Localization of urate transport in the nephron

of mongrel and Dalmatian dog kidney

RICHARD H. KESSLER,2 KLAUS HIERHOLZER3 AND
RUTH S. GURD4
Department of Physiolugy, Cornell University Medical College,
New Turk City
KESSLER, RICHARD H., KLAUS HIERHOLZER AND RUTH S.
GURD. Localisation of wale trans~ort in the nephron of the mongrel
and Dalmatian dog kidney. Am. J. Physiol. Ig7(3) : 601-603.
I g5g.-Localization of urate transport within the nephrons
of mongrel and Dalmatian dogs was studied by stop-flow
analysis. In mongrel dogs urate concentrations and clearance
ratios were lowest in the segment in which PAH was secreted.
Urate clearance ratios of 0.7 in free-flow samples were reduced
to about 0.3 in stop-flow samples from the proximal segment.
In the distal segment urate clearance ratios did not differ
significantly from ratios obtained in free-flow. Probenecid, in
,doses sufficient to block PAH secretion, inhibited urate re-
absorption thereby increasing urate clearance. In contrast to
these findings with mongrel dogs, the Dalmatians exhibited
weak but definite urate secretion within the proximal segment.
The action of probenecid in this strain of dogs was .to stop all
proximal secretory activity for urate thereby reducing urate
clearance. It was suggested that mongrel and Dalmatian dogs
transport urate by systems that are identical except for direc-
tion of urate movement.
T HE RENAL CLEARANCE of urate is considerably less
than the rate of glomerular filtration. As urate is com-
pletely filterable, its low clearance indicates reabsorp-
tion by tubule cells. Both reabsorption in dogs and a
reabsorptive Tm in human beings have been demon-
strated. In contrast to glucose, urate reabsorption is in-
complete, and even at low plasma concentrations some
urate appears in the urine. In this respect its tubular
transport is more akin to the reabsorptive mechanisms
of amino acids, sulfate and phosphate. Several agents
can increase urate excretion by diminishing its reabsorp-
tion. Among the most potent of these is probenecid
(Benemid) which, when used in maximally effective
dose, can increase urate excretion only 10-15 %. Even
Received for publication April 3, I 959.
l This study was aided by research grants from the National
Heart Institute, National Institutes of Health, and the Life
Insurance Medical Research Fund.
2 Senior Fellow of the Nathan Hofheimer Foundation.
3 New York Community Trust Fellow.
4 Life Insurance Medical Research Fund Fellow.
601
with this uricosuric drug the urate clearance is ap-
preciably less than glomerular filtration rate. An excep-
tion to these generalizations about the renal transport
of urate among the mammalia exists in the Dalmatian
coach hound. The Dalmatian lacks the ability to re-
absorb urate and exhibits urate clearances that approxi-
mate glomerular filtration rate. This study reports on the
localization of urate transport in the mongrel and Dal-
matian dogs. Using the stop-flow method, we have
observed urate reabsorption within the proximal seg-
ment of the mongrel dog kidney. The action of pro-
benecid in impeding urate reabsorption was likewise
placed within the proximal segment. The Dalmatians
not only failed to reabsorb 1 trate but exhibited modest
urate secretion within the proximal segment. This se-
cretory activity was abolished by probenecid.
METHODS
This report is based on 13 experiments performed on
seven dogs including two Dalmatians. Details of six ex-
periments will be presented. The technique employed
was the same as that described by Malvin et al. (I) as
modified in our laboratory (2). Animals were infused
intravenously with a creatinine and PAH solution and
with a solution of 20 % mannitol in saline. Uric acid in
concentrations of 0.5 or 1 .o gm/l. and either 0.7 %
sodium hydroxide or lithium carbonate were added to
each infusion. Resulting plasma urate levels ranged
from 3.0 to 14.0 mg % with filtered urate loads of from
0.8 to 4.0 mg/min/kidney.
Using a retroperitoneal approach, the right ureter
was catheterized with a polyethylene tube. Urinary
stasis was produced by clamping the catheter for a 4-min-
ute period. The volume of fluid trapped within the kid-
ney varied from 20 to 25 ml. On release of the clamp this
trapped fluid was rapidly ejected from the kidney and
collected in serial samples of about I ml each over the
ensuing 3 minutes. A single experiment consisted of this
procedure of stasis and serial collection preceded and
followed by conventional clearance samples. In all but
602 RICHARD H. KESSLER, KLAUS HIERHOLZER AND RUTH S. GURD
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FIG. z. Effect of Benemid (probenecid) on urate clearance
ratios, This interference with urate reabsorption may be confined
to proximal segment.
FIG. I. Localization of urate reabsorption to proximal segment.
Region of highest PAH clearance ratios mark proximal segment;
lowest Na clearance ratios mark distal segment.
one instance two consecutive studies were conducted
with each animal.
The concentration of urate in protein-free plasma
filtrates and in diluted urines was measured by the
arsenophosphotungstic acid method (3). Other chemical
methods and mode of presentation of data have been
published (2).
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RESULTS
Figure I illustrates the results of two consecutive ex-
periments performed on a single animal. The control
study is plotted with circles and the study during urate
infusion plotted with triangles. Data include U/PCr and
clearance ratios for PAH, urate and sodium. At the
lateral margins of the figure are the values for the con-
ventional clearance periods. Stop-flow results are pre-
sented in the center as percentage volume contained
within the clamped kidney. Initial or distal segment
samples appear to the right while later or proximal seg-
ment samples are shown to the left. At the top of the
figure a schematic nephron illustrates functional loci
based for the distal segment on lowest sodium ratios, for
the proximal segment on maximum PAH ratios and for
Benemid=r
II0 90 70 50 30 IO
% Volume wlthm Kidney
FIG. 3. Secretion of urate and its suppression by probenccid in
proximal segment of the Dalmatian.
LOCALIZATION OF URATE TRANSPORT IN NEPHRON IN DOG KIDNEY
603 c
the glomerulus on the appearance of inulin injected just
prior to collection of the serial samples.
During free flow, urate clearance ratios measured
0.6-0.7. Almost identical values were obtained in the
distal, stop-flow samples. Reabsorption of urate with
ratios approaching 0.3 appeared in those same samples
in which PAH ratios were highest. Urate was reabsorbed
within the proximal segment. Note also that the alkaline
infusion of urate delivered at 7 mg/min, did not alter
creatinine, PAH, or sodium ratios.
Figure 2 confirms these observations and demonstrates
the actions of probenecid. In this example both experi-
ments were performed during urate infusion before
(circles) and after (triangles) probenecid. The altera-
tions effected by this drug included inhibition of PAH
secretion and continued but diminished reabsorption
of urate. Clearance ratios for urate were higher in distal
as well as proximal segment samples. Blockade of urate
reabsorption at the proximal site would present to both
the distal segment and ureter more urate than that
delivered prior to probenecid. One need not invoke a
distal segment effect of probenecid to interpret the data.
Figure 3 illustrates consecutive experiments per-
formed on a Dalmatian before and after a probenecid
infusion. In the control study, shown by circles, urate
clearance ratios exceeded I .o in all specimens. These
ratios attained a value of 1.6 within the proximal seg-
ment. The data from this and a companion experiment
indicated modest secretion of urate in this strain of dog.
Following probenecid, urate secretion was abolished;
note that all ratios were less than I .o.
DISCUSSION
Urate reabsorption is incomplete even at low plasma
concentrations. Unlike glucose, urate appears in the urine
at filtered loads well below Tm. In this respect the mecha-
nism for urate reabsorption is similar to the tubular
transport of amino acids, sulfate and phosphate. There
are species differences in the capacity to reabsorb urate;
the transport maximum in the dog exceeds that of human
REFERENCES
I. MALVIN, R. L., L. P. SULLIVAN AND W. S. WILDE. Physidogist
1: 58, qj7*
2. PITTS, R. F., R. S. GURD, R. H. KESSLER AND K. HIERHOLZER.
Am. J. P&siol. 194: x25, 1958.
3. BUCHANAN, 0. H., W. D. BLOCK AND A. A. CHRISTMAN. 3.
Bid. Chem. 157: 181, 1957.
beings. Berliner and co-workers have been able to demon-
strate a Tm in man of I 5 mg/min/~ .73 m3 (4). In con-
trast to this, Friedman and Byers have reported urate
clearance ratios in dogs of less than 0.6 at a urate load
of 20 mg/min. (5). The relative insolubility of urate
limits the quantity that can be administered. As a result,
plasma levels of urate sufficient to reach Tm in the dog
have not been achieved.
Our data placed the site of urate reabsorption within
the proximal segment of the mongrel dog kidney. There
was no evidence for urate secretion. As might be pre-
dicted, the moderate inhibition of urate reabsorption
by probenecid occurred at this same site. Probenecid
blocked proximal secretion of PAH as it inhibited proxi-
mal urate reabsorption, However, there was less inhibi-
tion of the secretory system for urate than for PAH. This
is apparent in these data as well as in previous clear-
ance studies.
The Dalmatian is a curiosity among the mammalia.
This strain of dog excretes urate in quantities that suggest
the absence of a tubular reabsorptive system for urate.
Noted first by Benedict in I 915 (6), the high urate excre-
tion by the Dalmatian has been more precisely studied
by Friedman and Byers. These authors reported CUrate/
Ccr ratios of 0.89 to I. IO over a wide range of plasma
levels of urate (5). Weak secretion of urate in the Dal-
matian has been observed (personal communication
from A. B. Gutman). By the stop-flow technique exag-
geration of tubular transport has allowed localization of
this urate secretory mechanism to the proximal segment.
Probenecid exhibited a more pronounced action on urate
clearance ratios in the Dalmatian than in the mongrel
dog. Urate transport is in opposite direction in these two
strains, yet probenecid apparently interferes with urate
transport in both instances. Perhaps all mammals pos-
sess a common transport system for urate which in the
Dalmatian is directed from tubule cell to lumen. This
distinguishes the Dalmatian from other mammalia in
which the identical transport system directs urate from
lumen to tubule cell.
4. BERLINER, R. W.,J- G. HILTON, T. F. Yij AND T.J. KENNEDY,
JR. J. Clin. Invest. 29: 396, 1950.
5. FRIEDMAN, M. AND S. 0. BYERS. 3. Bid. Chem. 175: 727, 1948.
6. BENEDICT, S. R. Harvey Lect. I o : 346, I g 15-16.