 micrograms/kg/min. Although the mean drop in BP was 18 mm Hg after guanabenz infusion, at any given rate of Cl delivery, the reabsorption was less than that for the control period. Another group of studies utilizing the plasma repletion method to increase Cl excretion was used to evaluate collecting duct reabsorption. BP was lowered in rats by an aortic snare to approximately 95 mm Hg and papillary collecting duct (PCD) samples were obtained. The snare was then released and 100-400 micrograms/kg/min of guanabenz was infused and the rate adjusted to produce a drop in BP similar to that of the initial period. During the control period, PCD fluid to plasma inulin ratios were 23.1 +/- 4.3 and 37.3 +/- 4.6 at the base and tip, respectively. This ratio was decreased to 10.9 +/- 3.1 and 15.7 +/- 4.6 at the base and tip puncture sites, respectively, during guanabenz infusion. PCD chloride reabsorption during the initial period was 40.3% +/- 7.0% of the load delivered. During the guanabenz infusion, however, only 12.8% +/- 3.4% of the delivered load was reabsorbed (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)