The loop of Henle as the milestone of mammalian kindey concentrating ability: a historical review

Abstract

The first description of the renal tubules is attributed to Lorenzo Bellini in 1662 and four years later Marcello Malpighi described the glomerulus. In 1842 Sir William Bowman described the capsule that surrounds the Malpighian body and its connection with the renal tubule and introduced the "excretory" hypothesis of urine formation. In the same year, Carl Ludwig introduced the "filtration-reabsorption" hypothesis of urine formation. Bowman's hypothesis was accepted by the so-called "vitalists" and Ludwig's hypothesis by the so-called "mechanists". In the middle of this confliction, Jacob Henle described in 1862 the homonymous "U" shaped loop but his discovery has neglected. In 1942 Werner Kuhn, a physical chemist, proposed that the loop of Henle may be the natural analog of the hairpin countercurrent multiplication system which concentrates urine in mammalian kidneys. In 1951 Kuhn, Hargitay and Wirz showed experimentally that the loop of Henle was the most important part of the countercurrent multiplication system of urine-concentrating mechanism in mammalian kidneys. The new theory was accepted by English-speaking scientists later, in 1958, when Carl Gottschalk and Margaret Mylle published their experimental work and proved that Kuhn's theory was correct. Gottschalk summarized the evidence of the accumulated knowledge in 1962, three centuries after the first description of renal tubules and one century after description of Henle's loop.

Full text

7+(/2232)+(1/($67+(0,/(6721(
2)0$00$/,$1.,'1(<&21&(175$7,1*
$%,/,7<$+,6725,&$/5(9,(:
HENLEOVA PETLJA KAO PREKRETNICA
SPOSOBNOSTI KONCENTRIRANJA BUBREGA
SISAVACA: POVIJESNI PREGLED
(IVWDWKLRV.RXORXULGLV,RDQQLV.RXORXULGLV
S
The rst description of the renal tubules is attributed to Lorenzo Bellini in 1662 and four
years later Marcello Malpighi described the glomerulus. In 1842 Sir William Bowman de-
scribed the capsule that surrounds the Malpighian body and its connection with the renal
tubule and introduced the “excretory” hypothesis of urine formation. In the same year, Carl
Ludwig introduced the “ltration-reabsorption” hypothesis of urine formation. Bowman’s
hypothesis was accepted by the so-called “vitalists” and Ludwig’s hypothesis by the so-called
“mechanists”. In the middle of this coniction, Jacob Henle described in 1862 the homony-
mous “U” shaped loop but his discovery has neglected. In 1942 Werner Kuhn, a physical
chemist, proposed that the loop of Henle may be the natural analog of the hairpin countercur-
rent multiplication system which concentrates urine in mammalian kidneys. In 1951 Kuhn,
Hargitay and Wirz showed experimentally that the loop of Henle was the most important
part of the countercurrent multiplication system of urine-concentrating mechanism in mam-
malian kidneys. The new theory was accepted by English-speaking scientists later, in 1958,
when Carl Gottschalk and Margaret Mylle published their experimental work and proved
that Kuhn’s theory was correct. Gottschalk summarized the evidence of the accumulated

Nephrology Department, General Hospital of Corfu. Greece.

St. Elizabeth’s Medical Center, Boston. USA.
Corresponding author: Efstathios Koulouridis, MD. Nephrology Department. General
Hospital of Corfu. Greece. Spirou Rath . TK . Corfu, Greece. Tel. --
-. Fax: --. Electronic address: koulef@otenet.gr
Pregledni rad Acta med-hist Adriat 2014; 12(2);413-428
Review article UDK: 61(091):001.894


knowledge in 1962, three centuries after the rst description of renal tubules and one century
after description of Henle’s loop.
.H\ZRUGV: Loop of Henle, urine formation mechanism, vitalists, mechanists, countercur-
rent multiplication system.
I
At the end of the Mesozoic era, about 66 million years ago, mammals
migrated from the water to terrestrial life. As a consequence, they were de-
prived from free access to water and sodium. In order to survive in the new
environment, they had to develop an excretory organ with the capacity to
independently conserve salt and water. This organ was no other than the
kidney1.
We know now that lower vertebrates are capable to produce isotonic or
hypotonic urine. Nevertheless, only mammals and some birds are capable
to produce hypertonic urine. This capability is essential to conserve water
under conditions of environmental dryness and limited access to water [1].
The mechanism by which the mammalian kidney concentrates urine
is complex and relies upon the specialized architecture and sophisticated
function of certain nephron segments as well as accompanying blood ves-
sels. The fundamental structure of urine concentration is the “U” shaped
loop of Henle accompanied with the collecting duct and the vasa recta in
an obligatory manner unique only among species with the capacity of urine
concentration [2].
The loop of Henle was rst described by the German pathologist
Friedrich Gustav Jacob Henle in 1862, and presented with excellent accuracy
in his monograph with the title: “Zur Anatomie der Niere” Von J. Henle.
Gottingen. Verlag der Dieterichscen Buehhandlung. 1862. The monograph
is accompanied with marvelous hand made illustrations showing the thin
descending limb, the thick ascending limb and the transition from the thin
ascending to the thick ascending limb. (Figure: 1). It took almost a century
to recognize the importance of this structure in urine concentrating mech-
anism, for many years it was thought that loop of Henle has no functional
signicance and was considered only as an “incidence of organogenesis” [1,3].
The reason that Henle’s discovery remained buried for a long time before
physiologists recognize its importance in urine concentration is the lack of
knowledge upon the structure and function of the elemental kidney unit,


the nephron, as well as the lack of proper instruments for experimental work
upon renal function and proper estimation of plasma and urine constituents.
The rst description of renal tubule is attributed to the Italian anatomist
Lorenzo Bellini who in 1662 described the papillary ducts which took his
name. Four years later in 1666 the Italian physician and anatomist Marcello
Malpighi described the glomerulus in the renal cortex, which took the name
“Malpighian body”, and its connection with the eerent and aerent arter-
ies. He proposed also the possible connection with renal tubules but did not
prove it [4].
T   “V”  “M”.
After the above mentioned preliminary discoveries, it took about two
centuries until the English surgeon, histologist and anatomist Sir William
Bowman in 1842 described the capsule which surrounds the “Malpighian
body” and its connection with the kidney tubules. (Figure: 2). Bowman in-
vestigated also the epithelium of uriniferous tubules and he was impressed
with the similarities between this epithelium and the epithelium of excreto-
ry tubules of digestive glands and he arbitrarily inferred that tubular cells
Figure 1: Left: Friedrich Gustav Jacob Henle (1809-1885). German
physician, pathologist and anatomist. Right: Hand made drawings of the
homonymous loop showing with accuracy the thin descending limb, the
thick ascending limb and the transition from the thin ascending to the
thick ascending limb. (“Zur Anatomie der Niere”, 1862).
Slika 1. Lijevo: Friedrich Gustav Jacob Henle (1809.-1885.). Njemački liječnik,
patolog i anatom. Desno: Ručno rađeni crteži homonimne petlje, koji točno
pokazuju tanku silaznu granu, debelu uzlaznu granu i prelazak od tanke uzlazne
ka debeloj uzlaznoj grani. (“Zur Anatomie der Niere”, 1862.).


excrete the urine constituents and glomerulus produce only a stream of wa-
ter which washes out the excreted solutes from tubules [4].
In his seminal paper “On the structure and use of the Malpighian bodies
of the kidney, with observations on the circulation through that gland”, pre-
sented to the Royal Society of London, 17 February 1842, http://www.jstor.
org/stable/108143, he wrote:
“These tubes consists of an external tunic of transparent homogeneous tissue
(which I have termed the basement membrane), lined by epithelium. The Malpighian
bodies I saw to be rounded mass of minute vessels invested by a cyst or capsule of pre-
cisely similar appearance to the basement membrane of the tubes”.
“… I injected some kidneys through the artery, by this method, in order to notice
the nature of the vascular ramications in the Malpighian bodies. I not only found
what I sought, but the clearest evidence that the capsule which invest them is, in
Figure 2: Left: Sir William Bowman, 1st Baronet, (1816 - 1892). English
surgeon, histologist and anatomist. Right: Hand made drawings showing
the glomerulus the surrounding capsule and the uriniferous tubules
from many species including humans. (“On the structure and use of the
Malpighian bodies of the kidney”, 1842).
Slika 2. Lijevo: Sir William Bowman, I. Baron (1816. - 1892.). Engleski kirurg,
histolog i anatom. Desno: Ručno rađeni crteži koji pokazuju glomerus, okružujuću
kapsulu i urinoferusne tubule mnogih vrsta, uključujući i ljude. (“On the structure
and use of the Malpighian bodies of the kidney”, 1842.).


truth, the basement membrane of the uriniferous tube expanded over the tuft of
vessels”.
“It occurred to me that as the tubes and their plexus of capillaries were probably,
for reasons presently to be stated, the parts concerned in the secretion of that portion
of the urine to which its characteristic properties are due (the urea, lithic acid &c.),
the Malpighian bodies might be an apparatus destined to separate from the blood
the watery portion”.
“This abundance of water is apparently intended to serve chiey as a menstruum
for the proximate principles and salts which this secretion contains, and which,
speaking generally, are far less soluble than those of any other animal product”.
This arbitrary explanation was reinforced later, in 1874, when Rudolf
Heidenhain of Breslau established the “excretory” hypothesis in urine for-
mation which is known as the “Bowman-Heidenhain” hypothesis of “vital-
ists” [3,4].
In 1842, the same year that Bowman published his work, another brilliant
mind in Germany, Carl Ludwig, a young physiologist in the University of
Marburg, published his thesis in order to gain a senior degree. Carl Ludwing’s
thesis was a scientic work of 24 pages written in Latin “De viribus physics
secretionem urinae adjuvantibus” (On the physical forces that promote the
secretion of urine). Based upon his own experimental observations and lit-
erature available at that time, he introduced the hypothesis that glomerulus
acts as a sieving lter which produces an ultra ltrate of blood free of cells
and proteins and contains all the other constituents of the blood in the same
concentration without any modication by the glomerulus itself. He contin-
ued that the volume of the ltrate is inuenced by blood pressure variation
in the renal artery and that, as it passes through the renal tubules, it under-
goes reabsorption or secretion which alter the nal concentration of certain
substances in urine in relation to the blood [4,5]. (Figure: 3).
Somme details of his paper are as follow:
“… the membranes of the vessels in the glomeruli are subjected to high pressure,
resulting in a copious secretion from the delicate glomeruli. When kidneys were in-
jected with wax, I detected discharge of the wax from the glomeruli.
“The second physical process occurring in the kidney is an endosmotic action
between the solution of salts secreted and the partly altered blood retained in the
vessels. The rst and best proof of endosmosis is the fact that, given the same com-
position of the blood, the concentration of the urine depends on the urine ow rate.


It is clear that the process of expulsion of the urine is as follows: “When the blood
vascular system is lled with uid, pressure is exerted against the walls of the glom-
eruli, and the water in the blood leaves the glomeruli and is taken up by the urinifer-
ous ducts. It is here that endosmosis can occur as described above. The quantity of
urine secretion is accelerated when the blood vascular system is lled with uid, in
which case the pressure against the walls of the glomeruli is increased”.
In this work Ludwig introduced the hypothesis that the phenomena
of living organisms are inuenced from the laws of physics and chemis-
try and can be “the consequence of simple attractions and repulsions between a
limited numbers of chemical atoms”. With this revolutionary concept for his
era, Ludwig introduced the hypothesis of “ltration-reabsorption” in urine
formation which was accepted only from the so called “mechanists” and it
Figure 3: Left: Carl Friedrich Wilhelm Ludwig, (1816-1895). German
physician and physiologist. Right: Hand made drawings showing in
the upper panel a schematic representation of the glomerulus and the
uriniferous tubule with its blood supply and in the lower panel blood
hydrostatic pressure changes during its passage through the glomerular
capillaries. (“De viribus physics secretionem urinae adjuvantibus”, 1842).
Slika 3. Lijevo: Carl Friedrich Wilhelm Ludwig, (1816.-1895.). Njemački liječnik
i ziolog. Desno: Rukom rađeni crteži koji pokazuju u gornjem dijelu shematski
prikaz glomerusa i urinoferusne tubule sa pripadajućim sustavom opskrbe krvlju
te u donjem dijelu promjene hidrostatskog krvnog tlaka tijekom prolaska kroz
glomerualne kapilare. (“De viribus physics secretionem urinae adjuvantibus”, 1842.).


would be a matter of controversy between them and “vitalists” for the fol-
lowing 80 years [3-5].
At the middle of this conict, and with a lot of items of renal function un-
resolved, it was expected that Henle’s discovery would be neglected. At the
beginning of the 20th century, in 1922, Alfred Richards and his colleagues in-
troduced a new method in the experimental investigation of renal function
which is known as the “micropuncture technique” and proved that Ludwig
was quite right in his pioneer concept regarding the mechanism of urine for-
mation4. Somme details of his paper are as follow [4]:
“… it was possible to insert sharply pointed tubes into the space within Bowman’s
capsule and to abstract the uid which issues from the blood of the glomerular cap-
illaries ..”.
“The results showed that the glomerular uid is free from protein but contains
chloride and glucose, both of these being absent from the bladder urine. It is alka-
line, contains urea, and indeed every diusible constituent of plasma for which we
were able to make a test …”
“These results seem to me to leave little room for doubt that, in amphibia, the
glomerular urine actually has the same composition of a protein-free ltrate from
plasma, precisely as Ludwig had imagined ninety-three years ago”.
Thereafter the use of micropuncture technique and the measurement of
Glomerular Filtration Rate (GFR) with the use of the polysaccharide inulin
in animals as well as in man by Homer Smith and his colleagues in 1932 at
the New York University Medical College provided the scientic commu-
nity with a rapid increasing bulk of knowledge upon renal physiology and
the interest of researchers turned mainly to the ltration, reabsorption and
excretion of solutes along the nephron [6].
T  
During 1940-1944 Europe was almost devastated by the 2nd World Wa r
but Switzerland’s neutrality allowed some brilliant minds to continue their
experimental work and produce knowledge, one of them was Werner Kuhn,
Professor of Physical Chemistry in University of Basel, who worked upon
the enrichment of sugar in water using semi-permeable membranes and phe-
nol as an auxiliary liquid in a hairpin countercurrent system without any
other external force. He showed that at each bend of the hairpin counter-
current system solute concentration increased by a factor n which equals to


the length of the system divided by its width (n=L/W). Based upon these
observations Kuhn and his colleague Kaspar Ryel published, in 1942, a pa-
per in German and proposed that the “U” shaped loop of Henle may be the
natural analog of a countercurrent multiplication system capable to produce
urine concentration in mammalian kidney but the paper overlooked by renal
physiologists [7,8].
Although countercurrent exchangers and countercurrent multipliers
were known among engineers and utilized in many applications in indus-
try, mainly in heat exchange and solute concentration, the rst description
of the importance of heat exchange between arteries and veins in mammals
is attributed to Claude Bernard in 1876. Many years later in 1940’s Bazett
and his colleagues showed experimentally the heat exchange between deep
arteries and veins in the arms and the legs which prevents heat loss to the
environment and achieves blood warming before entrance to the central cir-
culation [1,9,10].
In 1950’s extensive experimental work showed that Arctic mammals and
birds utilizes a countercurrent heat exchange system between deep arter-
ies and veins in their legs in order to prevent freezing while standing on icy
ground or wading in icy water. It was also showed that some species utilizes
a specialized network of arteries and veins bundles capable to exchange heat
and gazes, known as “rete mirabile” which help them to regulate body tem-
perature, to exchange oxygen in sh gills and in the case of deep ocean shes
to store oxygen in swim bladder in high pressures exceeding in some cases a
hundred time the partial oxygen pressure of surrounding see water [1,9,10].
In early 1900’s Karl Peter in his book “Untersuchungen uber Bau und
Entwickelung der Niere” (Jena Fisher 1909, editor) rst pointed to the rela-
tion between length of Henle’s loop and urine concentrating ability among
some species. Later in 1944 Sperber pointed again to the relation between
length of Henle’s loop and urine concentration because animals with long
Henle’s loops exhibited the greater urine concentrating ability[7].
Meanwhile in 1946 Bart Hargitay, a young graduate of chemistry at the
University of Budapest, received a fellowship oered by the University of
Basel where he joined Werner Kuhn. As the Iron Curtail closed this year
he decided to stay in Switzerland and asked Professor Kuhn to accept him
as a graduate student to work in a thesis. Kuhn assigned Hargitay to prove
the hypothesis of countercurrent multiplication system of urine concentra-
tion in the kidney [8]. Hargitay contacted Dr Heinrich Wirz at the Physiology


Institute in order to obtain some knowledge about renal physiology and help
him in experiments with animals. Wirz enthused with the idea and started
promptly experiments with rat kidneys and later with Syrian hamster be-
cause the solitary papilla of this rodent protrudes in to the renal pelvis and
it is easier for micropuncture and collect urine sample from renal tubules.
Soon thereafter the two researchers encountered a serious problem: the
estimation of the chemical constituents had to be performed in a very scant
sample of urine, about 10−7 ml, obtained by micropuncture. Hargitay decided
to determine only the osmotic pressure of the samples by cryoscopic method
according to the formula:
Relative freezing point depression = 100 • Δx – Δisot / Δmax – Δisot
When Δx: the freezing point depression in x position in the kidney8.
In order to perform their calculations they needed a cryo-chamber with
temperature lower than -200 C. They used the cold room of the Burgerspital
hospital in Basel. The procedure needed to be carried out in the room, under
heavy clothes and furs in the middle of the summer, bringing with the mi-
cropipette urine samples and observing under polarized microscope the bi-
refringence of ice formed at the melting point of each sample. They gathered
multiple urine samples along the axis from the renal cortex to the papilla and
found that in all samples the osmotic pressure was equal at the same level but
it was gradually increasing at each deferent level from the renal cortex to the
papilla. The lowest pressure was observed in the renal cortex and estimated
to be 25 Atm while the greatest was observed in the papilla and was estimat-
ed to be 58 Atm [9]. (Figure: 4)
These ndings as well as experimental ndings from a mechanical hair-
pin model constructed by Hargitay and colleagues in his laboratory, prompt-
ed Hargitay and Wirz to consider that the “U” shaped loop of Henle is the
natural analog of a hairpin countercurrent multiplication system in the kid-
ney which by the antiparallel circulation of urine in the descending and as-
cending limb of the loop produces the maximum concentration of solutes at
the bending point of the loop in the deep renal medulla.
Although they were ignorant of the specialized properties of the descend-
ing and ascending limb of Henle’s loop concerning its water permeability
and active sodium chloride transport, they realized that the single eect, by
means of the leading process, in urine concentration mechanism could not
be a component of hydrostatic pressure dierence but an “electroosmosis”


phenomenon which they de-
scribed as follows: “It seems
much more likely that, in epitheli-
al cells, energy from metabolism
is used to establish a potential
eld and that in this potential
eld electroosmosis takes place”
[9].
In order to explain the se-
quence of events in urine con-
centration, they considered it
mainly as a process of water
absorption. They hypothe-
sized that the electroosmosis
phenomenon produces water
transport from the lumen of
descending limb to the inter-
stitial space and then to the
lumen of the ascending limb.
They said that the latter de-
livers diluted urine to the
distal convoluted tubule from
which water is transferred
to the blood. Hence, a nal
concentration of urine takes
place in the collecting duct as
it passes through the hypertonic medulla [9].
As we know now water permeability of the thin descending limb of
Henle’s loop is owing to the expression of aquaporin-1 (AQP-1) in its epithe-
lium. Thorough investigation of this nephron segment showed that short
looped nephrons do not express AQP-1 in their descending thin limb and
they are practically impermeable to water. Conversely AQP-1 is expressed in
the thin descending limb of long looped nephrons especially those extend-
ing deep in the medulla but AQP-1 expression is limited to the rst 40% of
their length and never beyond the last 2-2,5 mm before bending in the inner
medulla. The remainder 60% of their length is devoid of AQPs and hence
impermeable to water but it is permeable to urea and chloride ions because
of the expression of urea transporters and chloride channels. The thick
Figure 4: The original ndings from
experiments conducted by Bart Hargitay
and Heinrich Wirz showing the increase of
osmotic pressure from the renal cortex to
the tip of the renal papilla.
Slika 4. Izvorni nalazi iz opita Barta
Hargitaya i Heinricha Wirza koji pokazuju
povećanje osmotskog tlaka od bubrežnog
korteksa do vrha bubrežne papile.


ascending limb of Henle’s loop is impermeable to water but posses the capac-
ity of active transport of sodium, potassium and chloride via the Na+:K+:2Cl¯
cotransporter which transfer sodium chloride to the interstitium and con-
tributes signicantly to the hypertonicity of the renal medulla [12,13].
The work was rst presented in May 1951 by Hargitay to the Bunsen
Gesellschaft at the meeting for physical chemists in Gottingen and a few
weeks later by Wirz at the International Conference for Physiology in
Copenhagen. The physical chemists accepted the ndings by Hargitay and
Wirz with enthusiasm but the physiologists expressed their skepticism and
reluctance to accept the new theory. The work was published in German
in the same year and thereafter it became a mater of investigation among
German speaking scientists but not among English for at least the follow-
ing 7 years. Wirz continued his experiments by micropuncture but he never
managed to puncture with accuracy the lumen of Henle’s loop especially at
the tip of renal medulla [11,14,15].
During this period a considerable work upon urine concentration and
dilution was conducted by Karl Julius Ullrich and was published mainly in
German. Although during his contribution to Gottschalk’s laboratory in
Chapel Hill he published also some articles in English. Ullrich examined the
composition of interstitial uid in renal cortex and medulla and proved that
except electrolyte accumulation other osmolytes especially urea contributes
to the medullary hypertonicity of mammalian kidney [17]. He showed also
that glycerophosphocholine and inositol accumulate in the renal medulla
and act as osmolytes and that medullary collecting duct participates in urea
recycling [17].
The reluctance of English speaking scientists to accept the new theory
is in part attributed to the fact that the rst half of 20th century was predom-
inated by Homer Smith’s proposals in renal physiology. In his book “The
kidney: Structure and Function in Health and Disease”, published in 1951,
by drawing the nephron he omitted the loop of Henle and included only
a part of the descending limp as short straight tubule. Smith believed that
the urine concentration is accomplished at least by two mechanisms one at-
tributed to passive reabsorption of water in the proximal tubule and another
one attributed to active reabsorption of water in some parts of distal tubule
although no evidence of active water reabsorption mechanism had been
proved in any biological system. (Figure: 5).
He wrote exactly:


“… the reabsorption of water by the renal tubules involves at least two more or
less independent processes: 1. passive water reabsorption in the proximal tubule and
thin segment (proximal system), and, under appropriate circumstances, in the distal
tubule; and 2. active water reabsorption that is presumably conned to the distal
system, i.e., in the distal tubule and possibly in the collecting ducts also”.
When he asked by Carl Gottschalk what he believes about the counter-
current hypothesis he said: “The smart boys don’t believe in it” [7,18].
Meanwhile USA entered the 2nd World War in 1941 and Alfred Richard’s
laboratory stopped the experiments with micropuncture for almost a decade.
After the war Carl Gottschalk expressed the intention to revive renal micro-
puncture and asked Richard’s advice upon restarting kidney micropuncture
but for unknown reasons he discouraged him [18].
In 1952 Gottschalk joined the Department of Medicine at the University
of North Carolina and established in Chapel Hill his “Micropuncture
Laboratory” which was equipped with the Ramsey/Brown micro-osmome-
ter built especially for the Chapel Hill laboratory [18]. Gottschalk recruited
in his laboratory Margaret Mylle who was considered as “one of the most
Figure 5: Left: The rectilinear model of the nephron omitting the loop
of Henle proposed by Homer Smith in his book “The kidney: Structure
and Function in Health and Disease”. (London, Oxford University Press,
1951). Right: The countercurrent multiplication system with gradually
increasing osmolality from the cortex to the renal medulla proposed by
Kuhn, Hargitay and Wirz. (Das Multiplikationsprinzip als Grundlage der
Harnkonzentrierung in der Niere, 1951).
Slika 5. Lijevo: Rektilinearni model nefrona bez Henleove petlje predložen od
strane Homera Smitha u knjizi “The kidney: Structure and Function in Health
and Disease”. (London, Oxford University Press, 1951.). Desno: Protustrujni
multiplikacijski sustav sa postupno povećavajućom osmolarnošću od korteksa do
bubrežne medule po Kuhnu, Hargitayu i Wirzu. (Das Multiplikationsprinzip als
Grundlage der Harnkonzentrierung in der Niere, 1951.).


skilled micropuncturists in the word”. Gottschalk’s intention was to check
the hypothesis proposed by Robert Berliner that the urine at the tip of the
loop of Henle should be hypotonic [19].
After performing a series of brilliant experiments with Wistar rats, gold-
en hamsters, one kangaroo rat and Psammomys obesus, they collected urine
samples in nanoliter specimens from short looped nephrons, from the thin
limb and the bend of loop of Henle, collecting ducts as well as vasa recta.
They showed that uid from the bend of loops of Henle, collecting ducts and
vasa recta at the same level in the papilla were hyperosmotic and exhibited
almost equal osmotic pressure [7,18].
After that Gottschalk published a brief report of his ndings in an article
less than one page in Science in September 1958 with the title: “Evidence
that the mammalian nephron functions as a countercurrent multiplier sys-
tem” establishing by this way the validity of “the new theory” proposed by
Kuhn, Hargitay and Wirz [20]. According to Francois Morel’s declaration,
after personal communication with Gottschalk, he sent the data to Homer
Smith before full publication. Homer Smith was so impressed by these nd-
ings that he asked from Gottschalk to postpone the full publication until he
will make known his new opinion. After that Smith delivered a lecture with
the title “The fate of sodium and water in the renal tubules”, in October 17,
1958 at the Annual Postgraduate Week organized by the New York Academy
of Medicine and he recognized the importance of the “new theory” with re-
markable accuracy and humour [1,7]. In advance Gottschalk and Mylle pub-
lished their ndings in American Journal of Physiology the next year [21].
By extending their experiments they showed, by micropuncture in ham-
sters, that the water permeability of thin descending limb of Henle’s loop
greatly exceeded that of the thin ascending limb. Experiments in hamsters
with diabetes insipidus showed that the uid collected from the loop of
Henle and blood from the vasa recta, at the tip of the papilla, were hyperos-
motic in contrast to the uid of the adjacent collecting ducts which was hy-
po-osmotic. These experiments showed that water permeability and urine
concentration in the thin descending limb of Henle’s loop is independent of
the presence of ADH and that the nal concentration of urine takes place in
the medullary collecting duct [7].
Gottschalk summarized the evidence of the accumulated knowledge
upon the countercurrent hypothesis in a lecture with the title “Renal tubular
function: lessons from micropuncture” presented in “The Harvey Lectures”


(series 58) in 1962 three centuries after the rs description of renal tubules
and a century after Jacob Henle’s description of the homonymous loop in
mammalian kidney.
The above mentioned fundamental work was simply the beginning fol-
lowed by an enormous experimental investigation of renal physiology based
rst upon micropuncture and later upon microperfusion and patch clamp
technique which expanded our knowledge upon ion channels properties.
Genetic analysis of specic ion and solute transporters upon molecular level
as well as the use of specic gene knockout animals enabled researchers to
unravel step by step the mysteries of renal function [13,22,23].
Any further detailed analysis of the ongoing research upon this very in-
teresting topic is beyond the scope of this historical review but the adventure
is still in progress because “we have to go miles before sleep”.


R
. Smith HW. The fate of sodium and water in the renal tubules. Bull N Y Acad
Med ; : -.
. Soper Ch. The paradoxical urinary concentrating mechanism. Jurnal of
Creation ; : -
. Morel F. The loop of Henle, a turning-point in the history of kidney physiology.
Nephrol Dial Transplant ; : -.
. Richards AN. Physiology of the kidney. Bull N Y Acad. Med. ; : -.
. Davis JM, Thurau K, Haberle D. Carl Ludwig: the discoverer of glomerular l-
tration. Nephrol Dial Transplant ; : -.
. Smith HW, Goldring W, Chasis H. The measurement of the tubular excretory
mass, eective blood ow and ltration rate in the normal human kidney. J Clin
Invest ; : -.
. Gottschalk CW. History of the urinary concentrating mechanism. Kidney Int
; : -.
. Kuhn W, Ryel K. Herstellung konzentrienter Losungen aus verdunnten durch
blosse Membranwirkung. Ein Modellversuch zur Funktion der Niere. Hoppe-
Seyler’s Zeit Physiol Chem ; : -.
. Irving L, Krog J. Temperature of skin in the Arctic as a regulator of Heat. J Appl
Physiol ; : -.
. Scholander PF, Krog J. Countercurrent heat exchange and vascular bundles in
sloths. J Appl Physiol ; : -.
. Hargitay B, Kuhn W. The multiplication principle as the basis for concentrating
urine in the kidney. J Am Soc Nephrol ; : -.
. Zhai X-Y, Fenton RA, Andreasen A, Thomsen JS, Christensen EI. Aquaporin-
is not expressed in descending thin limbs of short-loop nephrons. J Am Soc
Nephrol ; : -.
. Pannabecker TL, Dantzler WH, Layton HE, Layton AT. Role of three-dimensi-
onal architecture in the urine concentrating mechanism of the rat renal medu-
lla. Am J Physiol Renal Physiol ; : F-F.
. Hargitay B, Kuhn W. Das Multiplikationsprinzip als Grundlage der
Harnkonzentrierung in der Niere. Z Electrochem Angew Phys Chem ; :
-.
. Wirz H, Hargitay B, Kuhn W. Lokalisation des Konzentreirungsprozesses in der
Niere durch direkte Kryoscopie. Helv Physiol Pharmacol Acta ; : -.


. Jarausch KH, Ullrich KJ. Studies on the problem of urine concentration and
dilution; osmotic behavior of renal cells and accompanying electrolyte accu-
mulation in renal tissue in various diuretic conditions. Pugers Arch. ;
():-.
. Murer H, Burckhardt G. Professor Karl Julius Ullrich – in memoriam. Kidney
Int. ; : -.
. Valtin H. Carl W Gottschalk’s contribution to elucidating the urinary concen-
trating mechanism. J Am Soc Nephrol ; : -.
. Schafer JA. Experimental validation of the countercurrent model of urinary
concentration. Am J Physiol Renal Physiol ; : F-F.
. Gottschalk CW, Mylle M. Evidence that the mammalian nephron functions as
a countercurrent multiplier system. Science ; : .
. Gottschalk CW, Mylle M. Micropuncture study of the mammalian urinary
concentrating mechanism: evidence for the countercurrent hypothesis. Am J
Physiol ; : -.
. Agree P, Preston GM, Smith BL, Jung JS, Raina S, Moon C et al. Aquaporin
CHIP: the archetypal molecular water channel. Am J Physiol Renal Physiol
; : F-F.
. Fenton RA, Knepper MA. Mouse models and the urinary concentrating mecha-
nism in the new millennium. Physiol Rev ; : -.
S
Prvi se opis bubrežnih tubula iz 1662. pripisuje Lorenzu Belliniju, a četiri je godine kasnije
Marcello Malpighi opisao glomerul. Godine 1842. je Sir William Bowman opisao kapsulu
koja okružuje malpigijevo tjelešce i njegovu vezu s bubrežnim tubulima te uveo “ekskretornu”
hipotezu o stvaranju urina. Iste je godine Carl Ludwig uveo je “ltracijsko-reasorpcijsku”
hipotezu stvaranja urina. Bowmanova je hipoteza bila prihvaćena od strane tzv. “vitalista”,
a Ludwigova hipoteza od strane tzv. “mehanicista”. U jeku tog sukoba Jakob Henle opisao je
1862. homonimne petlje u obliku slova “U”, ali njegovo je otkriće zanemareno. Godine 1942.
je Werner Kuhn, zikalni kemičar, predložio ideju da je Henleova petlja možda prirodni
analogon kopče protustrujnog multiplikacijskog sustava koji koncentrira urin u bubrezi-
ma sisavaca. Godine su 1951. Kuhn, Hargitay i Wirz eksperimentalno pokazali da da je
Henleova petlja najvažniji dio protustrujnog multiplikacijskog sustava mehanizma za kon-
centriranje urina u bubrezima sisavaca. Nova je teorija prihvaćena od strane anglofonih
znanstvenika kasnije, 1958. godine, kada su Carl Gottschalk i Margaret Mylle objavili svoj
eksperimentalni rad i dokazali da je Kuhnova teorija bila točna. Gottschalk je sažeo dokaze
sakupljenog znanja 1962., tri stoljeća nakon prvog opisa bubrežnih tubula i jednog stoljeća
nakon opisa Henleove petlje.
.OMXĀQHULMHĀL: Henleova petlja, mehanizam formiranja urina, vitalisti, mehanicisti, protu-
strujni multiplikacijski sustav.
:
