Dopamine production by the isolated perfused rat kidney

1984 ◽  
Vol 62 (3) ◽  
pp. 272-276 ◽  
Author(s):  
Andrew D. Baines ◽  
Rosa Drangova

We used isolated perfused rat kidneys to examine dopamine (DA) production and its relation to renal function. Both innervated and chronically surgically denervated kidneys perfused with a solution containing neither albumin nor tyrosine, excreted 0.2 ± 0.1 ng DA∙min−1∙g wet weight−1 during the 10-min collection period between 30 and 40 min after starting perfusion. When perfused with 6.7% albumin, without tyrosine, innervated kidneys excreted 1.0 ± 0.06 ng DA∙min−1∙g−1 and denervated kidneys excreted 1.0 ± 0.07 DA∙min−1∙g−1. When 0.03 mM tyrosine was included in the albumin perfusate, innervated kidneys excreted 1.2 ± 0.1 ng DA∙min−1∙g−1 (p < 0.1). Under these conditions DA excretion continued for at least 100 min at which time it was 0.6 ng∙min−1∙g−1 and 86 ng/g kidney weight had been excreted. Denervated kidneys perfused with albumin + tyrosine excreted 0.9 ± 0.13 ng DA∙min−1∙g−1. Renal stores of free DA, conjugated DA, and dihydroxyphenylalanine (DOPA) could have provided at the most 30 ng/g of DA. Carbidopa inhibited DA excretion completely. DA excretion did not correlate with renal vascular resistance, inulin clearance, or fractional sodium excretion. In summary, nonneural tissue in isolated perfused kidneys produced DA at the same rate as denervated kidneys in vivo. Less than one-third of the DA produced by isolated kidneys could have come from intrarenal stores of DOPA, free DA, and conjugated DA; the rest was synthesized from unknown precursors. Circulating DOPA and tyrosine were not the DA precursors, but albumin was required to obtain production rates similar to those in vivo. Nonneuronal DA production did not influence renal hemodynamics, glomerular filtration rate (GFR), or sodium excretion.

1987 ◽  
Vol 252 (2) ◽  
pp. F331-F337 ◽  
Author(s):  
W. Lieberthal ◽  
M. L. Vasilevsky ◽  
C. R. Valeri ◽  
N. G. Levinsky

Interactions between antidiuretic hormone (ADH) and renal prostaglandins in the regulation of sodium reabsorption and urinary concentrating ability were studied in isolated erythrocyte-perfused rat kidneys (IEPK). In this model, hemodynamic characteristics are comparable to those found in vivo, and tubular morphology is preserved throughout the period of perfusion. [Deamino]-D-arginine vasopressin (dDAVP) markedly reduced fractional sodium excretion (FE Na) in the IEPK from 3.5 +/- 0.6 to 0.45 +/- 0.14%. After indomethacin, FE Na fell still further to 0.08 +/- 0.02%. In the absence of dDAVP indomethacin had no effect on sodium excretion; FE Na was 2.4 +/- 0.6% in control and 2.0 +/- 0.4% in indomethacin-treated groups. dDAVP increased urine osmolality in the IEPK to 741 +/- 26 mosmol/kg. When prostaglandin synthesis was blocked with indomethacin, urinary osmolality increased further to 1,180 +/- 94 mosmol/kg. In isolated kidneys perfused without erythrocytes (IPK), dDAVP decreased FENa from 14.5 +/- 1.8% to 9.6 +/- 1.2%; addition of indomethacin had no further effect. dDAVP increased urine osmolality only modestly to 350 +/- 12 mosmol/kg in the IPK and indomethacin did not increase concentrating ability further (342 +/- 7 mosmol/kg). Thus the IEPK (unlike the IPK) can excrete a markedly hypertonic urine in response to ADH. ADH also enhances tubular reabsorption of sodium in the IEPK. Prostaglandins inhibit both these actions of ADH but do not directly affect sodium excretion in the absence of the hormone.


1986 ◽  
Vol 64 (11) ◽  
pp. 1427-1433 ◽  
Author(s):  
Daniel B. Ornt

Renal adaptation for potassium (K) conservation has been demonstrated in isolated perfused kidneys from rats within 3 days of K depletion and appears to be independent of aldosterone and sodium excretion. This study was designed to investigate whether the renal adaptation for K conservation is independent of ambient [K] and renal tissue levels of K and whether ouabain may have effects on K excretion, which are in constrast to the effects on K excretion in normal animals, in the first study, rats K depleted for 3 days received 2500 μequiv. KCl intraperitoneally, while other K-depleted rats and a group of control diet animals received intraperitoneal H2O alone to determine whether simple restoration of K deficits would reverse the renal adaptation for K conservation. Intraperitoneal KCl increased plasma [K] and kidney tissue K significantly within 3 h in the K-repleted group compared with the K-depleted rats. Isolated kidneys were perfused from the three groups of rats 3 h after intraperitoneal injection. Despite K repletion in vivo, perfused kidneys from the K-repleted group still had significantly decreased K excretion (1.28 ± 0.085 μequiv./min) compared with controls (2.05 ± 0.291 μequiv./min), and K excretion was still not different from the K-depleted group (0.57 ± 0.134 μequiv./min). However, fractional K excretion by the kidneys from K-repleted rats was increased above K-depleted kidneys (0.48 ± 0.051 vs. 0.18 ± 0.034, p < 0.01). Despite the increased renal tissue K in K-repleted kidneys at the start of perfusion (285 ± 5.1 vs. 257 ± 5.4 μequiv./g), by the end of the perfusion tissue K in perfused kidneys was identical in all three groups. In the second study, isolated kidneys were perfused from 3-day K-depleted or control rats with either 2 or 6 mM [K] in the perfusate. Isolated kidneys adapted to 3 days of K depletion excreted less K at both 2 and 6 mM [K] compared with controls at the same ambient [K]. The linear relationship of K excretion to perfusate [K] was significantly different in controls compared with low K adapted kidneys (p < 0.001). Finally, when 10−4 M ouabain was added after 60 min of perfusion in kidneys from control diet rats, there was a sodium diuresis and fractional K excretion decreased significantly (0.55 ± 0.043 to 0.32 ± 0.044, p < 0.01). However, in low K adapted kidneys, ouabain had no effect on fractional K excretion (0.020 ± 0.051 to 0.18 ± 0.038) despite a similar increase in sodium excretion. Perfusions of kidneys from 3-day K-depleted rats at 4 × 10−3 M ouabain gave similar results, showing no change in fractional K excretion. Low K adaptation to K depletion developed within 3 days and was not totally abolished by acute K repletion. Maneuvers that favored either a decrease in renal tissue K or an increase in tissue K did not reverse low K adaptation, although renal tissue K levels did alter the rate of K excretion in both controls and K-depleted kidneys. Therefore, a reduction in tissue K was clearly not the sole mediator of renal K conservation. Finally, the markedly different response of low K adapted kidneys to ouabain compared with controls strongly suggests a mechanism for K reabsorption that developed within 3 days of K depletion and is ouabain sensitive.


1990 ◽  
Vol 258 (5) ◽  
pp. F1266-F1272 ◽  
Author(s):  
W. Lieberthal ◽  
A. M. Sheridan ◽  
C. R. Valeri

We have examined the effect of atrial natriuretic factor (ANF) administered with and without mannitol on renal function following ischemic injury in both the isolated erythrocyte-perfused rat kidney and in the rat in vivo. ANF, administered alone after 25 min ischemia in the isolated kidney, reversed postischemic vasoconstriction but did not improve glomerular filtration rate (GFR). Mannitol alone had no effect on either renal vascular resistance or GFR. However, in isolated kidneys treated with the combination of both ANF and mannitol following reflow, GFR (0.65 +/- 0.04 ml.min-1.g-1) was markedly improved compared with GFR in the untreated ischemia group (0.20 +/- 0.04 ml.min-1.g-1) and was not different from GFR in the nonischemic controls (0.68 +/- 0.05 ml.min-1.g-1). Comparable results were obtained in studies performed in vivo. In rats subjected to 45 min ischemia, GFR (0.15 +/- 0.05 ml/min) was reduced compared with the GFR in sham-operated animals (0.95 +/- 0.07 ml/min). ANF or mannitol administered alone following ischemia and reflow did not improve GFR compared with the untreated ischemic group. However, in rats subjected to ischemia and treated with a combination of ANF and mannitol postreflow, GFR (0.69 +/- 0.10 ml/min) was 4.6-fold higher than GFR in the untreated ischemic group. Thus the combination of ANF and mannitol appear to act synergistically to improve GFR following ischemic injury.


1984 ◽  
Vol 246 (4) ◽  
pp. F447-F456 ◽  
Author(s):  
M. J. Camargo ◽  
H. D. Kleinert ◽  
S. A. Atlas ◽  
J. E. Sealey ◽  
J. H. Laragh ◽  
...  

The effects of rat atrial tissue extract on renal hemodynamics and fluid and electrolyte excretion were investigated in the isolated perfused rat kidney (IK). IK were perfused at a constant effective perfusion pressure of about 90 mmHg. After control clearance periods (C), extracts of rat atria (AE) or ventricles (VE) were added to the perfusate and three 10-min experimental periods followed. AE, but not VE, significantly increased (P less than 0.001) renal vascular resistance (RVR) to 133 +/- 8% of C, GFR to 201 +/- 34%, filtration fraction to 245 +/- 41%, urine flow (V) to 675 +/- 131%, fractional excretion (FE) of H2O to 336 +/- 29%, absolute Na excretion (UNaV) to 1,259 +/- 290%, FENa to 642 +/- 129%, UKV to 2,226 +/- 1,237%, and FEK to 542 +/- 119%. Despite the marked natriuresis, since GFR doubled, Na reabsorption rose from 78.3 +/- 36.3 in C to 132 +/- 36.3 mueq/min after AE. The effects of AE were immediate and lasted to the end of the perfusion. The lower the initial control GFR, the larger was the AE-induced increase in GFR. Perfusion with low [Ca] (0.2 mM) or verapamil (10(-5) M) severely blunted the hemodynamic, diuretic, kaliuretic, and natriuretic effects of AE. AE decreased rather than increased the RVR when IK were perfused with vasoconstrictors such as angiotensin II, norepinephrine, or vasopressin. The results demonstrate that AE acts directly on the kidney, eliciting powerful Ca-dependent hemodynamic and natriuretic responses. The natriuresis induced by AE can be accounted for, at least in part, by its renal hemodynamic effects rather than by the presence of a putative tubular natriuretic factor. The hypothesis is advanced that AE contains a substance(s) which behaves as a functional agonist/antagonist of endogenous vasoconstrictors with a preferential site of action on the efferent arterioles of the renal vasculature.


1985 ◽  
Vol 229 (2) ◽  
pp. 545-549 ◽  
Author(s):  
M Lowry ◽  
D E Hall ◽  
J T Brosnan

Isolated perfused rat kidneys removed considerable quantities of glycyltyrosine, glycylhydroxyproline, tetraglycine and prolylhydroxyproline from the perfusate. The component amino acids are released into the perfusate and, in the case of the glycine-containing peptides, there is increased synthesis of serine. Removal of peptides was more than could be accounted for on the basis of filtration, so antiluminal metabolism is indicated. Metabolism of such peptides by the kidney may contribute to renal serine synthesis in vivo.


2008 ◽  
Vol 68 (1) ◽  
pp. 149-154 ◽  
Author(s):  
AF. Carvalho ◽  
MS. Santos-Neto ◽  
HSA. Monteiro ◽  
SM. Freitas ◽  
L. Morhy ◽  
...  

Guanylin and uroguanylin are small cysteine-rich peptides involved in the regulation of fluid and electrolyte homeostasis through binding and activation of guanylyl cyclases signaling molecules expressed in intestine and kidney. Guanylin is less potent than uroguanylin as a natriuretic agent and is degraded in vitro by chymotrypsin due to unique structural features in the bioactive moiety of the peptide. Thus, the aim of this study was to verify whether or not guanylin is degraded by chymotrypsin-like proteases present in the kidney brush-border membranes. The isolated perfused rat kidney assay was used in this regard. Guanylin (0.2 µM) induced no changes in kidney function. However, when pretreated by the black-eyed pea trypsin and chymotrypsin inhibitor (BTCI - 1.0 µM; guanylin - 0.2 µM) it promoted increases in urine flow (deltaUF of 0.25 ± 0.09 mL.g-1/min, P < 0.05) and Na+ excretion (% delta ENa+ of 18.20 ± 2.17, P < 0.05). BTCI (1.0 µM) also increased %ENa+ (from 22.8 ± 1.30 to 34.4 ± 3.48, P < 0.05, 90 minutes). Furthermore, BTCI (3.0 µM) induced increases in glomerular filtration rate (GFR; from 0.96 ± 0.02 to 1.28 0.02 mL.g-1/min, P < 0.05, 60 minutes). The present paper strongly suggests that chymotrypsin-like proteases play a role in renal metabolism of guanylin and describes for the first time renal effects induced by a member of the Bowman-Birk family of protease inhibitors.


1981 ◽  
Vol 240 (4) ◽  
pp. F306-F310
Author(s):  
M. H. Garvey ◽  
D. L. Maude

We measured titratable acid (TA) and NH4 excretion by isolated rat kidneys perfused either with conventional bicarbonate-containing solutions or with solutions in which bicarbonate was replaced by propionate. Rates of TA excretion by bicarbonate-perfused kidneys were similar to in vivo values, 0.27 +/- 0.04 mueq.ml GF-1 (0.21 mueq.min-1.g-1), and increased significantly under bicarbonate-free conditions to 0.70 +/- 0.12 mueq.ml GF-1 (0.42 mueq.min-1.g-1). At the same time the perfusate/urine pH difference (delta pH) increased significantly, from 0.63 +/- 0.06 to 0.92 +/- 0.06. Carbonic anhydrase inhibition by 5 X 10(-4) M acetazolamide alkalinized the urine of bicarbonate-perfused kidneys, while in the bicarbonate-free preparation the urine remained acid (delta pH = 0.27 +/- 0.04) and titratable acid continued to be excreted, though at a reduced rate, 0.19 +/- 0.04 mueq.ml GF-1. Under these same bicarbonate-free carbonic anhydrase-inhibited conditions, lowering the perfusate pH from 7.4 to 7.1 increased delta pH to 0.36 +/- 0.02 and caused total acid excretion (TA + NH4) to rise from 0.29 +/- 0.04 to 0.45 +/- 0.06 mueq.ml GF-1, and increasing the perfusate [HPO4] from 2.4 to 9.6 mM increased TA to 0.80 +/- 0.09 mueq.ml GF-1.


1977 ◽  
Vol 232 (4) ◽  
pp. F377-F382 ◽  
Author(s):  
J. S. Fray

These experiments were designed to test whether changing perfusate calcium or magnesium concentrations affected renin release in the isolated perfused rat kidney, and whether kidneys removed from sodium-loaded or sodium-deprived rats released the same amount of renin in response to identical stimuli. Kidneys were perfused with Kreb-Henseleit solution containing albumin. Renin release was inversely related to perfusate calcium concentration, whereas renin release was directly related to perfusate magnesium. Although a low calcium medium or low perfusion pressure (50 mmHg) stimulated renin release, the release was substantially greater in the sodium-deprived rats. Increasing the perfusate sodium concentration from 85 to 206 mM increased excretion, but did not alter renin release. It is concluded that a) low perfusate calcium and high magnesium concentrations stimulate renin release, b) kidneys removed from sodium-deprived rats released substantially more renin thatn those from sodium-loaded rats, and c) changing perfusate sodium concentration alters sodium excretion, but does not affect renin release.


1963 ◽  
Vol 18 (6) ◽  
pp. 1239-1246 ◽  
Author(s):  
Arthur W. Bauman ◽  
Thomas W. Clarkson ◽  
Ellen M. Miles

A technique for perfusing the isolated rat kidney with heparinized whole blood is described. Numerous aspects of renal function are evaluated. Blood and urine flows approximated normal limits occasionally but diminished and increased, respectively, with time. The reabsorption rates of osmoles, sodium, chloride, potassium, lactate, and phosphate were proportional to the filtered loads. Variations in filtered load were due mainly to changes in glomerular filtration rate (GFR). Tubular secretion of p-amino hippuric acid varied with the GFR, and TmPAH per milliliter GFR was 60% of in vivo values. Glucose reabsorption was practically complete. Urine osmolalities, initially in the range 400–1,000 mosmoles/liter, fell during a perfusion, often to values well below plasma levels. Postperfusion histologic studies usually showed slight to moderate tubular dilatation. It was concluded that: 1) The handling of individual ions approximated in vivo function. 2) Changes in GFR and in the rate of excretion of PAH represented changes in the number of active nephrons. 3) Low urine osmolalities were due to decreasing ability of the nephrons to reabsorb water. blood and urine flow; electrolyte excretion Submitted on February 25, 1963


1995 ◽  
Vol 269 (4) ◽  
pp. C917-C922 ◽  
Author(s):  
K. Yoshioka ◽  
J. W. Fisher

We have previously reported that nitric oxide (NO) and guanosine 3',5'-cyclic monophosphate (cGMP) may be involved in the regulation of erythropoietin (Epo) production in response to hypoxia both in vivo and in vitro (20). In the present studies, we have used the isolated perfused rat kidney to assess the role of NO in oxygen sensing and Epo production. When arterial PO2 was reduced from 100 mmHg (normoxemic) to 30 mmHg (hypoxemic) in the perfusate of this system, perfusate levels of Epo were significantly increased. This hypoxia-induced increase in Epo production was significantly decreased by the addition of NG-nitro-L-arginine methyl ester (L-NAME; 1 mM) to the perfusates. Hypoxemic perfusion also produced a significant increase, and L-NAME significantly inhibited this increase, in intracellular cGMP levels in the kidney when compared with normoxemic perfused kidneys. Quantitative reverse transcription-polymerase chain reaction also revealed that hypoxemic perfusion produced significant increases in Epo mRNA levels in the kidney, which was blocked by L-NAME. Our findings further support an important role for the NO/cGMP system in hypoxic regulation of Epo production.


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