Tubuloglomerular feedback control in kidneys of adrenalectomized rats

The technique of orthograde tubular microperfusion has been used to test the functional integrity of tubuloglomerular feedback control in adrenalectomized rats. Kidneys of these animals display high renin contents, as well as an impairment of electrolyte transport in the loop of Henle, which was demonstrated by monitoring the electrical conductivity of fluid entering the distal tubule over a perfusion range of 10–50 nl/min. The calculated electrolyte concentration of the perfusion fluid leaving the loop of Henle was significantly higher in adrenalectomized rats than in controls. Intratubular stop flow pressure (SFP) during perfusion with a modified endproximal Ringer's solution was recorded continuously in early proximal segments at rates of 0–40 nl/min. Direct pressure transmission from perfused endproximal tubular segments to the pressure recording capillary was prevented by paraffin blockade of the intermediate proximal segment. An intact feedback response in each of the 32 tubules of the adrenalectomized rats studied was observed, which was reversible and could be demonstrated repetitively in individual tubules. Thus SFP at 40 nl/min was –11.2±1.3 mm Hg in adx rats as compared to –8.1±1.3 in control rats. An increased freedback sensitivity was observed in dexamethasone treated adrenalectomized rats despite normal renal renin content. Here SFP at 40 nl/min was –15±1.1. Thus, the feedback signal can be mediated in the apparent absence of adrenal steroid hormones and a high sensitivity of tubuloglomerular feedback control is not necessarily paralleled by high renin content of renal tissue.Index of Abbreviations adx adrenalectomized - AI angiotensin I - b.w. body weight - k conductivity (Siemens cm^–1) - EPFR early proximal flow rate - FFP free flow pressure - P_tub intratubular pressure - P mean arterial pressure - SFP stop, flow pressure - SNGFR single nephron glomerular filtration rate - V_u urine flow rate - ø perfusion rate through the loop of Henle Supported by the Deutsche Forschungsgemeinschaft (Hi 97/14)     

Modification of feedback influence on glomerular filtration rate by acute isotonic extracellular volume expansion

The behavior of the feedback mechanism, that causes glomerular capillary pressure and filtration rate to decrease when tubule fluid flow rate through the loop of Henle of the same nephron is increased, was examined in rats before and during isotonic extracellular fluid volume expansion. The loop of Henle was perfused from the late proximal tubule at either 10 or 40 nl/min while proximal fluid was collected to measure single nephron filtration rate (SNGFR), while proximal stop-flow pressure (P_SF) was measured, or while fluid was collected from the early distal tubule to assess reabsorption of fluid and electrolytes by the loop of Henle. During control periods increasing loop perfusion caused SNGFR to decrease 37%, P_SF to decrease 19%, and absorption of fluid, sodium and chloride by the loop of Henle to increase. After 1 h of infusion of isotonic NaCl solution the same change in loop flow causes a 19% decrease in SNGFR and an 8% decrease in P_SF. Fluid absorption by the loop of Henle did not increase with increased loop perfusion. Increases in Na and Cl absorption were similar to the increases in control periods. The smaller decreases in SNGFR and P_SF indicate that acute volume expansion decreases the sensitivity of the feedback response. The mechanism of this decrease in gain could involve interference with local generation or action of angiotensin, or a change in the composition or pressure of interstitial fluid tending to dilate the pre-glomerular resistance vessels.     

Chronic salt loading: Effects on plasma volume and regulation of glomerular filtration rate in Wistar rats

Summary Experiments were carried out in Wistar rats to determine whether the loss of sensitivity of the tubuloglomerular feedback mechanism (TGF) which is known to occur in volume expansion is due to a change in the functional characteristics of the juxtaglomerular apparatus or to a change in some property of the tubular fluid which influences the feedback signal at the macula densa. Proximal tubular fluid was collected by means of a microperfusion/suction pump from Wistar rats maintained for a minimum of 10 days on a high salt diet and also from rats fed a control low salt diet. Both fluids were then used to perfuse loops of Henle in rats from both groups and the feedback response assessed from the change in early proximal tubular flow rate (EPF). In high salt rats, perfusion of the loop of Henle with homologous tubular fluid confirmed the loss of sensitivity of the TGF mechanism in volume expansion, the response of EPF was practically absent. In contrast, the low salt rat responded with a 50% decrease in EPF to loop perfusion at 40 nl/min with its homologous fluid. On the other hand, when the loop of Henle in high salt rats was perfused at 40 nl/min with heterologous (low salt) tubular fluid, EPF again decreased by some 50% whereas EPF in low salt rats failed to respond to loop perfusion with high salt fluid. From these results it is concluded that in rats chronically volume expanded by a high salt diet an unknown inhibitory principle occurs in the proximal tubular fluid which reduces the sensitivity of the tubuloglomerular feedback mechanism.     

Resetting of tubuloglomerular feedback in acute volume expansion in rats

Loss of sensitivity or “resetting” of tubuloglomerular feedback has been reported after both acute and chronic volume expansion in rats. In chronic volume expansion due to dietary salt loading, resetting was found to result from the appearance of an inhibitory factor in tubular fluid. The aim of the present study was to test the possibility that resetting after acute isooncotic volume expansion may also be due to such an inhibitor. Rats were acutely volume expanded (4.5% of body weight) by infusion of a solution of fresh plasma and Ringer's solution. Tubuloglomerular feedback activity was assessed in expanded and control animals by measuring early proximal flow (EPF) rate during perfusion of the loop of Henle at varying rates with proximal tubular fluid harvested from the control (control TF) and expanded animals (AVE TF). When loops of Henle in control animals were perfused with control TF at 10, 20 or 40 nl min⁻¹, EPF fell from (mean ±SD) 29.8±5.6 at zero loop flow to 27.5±7.5, 21.1±4.2 and 15.5±4.5 nl min⁻¹ gKW⁻¹ respectively. Perfusion at the same rates with control TF in expanded animals reduced EPF from 39.5±9.6 (at zero loop flow) to 35.9±11.3, 31.6±4.3 and 22.9±6.8 nl min⁻¹ gKW⁻¹ respectively. When loops of Henle in control animals were perfused with AVE TF, EPF fell from 28.6±9.5 (zero loop flow) to 23.5±8.6, 19.9±8.2 and 15.6±6.5 nl min⁻¹ gKW⁻¹ respectively. Perfusion at these rates with AVE TF in the expanded animals depressed EPF from 36.7±7.8 (at zero loop flow) to 33.6±7.3, 28.6±7.6 and 22.7±8.0 nl min⁻¹ gKW⁻¹ respectively. Since the responses to the two perfusion fluids were the same in each group, it is concluded that there is no inhibitory factor present in AVE TF. Although EPF at each perfusion rate was significantly higher in the expanded animals than in control, the change in EPF per unit change in loop perfusion rate was the same in both groups from which it is concluded that no resetting of tubuloglomerular feedback occurred in the present study. Some of the work described here was presented to the German Physiological Society at its 59th Meeting, Dortmund, March 26–30, 1984 and to the 17th Congress of the Gesellschaft für Nephrologie, Mainz, Sept. 22–25, 1985 and appears in abstract form in Pflügers Arch 400:R21 (1984), and Kidney Int 29:1253 (1986) respectively     

Evidence for tubuloglomerular feedback in juxtamedullary nephrons of young rats

To determine whether the filtration rate of juxtamedullary nephrons is regulated by tubuloglomerular feedback (TGF), we developed two micropuncture techniques suitable for the papilla of young rats. One consisted of measuring the tubular flow in descending limbs of Henle loops (VDLH) while the ascending limbs of the loop of Henle (ALH) were perfused at various rates with three different solutions: modified Ringer, artificial Henle loop solution, and Ringer containing 10(-4) M furosemide. SNGFR was also measured in several juxtamedullary nephrons. The other protocol consisted of measuring the tubular stop-flow pressure (PSF) in descending limbs of the loop of Henle upstream to a wax block. Distal to the block Ringer was perfused at various rates through ALH. Our results provide the first evidence of a TGF response in juxtamedullary nephrons. VDLH and SNGFR decreased during Ringer perfusion to 42 +/- 4 and 44 +/- 4% of their values at zero perfusion. The same pattern was observed using Henle loop solution as perfusate, whereas with furosemide VDLH did not change. The maximal decrease in PSF was 14.1 +/- 1.4 mmHg. The perfusion rate necessary to induce a half-maximal PSF decrease was 9.1 +/- 0.9 nl/min. Similar micropuncture techniques were used in proximal tubules of surface nephrons in these rats, which in comparison to the deep nephrons showed smaller feedback responses. The mechanism seems to be active at physiological nephron flow rates in both nephron populations. Thus, TGF can exert its effect on GFR of the whole kidney by acting in both deep and surface nephrons.     

Micropuncture method for the determination of nephron filtration rate

Summary Two possible artifacts may explain the phenomenon that nephron GFR (N-GFR) measured by distal tubular puncture is smaller than that measured by proximal tubular puncture: a loss of the inulin-like substance used in this laboratory (polyfructosan) from the tubular lumen or unreliable distal punctures. To test these possibilities (a) known amounts of polyfructosan were injected into the proximal tubule and the percentage recovery from the distal tubule measured, (b) N-GFR was measured by distal puncture, subsequently by recollection from the same site and finally by a proximal puncture.On the average, 98.5±7.5% of the proximally injected polyfructosan was recovered from the distal tubule. This is not significantly different from 100% (p>0.1) and demonstrates that proximal tubule and loop of Henle are impermeable to polyfructosan. The ratio between the N-GFR measured by a distal puncture and that measured by subsequent recollection was 1.016±0.096 and not significantly different from 1.000 (p<0.20), demonstrating the reliability of distal tubular puncture. The mean distal N-GFR of 27.9±5.3 nl/min was significantly smaller (p<0.001) than the proximal N-GFR of 35.1±8.0 nl/min. The existence of the proximal-distal N-GFR difference thus is confirmed and two possible artifacts eliminated. The best explanation remains the operation of a tubulo-glomerular feedback mechanism.A current point of dispute is the effect of alterations in intratubular pressure (ITP) on N-GFR. Collection of tubular fluid at ITPs below the previously measured free flow pressure (FFP) resulted in a change of N-GFR of 0.45 nl/min· cm H₂O. In contrast, fluid collection at ITPs greater than the FFP resulted in a change of N-GFR of 1.48 nl/min· cm H₂O. We conclude that although N-GFR is sensitive to ITP changes in both directions, pressure decreases are of little practical importance for the determination of N-GFR whereas intratubular pressure increases are to be avoided.This work was supported by the Deutsche Forschungsgemeinschaft.On leave from the University of Melbourne as a recipient of a scholarship from the Deutscher Akademischer Austauschdienst.     

The existence of a tubulo-glomerular feedback mechanism in the Amphiuma nephron

A single nephron tubulo-glomerular feedback control of the glomerular filtration rate, which is known in mammlian animals, could be one way by which amphibians regulate the glomerular filtration rate (GFR). To investigate whether theAmphiuma means shows any sign of a tubuloglomerular feedback control, micropuncture experiments were carried out. Six different series of experiments were performed.In the first series, tubular stop-flow pressure (SFP) was measured during distal tubular perfusion with amphibian Ringer solution at a rate of 10, 25 and 50 nl/min. A significant decrease of SFP was found at the three perfusion rates compared to the controls. In the second group, single nephron glomerular filtration rate (SNGFR) was measured, while the distal tubule was perfused at 10, 25 and 50 nl/min. At a perfusion rate of 10 nl/min the SNGFR did not decrease, whereas at 25 and 50 nl/min it decreased significantly. In the third group the perfusion pipette was located in the proximal tubule and the nephron was perfused at 10, 25 and 50 nl/min, while at the same time the proximal tubular stop-flow pressure was measured. No reduction of SFP was found at a perfusion rate of 10 nl/min, while significant reductions were noted at rates of 25 and 50 nl/min. In the fourth group the SNGFR was measured in the distal tubule beyond the macula densa and in Bowman's space of the same nephron. No significant difference was found. In the fifth group, the glomerular capillary pressure (GCP) was measured before and after blockade of the tubular fluid flow. No significant difference was found between these two measurements.The sixth series deals with the changes occuring at the single nephron level by the tubulo-glomerular feedback control. The single nephron filtration fraction (FF) was determined from efferent arteriolar protein concentration with and without a feedback-induced reduction of the SNGFR. The FF values were not significantly different from one another. From these results and data from the other series, the afferent (R _aff) and efferent (R _eff) arteriolar resistances were calculated.R _eff did not change, whileR _aff increased significantly when a feedback stimulus was applied.These experiments indicate the existence of a tubuloglomerular feedback control which depresses the SNGFR and SFP by contracting the afferent arteriole.     

Renal response to volume depletion and expansion in Milan hypertensive rats

In previous studies on Milan hypertensive (MHS) rats, we found an impaired tubuloglomerular feedback (TGF) response before, during and after development of hypertension. In the present study MHS rats and rats of the Milan normotensive strain (MNS) were investigated after 24 hours of volume depletion (VD) and subsequently after 5% isotonic volume expansion (VE) with respect to whole kidney function, interstitial hydrostatic (P_int) and oncotic (II_int) pressures, stop-flow pressure characteristics of TGF and changes in early proximal flow rate in response to increased loop of Henle flow. MHS rats had higher mean arterial blood pressure (P_a) than MNS rats (129 vs. 101 mmHg) both after VD and after subsequent VE. No difference in glomerular filtration rate (GFR) was found. Both strains had a low urine flow rate (1.5 μl min^-1) during VD, which increased fourfold after VE. The interstitium was significantly more dehydrated in MHS, as indicated by a more negative net interstitial pressure (P_int–±_int t than in MNS (-1.3 ± 0.3 vs. ± 0.0 ± 0.5 mmHg) after VE. The TGF mechanism was more activated in MHS during volume depletion, as indicated by a larger drop in stop-flow pressure (P_sf) in response to loop of Henle perfusion (7.1 ± 0.7 vs. 4.7 ± 0.2 mmHg, P < 0.05). However, during VD the loop of Henle flow that elicited half maximal response in P_sf, the turning point (TP), was equally low in MHS and MNS (13.5 ± 0.6 and 14.3 ± 0.4, respectively). After VE, however, TP increased significantly more in MNS to (32.6 ± 2.1 nl min^-1) then in MHS (to 21.8 ± 0.9 nl min^-1, P < 0.05). It is concluded that the blunting of the TGF resetting in response to VE in MHS rats may well be of importance in the development of hypertension in the MHS strain.     

Inhibitory effect of methylxanthines on feedback control of glomerular filtration rate in the rat kidney

Summary Microperfusion experiments were performed in rats to assess the effect of luminal application of theophylline and the more lipophilic 3-isobutyl-1-methylxanthine (IBMX) on feedback regulation of glomerular filtration rate. Elevation of loop of Henle flow from 0 to 40 nl/min caused a 20.3±4.5% reduction of stop flow pressure (SFP) and a 32.2±3.7% reduction of early proximal flow rate (EPFR) when the perfusate was a 140 mM NaCl solution. When theophylline was added in a concentration of 5 mM SFP fell by only 5.3±1.8% and EPFR by 7.9±3.3%, changes which were significantly smaller than in the control perfusions (P<0.01). An identical change of loop of Henle flow in the presence of IBMX in concentrations of 1 and 5 mM was associated with a 4.5±3.9% decrease and a 12.1±2.7% increase of EPFR. In orthograde perfusion experiments full inhibition of the feedback response was noted at an IBMX concentration of about 1 mM while during retrograde perfusion a concentration of 0.4 mM was sufficient to produce the same effect. This indicates that methylxanthines diffuse out of the loop of Henle to a considerable extent. Methylxanthines reduced absolute and fractional water absorption along the loop of Henle to some extent while Cl absorption rates and early distal Cl concentrations were not significantly altered. Cyclic AMP applied from the luminal side in a concentration of 10 mM did not affect the feedback response of EPFR to flow elevation from 0 to 40 nl/min. Luminal application of dibutyryl cyclic AMP at 10 mM induced a small, but significant reduction of the feedback response when tested by pairedt-test (P<0.05). Our results show that luminal application of methylxanthines strongly interfere with feedback regulation of glomerular filtration rate. It is unclear at present whether this effect is related to inhibition of cyclic nucleotide phosphodiesterase and a rise in tissue cyclic AMP levels or to interference with a mechanism involving the local action of adenosine or 5-AMP.Supported by the Deutsche Forschungsgemeinschaft     

Proximal tubular lithium/plasma concentration ratio depends on tubular fluid flow rate

Previous studies have given slightly different values of proximal tubular fluid/plasma lithium concentration ratios (TF/P)_Li. The aim of the present study was to investigate whether the technique of tubular fluid collection by micropuncture might contribute to this difference. Early distal and late proximal tubular fluid was collected by micropuncture during recording of early proximal hydrostatic pressure with varying pressures applied to the collection pipette. Distal fluid collection with 2–3 mmHg suction was unable to influence distal flow, proximal tubular pressure, nephron filtration rate (SNGFR), (TF/P)_Li or (TF/P)_Na significantly, as compared with values obtained without suction. In contrast, during late proximal collections, tubular pressure could be controlled. Within the range 2 mmHg below to 6 mmHg above free flow pressure maintained during collection, proximal flow rate varied from 9 to 44 nL min^-1. (TF/P)_Li was positively and significantly correlated to late proximal flow rate (P<0.005, r=0.65), independent of tubular pressure. (TF/P)_Na was 1.03 and independent of flow rate and pressure. With collection adjusted to a late proximal flow of 24 nL min^-1, (TF/P)_Li would be about 1.15. At the more physiological flow rate of 12–14 nL min^-1, (TF/P)_Li was not significantly different from 1.00. It is concluded that proximal Li transport is distinct from Na transport and lags behind Na and water transfer only when flow rate is increased above normal. The result emphasizes the importance of the micropuncture fluid collection technique for estimates of proximal (TF/P)_Li.     

Further studies on oscillating tubulo-glomerular feedback responses in the rat kidney

Free flow microperfusion (closed feedback loop) experiments in halothane-nitrous oxide-anaesthetized rats demonstrate two distinct oscillating TGF pressure responses to pertubations in early distal tubular flow. Flow rate changes of 2-8 nl min-1 induced a 0.02-0.03 Hz rhythm, while changes above 25 nl min-1 often induced a 0.17-0.18 Hz rhythm in proximal tubular pressures. Following interruption of microperfusion, the intratubular pressure showed an initial undershoot and a gradual oscillating return to the control level. Each nephron responded individually to systemic changes with oscillations differing in frequency, phase and amplitude from that of its neighbour. Postglomerular vascular pressure (PSV) oscillated synchronous with the proximal luminal pressure (PProx) of the same nephron; the amplitude of the PSV might be larger than that of the corresponding PProx. The PSV decreased by 2-3 mmHg as Henle loop flow was increased by 7 nl min-1. Furosemide (0.1-0.2 mM) in the microperfusate abolished the oscillations, and caused PSV and PProx to increase by 3-4 mmHg. This effect was rapidly reversible.     

Further evidence for an inverse relationship between macula densa NaCl concentration and filtration rate

It has been concluded that tubulo-glomerular feedback mechanism is triggered by changes in NaCl concentration ([NaCl]) at the macula densa. This conclusion is based on the demonstration that changes in filtration rate produced during retrograde perfusion of the loop of Henle depend upon the perfusate [NaCl]. Experiments were performed to evaluate whether the effect on glomerular function of orthograde perfusion of the loop of Henle is consistent with this conclusion. Early proximal flow rate ( ), stop-flow pressure (P _SF), early distal chloride concentration ([Cl]), and flow rate were measured during perfusion of the loop of Henle with mannitol solution (300 mosm kg^–1), 30 mM NaCl+mannitol (300 mosm kg^–1), 140 mM Na isethionate and artificial tubular fluid. When distal flow exceeded 10 nl min^–1, the magnitude of the glomerular response was predictable from the [Cl]. The linear regression line, , did not differ from that obtained previously with the retrograde technique. Retrograde perfusion with 140 mM Na isethionate was without effect on . We conclude that the effect on glomerular function of perfusion of the loop of Henle in either an orthograde or a retrograde direction with these solutions depends upon the chloride concentration at the macula densa.     

Technical problems in the micropuncture determination of nephron filtration rate and their functional implications

Summary The theory of a functional coupling between distal tubular fluid composition and glomerular filtration rate implies that the blockade of flow at a proximal site should lead to a marked increase of GFR. This potential alteration of steady state GFR was studied by comparing the influence of sampling from distal or proximal sites on the filtration rate of identical nephrons. During antidiuresis an average GFR of 25.2 nl/min±7.5 S.D. was found in distal collections, while proximally collected samples gave an average GFR of 34.5 nl/min±8.4 S.D. This difference of 9.3 nl/min is highly significant (p<0.001). During saline diuresis a mean nephron GFR of 41.6 nl/min±5.0 was found by distal sampling and of 45.3 nl/min±5.4 by proximal sampling (p>0.05). The proximal-distal difference in nephron GFR is interpreted to indicate the operation of a tubulo-glomerular feedback control system. Thus, a true steady-state GFR probably cannot be obtained by proximal fluid collection.Even in the presence of high intratubular pressures and unusually short oil blocks no evidence of sample contamination by retrograde fluid flow past an injected oil block was obtained.The application of a counter-pressure to the sampling pipette which has been recommended by Gertzet al. [5] as a means to standardize fluid collections, was found to lead to abnormally high intratubular pressures. The reason for this finding appears to be an unexpectedly high and inconstant tip resistance to flow during fluid flow into the pipette.     

Influence of water-diuresis or saline volume expansion on deep nephron tubuloglomerular feedback

We have recently demonstrated the existence of a tubuloglomerular feedback mechanism in juxtamedullary nephrons in rat kidneys during antidiuresis. In the present experiments, we have investigated the influence of water-diuresis on Munich Wistar rats and in homozygote Brattleboro rats. We have also observed the effect of saline volume expansion on the tubuloglomerular feedback of juxtamedullary nephrons in Munich Wistar rats. For comparison, the feedback mechanism was also studied in surface nephrons during water-diuresis in Munich Wistar rats. Measurements of flow rate in the descending limb of Henle and single nephron glomerular filtration rate (GFR) using micropuncture at the renal papilla were performed, while the ascending limb was microperfused at varying perfusion rates with a modified Ringer solution. In surface nephrons early proximal flow rate, single nephron GFR and stop-flow pressure was measured during microperfusion of the end-proximal loop. No significant changes were measured in surface nephrons during water-diuresis: the tubular flow rates, feedback responses, stop-flow pressure and stop-flow pressure changes were similar to those during anti-diuresis. In juxtamedullary nephrons, Henle loop flow rate increases during water-diuresis but the feedback-mediated flow and single nephron GFR response curves obtained during microperfusion were unaltered compared to controls. Together, these results indicate that the feedback could be more activated during water-diuresis than during control conditions. On the other hand, during saline volume expansion, reduced tubuloglomerular feedback sensitivity was found as shown earlier for surface nephrons. The reduction of tubuloglomerular feedback sensitivity therefore seems to be important in maintaining salt-, but not water-balance.     

Tubuloglomerular feedback in rat kidneys of different renin contents

Variations in flow rate through the loop of Henle in the range of 0--50 nl/min were induced using pressure controlled microperfusion. Simultaneously, with the aid of a second pressure-microperfusionsystem, the glomerular function of the same nephron was studied by continuous measurement of two parameters, early proximal flow rate (EPFR) and/or stop flow pressure (SFP). Elevation of loop perfusion above physiological values (40 nl/min) resulted in a drop of EPFR and SFP, whereas lowering perfusion rates had no effect. This feedback behaviour was studied in kidneys with different renin contents to test the role of the renin-angiotensin system in the mediation of the macula densa signal to the adjacent glomerular vessels. Renal renin content, measured after micropuncture experiments by incubation with substrate followed by radioimmunoassay of angiotensin I, was unaltered in control (Ia) and heminephrectomized rats (Ib), lowered in contralateral kidneys of 2 kidneys Goldblatt hypertensive rats (IIa), in DOCA- and salt-loaded rats (IIb), and in DOCA-, salt-loaded and heminephrectomized rats (IIc), and it was evaluated in clipped kidneys of Goldblatt hypertension rats (IIIa). Micropuncture evaluation of the tubuloglomerular feedback behaviour in these experimental groups revealed the following results: 1. a feedback response under all conditions independent of the widely varying renin contents (1000-fold), 2. an asymmetrical behaviour of the feedback response in all kidneys as demonstrated by suppression of EPFR and SFP at elevated loop flow rates, but no change of these parameters when loop flow was interrupted. 3. compared to controls the decrease of each GFR parameter between 0 and 40 nl/min loop perfusion was lower in DOCA- and salt-loaded rats (IIb, IIc). Additional heminephrectomy (IIc) had no further influence on the reduced feedback response in DOCA- and salt-loaded rats, whereas this maneuver reduced the renal renin content drastically. A somewhat higher response than in controls was found in heminephrectomized rats (IIb) and in clipped kidneys of Goldblatt hypertensive rats (IIIa). These different magnitudes of feedback responses do not correlate with the renal renin content. It has been concluded, therefore, that renal renin activity is not the sole determinant of the effectiveness of the tubuloglomerular feedback response.     

Evidence for feedback mediated reduction of glomerular filtration rate during infusion of acetazolamide

Systemic administration of acetazolamide (ACZ) causes glomerular filtration rate (GFR) to fall. Clearance and micropuncture experiments were done to define the mechanism of this drug effect. When rats were infused with ACZ intravenously, kidney GFR fell by 30% and single nephron (SN) GFR (measured by collecting distal tubule fluid) fell by 23%. Changes in arterial blood pressure, arterial pH, extracellular fluid volume, and proximal tubule pressure were not sufficient to account for the decrease in GFR. When SNGFR was measured by collecting proximal tubule fluid, with the loop of Henle having been blocked for 2–5 min, SNGFR was higher than the distally measured value and was not different than control. The results are consistent with the fall in GFR being caused by activation of the tubulo-glomerular feedback mechanism.     

Autoregulation of the glomerular filtration rate and the single-nephron glomerular filtration rate despite inhibition of tubuloglomerular feedback in rats chronically volume-expanded by deoxycorticosterone acetate

Tubuloglomerular feedback (TGF) function and autoregulation (renal blood flow RBF; glomerular filtration rate, GFR; single-nephron glomerular filtration rate, SNGFR) were examined in rats chronically treated with deoxycorticosterone acetate (DOCA) and given isotonic saline to drink. DOCA treatment depressed arterial plasma renin activity, expanded plasma volume by 25% and increased arterial blood pressure. Autoregulation of RBF and GFR was maintained in the DOCA animals above 90 mm Hg and 110 mm Hg respectively, whereby both GFR and RBF were lower than in controls. Micropuncture experiments demonstrated the absence of TGF in the DOCA animals. There was no difference between SNGFR values measured in the distal and proximal tubules, nor was there a significant response of SNGFR when loops of Henle were perfused with Ringer's solution at 20 nl/min. Loop perfusion in control rats with tubular fluid collected in DOCA rats elicited a normal TGF response, showing that TGF inhibition in the DOCA animals is due to changes in the function of the juxtaglomerular apparatus. In contrast to control rats, proximal SNGFR was perfectly autoregulated. These results suggest that TGF is not primarily responsible for autoregulation and that the vasodilatation normally resulting from acute TGF interruption is therefore compensated by some other mechanism such that RBF and GFR are lower than in controls.     

Micropuncture studies on the filtration rate of single superficial and juxtamedullary glomeruli in the rat kidney

Summary Single nephron filtration rates of superficial and juxtamedullary nephrons were determined in high and low sodium rats. Single nephron GFR was calculated from TF/P inulin and tubular flow rate in superficial nephrons and single juxtamedullary GFR from corresponding data in long loops of Henle. In low sodium rats superficial nephron GFR was 23.5±6.4 (SD)×10^–6 ml/min×g KW, juxtamedullary nephron GFR was 58.2±13.6 and total kidney GFR (C _In) was 0.94±0.16 ml/min×g KW. Using these single nephron values, total kidney GFR and a total number of 30,000 glomeruli per kidney, the number of superficial and juxtamedullary glomeruli was calculated to be 23,267 and 6,733, respectively. During high sodium diet superficial nephron GFR increased to 38.1±11.3 and single juxtamedullary GFR decreased to 16.5±6.6, total kidney GFR increasing to 1.01±0.24. Calculation again revealed the same distribution of the two nephron types. End-proximal TF/P inulin in superficial nephrons was 2.36±0.36 in low sodium and 2.31±0.28 in high sodium rats. Loops of Henle TF/P inulin and intratubular flow rate were inversely related: the highest TF/P inulin values and lowest intratubular flow rates were found in the descending limb. These data quantify the distribution of superficial and juxtamedullary nephrons on a functional basis and suggest a mechanism by which the kidney adjusts sodium excretion by altering the contribution of each nephron type to total kidney GFR.Supported by the Deutsche Forschungsgemeinschaft and the U.S. Department of the Army, through its European Research Office.     

Beschleunigte Natriurese und Diurese beim Hochdruck: Folge einer Resorptionshemmung in der Henleschen Schleife

Summary Exaggerated natriuresis and diuresis in chronic hypertension: Result of impaired fluid reabsorption in the loop of Henle.Micropuncture studies were performed in normotensive and chronically hypertensive rats (unilateral renal artery stenosis). Excretory and tubular function of the untouched kidney exposed to the high blood pressure (185 mm Hg) was investigated before and after i.v. 2.5% sodium chloride loading (0.1 ml/min) and compared to that of normotensive rats (98 mm Hg). The onset of diuresis and natriuresis was more rapid in the hypertensive rats than in the control group. Fractional sodium excretion of the hypertensive rats rose to 11% of the filtered load, while it reached only 3% in the controls. Total GFR and superficial single nephron GFR were elevated in both groups, but the increase in filtration rate occured earlier in the hypertensive group. Fractional sodium and water reabsorption in the proximal tubule decreased in both groups to the same extent. In the hypertensive rats, fractional water reabsorption along the loop of Henle was less than half of normal. The reduced fractional reabsorption in Henle's loop was already present in the control period and is thought to be the result of an increased medullary blood flow caused by the high blood pressure. Early distal TF/P-Inulin ratio declined from 2.8 to 1.9 in the hypertensive group and from 4.7 to 2.45 in the control group. In the normotensive group the reduced proximal reabsorption was partly compensated by an increased fractional reabsorption in distal tubules and collecting ducts. In contrast, there was a marked reduction of fractional water reabsorption in distal tubules of the hypertensive group. The combined inhibition of fractional reabsorption in the proximal tubule, in Henle's loop and in the distal tubule leads to the exaggerated natriuresis and diuresis in the hypertensive rats.

     

Functional consequences at the single-nephron level of the lack of adenosine A1 receptors and tubuloglomerular feedback in mice

Mice deficient for adenosine A1 receptors (A1AR) lack tubuloglomerular feedback (TGF). In vivo micropuncture experiments were performed under anesthesia in A1AR-deficient and wild-type littermate mice to study the effects of chronic absence of A1AR on fluid and Na⁺ reabsorption along the nephron, as well as the functional consequences at the single-nephron level of the lack TGF. Evidence is provided for an A1AR-mediated tonic inhibition of Na⁺ reabsorption in a water-impermeable segment of the loop of Henle, possibly the thick ascending limb. In contrast, proximal tubular reabsorption of fluid, Na⁺ and K⁺ was unaffected by the chronic absence of A1AR. Experiments in which artificial tubular fluid was added to free-flowing late-proximal tubules demonstrated an essential role of A1AR/TGF in the stabilization of fluid and Na⁺ delivery to the distal nephron. Further, the occurrence of spontaneous oscillations of hydrostatic pressure in proximal tubule (P _PT) at a frequency of about 32 mHz depended on intact A1AR/TGF. In comparison, the normal, stabilizing reduction in P _PT following the initial rise in P _PT during sustained small increases in proximal tubular flow rate does not require A1AR/TGF; TGF-independent mechanisms appear to compensate in this regard for a lack of TGF under physiological conditions and the lack of TGF is unmasked only when supraphysiological flow rates overwhelm TGF-independent compensation.