Structure and Function of the Thin Limbs

of the Loop of Henle

Thomas L. Pannabecker*1

ABSTRACT
The thin limbs of the loop of Henle, which comprise the intermediate segment, connect the prox-
imal tubule to the distal tubule and lie entirely within the renal medulla. The descending thin
limb consists of at least two or three morphologically and functionally distinct subsegments and
participates in transepithelial transport of NaCl, urea, and water. Only one functionally distinct
segment is recognized for the ascending thin limb, which carries out transepithelial transport of
NaCl and urea in the reabsorptive and/or secretory directions. Membrane transporters involved
with passive transcellular Cl, urea, and water fluxes have been characterized for thin limbs; how-
ever, these pathways do not account for all transepithelial fluid and solute fluxes that have been
measured in vivo. The paracellular pathway has been proposed to play an important role in
transepithelial Na and urea fluxes in defined thin-limb subsegments. As the transport pathways
become clearer, the overall function of the thin limbs is becoming better understood. Primary
and secondary signaling pathways and protein-protein interactions are increasingly recognized
as important modulators of thin-limb cell function and cell metabolism. These functions must be
investigated under diverse extracellular conditions, particularly for those cells of the deep inner
medulla that function in an environment of wide variation in hyperosmolality. Transgenic mouse
models of several key water and solute transport proteins have provided significant insights into
thin-limb function. An understanding of the overall architecture of the medulla, including juxta-
positions of thin limbs with collecting ducts, thick ascending limbs, and vasa recta, is essential
for understanding the role of the kidney in maintaining Na and water homeostasis, and for un-
derstanding the urine concentrating mechanism.  C 2012 American Physiological Society. Compr

Physiol 2:2063-2086, 2012.

Introduction several functionally distinct subsegments of the DTLs. No-

The descending thin limbs (DTLs) and ascending thin limbs
(ATLs) of the loop of Henle connect the proximal nephron
segments with the distal segments, and in so doing, pass
through an interstitial fluid that is low in oxygen and that
has an osmolality substantially higher than that of blood
plasma. The cells of thin limbs have few mitochondria and
carry out a relatively low level of metabolic activity, relative
to other nephron segments. Transepithelial fluid flows and
solute fluxes do occur in thin limbs in substantive ways, and
these fluxes are considered to be driven primarily by passive
mechanisms. These properties set the stage for understanding
the primary functional roles of thin limbs of the loop of Henle
in the renal medulla.
The thin limbs are believed to play an important role in
generating the high interstitial osmolality that can be attained
in the renal inner medulla (IM), and therefore, the magnitudes
of thin-limb cell and transepithelial fluid and solute fluxes are
of considerable interest in understanding the integrative role
of the kidney in the processes of Na and water homeostasis
and the urine concentrating mechanism.
A number of proteins that are relevant to fluid and so-
lute transport across the thin-limb cells and epithelia have
been identified. Expression patterns of these proteins, and
axial variations in fluid and solute permeabilities, reveal
tably, these include aquaporin 1 (AQP1)-positive and AQP1-
negative DTL segments. On the basis of morphology and
physiology, little or no evidence has been found for functional
subsegments of the ATL. DTLs and ATLs are anatomically
and physiologically integrated in a complex, yet poorly un-
derstood fashion, with the multiple subsegments of medullary
collecting ducts (CDs) and vasa recta. It is generally believed
that these structural and functional interrelationships play crit-
ical roles in accomplishing coordinated fluid and solute flows
and solute distribution through the different medullary zones.
The three-dimensional architecture of thin-limb segments
and their positional relationships to other structures as they de-
scend or ascend the corticopapillary axis in the outer medulla
and inner medulla have become much more clearly defined in
recent years. The interaction of thin limbs of the loop of Henle
with neighboring structures is important for understanding
overall medullary function; therefore, medullary architecture
* Correspondence to pannabec@u.arizona.edu
1 Department of Physiology, University of Arizona Health Sciences
Center, Tucson, Arizona
Published online, July 2012 (comprehensivephysiology.com)
DOI: 10.1002/cphy.c110019
Copyright 
C American Physiological Society

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Structure and Function of the Thin Limbs of the Loop of Henle Comprehensive Physiology

is given broad coverage in the following review of thin-limb

structure and function.
This article reviews the structure and function of the adult
mammalian thin limbs of the loops of Henle, and focuses Cortical labyrinthe
primarily on rat, mouse, hamster, and chinchilla. The thin
limb of the loop of Henle is also present in the kidney of birds,
the only other animal class possessing this segment. Birds are
the only other vertebrates capable of producing concentrated
urine; however, in birds, the thin limb exhibits structural and
functional features that are markedly different from those of
mammals (15, 121). Medullary ray

Outer
stripe
Outer medulla
Morphology, Cytology, and 2
Ultrastructural Characteristics of the DTLupper
2

stripe
Inner
1
DTLupper
Thin Limb of the Loop of Henle 2
DTLupper
The loop of Henle consists of the proximal straight tubule,
DTL, ATL (present in the long-loop nephron only), and the
medullary and cortical portions of the thick ascending limb.
4 4 4
The proximal straight tubule, also known as the pars recta,

Inner medulla
3
consists of the terminal portion of the S2 segment and entire DTLlower
S3 segment of the proximal tubule. The DTL and ATL seg-
ments comprise what is known as the intermediate tubule (82). 3
The terminal portion of the proximal straight tubule lies in the DTLlower
outer stripe of the outer medulla. At the boundary between
the outer and inner stripes of the outer medulla, the proximal
straight tubule abruptly becomes the thin descending limb of 3
the loop of Henle. Nephrons are categorized into two types, DTLlower

determined by the depths at which the loops of Henle form

their bends. These two types are the short-loop nephrons,
which, in most mammalian species, form their bends in the
outer medulla, and the long-loop nephrons, which form their
bends in the inner medulla (Figs. 1 and 2). Variations occur,
with nephrons from human, beaver, and some rabbits form-
ing cortical bends, whereas all loops from some carnivores
form bends in the inner medulla (8). The ratio of short-loop
nephrons to long-loop nephrons is about 82:18 in mice, 70:30
in rats, and 85:15 in humans (8, 184).
Short-loop nephron
Short-loop nephrons have a thin descending segment but no
thin ascending segment; their ascending segment consists only
of the thick ascending limb of the loop of Henle. The bends
of the short loops of most species lie at approximately the
same axial level of the inner stripe, at the junction of the
outer medulla and inner medulla. In humans and several other
species, short loops form their bends at any level of the outer
medulla and in the cortex (63). In the mouse, short loops form
their bends at all levels between the middle level of the in-
ner stripe and the outer medullary-inner medullary boundary
(184). The transition from the DTL-type epithelium to the
thick ascending-type epithelium occurs some distance before
the bend or at the bend in short loops of rat and hamster (5,63).
Figure 1 Segmentation of thin limbs of the loop of Henle. The short-
loop nephron (right, belonging to a superficial glomerulus) has a
descending thick limb (pars recta of proximal tubule; hatched), a de-
scending thin limb (DTL) (turning back near the outer medullary-inner
medullary boundary), and a thick ascending limb (cross hatched),
which passes into the distal convoluted tubule a short distance be-
yond the macula densa (shown in black). The long-loop nephron (sec-
ond from right, belonging to a juxtamedullary glomerulus) contains a
DTL subdivided into two parts, type 2 and type 3 epithelium; and an
ascending thin limb (ATL), type 4 epithelium; the bend is located in
the inner medulla. Two additional long-loop nephrons (incompletely
drawn) demonstrate heterogeneity among long-loop nephrons, which
turn back at different levels within the inner medulla. Numbers 1-4 re-
fer to type of epithelium encountered in corresponding thin limb part:
type 1, DTL of short loops; type 2, upper part of DTL of long loops; type
3, lower part of DTL of long loops; type 4 (beginning short distance
before bend), ATL. Aquaporin 1 (AQP1)-negative segments/yellow;
AQP1-positive segments/red; ClC-K1-positive segments/green. Figure
based on data adapted, with permission, from references 27, 63, 125,
and 185.
Three types of bends occur in C57/BL/6J mouse short
loops (Fig. 2), each type being defined by the extent to which
the bend comprises DTL epithelium (184). In type 1 bends
(21% of total short-loop nephrons) the DTL-type epithe-
lium continues beyond the bend into the ascending limb (the

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Comprehensive Physiology
Cortex
OSOM
SLN1 SLN2
ISOM
SLN3
LLNt
Transitional zone
IM
LLN
Proximal tubule (PT)
Thin limb (TL)
Thick ascending limb (TAL)
Distal convoluted tubule (DCT)
Connecting tubule (CNT)
Initial connecting tubule (ICT)
Collecting duct (CD)
Figure 2 Segmentation of C57/BL/6J mouse short-loop and long-
loop nephrons. Tortuous descending thin limb (DTL) of long-loop
nephron (LLN; large arrow); winding course of thick ascending limb
(red arrowhead) of short-loop nephron (SLN) and CD (black arrow-
head); a piece of TAL inserted in the DTL of LLN (LLNt; small arrow),
which forms its bend just beneath the “transitional zone” within the
inner medulla; and three different types of SLN bends (SLN1, SLN2,
and SLN3). Outer stripe of outer medulla (OSOM), inner stripe of outer
medulla (ISOM), and inner medulla (IM). Figure adapted, with permis-
sion, from reference 184.
axial length of DTL epithelium on the ascending side of the
bend is less than 60 μm); in type 2 bends (33% of total)
the DTL epithelium transforms into the thick ascending limb
(TAL) within the bend itself; and in type 3 bends (47% of
total) the DTL epithelium ends on the descending side of
the loop at a point 50 to 450 μm before the bend, prior to
forming a thick ascending limb type of epithelium that con-
tinues into and through the ascending segment. Bend types
1-3 correspond to the positions of their glomeruli, which lie
at progressively deeper superficial and middle cortical lev-
els, respectively (184). Near the bends of most short-loop
nephrons in mouse, DTL and ATL are separated by approx-
imately 20 μm (184). Broad and flat bends with limbs that
are separated by a distance of 40 to 50 μm are occasionally
identified among all three bend types.
Volume 2, July 2012
Structure and Function of the Thin Limbs of the Loop of Henle
Long-loop nephron
Long-loop nephrons have a thin descending segment and a
thin ascending segment, the latter becoming the thick ascend-
ing limb at the outer medullary-inner medullary boundary.
The long-loop DTL of several mammalian species is known
to consist of two morphologically distinct segments. These
two segments are designated as DTLupper , for those descend-
ing segments lying in the inner stripe and outer inner medulla,
and DTLlower , for those segments lying in the innermost re-
gion of the inner medulla. The bends of long loops form at
all levels of the inner medulla in a cascading fashion (Fig. 1);
as a result, the number of loops at progressively deeper trans-
verse levels declines in number. The pattern of this decline
has been shown to approximate an exponential rate of decline
(71, 93, 132). In the Munich-Wistar rat approximately 40%
of all long loops form their bends within the first millimeter
below the outer medullary-inner medullary boundary (132).
In nearly all species that have been investigated to
date, the DTL terminates with a prebend segment that has
ultrastructural characteristics of the ATL. Prebend segments
with these characteristics have a mean length of about
700 μm in mouse (184) and range from about 50 to 140 μm
in the Sprague-Dawley rat (153) and from 0 to 140 μm in
rabbit (62) (functional aspects of the prebend segment are
discussed later in Section “Fluid and solute transport in the
descending thin limb of the loop of Henle”). In chinchilla, a
unique papillary segment beginning 2000 μm or less prior
to the bend and terminating some distance after the bend is
morphologically distinct from the ATL (see later) (27). A
segment exhibiting some similarity to the chinchilla papillary
segment is present in the desert sand rat, Psammomys (10),
a species which can produce a urine having a maximal
osmolality similar to that of the mouse and chinchilla (about
6000 mOsm/kg H2 O) (12). In most species, the bends of
long-loop nephrons resemble U-shaped hairpins. However, in
the terminal 500 μm region of the Munich-Wistar rat papilla,
some loop bends exhibit 5- to 10-fold greater transverse
length than the average transverse length of hairpin bends
(Fig. 3) (130). In the mouse kidney, long-loop DTLs arising
from glomeruli that originate deeper than mid-cortical levels
have a convoluted portion in the inner stripe that adds about
30% to their total DTL length (Fig. 2) (184).
In the mouse kidney, a transitional zone spans a narrow
region between the inner stripe and the initial 100 μm below
the outer medullary-inner medullary boundary (Fig. 2) (184).
In this transitional zone, which is known only in the mouse,
almost no loop bends are found, either from short or long
nephrons. Very few bends from short-loop nephrons reach
the innermost region of the inner stripe, and these short-loop
nephrons are characterized by a winding terminal loop and
a type 3 bend. A few long-loop nephrons form their bends
immediately beneath the transitional zone. These long-loop
nephrons are structurally similar to the other long-loop
nephrons, except for an approximately 100-μm-long segment
characterized by thick ascending limb-type epithelium, which
is inserted into the DTL at the level of the transitional zone.
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Structure and Function of the Thin Limbs of the Loop of Henle Comprehensive Physiology

Cilium

(A)

Figure 3 Immunolocalization of wide-bend thin limbs of loops of

Henle (arrows), ascending thin limb (ATLs), and CDs in Munich-
Wistar rat papilla. ATLs and wide-bend loops/ClC-K1/green (struc-
ture/immunogen/color), and CDs/aquaporin 2 (AQP2)/blue in a trans-
verse section that lies 70 μm above the tip of the papilla. Scale bar,
100 μm. Figure adapted, with permission, from reference 130.
(B)

Ultrastructure of the thin limb of the

loop of Henle
The ultrastructural characteristics of cells of thin limbs of
the loop of Henle have been reviewed in detail (5, 57, 63,
81, 152, 153, 184). Some of the distinguishing ultrastructural
characteristics of cells present in different subsegments are (C)
summarized in Table 1.
DTLs of short loops in all mammalian species that Figure 4 Type 1 epithelium of rat short-loop descending thin limb
have been investigated, including rat, mouse, hamster, (DTL). (A) Overview of cross-sectional profile, x ∼ 4000. (B) Simple
type 1 epithelium, x ∼ 12,000. (C) Complex tight junction in freeze-
chinchilla, rabbit, Psammomys, and Perognathus (63) consist fracture replica, x ∼ 71000. Intramembrane particle clusters (* ) seen on
of a simple (type 1) epithelium lying on a thin basement P-face of basolateral membrane correspond to desmosomes. Figure
membrane (Fig. 4). The epithelium is composed of relatively modified, with permission, from reference 63.
flat cells of simple structure that contain few cytoplasmic
organelles. These cells exhibit no interdigitation with each
other and are joined by a tight junctional belt of intermediate

Table 1 Summary of Structural Characteristics Typical of Thin-Limb Segments and Types 1-4 Thin-Limb Epithelia for Rat, Mouse, Hamster, and
Chinchilla (5, 27, 57, 63, 81, 152, 153, 184)

Feature Type 1 Type 2 Type 3 Type 4 Papillary type

Tubule outer diameter (μm) 19-27 15-45 16-19 19-21 ∼17-24

Cell height (μm) 0.1-0.3 0.3-2.1 0.3-2.1 0.3-0.5 ∼1-2
Microvilli projection + +++ + – ++
Interdigitation Diminished High Diminished High Highly
Number of tight junctions (per cross section) + +++ + +++ +++
Apicobasal depth of tight junction Deep Shallow Deep Very shallow Shallow
Intercellular space Narrow Narrow Narrow Widen toward basement membrane Narrow
Narrow basilar slits Yes Yes Yes No Yes
Basal infoldings Yes Yes Yes No Yes
Basement membrane – – – Multilaminated Multilaminated

The papillary-type epithelium has been found only in chinchilla; chinchilla tubule outer diameter and cell height are estimated from reference 27.
Relative intercellular junction solute permeability, based on degree of cellular interdigitation, number of tight junctions, and apicobasal depth of
tight junctions, is either tight (types 1 and 3) or leaky (types 2, 4, and papillary type). Table modified from reference 27.

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Comprehensive Physiology Structure and Function of the Thin Limbs of the Loop of Henle

apical-to-basal depth. This tight junction consists of several

junctional strands (3.75 on average in rat short loops (152))
and many desmosomes. A luminal cilium may be present in
the cells of the short-loop DTL (Fig. 4) (63).
Long-loop DTLs of most species that have been studied
have two subsegments that can be distinguished on the ba-
sis of cell structure (63): the type 2 segment (DTLupper ) that
abruptly follows the proximal tubule, and the type 3 segment
(DTLlower ) that lies distal to the type 2 segment (Table 1). The
type 3 segment continues almost to the bend of the loop of
Henle for all species studied, except for chinchilla. The type
2 epithelium is characterized by a high degree of interdigita-
tion along the cells’ lateral aspects between their apical and
basal poles, and by numerous shallow tight junctional profiles
consisting of one, or at most two, junctional strands [1.00 on
average in rat long loops (152)] (Fig. 5). These characteris-
(A)
tics of type 2 segments are similar for rat, mouse, hamster,
chinchilla, Psammomys, and Perognathus. However, the type
2 segment epithelial cells of rabbit, guinea pig, and prob-
ably human lack the high degree of cellular interdigitation
and are joined by deeper tight junctions consisting of several
junctional strands (81). Type 2 DTLs also exhibit prominent
microvilli on the luminal surface and a thin basement mem-
brane (63, 152).
Most long DTLs in the outer half of the inner stripe consist
of type 2 epithelia; however, a clearly heterogeneous and (B)
species-specific distribution of types 2 and 3 DTLs exists
at deeper levels of the medulla. In Psammomys and mouse,
most long DTLs at the beginning of the inner medulla consist
of type 2 epithelium (63), whereas in the rat most DTLs at
this level are lined with type 3 epithelium (153). Taking a
quantitative approach, Schwartz and Venkatachalam divided
the inner medulla into six equal levels and reported the number
of DTLs lined with types 2 and 3 epithelia at each level
beginning at the outer medullary-inner medullary boundary
(153). The percentages of type 2 epithelium at each of the six
progressively deeper levels were approximately 30, 16, 6, 3,
0, and 0. Similarly, in the outer 25% of the inner medulla of
the chinchilla kidney, somewhat less than 50% of DTLs are (C)
lined with type 2 epithelia, and in the terminal 2 mm of the
chinchilla papilla virtually all DTLs consist of the papillary
Figure 5 Type 2 epithelium of rat long-loop descending thin limb
type epithelium (see later) (27). The transition from type 2 (DTL) (DTLupper ). (A) Overview of tubular profile; note many tight junc-
to type 3 DTL occurs relatively abruptly for both rat and tions (arrows), x ∼ 4000. (B) Longitudinal section through epithelium;
chinchilla (25, 153). numerous tight junctions (arrows) indicate extensive cellular interdigita-
tion. Note basolateral “labyrinth,” x ∼ 18,000. (C) Flat section through
The type 3 epithelium, which comprises the lower seg- epithelium showing star-like shape of epithelial cells responsible for
ments of long DTLs consists of flat simple cells that appose cellular interdigitation. Note microfilament bundles (* ) in basal epithe-
each other with no interdigitations (Fig. 6). These cells lie on lium, x ∼ 13,500. Figure modified, with permission, from reference
63.
a thin basement membrane (63). The tight junctions found in
type 3 epithelia are deeper than those in type 2 and consist of
about 3 anastomosing junctional strands (152). The luminal
membrane has few microvilli and a thick electron-dense gly- descending segment, and in the entire ATL. Type 4 epithelia
cocalyx covering its surface (153). The basolateral membrane are characterized by flat, heavily interdigitated cells, but in-
forms basal infoldings. terdigitation is less than that occurring in type 2 DTLs (63)
Type 4 epithelia are present in segments that lie distal to (Fig. 7). Interdigitation occurs around the entire cell, produc-
those segments containing type 3 epithelia, and are found in ing a lengthy junctional belt. Tight junctions are shallow and
the prebend segment that lies at the terminal portion of the generally consist of a single junctional strand (152).

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Structure and Function of the Thin Limbs of the Loop of Henle
(A)
(B)
(C)
Figure 6 Type 3 epithelium of rat long-loop descending thin limb
(DTL) (DTLlower ). (A) Overview of cross-sectional profile. Only two tight
junctions (arrows) are encountered, x ∼ 3750. (B) Simple type 3 ep-
ithelium, x ∼ 16,500. (C) Freeze-fracture electron microscopy shows
complex tight junction, x ∼ 52,000. Figure modified, with permission,
from reference 63.
The chinchilla has a single type of thin-limb segment in
the terminal 20% of the papilla (Table 1) (27). This “papillary
type” epithelium has a length as long as 2 mm on the descend-
ing side of the loop and extends some distance into the ATL
before changing to the type 4 epithelium. All thin limbs in this
region of the chinchilla papilla, both DTLs and ATLs, exhibit
structural characteristics distinct from those found in types
1-4. The papillary type segment has relatively tall cells and a
complex cellular organization with extensive interdigitation,
numerous shallow tight junctions, and microvilli.
The distinct cytology of cells in types 2 and 4 epithelia, as
viewed in vitro with differential interference contrast optics
or by light-level microscopy, can be useful for distinguishing
the DTLupper from the ATL (Fig. 8) (16, 25-27, 50, 133). Cells
of type 3 epithelia, which closely resemble those of type 4
epithelia, when viewed with differential interference contrast
optics, can be distinguished as the continuation of type 2, or as
2068
Comprehensive Physiology
(A)
(B)
(C)
Figure 7 Type 4 epithelium of rat long-loop ascending thin limb
(ATL). (A) Overview of cross-sectional profile, x ∼ 3000. Note many
tight junctions (arrows). (B) Longitudinal section through epithelium
showing high degree of cellular interdigitation (junctions marked by
arrows), x ∼ 11,500. (C) Freeze-fracture electron micrograph showing
luminal aspect of two interdigitating cells, x ∼ 12,000. Note two differ-
ent types of intramembrane textures. Figure modified, with permission,
from reference 63.
the segment immediately preceding the prebend segment. A
successful method for ensuring correct segment identification
in vitro has been to retrieve tubules after experimental manip-
ulation and determine cell types of those tubules with electron
microscopy (27,49); however, even with electron microscopy
confirmation, considerable scatter in data values can persist
(54), underscoring an enduring difficulty in clearly identify-
ing the subsegments of thin limbs of the loop of Henle in
vitro.
About 50% of Munich-Wistar rat inner medullary thin
limbs, lying at positions distinctly above the bend, have been
shown to have segments exhibiting structural characteris-
tics of type 2 DTL (AQP1-positive) that are contiguous to
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Comprehensive Physiology Structure and Function of the Thin Limbs of the Loop of Henle

(A)

(B)

Figure 8 Photomicrographs of Munich-Wistar rat thin limbs of the loop of Henle from the
inner medullary outer zone (OZ; see Fig. 13), viewed with differential interference contrast
optics. (A) Type 2 epithelium [descending thin limb (DTLupper )] and (B) type 4 epithelium [as-
cending thin limb (ATL)]. Note cells with nuclei protruding into lumen in type 2 epithelium and
cells with large, round, flat nuclei in type 4 epithelium. Scale bar, 100 μm. Figure adapted,
with permission, from 127.

segments exhibiting structural characteristics of type 4 ATL
(ClC-K1-positive) (16, 127). Multiple ATL-type segments of
variable length are interspersed along a single straight por-
tion of these “mixed-type” thin limbs of the loop of Henle,
likely on the descending side of the bend. Inner medullary
thin limbs with repeating, sequential expression of DTL-type
and ATL-type regions are also present in Sprague-Dawley
rats, mice, and rabbits. Reverse-transcribed polymerase chain
reaction (RT-PCR) of microdissected segments showed that
the water channel AQP1 and the urea transporter UT-A2 are
expressed in pure DTL, but not in pure ATL, and in DTL-
type, but not in ATL-type regions of mixed-type thin limbs.
Immunocytochemistry revealed expression of AQP1 in cells
of DTL-type, but not ATL-type regions of mixed-type thin
limbs. In contrast, the chloride channel ClC-K1 is expressed
in ATL-type, but not DTL-type regions of mixed-type thin
limbs. A functional role for mixed-typed thin limbs would be
critically dependent on the number, length, position within the
papilla, and overall architectural arrangement of the segments
of each type; however, no role for mixed-type thin limbs has
been identified (127).
Histotopography of the Thin Limb
of the Loop of Henle and
Medullary Zonation
Considerable attention has been placed on understanding the
patterned organization of medullary tubules and vessels, and
their geometrical relationships to each other. The proximity
of one structure to another, and the expression of segment-
specific cell-membrane transport pathways largely determine
both axial flows as well as the highly organized lateral flows of
fluid and solutes that occur between tubules and vessels. These
flows are critical for understanding solute exchange, cycling,
and sequestration patterns related to the urine concentrating
mechanism, as well as for understanding medullary and re-
nal function more generally. Medullary histotopography and
zonation, arising from the geometrical relationships found in
the rat, and in the mouse, where noted, are summarized in this
section.
The entire medulla consists of multiple, spatially distinct
zones and regions distributed across the axial (corticopap-
illary) and transverse (perpendicular to the corticopapillary
axis) dimensions, respectively. Regional and zonal designa-
tions are based on architecture of loops of Henle, CDs, and
blood vessels. The outer medulla is generally considered to
consist of two zones in the axial dimension and two zones in
the transverse dimension. The axial zones include the outer
stripe and the inner stripe; the transverse regions include the
vascular bundle and the interbundle regions (8, 63). The inner
medulla can be considered to consist of four axial zones and
two transverse regions. The axial zones include outer zones
1 and 2, and inner zones 1 and 2; the two transverse regions
include the intracluster and intercluster regions. Structural
characteristics of the thin limbs of Henle that reside in the
inner stripe and inner medullary zones are summarized in
Table 2 (131).
In the outer stripe of the outer medulla, the histotopogra-
phy is dominated by the straight proximal tubule segments,
which make the most abundant contribution in terms of tissue
mass. In the inner stripe the thick ascending limbs dominate
the histotopography. In the outer medulla, the vascular bundle
region can be viewed as the central organizing feature in the
transverse dimension (97) (Fig. 9). The vascular bundles con-
sist of descending and ascending vasa recta, and, in the inner
stripe of mouse and rat, short-loop DTLs. The interbundle

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Structure and Function of the Thin Limbs of the Loop of Henle Comprehensive Physiology

Table 2 Characteristics of Rat Thin Limbs that Reside in 5 Medullary Axial Zones

OM Inner inner medulla inner medulla inner medulla inner medulla

Zone Characteristic stripe OZ-1 OZ-2 IZ-1 IZ-2

√
Short-loop DTLs with first outer medullary segment 10%
AQP1-positive, last 90% AQP1-negative √ √
Long-loop DTLs completely lacking inner medullary AQP1 √ √ √ √ √
Long-loop DTLs with first inner medullary segment 40%
AQP1-positive, last 60% AQP1-negative √ √
Only AQP1-negative portions of long-loop DTLs* remaining √ √ √ √
ATLs with ClC-K1-positive, narrow bend √
ATLs with ClC-K1-positive, wide bend

* DTLs with first inner medullary segment 40% AQP1-positive, last 60% AQP1-negative.
For the outer medulla (OM), the outer stripe extends for 1 mm below the cortico-medullary boundary; the inner stripe extends from about 1 mm
below the cortico-medullary boundary to about 2 mm below the cortico-medullary boundary. For the inner medulla (IM), outer zone 1 (OZ-1)
extends for 1 mm below the outer medullary-inner medullary boundary; outer zone 2 (OZ-2) extends from 1 mm below the outer medullary-inner
medullary boundary to 3.0 to 3.5 mm below the outer medullary-inner medullary boundary; inner zone 1 (IZ-1) extends from 3 to 3.5 mm below
the outer medullary-inner medullary boundary to 4.5 mm below the outer medullary-inner medullary boundary; inner zone 2 (IZ-2) extends from
4.5 mm below the outer medullary-inner medullary boundary to 5 mm below the outer medullary-inner medullary boundary. Modified from
reference 132.

region consists of thin and thick limbs of the loop of Henle,
CDs, and interconnecting capillaries.
Throughout the inner stripe of the outer medulla of nonro-
dent mammalian species, the thin descending segments of the
short loops lie within the interbundle region, distant from the
vascular bundle, and maintain a relatively constant distance
from the vascular bundle in their descent (81). The descending
segments of the longest long-loop nephrons of these species
lie nearer to the vascular bundle than descending segments
of shorter long-loop nephrons. As indicated previously, ro-
dent medullary architecture shows contrasting variations to
this pattern; upon entering the inner stripe, thin descending
segments of short-loop nephrons of the rat approach the vas-
cular bundle and then descend alongside the perimeter of the
bundle (81,84). In mouse, thin descending segments of short-
loop nephrons approach and then enter the vascular bundle,
whereupon they continue their descent within the bundle. The
Figure 9 Immunolocalization of tubules and vessels in the inner
stripe of the Munich-Wistar rat outer medulla. CDs/aquaporin 2
(AQP2)/yellow; long-loop descending thin limb (DTL)/aquaporin 1
(AQP1)/white; short-loop DTL/UT-A2/blue; thick ascending limb/ClC-
K2/orange; descending vasa recta/UT-B/green. Ascending vasa recta,
capillaries, and AQP1-negative long-loop DTLs are not shown. Overlay
of two adjacent transverse sections, 1 μm apart. Scale bar, 250 μm.
Unpublished figure, Thomas Pannabecker.
close proximity between the DTLs of short-loop nephrons and
the descending and ascending vasa recta that is brought about
by this repositioning within the inner stripe is considered to
serve as a significant functional adaptation that contributes
to the relatively high urine concentrating ability of rodents
(8, 72, 97, 176). The combination of low transepithelial water
permeability and significant transepithelial urea permeability
in the terminal portions of DTLs of short-loop nephrons have
been shown to support urea cycling in mathematical models
of the urine concentrating mechanism (89).
In the inner medulla, the CDs coalesce as they descend the
corticopapillary axis, forming organized CD clusters that can
be viewed as the central organizing feature; these CD clus-
ters define two transverse regions (128) (Fig. 10). The “in-
tracluster” region includes the area encompassed by a group
of coalescing CDs, the “intercluster” region consists of the
region that lies at the periphery and outside the CD cluster
(Fig. 10). Distinguishing features of the intracluster region are
the four or more symmetrically arranged capillaries that abut
and run alongside every CD in the corticopapillary dimen-
sion. In the transverse dimension, particularly noticeable in
the rat, inner medullary AQP1-positive DTLs are distributed
non-uniformly, whereas ATLs (and prebend segments) are
distributed in a much more uniform pattern (Fig. 11). Three-
dimensional reconstructions of rat inner medulla have shown
that AQP1-positive DTLs lie in the intercluster region, out-
side and at the periphery of CD clusters, throughout most of
their descent along the corticopapillary axis (Fig. 12). There-
fore, AQP1-positive DTLs are anatomically separated from
CDs, lying in a spatially distinct compartment. In contrast,
ATLs of the inner medulla lie in both the intercluster and
intracluster regions (Fig. 12); some, but not all ATLs lie in
the compartment that is spatially distinct from that occupied
by CDs. Some of these arrangements have been shown for
mouse inner medulla (184), including a similar separation of
AQP1-positive DTLs from CDs (Pannabecker, unpublished
observations).

2070 Volume 2, July 2012

Comprehensive Physiology
Figure 10 Immunolocalization of CDs and associated thin limbs of
the loop of Henle in the Munich-Wistar rat inner medulla. The “intra-
cluster” region is bounded by the red borders, the “intercluster” region
lies between the red and white borders. Transverse section is from
about 400 μm below the outer medullary-inner medullary boundary
(outer zone 1; see Fig. 13). Five primary CD clusters are outlined by
white borders; borders were determined by the Euclidean distant map
technique (132). These five primary clusters make up a single sec-
ondary CD cluster that consists of 31 CDs near the outer medullary-
inner medullary boundary. Descending thin limb (DTL)/aquaporin
1 (AQP1)/red, CD/aquaporin 2 (AQP2)/blue, ascending thin limb
(ATL)/ClC-K1/green. Scale bar, 100 μm. Figure modified, with per-
mission, from reference 132.
In the inner medulla of the Munich-Wistar rat, four axial
zones have been distinguished (131, 132) (Fig. 13): (1) an
outermost zone (OZ1) of about 1 mm thickness, just below
the outer medulla, where loops expressing negligible or no
AQP-1 have their bends (128); (2) a larger outer zone (OZ2),
just below the outermost zone, 2 – 2.5 mm in thickness, which
contains well-organized CD clusters where tubules and ves-
sels are tightly packed and where loops bend within the central
portions of the clusters; (3) an “outer” inner zone (IZ1) where
the organization of the CD clusters is diminishing and nearly
all vasa recta are fenestrated; and (4) an innermost zone (IZ2)
where CD clusters can no longer be distinguished, where the
CDs appear to dominate all other structures, where nearly
all vasa recta are fenestrated, and where a large fraction of
loops exhibit substantial transverse-running segments (Fig. 3)
(130). The two inner zones make up the approximately 1.5 to
2 mm of the Munich-Wistar rat papilla. The unique papillary
loop of Henle segment of chinchilla (27) and a comparable
segment hypothesized to exist in Psammomys obesis (10, 27)
lie in the inner zone.
Short long-loop nephrons that have DTLs which express
undetectable AQP1 in their inner medullary segments, and
Volume 2, July 2012
Structure and Function of the Thin Limbs of the Loop of Henle
(A)
(B)
Figure 11 Immunolocalization of thin limbs of the loop of Henle
in the Munich-Wistar rat inner medulla. Transverse sections showing
(A) nonuniform distribution of aquaporin 1 (AQP1)-positive descending
thin limb (DTL)/AQP1/red and (B) near uniform distribution of prebend
segments and ascending thin limb (ATL)/ClC-K1/green. Sections lie
within 1300 μm below the outer medullary-inner medullary boundary
(outer zone 2, see Fig. 13). AQP1-negative DTLs are not shown. Scale
bars, 100 μm. Figure modified, with permission, from reference 129.
reach to no more than about 1 mm into the Munich-Wistar
rat inner medulla, lie at the periphery of CD clusters in the
transverse dimension (Fig. 14) (128). All ATLs of short long-
loop nephrons, and for the most part, only ATLs of short
long-loop nephrons, lie within the OZ 1 intracluster region
(128). Thus, short long-loop nephrons are a defining feature
of OZ1 in the rat kidney. Long long-loop nephrons of the
rat kidney reach more than 1 mm into the inner medulla, and
label for AQP1 only in the upper 40% of their DTLs; the lower
60% do not express detectable AQP1 (Fig. 15). These loops
usually enter CD clusters before the bend, and then ascend
within the cluster for variable distances, before exiting into
the intercluster region (Fig. 16) (124). Little or no detectable
AQP1 is expressed in any DTL segment in the final 2.0 to
2.5 mm of the rat and chinchilla inner medulla (27, 130).
These expression patterns of AQP1 suggest that DTL luminal
fluid may not equilibrate with interstitial fluid by way of water
reabsorption in the lower, type 3 segments.
The rate of change in numbers of AQP1-positive
DTLs, AQP1-negative DTLs, and ATLs at each axial
level in the rat inner medulla exhibits a complex pattern
(132). The total number of thin limbs at progressively
deeper axial levels declines at a rate that closely approxi-
mates an exponential rate of change (Fig. 17) (43, 91-93).
The AQP1-positive DTLs decline in number at a nearly
linear rate, with few or none present below approxi-
mately 1700 μm from the outer medullary-inner medullary
boundary (Fig. 17). Throughout this initial 1700 μm below
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Structure and Function of the Thin Limbs of the Loop of Henle Comprehensive Physiology

(A) (B)

Collecting duct Collecting duct

Descending thin limb Ascending thin limb

Descending vasa recta

Figure 12 Three-dimensional reconstruction of a primary CD cluster and associated tubules and ves-
sels in the Munich-Wistar rat inner medulla. (A) Descending thin limbs (DTLs) and descending vasa recta
that are associated with a primary CD cluster lie at the periphery of or outside of the cluster along the
entire axial length of the cluster. Aquaporin 1 (AQP1)-positive DTLs/AQP1/red; AQP1-negative DTLs/α-B
crystalline/gray; descending vasa recta/UT-B/green; CDs/aquaporin 2 (AQP2)/blue. (B) Ascending thin
limbs (ATLs) and prebend segments associated with a primary CD cluster lie at the periphery of, outside of,
or amongst the CDs along the entire axial length of the cluster. ATLs and prebend segments/ClCK/green;
CDs/AQP2/blue. The upper edge of the image is positioned near the outer medullary-inner medullary
boundary. Scale bars, 250 μm; inset scale bars, 500 μm. Figure modified, with permission, from refer-
ences 128 and 129.

the outer medullary-inner medullary boundary, the number
of AQP1-negative DTLs is relatively constant. This results
from a combination of two factors: (1) the DTLs of short
long-loop nephrons that form their bends within 1 mm be-
low the outer medullary-inner medullary boundary and ex-
press no detectable AQP1 (∼40% of all DTLs) reduce the
number of AQP1-negative DTLs through attrition as they
form their bends; and (2) the DTLs of AQP1-positive long
long-loop nephrons gradually become AQP1-negative as they
descend to deeper papillary levels, and these loops add to
the existing AQP1-negative DTL population through a one-
for-one replacement of AQP1-positive DTLs. This tends to
offset the number of AQP1-negative segments lost by at-
trition. After the point where AQP1-positive DTLs disap-
pear entirely (∼1700 μm below the outer medullary-inner
medullary boundary in the rat), the number of AQP1-negative
DTLs declines at an exponential rate similar to that of other
limbs. The changes in numerical density of AQP1-positive
and AQP1-negative DTLs along the corticopapillary axis cor-
respond to axial changes in ratios of types 2 and 3 DTLs
mentioned earlier (153). The transition from AQP1-positive
to AQP1-negative DTL, for each tubule, occurs over a dis-
tance of about 100 to 200 microns (125), corresponding to a
relatively abrupt transition from type 2 to type 3 epithelium
(25, 153).
CDs in combination with some of the ascending vasa recta
and ATLs, when viewed in transverse sections, are arranged
so as to form interstitial compartments or microdomains.
These compartments have been called interstitial nodal spaces
(INSs) (Fig. 18). The INSs are bordered on one side by a CD,
on the opposite side by one or more prebend segments or
ATLs, and on the other two sides by ascending vasa recta or
capillaries (128, 129). INSs are arrayed at structured intervals
throughout the inner medulla, primarily within CD clusters,
and could play an important role in generating the cortico-
papillary osmotic gradient, possibly by serving as compart-
mentalized solute mixing chambers (92). The entire length of
each prebend and postbend equivalent length ATL segment
lies in contact with one or more INSs (124). This placement of
NaCl-permeable loop bends to INSs may function specifically
to raise the osmolality of CD tubular fluid by facilitating the
targeted delivery of NaCl from thin limbs to the CD clusters
(131). The number of contacts made by the total population
of prebend and postbend segments exhibits a marked, but
variable, periodic increase and decrease at regular intervals
of medullary depth; this number of contacts correlates with
variable, periodic changes in prebend and postbend number
that occur at regular intervals of medullary depth (124). The
impact of periodic changes in prebend and postbend number
on renal function is not known.

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Comprehensive Physiology
OZ1
OZ
OZ2
IZ1
IZ
IZ2
Figure 13 Four subsections, or zones, of the rat inner medulla;
based on data from the Munich-Wistar rat (132): (1) an outer-most
zone (OZ1) of about 1 mm thickness, just below the outer medulla, in
which loops expressing negligible or no inner medullary aquaporin 1
(AQP-1) have their bends; (2) a larger outer zone (OZ2), just below
the outermost zone, 2 to 2.5 mm in thickness, which contains well-
organized CD clusters in which tubules and vessels are tightly packed
and in which loops bend within the central portions of the clusters; (3)
an outer inner zone (IZ1) in which the organization of the CD clusters is
diminishing and nearly all vasa recta are fenestrated; and (4) an inner-
most zone (IZ2) in which CD clusters can no longer be distinguished,
the CDs appear to dominate all other structures, nearly all vasa recta
are fenestrated, and a large fraction of loops have transversely running
segments. The two inner zones make up approximately 1.5 to 2 mm of
the papilla. CD clusters/blue coalesce into single CDs. AQP1-positive
DTLs/red, AQP1-negative descending thin limbs (DTLs)/yellow, and
ascending thin limbs (ATLs) and prebend segments/green. Scale bar,
1 mm along the axial dimension; lateral dimensions are not to scale.
Figure adapted, with permission, from reference 90.
Fluid and Solute Transport in the
Thin Limb of the Loop of Henle
Water, NaCl, and urea flows and distributions within the
medulla are important determinants of sodium and water
balance and of long-term regulation of arterial pressure.
Moreover, the flows and distributions of water, NaCl, and
urea play critical roles in the urine concentrating mechanism
(28, 123). Loops of Henle are involved with the redistribution
of medullary fluid and solutes among the interstitial, tubu-
lar, and vascular compartments. The processes of fluid and
solute secretion and reabsorption by thin limbs of the loop
of Henle have been investigated using micropuncture tech-
niques, in vivo (88, 107, 134); however, defining net fluid and
solute transepithelial flux in vivo is complicated by poor ac-
cessibility to the loops in their native configurations. Another
approach to defining net flux, in vivo, is to determine transep-
ithelial permeabilities for water, Na, Cl, and urea in the loops
of Henle by way of in vitro microperfusion, and then make
assumptions about in vivo transepithelial solute gradients that
drive these fluxes (52, 53, 96). With this approach, estimates
of fluid and solute movements within and between compart-
ments can be determined. Mathematical modeling has played
a large role in attempting to gain comprehensive understand-
ing of these fluid and solute movements in the kidney (145).
Volume 2, July 2012
Structure and Function of the Thin Limbs of the Loop of Henle
In all thin-limb segments, transepithelial flux of major
solutes is generally considered to occur primarily by diffusion.
There is very little evidence that active transport contributes
to substantial, transcellular solute flux in any of the thin limbs
of the loop of Henle (25, 26, 47, 50, 158).
Defined axial zonation of the inner medulla of chinchilla
(27) and rat (Fig. 13) (111, 125, 128) provides a method-
ology for isolating AQP1-positive and AQP1-negative DTL
segments (types 2 and 3 epithelia, respectively) (29). AQP1-
positive DTLs of the rat can be teased from the outer 2.5 to
3 mm of the inner medulla (outer zone), selecting only DTL
segments from loops that extend well beyond the first 1 mm
of the inner medulla because the short long loops that turn
within this first mm express no detectable AQP1. Although
the longer DTL segments express AQP1, they probably in-
clude a portion lacking AQP1. AQP1-null DTL segments can
be teased from the inner 2 to 2.5 mm of the inner medulla (in-
ner zone) in which there is no detectable expression of AQP1.
The terminal 250 μm above the loop bend on the descending
side of the loop is removed to eliminate the prebend region.
Functional studies of fluid and solute transport in thin
limbs of the loop of Henle are most complete for the rat,
hamster, chinchilla, and rabbit (25, 26, 51, 52, 57, 83).
Fluid and solute transport in the descending
thin limb of the loop of Henle
Short DTLs (type 1) of hamster were reported to have sub-
stantial water permeability in the single published study of
short loop water permeability (49). However, in their study
of hamster short DTLs, Imai and colleagues acknowledged
the possibility of failing to distinguish between types 1 and
2 segments (49), so the high water permeability values of
short loop DTLs may be significantly lower than those re-
ported from that study. The absence of detectable AQP1 in
short loop DTLs of mouse, rat, and human, determined with
immunohistochemistry (185), as well as the high osmotic wa-
ter permeability in long-loop DTLs of AQP1 knockout mice
(discussed later) suggest the possible existence of alternate
pathways for transepithelial water flux.
The water permeability of type 1 segments is lower than
the very high permeabilities measured in segments of the
long-loop DTLupper (type 2; Table 3) from hamster (49,52,53),
rabbit (74), chinchilla (25), rat (29, 47), and mouse (23). The
osmotic water permeability along the axis of long-loop DTLs
declines significantly with increased depth below the outer
medullary-inner medullary boundary in the rat and chinchilla,
the only species for which measurements are available for both
upper and lower DTL segments (Table 3).
The long-loop DTLs are functionally heterogeneous;
tubule segments composed of cells with structural charac-
teristics typical of types 2 and 3 epithelia (Fig. 1) exhibit
high and low osmotic water permeability, respectively in
chinchilla (27), and likely do so in rat where high and low
osmotic permeability (29) correlate with presence and ab-
sence of AQP1 protein expression, respectively (125, 128).
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Structure and Function of the Thin Limbs of the Loop of Henle Comprehensive Physiology

(A)

(B)

(C) (D) (E) (F)

Figure 14 Organization of a primary CD cluster and thin limbs of the loop of Henle in the Munich-
Wistar rat inner medulla. (A, B) A single transverse section from near the outer medullary-inner
medullary boundary (outer zone 1; see Fig. 13) showing profiles of CDs and associated thin limbs.
Aquaporin 1 (AQP1)-positive descending thin limbs (DTLs)/AQP1/filled red; AQP1-negative DTLs/α-B
crystallin/unfilled red; ATLs and prebend segments/ClCK/green and white; CDs associated with the
primary cluster/aquaporin 2 (AQP2)/dark blue; CDs not associated with the primary cluster are shown
in light blue. (A) Thin limbs of short long-loop nephrons associated with the CD cluster. These thin
limbs form their bends within 1 mm below the outer medullary-inner medullary boundary. AQP1-
negative DTLs lie at the edge of the CD cluster and their connecting prebend segments and ATLs lie
in the intracluster region. (B) Thin limbs of long long-loop nephrons associated with the CD cluster.
These thin limbs form their bends between approximately 2 to 3 mm below the outer medullary-inner
medullary boundary. AQP1-positive DTLs and their connecting ascending thin limbs (ATLs) lie in the
intercluster region, distant from CDs. (C-F) Three-dimensional reconstruction of primary cluster CDs
and associated thin limbs that are shown in A and B. AQP1-positive/red; AQP1-negative/yellow;
ATLs and prebend segments/green; CDs/blue. The outer medullary-inner medullary boundary lies
near the top edge of the figure. (C) DTLs of nephrons that form their bends within 1 mm below
the outer medullary-inner medullary boundary. (D) DTLs of nephrons that form their bends between
approximately 2 to 3 mm below the outer medullary-inner medullary boundary. (E) ATLs of nephrons
that form their bends within 1 mm below the outer medullary-inner medullary boundary, and (F) ATLs
of nephrons that form their bends between approximately 2 to 3 mm below the outer medullary-inner
medullary boundary. Scale bars, 100 μm. Figure modified, with permission, from reference 128.

Chinchilla DTLs have an additional papillary segment inter-
vening between types 3 and 4 (27). The isolated, perfused
papillary type thin limbs of chinchilla exhibit very low but
nonzero water permeability (68 ± 9 μm/s). Variations in the
imposed osmotic driving force in isolated tubule experiments,
at least within the range of several hundred mOsmol, do not
significantly alter the very low fluid permeability from DTLs
of the deep papilla of chinchilla (25).
It has been shown for the moderately antidiuretic rat and
hamster, that luminal fluid at the bend of the loops that lie
near the tip of the papilla is in approximate osmotic equilib-
rium with capillary fluid (42,60,108,109,135), and therefore,
is likely in equilibrium with interstitial fluid. Transepithelial
water permeability data for rat, hamster, and chinchilla thin
limbs (discussed earlier; Table 3) have shown that signifi-
cant osmotic equilibration by water flux certainly occurs in
the upper DTL, whereas studies with rat and chinchilla have
shown that little or no osmotic equilibration by water flux
occurs in the lower DTL. Equilibration may occur by solute
flux, to varying degrees, in both the upper and lower DTL of
rat, hamster, chinchilla, and Psammomys (91). The collective
histotopography of inner medullary AQP1-positive DTLs, on
one hand, and AQP1-negative DTLs on the other, and their
different water permeabilities, are likely related to distinct
contributions of these segments to production of the axial
osmotic gradient of the inner medulla (92).
Specific transepithelial transport pathways for Na, K, and
Cl are poorly understood for the DTL; the Na flux is be-
lieved to be largely paracellular (54). In short DTLs of rat
and hamster, Na:Cl and K:Cl permeability ratios are nearly
equal, whereas in long DTLupper , the Na and K permeabili-
ties exceed that for Cl (i.e., Na:Cl and K:Cl ratios exceed 1)
(48). In chinchilla long DTLupper , the Na permeability exceeds
that for Cl (Na:Cl ratio = 2.3), as in the hamster; however,

2074 Volume 2, July 2012

Comprehensive Physiology Structure and Function of the Thin Limbs of the Loop of Henle

(A) (B) (C) (D) (E)

Figure 15 Three-dimensional reconstruction of Munich-Wistar rat thin limbs of the loop of Henle
that form bends at four different levels below the outer medullary-inner medullary boundary. (A)
Thin limbs that form bends within the first mm below the outer medullary-inner medullary boundary.
Descending thin limbs (DTLs) lack detectable aquaporin 1 (AQP1). ClC-K1 is expressed continuously
along the prebend segment and the ascending thin limb (ATL). (B-D) Thin limbs that form bends below
the first millimeter of the inner medulla. AQP1 is expressed along the initial 40% of each DTL (type 2
epithelium), and is absent from the terminal 60% (type 3 epithelium). ClC-K1 is expressed continuously
along the prebend segment and the ATL. Boxed area is enlarged in E. (E) Enlargement of near-bend
regions of four thin limbs from box in D. ClC-K1 expression begins, on average, approximately 170 μm
before the bend (arrows). AQP1-positive DTLs/AQP1/red; AQP1-negative DTLs/α-B crystallin/gray;
ATLs and prebend segments/ClCK/green. Scale bars, (A-D) 500 μm; (E) 100 μm. Figure modified,
with permission, from reference 91.

the Na:Cl permeability ratio declines at progressively deeper
levels, reaching 0.55 in the long DTLlower (26). The Na per-
meabilities of the long DTLupper are roughly comparable in
hamster and chinchilla (Table 3); however, in chinchilla, the
Na permeability of the long DTLlower becomes substantially
higher than that of the long DTLupper .
The terminal portion of the descending side of the thin
limb of the loop of Henle (the prebend region) consists of
type 4 epithelium, expresses the Cl channel ClC-K1, and lacks
expression of AQP1, indicating a functional resemblance to
the ATL (27,111,125,153). The prebend segment is generally
assumed to be permeable to NaCl and impermeable to water
(27, 91, 96, 111, 125); however, some uncertainty remains as
no functional studies of this segment have been reported in
the literature.
It has been hypothesized that significant net NaCl efflux
from the prebend segment occurs primarily along its short ax-
ial distance and along an equivalent length of the ATL. Above
this region, along the ascending limb, the driving force for
NaCl diminishes as the ATL approaches the outer medulla
(91, 94). Mathematical models indicate that such delivery of
NaCl along a narrow axial zone would be most effective for
producing highly concentrated urine (95). The transverse-
running segments (wide-bend loops of Henle) that lie in the
terminal 500 μm of the Munich-Wistar rat papilla (Fig. 3)
(130) provide for a high NaCl-reabsorbing surface area, es-
pecially over the final 250 μm of the papilla tip, in contrast to
that which would occur if all loops had narrow bends. Of per-
haps even greater significance, the occurrence of wide-bend
loops leads to a striking increase in the apparently critical
loop NaCl-reabsorbing surface area relative to the CD urea-
and water-reabsorbing surface area over only 50 μm in axial
distance (from 25 μm to 75 μm above the papilla tip) (130).
Thus, the wide-bend architecture may well underlie the very
high osmolality of the deep papilla in the Munich-Wistar
rat.
In the Munich-Wistar rat (Dantzler, Evans, and
Pannabecker, unpublished) and chinchilla (Table 3) (25, 27)
long-loop DTL, urea permeability increases with increasing
depth below the outer medullary-inner medullary boundary,
qualitatively opposite to the axial decline in water permeabil-
ity. These transepithelial urea fluxes in inner medullary DTL
segments appear to be nonsaturable and are not inhibited by
phloretin, and therefore are unlikely to be mediated by any of
the known phloretin-sensitive renal urea transporters (UT-A1,
UT-A2, and UT-A3) (145). Two hypothetical pathways for
these urea movements include an unknown urea transporter
and/or the paracellular pathway.

Volume 2, July 2012 2075

Structure and Function of the Thin Limbs of the Loop of Henle
Figure 16 Schematic diagram of Munich-Wistar rat thin limb of loop
of Henle architecture along the corticopapillary axis (124, 131). In the
outer medulla, vascular bundles can be considered to be the central
organizing elements around which DTLS of long-loop nephrons, TALs,
CDs, and capillaries are systematically arranged; in the inner medulla,
the CD clusters can be considered to be the central organizing ele-
ments. DTLs of nephrons that form their bends within 1 mm below
the outer medullary-inner medullary boundary express no detectable
AQP1 in their inner medullary segments and lie near the interface of
the intercluster and intracluster regions; their ATLs lie within the intra-
cluster region (cf. Fig. 14). Descending thin limbs (DTLs) of loops that
form their bends deeper than 1 mm below the outer medullary-inner
medullary boundary pass from the intercluster region into the intraclus-
ter region above the prebend segment, and their ascending thin limbs
(ATLs) exit into the intercluster region above the equivalent postbend
length. ATLs may be positioned either adjacent to or distant from their
contiguous DTLs. The depicted symmetry between bundle and cluster
regions of the outer medulla and inner medulla, respectively, has not
been demonstrated.
Micropuncture studies have shown a large net secretion
of urea into rat and hamster short-loop and long-loop DTLs
(88, 107, 134). The UT-A2 urea transporter is one possi-
ble pathway for urea secretion into the DTL, at least for
those DTL segments lying near the outer medullary-inner
medullary boundary. Most ascending vasa recta that leave the
inner medulla exit by way of vascular bundles that span the
outer medullary-inner medullary boundary, and continue to
the cortex (8, 113, 123, 142). Short DTLs lie within these vas-
cular bundles in the rat and the mouse. Urea diffusing from
the highly fenestrated and highly urea-permeable ascending
vasa recta likely diffuses into the short DTLs, possibly by way
of UT-A2. This process could serve as a countercurrent ex-
change process that would return urea to the inner medulla via
more distal tubule segments; in this manner, countercurrent
exchange could add significant amounts of urea to the inner
medullary interstitium. A population of fenestrated capillar-
ies that ascend from the inner medullary intracluster region
passes directly into interbundle regions of the outer medulla
and thereby bypasses the outer medullary vascular bundles
2076
Comprehensive Physiology
100
Inner medulla Outer medulla
AQP1-pos DTL
AQP1-neg DTL
Total DTLs
80 ATL
Number of segments
Loop decay rate
60
40
20
0
0 1000 2000 3000 4000 5000
Distance below inner medullary base (microns)
Figure 17 Number of Munich-Wistar rat thin-limb subsegments
along the inner medullary corticopapillary axis. The plot shows the
numbers of aquaporin 1 (AQP1)-positive and AQP1-negative descend-
ing thin limbs (DTLs) and contiguous ascending thin limbs (ATLs) as-
sociated with a single secondary CD cluster at successive transverse
levels. Prebend segments were not included in the DTL count, and
as a result, the number of ATLs exceeds the number of DTLs. The
thin-limb population declines with increasing depth below the outer
medullary-inner medullary boundary, at the exponential rate defined
in previous studies (loop decay rate) (43, 91, 93). Inset: DTL and ATL
segments for two inner medullary nephrons. AQP1-positive DTL/red;
AQP1-negative DTL/yellow; prebend and ATL/green. Figure modified,
with permission, from reference 132.
(67). These capillaries possibly carry significant amounts of
urea into the interbundle region (92). Urea secretion from
these capillaries into the interbundle long DTLs by way of
UT-A2 could therefore represent another significant urea re-
cycling pathway that would return urea directly to the inner
medulla.
Figure 18 Interstitial nodal spaces in Munich-Wistar rat inner
medulla, as seen with electron microscopy. Electron micrograph shows
a transverse section from outer zone 2 (OZ2; see Fig. 13), showing
ascending thin limbs (ATLs) and ascending vasa recta (AVR) arranged
around a single CD. Interstitial nodal spaces are marked with X. Scale
bar, 10 μm. Figure adapted, with permission, from reference 129.
Volume 2, July 2012
Comprehensive Physiology Structure and Function of the Thin Limbs of the Loop of Henle

Table 3 Permeability Properties of Subsegments from the Long-Loop Thin Limb of the Loop of Henle

Rabbit Hamster Rat Chinchilla Mouse

Permeability
Segment Origin Pf Urea Na Cl Pf Urea Na Cl Pf Urea Na Cl Pf Urea Na Cl Pf Urea Na Cl

DTL
Outer Medulla
Inner Stripe 2530 1.5 1.6 2430 2.0 20.8 2187 2584 3.3 25.7
(74) (75) (74) (53) (52) (52) (47) (25) (26) (26)
Inner Medulla
Upper level 3809 13.5 3.5 4.2 1670 1520 16.8 28.7 2600
(52) (52) (52) (51) (29) (25) (26) (26) (23)
Lower level 397,0 13.0 33.9 50 47.6 74.8
(25, 29) (116) (115) (25) (26) (26)
ATL
Inner Medulla
Upper level 12.5 7.0 25.5 117 28.5 18.5 87.6 196 24.3 22.8 79.6 184
(56) (51) (47) (47) (47) (56)
Lower level 3 171 238
(25) (26) (26)

References are in parentheses below their values.

Units: Pf (water permeability, μm s−1 ); urea, Na, Cl permeability (10−5 cm s−1 ).
Pf values for chinchilla DTL from the outer medulla-inner stripe, inner medulla upper, and inner medulla lower are comparable to values reported
for chinchilla types 2, 3, and novel papillary DTL segments, respectively (27).
Modified from references 22 and 56.

The rodent renal papilla undergoes rhythmic contractions
that have been postulated to significantly impact the urine con-
centrating mechanism by influencing axial and transepithelial
fluid flows in nephrons, CDs, and vasa recta (141, 149). The
mechanical energy of these contractions might lower intersti-
tial pressure sufficiently so as to drive water efflux from the
DTL (and other water permeable structures), thereby increas-
ing the luminal osmolality. These contractions may involve
hyaluronan acting as a mechanico-osmotic transducer (73).
Fluid and solute transport in the ascending thin
limb of the loop of Henle
ATLs from all mammalian species investigated have repeat-
edly been shown to have little or no transepithelial osmotic
water permeability (25, 29, 47, 50, 102, 182).
Studies of Na and Cl permeabilities in rat and hamster
ATLupper using both isotopic and electrophysiological
methods showed that Cl permeability is nearly twice the
permeability of Na (47). The chinchilla ATL is likewise more
permeable to Cl than to Na (26). The paracellular pathway is
likely the dominant pathway for Na reabsorption in the ATL
(26, 79).
ClC-K1 is the primary pathway for transepithelial Cl flux
in the ATL. ClC-K1 protein expression in rat inner medulla
increases with water restriction (168, 174), consistent with
the role of ClC-K1 in antidiuresis. Reduction in extracellular
Ca2+ inhibits Cl transport with no effect on Na transport in
isolated, perfused tubules of hamster (78). Since Na trans-
port appears to continue unabated (78), it is plausible that a
second counteranion may exist, or alternatively, a Na/cation
exchanger may exist, so as to maintain electroneutrality of
transported solutes.
The chinchilla ATL has a very high transepithelial urea
permeability (26), substantially higher than urea permeabili-
ties reported for ATLs from hamster and rat (47) (Table 3). As
in the DTL, the transepithelial urea fluxes shown to occur in
chinchilla ATL are not inhibited by phloretin, and therefore,
are unlikely to be mediated by any of the known phloretin-
sensitive renal urea transporters (145). Also, as in the DTL,
two hypothetical pathways for these urea movements include
an unknown urea transporter and/or the paracellular pathway.
The high urea permeability of chinchilla ATL, combined with
its high NaCl permeability, would enable rapid osmotic equili-
bration of luminal loop of Henle fluid to occur as fluid ascends
toward the outer medulla. This rapid equilibration may serve
to retain solute in the inner medulla and sustain the medullary
axial solute gradient.
Water, Chloride, and Urea Transport
Proteins Expressed in the Thin Limb
of the Loop of Henle
Several key membrane proteins involved with transepithelial
fluid and solute flux in thin limbs of the loop of Henle
have been cloned and expressed in heterologous systems.
These studies have provided some important insights into
long standing, but poorly understood issues of thin-limb cell
and epithelial function related to the urine concentrating

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Structure and Function of the Thin Limbs of the Loop of Henle
mechanism. These proteins include the water channel AQP1
(138), the Cl channel ClC-K1 (168), and the urea transporter
UT-A2 (183).
Water channels
A specific pathway for transepithelial osmotic water flux in
DTLs was identified with the cloning of the first water chan-
nel, AQP1 (reviewed in reference 118). This constitutively
active water channel is weakly expressed along initial seg-
ments of the short-loop DTLs, immediately subsequent to the
proximal tubule (185), and is abundantly expressed in long-
loop DTLs (119, 185). AQP1 is expressed in both apical and
basolateral plasma membranes in cells of DTLs (119), con-
sistent with a role for AQP1 in the movement of water across
both surfaces of the cells.
Zhai and colleagues reported a complete absence of AQP1
along the entire length of the lower 90% of mouse short-loop
DTLs (185). Comparable patterns of AQP1 expression were
seen in rat and human kidneys, which exhibited an abrupt
transition from AQP1-positive proximal tubules to AQP1-
negative DTLs of short-loop nephrons (185) (see Section
“Fluid and solute transport in the descending thin limb of
the loop of Henle”).
The absolute abundance of AQP1 in individual mi-
crodissected renal tubule segments of Sprague-Dawley rats
was measured with a fluorescence-based enzyme-linked im-
munosorbant assay (ELISA) (105). This assay showed that
proximal tubule segments S1 and S2, and short-loop DTLs
(type 1 epithelium) express nearly equivalent amounts of
AQP1, relative to tubule length; proximal S3 segments ex-
press nearly 2-fold higher protein levels. In contrast, DTLs
with type 2 epithelium express almost 7-fold higher protein
levels and DTLs with type 3 epithelium express about 3.3-fold
higher levels than short-loop DTLs (105). As immunohisto-
chemistry showed no detectable AQP1 in the lower 90% of
the short DTL (185), the levels of AQP1 protein measured
with ELISA may represent protein which is expressed along
the initial 10% of this segment.
DTLs with type 2 epithelium express a high density of in-
tramembranous particles (IMPs) in the apical and basolateral
membranes of thin-limb segments from Wistar rat kidney (46)
and Sprague-Dawley rat kidney (152). Freeze-fracture elec-
tron microscopy studies of rat and mouse kidney have shown
that the IMPs in inner medullary long-loop DTLs are pre-
dominantly AQP1 water channels (23, 175). The density of
IMPs in DTLs extending through the outer medulla and into
the initial third of the inner medulla is quantitatively greater
than the density of IMPs in DTLs from the lower two-thirds
of the inner medulla (153). The density of IMPs in DTLs
from the lower two-thirds of inner medullary DTLs is quanti-
tatively similar to the density of inner medullary ATL IMPs.
These studies are consistent with a significant reduction in
abundance of water channels in the lower two-thirds of inner
medullary DTLs. In the ATL, IMPs likely reflect expression
of proteins unrelated to AQP1 (175).
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Comprehensive Physiology
Chloride channels
The rodent Cl channel ClC-K1 and the human orthologue,
ClC-Ka, are expressed in the ATL (66). In all thin limbs of the
Munich-Wistar rat inner medulla, expression of the chloride
channel ClC-K1 begins abruptly in the AQP1-negative portion
of the DTL about 170 μm on average above the hairpin bend,
and continues, apparently uniformly along the entire length of
the ATL within the inner medulla (125). Prebend expression
of ClC-K1 continues for about 170 μm on average in all
long loops of Henle, regardless of the depth at which the
bend is formed. ClC-K1 has been shown to be expressed in
both the apical and basolateral membranes (168) of the ATL
in the rat kidney, where it provides the dominant pathway
for transepithelial Cl flux associated with NaCl reabsorption.
A closely related protein, ClC-K2 (ClC-Kb in human), is
expressed in the thick ascending limb and distal segments
(66).
ClC-K1 (ClC-Ka) is expressed along with its accessory
protein barttin, the gene product of BSND (14), which is
a relatively small protein that forms heteromers with ClC-
K1 (ClC-Ka) in the ATL. Barttin is required for normal and
complete function of ClC-K1 (ClC-Ka) in heterologous ex-
pression systems, influencing surface expression as well as
permeation and gating of Cl channels (33, 151, 177). When
ClC-Ka and barttin are coexpressed in heterologous systems,
anion current is enhanced with increased extracellular Ca2+
and inhibited by low extracellular pH (33), effects that are
consistent with ClC-K1 function in heterologous expression
(169) and with transepithelial Cl transport in isolated perfused
rat ATL (77, 78). Excision of membrane patches from whole
cell heterologous expression systems leads to ClC-K/barttin
channel closure, suggesting that a cellular component, pos-
sibly an intact cytoskeleton, is required for normal barttin
gating (38).
Water deprivation increases inner medullary CLC-K1 and
barttin mRNA expression approximately 50% and increases
total kidney ClC-K1 and barttin protein expression in C57BL6
mice (11, 168, 174). Six-day treatment with furosemide de-
creases ClC-K1 and barttin mRNA expression approximately
50%, and decreases protein expression in the inner medulla
of the Sprague-Dawley rat. The effects on ClC-K1 and barttin
were limited to the ATLs (180). Wolf et al. have proposed that
interstitial tonicity may mediate transcriptional regulation of
ClC-K1 and barttin in the inner medulla (180). In humans, sin-
gle nucleotide polymorphisms of the renal ClC-Ka gene are
significantly associated with blood pressure change following
Na loading, supporting ClC-Ka as a susceptibility gene for
salt sensitivity (9). Thus, functional mutations within ClC-K1
(ClC-Ka) or the BSND gene may increase NaCl reabsorption,
leading to a form of volume-dependent and salt-sensitive hy-
pertension.
Although specific channel blockers for ClC-K channels
have not been fully characterized, ClC-Ka channels expressed
in Xenopus oocytes are inhibited by phenyl-benzofuran
carboxylic acid derivatives with an affinity less than
10 μmol/L (100).
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Comprehensive Physiology
Urea transporters
In the kidney, the urea transporter UT-A2 is expressed only in
the DTL. Expression of mRNA for UT-A2 was observed in
the lower portion of type 1 epithelia of the Sprague-Dawley
rat short-loop DTL and in the long-loop DTL (154). The abun-
dance of the UT-A2 transcript is increased in Sprague-Dawley
rats in response to water restriction (155), and is increased in
Brattleboro rats (a strain with impaired vasopressin synthesis
and release) by 1-deamino-8-d-arginine vasopressin (dDAVP)
infusion (139).
Strong immunohistochemical labeling for UT-A2 protein
was observed in the apical and basolateral plasma membranes,
as well as the cytoplasm of the lower portion of type 1 epithelia
of the rat short-loop DTL (68,101,120,176) and in the mouse
short-loop and long-loop DTL (37, 176). Weak labeling for
UT-A2 was also observed in the outer and inner medullary
segments of rat long-loop DTLs close to the outer medullary-
inner medullary boundary (68, 101).
In Brattleboro rats, UT-A2 antibody labels the lower
portion of descending limbs from short-loop nephrons and
this labeling is increased in response to treatment with the
vasopressin analog dDAVP (176). Increased labeling with
dDAVP administration is also seen in Brattleboro rat descend-
ing limbs of long-loop nephrons near the outer medullary-
inner medullary boundary (176). Tissue levels of total
UT-A2 protein, both in the outer medulla and inner medulla,
were increased with dDAVP infusion, as determined with im-
munoblotting. Further studies are needed to determine if the
increased levels of UT-A2 expression with dDAVP reflect an
increased length of expression along the entire tubule axis.
Water restriction for 3 days increased UT-A2 immunoreactiv-
ity intensities in the plasma membrane and cytoplasm of rat
loop of Henle types 1, 2, and 3 epithelia (101). UT-A2 ex-
pression decreased in hydrated rats given 3% sucrose in water
for 3 days (101) and in rats treated with furosemide for up to
7 days (98). These studies indicate that changes in hydration
status over a 3- to 7-day period can affect urea transporter pro-
tein expression without changing its subcellular distribution.
In mice fed a low-potassium diet for 2 weeks, UT-A2 expres-
sion in the DTL of the inner stripe and outer inner medulla is
increased (61).
Cytoskeletal Proteins and
Intercellular Junction Proteins in the
Thin Limb of the Loop of Henle
Freeze-fracture studies have shown that the tight junction
structure varies along the axis of the short and long DTLs and
ATLs (5, 57, 63, 81, 152, 153, 184), consistent with variable
paracellular solute permeability for ions and small, uncharged
solutes along the tubule axis. Paracellular pathways provide
transepithelial routes for substantial Na and urea fluxes in
DTLs and ATLs (26, 79, 80, 96, 103, 144); although, the phys-
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Structure and Function of the Thin Limbs of the Loop of Henle
iology of solute movement through paracellular pathways of
these segments is poorly understood.
Members of the claudin family of proteins are princi-
ple components of tight junction structure; thus, claudins are
viewed as critical in formation of tight junction strands that
are associated with charge- and size-selective paracellular so-
lute permeability. Accumulating evidence suggests that the
framework of strands is determined by the total amount of ex-
pressed claudins as well as by the ratios of different claudins
(40, 166). Therefore, expression of one or more claudins at
the tight junction can give rise to either a pore or a barrier
function and serve as a significant determinant of paracellular
solute flux (3, 172).
The many components that make up the tight junctional
complex can be considered as members of three groups; these
include: (1) the integral tight junction proteins that bridge
the apical intercellular space such as the claudins and oc-
cludins, (2) the tight junction plaque proteins that serve as
links between the integral tight junction proteins and the actin
cytoskeleton, and that are associated with cytosolic molecules
implicated in cell signaling, and (3) a miscellaneous group of
cytosolic and nuclear proteins, including regulatory proteins,
tumor suppressors, and transcriptional and posttranscriptional
factors (40, 150). In addition to the claudins and occludins
(discussed later), a number of these proteins are expressed
in thin limbs of the loop of Henle. These include the mul-
tifunctional junction protein Ksp-cadherin, which is present
along the basolateral membrane of the thin limb of the loop
of Henle (162), as well as fodrin, ankyrin 3, and E-cadherin,
all of which have been shown by immunohistochemistry to
be expressed at low, but detectable levels in the mouse DTL
and ATL of the loop of Henle, near the outer medullary-inner
medullary boundary (136).
Claudins
Claudin 2 expression, shown by immunolabeling, occurs in
some but not all AQP1-positive DTLs of the mouse (69).
Claudin 7 and 8 exhibit little or no expression in the outer
medullary DTL, but are expressed in the inner medullary
AQP1-positive DTLs at the tight junction and basolateral
membranes, respectively (40, 99).
Claudin 3 and 4 are expressed in the mouse ATL (69), and
claudin 16 and 19 are strongly expressed in the mouse ATL,
but only in a minority of cells (4).
In the mouse inner medulla, claudin 10 is weakly ex-
pressed in the DTL and strongly expressed in the ATL (173).
Nearly all the claudins expressed in thin limbs have been
shown to colocalize with the tight junction protein ZO1.
Gap junctions
Connexin 30.3 is present in mouse and rabbit ATL, where it
is expressed in a punctuate pattern typical of gap junctions
(mouse) or in a continuous pattern in the apical membrane
(rabbit) of AQP1-negative thin limbs (44).
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Structure and Function of the Thin Limbs of the Loop of Henle
ZO1 and occludin
ZO1 and occludin are present in DTLs and ATLs (4,41,69,86).
ZO1 expression in mouse resembles the complex architecture
typically seen for junctional strands that are known to lie
along the highly interdigitated lateral membranes of the type
4 epithelium (4).
Regulation of the Thin Limb of the
Loop of Henle
Vasopressin
There is generally good agreement as to the axial hetero-
geneity of permeabilities for NaCl, urea, and water in CDs
as well as to permeability changes elicited by vasopressin in
the antidiuretic state (146, 147). Thus, vasopressin is a key
regulator of the urine concentrating mechanism (145). How-
ever, there are few reports of fluid and solute flux regulation
by vasopressin (or by any hormone) in thin limbs of the loop
of Henle. There is no evidence of regulated AQP1 expression
or water flux in DTLs, although AQP1 expression in IMCD3
cells can be induced by hypertonicity, a response augmented
by vasopressin (58, 171). The V2 receptor agonist dDAVP
upregulates UTA2 expression in DTLs of the inner stripe and
inner medulla; however, this apparently is an indirect effect
as no vasopressin receptors have been shown to be expressed
in DTLs (34, 139, 176). Increased expression of UTA2 poten-
tially leads to increased urea transport.
Vasopressin stimulates Cl transport in the isolated, per-
fused hamster ATL, by increasing passive Cl flux through
Cl channels (159), an effect favorable to urine concentration
following the passive model (76, 156). Studies have shown
that dehydration increases inner medullary ClC-K1 expres-
sion (168, 174) and that decreased inner medullary tonicity
with furosemide decreases ClC-K1 expression (180), further
suggesting that NaCl transport in the ATL may be a regulated
process.
Evidence for plasma membrane receptor-mediated signal
transduction pathways in ATLs of the rat inner medulla in-
cludes vasopressin-activated adenylate cyclase activity (114).
The calmodulin-sensitive adenylate cyclase type 3 (45) and
ryanodine receptor type 3 (24) are expressed in rat thin limbs
(DTL and/or ATL), as shown by immunohistochemistry, sug-
gesting possible roles for calcium signaling pathways.
Endothelin
As endothelin plays an essential role in control of blood pres-
sure and sodium homeostasis, there is some interest in un-
derstanding a potential role for this family of peptides in reg-
ulation of thin-limb function. Although functional roles for
endothelin in thin limbs have not been characterized, there is
evidence that the endothelin receptor does exist in thin limbs.
The endothelin ETA and ETB receptors were detected in the
rat long-loop DTL by RT-PCR, but were not detectable in
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Comprehensive Physiology
the short-loop DTL or the ATL (7). Exogenous endothelin
produced an increase in intracellular free [Ca2+ ] in DTLs of
several rat strains, and this effect was inhibited by an ETA
receptor antagonist, but not by an ETB antagonist (7). The
ETA -induced calcium signaling was impaired in two strains
of genetically hypertensive rats.
Other studies have shown ET-1 mRNA expression in rat
DTL (117), ET-1 protein expression in human DTL (122),
and ET-3 receptor mRNA expression in rat thin limbs (161).
In contrast, other studies have failed to detect ET-1 mRNA
(19) or protein (179) in rat DTLs, and have failed to detect
mRNA in human DTLs (140).
Guanylin and uroguanylin
Guanylin and uroguanylin are two peptides that affect water
and electrolyte transport in the intestine and kidney. Messen-
ger RNA for both peptides has been detected by RT-PCR in
microdissected rat thin limbs of the loop of Henle (20), sug-
gesting that intrarenally produced peptide may be involved
with regulation of renal tubule transport. The mRNA for as-
sociated signal transduction molecules guanylyl cyclase-C
(GC-C) receptor and the cyclic guanosine monophosphate
(cyclic GMP)-dependent protein kinase II were also detected
in microdissected thin limbs (20).
Renin angiotensin
In ACE.2 transgenic mice, animals that exhibit only 33% of
wild-type plasma angiotensin converting enzyme (ACE) and
no tissue ACE, inner medullary AQP1 and ClC-K1 protein
are reduced to 29% and 6%, respectively, of wild-type values
(70). The maximal urine concentrating capacity of ACE.2
mice is reduced to a level comparable to that of the CLC-K1-
null mouse. These results suggest a potential involvement of
tissue ACE in regulating ClC-K1 expression in ATLs.
Chronic treatment with the ACE-inhibitor ramipril and
the angiotensin AT1-receptor antagonist irbesartan has been
shown to induce expression of the bradykinin B1 receptor
mRNA in inner medullary thin limbs of normotensive mice.
The B1 receptor agonist des-Arg9 [BK] increased release of
prostaglandin-E2 (PGE2 ) from this segment in the treated ani-
mals, indicating the induction of B1 receptor protein had taken
place (106). Possibly, thin-limb B1 receptors contribute to the
cardioprotective effects of ACE inhibitors and angiotensin
receptor antagonists (165).
Nitric oxide
Nitric oxide synthase enzymatic activity and mRNA, deter-
mined by PCR, have been localized to microdissected rat
thin limbs, albeit at low levels relative to other medullary
structures (160, 181). Soluble guanylate cyclase mRNA is
expressed in inner medullary thin limbs (160). Evidence for
a soluble cyclic GMP signal transduction pathway in the thin
Volume 2, July 2012
Comprehensive Physiology
limbs supports the idea that nitric oxide could have paracrine
effects on tubule function.
Cell Homeostasis in the Thin Limb
of the Loop of Henle
Na-K-ATPase
Antibodies raised against either holoenzyme, α, or γ sub-
units of Na-K-ATPase detect little or no protein expres-
sion in cells of all segments of thin limbs and functional
studies have shown only very low Na-K-ATPase activity
(64,104,137,143,157,178). Where present, Na-K-ATPase im-
munoreactivity has been reported to be more abundant at baso-
lateral membranes compared to apical membranes (137,157).
Cells of thin limbs of the loop of Henle from the innermost
third of the inner medulla show stronger expression of γ sub-
unit and splice variant γa than in the first and second third
(137). In thin limbs of the loop of Henle, Na-K-ATPase may
be involved with intracellular Na+ , K+ , and volume mainte-
nance (104).
Intracellular pH regulation
Plasma of the rat papillary vasa recta is acidic (65), likely
reflecting the pH of interstitial fluid. Studies with isolated,
perfused and nonperfused thin limbs from the moderately an-
tidiuretic rat have shown that resting intracellular pH is acidic,
and is more acidic in DTL than in ATL, regardless of the pH
of the external bathing medium (30,126). The differences be-
tween DTLs and ATLs in resting intracellular pH likely reflect
an intrinsic difference between cell types. In vivo measure-
ments indicate that the pH of the blood in the ascending vasa
recta rises significantly (to ∼6.9) when rats are made diuretic
(39). If thin limbs maintain their intracellular pH despite a
change in the pH of the surrounding medium, as suggested by
in vitro studies, then regulatory mechanisms associated with
transepithelial flux of H+ and HCO3 in thin limbs of the loop
of Henle may be pertinent to countercurrent flows in the pap-
illary interstitium, and to the urine concentrating mechanism
(30). Interestingly, resting intracellular pH was reported as be-
ing slightly but significantly higher in both DTLs and ATLs
in high osmolality (670 mOsmol/kg H2 O) than in low osmo-
lality (290 mOsmol/kg H2 O) medium, but not when sucrose
replaced urea, suggesting that urea plays a role in determining
resting intracellular pH (126).
The vacuolar H+ -ATPase was shown by immunohisto-
chemistry to be expressed in the initial portion of the long-
loop DTL (type 2 epithelium) of rat kidney, immediately distal
to the S3 segment of the proximal tubule (17, 126). Vacuolar
H+ -ATPase protein was expressed in basolateral and apical
membranes. The gastric H+ -K+ -ATPase α-subunit, and the
Na/H+ exchangers NHE1 and NHE3 (the latter present at the
basolateral and apical membranes, respectively) have been
detected in the Munich-Wistar rat long-loop thin limbs of the
Volume 2, July 2012
Structure and Function of the Thin Limbs of the Loop of Henle
inner medulla (DTL and/or ATL) by immunocytochemical
localization (13, 126). The Cl/HCO3 anion exchanger, AE2,
is readily detected in basolateral membranes of the rat outer
medullary long-loop DTLs that lie outside the vascular bun-
dles; however, short-loop DTLs that lie within vascular bun-
dles show little or no AE2 staining (2). Carbonic anhydrase
isozyme C has been shown by immunohistochemistry to be
in the initial portion of the long DTL (type 2) of rat kidney,
immediately after the S3 segment of the proximal tubule (18).
Expression was most intense in the outer medulla, very weak
in the outer zone of the inner medulla, and rarely seen in the
inner zone of the inner medulla. No expression was seen in
the ATL.
Functional studies of intracellular pH regulation in per-
fused or nonperfused long-loop thin limbs have been con-
ducted in rat, hamster, and rabbit (30, 39, 85, 126, 158). In
DTLs and ATLs, the rate of recovery of intracellular pH fol-
lowing additional acidification with an NH4 Cl pulse was re-
duced by Na+ removal from the medium and by the addition
of 60 μmol/L HOE642 (an inhibitor of the Na+ /H+ exchanger,
NHE1), 55 μmol/L S1611 (inhibitor of Na+ /H+ exchanger,
NHE3), 1 μmol/L bafilomycin A1 (an inhibitor of vacuo-
lar H+ -ATPase), or 20 μmol/L Schering 28080 (an inhibitor
of H+ -K+ -ATPase) to the bathing medium (126). Resting
intracellular pH was also reduced by 60 μmol/L HOE642,
55 μmol/L S1611, and 20 μmol/L Schering 28080. Regula-
tion of intracellular pH in response to perturbations apparently
involves not only the expected basolateral Na+ /H+ exchanger,
NHE1, but also the luminal Na+ /H+ exchanger, NHE3, and
the primary active H+ transporters, vacuolar H+ -ATPase and
H+ -K+ -ATPase.
Organic osmolytes and osmoprotective proteins
Organic osmolytes play a role in balancing the intracellu-
lar osmolality of medullary tubular epithelial cells with the
extracellular environment, and in doing so can preserve cell
integrity in the face of high osmotic stress. Osmotically ac-
tive organic solutes that localize almost exclusively to the
inner medulla of rat and rabbit kidneys include large amounts
of glycerophosphorylcholine, betaine, sorbitol, and inositol
(6). A number of enzymes and membrane transporters are
involved with regulating the level of these organic solutes in
the cytoplasm of thin-limb cells. The sodium/myo-inositol
cotransporter mRNA is expressed in rat thin limbs, and ex-
pression is increased with dehydration (21). Aldose reductase
catalyzes the production of sorbitol from glucose. Aldose re-
ductase activity is found in the inner medullary DTLs and
ATLs, with progressively higher activities at deeper levels
(148). Activities of the deepest papillary segments were nor-
malized to cell volume, showing that equivalent amounts were
seen in DTLs, ATLs, and CDs in this region. Aldose reductase
activity from outer medullary short and long loop DTLs was
undetectable (148).
The transcriptional regulator TonEBP (tonicity-
responsive enhancer binding protein) is an essential regulator
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Structure and Function of the Thin Limbs of the Loop of Henle
of the urine concentrating mechanism (87), and plays a major
role in determining intracellular levels of organic osmolytes
by promoting transcription of membrane transporters and
rate-limiting biosynthetic enzymes. Outer medullary TonEBP
protein expression is highest in nuclei and cytoplasm of
DTLs, as shown by immunohistochemistry (21). All thin
limbs of the loops of Henle are labeled intensely for TonEBP
expression throughout the inner medulla (21). The relative
levels of TonEBP protein expression in inner medullary
thin-limb cells are low in nuclei and high in cytoplasm, in
water-loaded rats (21). This pattern is markedly reversed in
thin limbs of the dehydrated rat.
GLUT1 protein has been localized by immunohistochem-
istry to the basolateral membrane of cells in thin limbs of the
inner stripe of the outer medulla (163, 164). GLUT1 expres-
sion possibly correlates with the glycolytic activity of thin
limbs, as in other nephron segments (164,167). Its expression
suggests that GLUT1 may be involved with providing glucose
as a source of metabolic energy in thin limbs (164).
In vivo microperfusion and microinfusion studies with
Munich-Wistar rat inner medulla have suggested that acidic,
basic, and neutral amino acids are recycled between the vasa
recta and thin limbs (31, 32). In vitro studies with isolated
perfused tubules have shown that high unidirectional secretory
and reabsorptive amino acid fluxes in thin limbs are neither
inhibitable nor saturable (16, 133). These properties suggest
that transepithelial movement of amino acids in thin limbs of
the loop of Henle may occur via the paracellular pathway.
Messenger RNA for the heat-shock protein and molecular
chaperone αB-crystallin is abundantly expressed in the renal
medulla and the corresponding protein has been localized to
cells of DTLs and ATLs in the rat outer medulla and inner
medulla (55,112,125). The αB-crystallin protein is expressed
along the entire length of the rat inner medullary DTL and
ATL (125).
Transgenic Mouse Models of the Thin
Limb of the Loop of Henle
Genetic models have been influential in understanding physi-
ological processes associated with the thin limb of the loop of
Henle and medullary function (1, 23, 36, 103, 110, 170). Thin
limb of the loop of Henle models include mouse knockouts
of AQP1, ClCK-1, and UT-A2.
AQP1-knockout mouse
Transepithelial osmotic water permeability (Pf ) was measured
in isolated perfused segments of DTLs from wild-type, AQP1
heterozygous, and AQP1 null mice (23). Pf was significantly
higher in wild type (2600 μm/s) compared to heterozygous
(2100 μm/s) and null mice (310 μm/s). The identity of a path-
way for transepithelial water flux in the AQP1-null mouse
DTL is unknown; three possibilities include an unidenti-
2082
Comprehensive Physiology
fied AQP, nonzero paracellular water flux, and nonzero flux
through apical and basolateral cell-membrane lipids (23).
Urine osmolality in response to 36-h water deprivation in-
creased to values exceeding 3000 mOsm in wild-type and
heterozygous mice, but was unchanged in AQP1-null mice
(23). Freeze-fracture electron microscopy of kidney medulla
showed that intramembrane particle densities were high in
wild-type DTL (5880 μm−2 ), reduced slightly in heterozy-
gous mice (5780 μm−2 ), and reduced markedly in AQP1-null
mice (877 μm−2 ). This study demonstrates that AQP1 is the
principal water channel in the DTL and supports the view that
osmotic equilibration by water transport in the DTL plays a
key role in the renal countercurrent concentrating mechanism.
Comprehension of the functional role of AQP1 in the thin
limbs of the knockout mouse is complicated by the marked
increase in fluid delivery that arises from reduced water re-
absorption in the proximal tubule, which, in the wild-type,
expresses abundant AQP1.
ClC-K1 knockout mouse
Knockout of the ATL-specific protein ClC-K1 (59) leads to
lowered urine osmolality in the mouse (110). Intraperitoneal
injection of the vasopressin V2 agonist dDAVP into the ClC-
K1 null mouse produced a minimal increase in urine osmo-
lality, indicating nephrogenic diabetes insipidus. The ATL Cl
efflux coefficient was reduced from about 188 × 10−5 cm s−1
in the wild type to about 17 × 10−5 cm s−1 in the ClC-K1
null, whereas the Na efflux coefficient was unchanged; the
Cl:Na permeability ratio declined from about 4 to about 0.6
(103). Akizuki et al. demonstrated that the inner medullary
urea accumulation (and a concentrating effect) is impaired in
the ClC-K1 null mouse (1). This directly implies that inner
medullary NaCl and urea hypertonicity and the axial solute
gradient, essential to the concentrating mechanism, arise from
interacting transport systems, such as countercurrent systems.
UT-A2 knockout mouse
As noted previously, UT-A2 is expressed in segments of the
short- and long-loop DTLs that lie near the outer medullary-
inner medullary boundary. Deletion of the urea transporter
UT-A2 from medullary DTLs had no effect on urine output
for mice raised on a normal protein diet consisting of 20% pro-
tein, under normal or dehydrating conditions (170). Medullary
urea accumulation also was unimpaired in the UT-A2-null an-
imals. In contrast, UT-A2-null mice raised on a low-protein
diet, exhibited a modest, but significantly impaired urine con-
centrating ability and reduced medullary urea content under
dehydrating conditions. These studies indicate that DTL UT-
A2 expression plays a modest role in maintaining a high
concentration of urea in the inner medulla when urea supply
to the kidney is limited (170). The exact role of UT-A2 in thin
limbs needs further investigations.
Volume 2, July 2012
Comprehensive Physiology
Animal Models of Disease of the Thin
Limb of the Loop of Henle
Mice lacking ClC-K1 (the murine homologue of ClC-Ka) in
the ATL, manifest nephrogenic diabetes insipidus with de-
fective urine concentrating ability (110). A more severe renal
salt wasting syndrome (Bartter’s syndrome type 4) is caused
by mutations in an unrelated protein, barttin (14), which is
needed for correct plasma membrane expression of ClC-Ka
and ClC-Kb (33, 151, 177) (see previous text).
Conclusions
The morphological distinction of DTL segments from long-
loop nephrons into at least two functionally distinct types of
epithelia has been long recognized, and is increasingly sup-
ported by segment-specific protein expression patterns and
functional studies of fluid and solute transepithelial transport.
The complex individual and collective functional interactions,
notably, solute and water movements between thin limbs and
each of the other medullary structures, must be more clearly
defined to better understand compartmentation arising from
dynamic flow of fluid and solutes between these structures.
The constitutive, pipe-like functional concept of thin limbs
appears increasingly inappropriate with the growing number
of regulatory elements and associated target proteins that are
known to be expressed in thin limbs of the loops of Henle.
More complete understanding of how medullary compart-
mentation is regulated is essential for understanding the role
of the renal medulla in processes associated with fluid and
solute flow dynamics, urine concentration [a process consid-
ered to involve either some form of the passive mechanism
(76, 131, 156), or alternative mechanisms (35, 36)], and the
long-term control of arterial pressure (28, 123).
Acknowledgements
Research in the author’s laboratory is supported by the Na-
tional Institutes of Health: National Institute of Diabetes and
Digestive and Kidney Diseases, grant DK083338, and by the
National Science Foundation, grant IOS-0952885. This article
was based, in part, on a previous review by Brigitte Kaissling
and Wilhelm Kriz, which was published in the Handbook of
Physiology (63).
References
1. Akizuki N, Uchida S, Sasaki S, Marumo F. Impaired solute accumu-
lation in inner medulla of Clcnk1-/- mice kidney. Am J Physiol Renal
Physiol 280: F79-F87, 2001.
2. Alper SL, Stuart-Tilley AK, Biemesderfer D, Shmukler BE, Brown
D. Immunolocalization of AE2 anion exchanger in rat kidney. Am J
Physiol Renal Physiol 273: F601-F614, 1997.
3. Angelow S, Ahlstrom R, Yu ASL. Biology of claudins. Am J Physiol
Renal Physiol 295: F867-F876, 2008.
Volume 2, July 2012
Structure and Function of the Thin Limbs of the Loop of Henle
4. Angelow S, El-Husseini R, Kanzawa SA, Yu ASL. Renal localization
and function of the tight junction protein, claudin-19. Am J Physiol
5.
Renal Physiol 293: F166-F177, 2007.
Bachmann S, Kriz W. Histotopography and ultrastructure of the thin
limbs of the loop of Henle in the hamster. Cell Tissue Res 225: 111-127,
1982.
6. Bagnasco S, Balaban R, Fales HM, Yang Y-M, Burg M. Predominant
osmotically active organic solutes in rat and rabbit renal medullas. J
Biol Chem 261: 5872-5877, 1986.
7. Bailey MA, Haton C, Orea V, Sassard J, Bailly C, Unwin RJ, Imbert-
Teboul M. ETA receptor-mediated Ca2+ signaling in thin descending
limbs of Henle’s loop: Impairment in genetic hypertension. Kidney Int
63: 1276-1284, 2003.
8. Bankir L, De Rouffignac C. Urinary concentrating ability: Insights from
comparative anatomy. Am J Physiol 249: R643-R666, 1985.
9. Barlassina C, Dal Fiume C, Lanzani C, Manunta P, Guffanti G, Ruello
A, Bianchi G, Del Vecchio L, Macciardi F, Cusi D. Common genetic
variants and haplotypes in renal CLCNKA gene are associated to salt-
sensitive hypertension. Hum Mol Genet 16: 1630-1638, 2007.
10. Barrett JM, Kriz W, Kaissling B, De Rouffignac C. The ultrastructure
of the nephrons of the desert rodent (Psammonys obesus) kidney. II.
Thin limbs of Henle of long-looped nephrons. Am J Anat 151: 499-514,
1978.
11. Bergler T, Stoelcker B, Jeblick R, Reinhold SW, Wolf K, Riegger
GAJ, Kramer BK. High osmolality induces the kidney-specific chloride
channel CLC-K1 by a serum and glucocorticoid-inducible kinase 1
MAPK pathway. Kidney Int 74: 1170-1177, 2008.
12. Beuchat CA. Structure and concentrating ability of the mammalian
kidney: Correlations with habitat. Am J Physiol Regul Integr Comp
Physiol 271: R157-R179, 1996.
13. Biemesderfer D, Rutherford PA, Nagy T, Pizzonia JH, Abu-Alfa AK,
Aronson PS. Monoclonal antibodies for high-resolution localization of
NHE3 in adult and neonatal rat kidney. Am J Physiol Renal Physiol
273: F289-F299, 1997.
14. Birkenhager R, Otto E, Schurmann MJ, Vollmer M, Ruf EM, Maier-
Lutz I, Beekmann F, Fekete A, Omran H, Feldmann D, Milford DV,
Jeck N, Konrad M, Landau D, Knoers NVAM, Antignac C, Sudbrak R,
Kispert A, Hildebrandt F. Mutation of BSND causes Bartter syndrome
with sensorineural deafness and kidney failure. Nat Genet 29: 310-314,
2001.
15. Braun EJ, Dantzler WH. Vertebrate renal system. In: Handbook of Phys-
iology: Comparative Physiology, Vol. 1. 1997. American Physiological
Society; Oxford University Press, NY.
16. Brokl OH, Dantzler WH. Amino acid fluxes in rat thin limb segments
of Henle’s loop during in vitro microperfusion. Am J Physiol 277:
F204-F210, 1999.
17. Brown D, Hirsch S, Gluck S. Localization of a proton-pumping ATPase
in rat-kidney. J Clin Invest 82: 2114-2126, 1988.
18. Brown D, Kumpulainen T, Roth J, Orci L. Immunohistochemical lo-
calization of carbonic-anhydrase in postnatal and adult-rat kidney. Am
J Physiol 245: F110-F118, 1983.
19. Bruzzi I, Corna D, Zoja C, Orisio S, Schiffrin EL, Cavallotti D, Re-
muzzi G, Benigni A. Time course and localization of endothelin-1 gene
expression in a model of renal disease progression. Am J Pathol 151:
1241-1247, 1997.
20. Carrithers SL, Taylor B, Cai WY, Johnson BR, Ott CE, Greenberg RN,
Jackson BA. Guanylyl cyclase-C receptor mRNA distribution along the
rat nephron. Regul Pept 95: 65-74, 2000.
21. Cha JH, Woo SK, Han KH, Kim YH, Handler JS, Kim J, Kwon HM. Hy-
dration status affects nuclear distribution of transcription factor tonicity
responsive enhancer binding protein in rat kidney. J Am Soc Nephrol
12: 2221-2230, 2001.
22. Chou CL, Knepper MA, Layton HE. Urinary concentrating mechanism
- the role of the inner medulla. Semin Nephrol 13: 168-181, 1993.
23. Chou CL, Knepper MA, Van Hoek AN, Brown D, Ma T, Verkman AS.
Reduced water permeability and altered ultrastructure in thin descend-
ing limb of Henle in aquaporin-1 null mice. J Clin Invest 103: 491-496,
1999.
24. Chou CL, Yip KP, Michea L, Kador K, Ferraris JD, Wade JB, Knep-
per MA. Regulation of aquaporin-2 trafficking by vasopressin in the
renal collecting duct - roles of ryanodine-sensitive Ca2+ stores and
calmodulin. J Biol Chem 275: 36839-36846, 2000.
25. Chou C-L, Knepper MA. In vitro perfusion of chinchilla thin limb
segments: Segmentation and osmotic water permeability. Am J Physiol
263(3): F417-F426, 1992.
26. Chou C-L, Knepper MA. In vitro perfusion of chinchilla thin limb
segments: Urea and NaCl permeabilities. Am J Physiol 264: F337-
F343, 1993.
27. Chou C-L, Nielsen S, Knepper MA. Structural-functional correlation in
chinchilla long loop of Henle thin limbs: A novel papillary subsegment.
Am J Physiol Renal 265: F863-F874, 1993.
28. Cowley AW, Jr. Role of the renal medulla in volume and arterial pres-
sure regulation. Am J Physiol Regul Integr Comp Physiol 273: R1-R15,
1997.
2083
Structure and Function of the Thin Limbs of the Loop of Henle
29. Dantzler WH, Evans KK, Pannabecker TL. Osmotic water permeabil-
ities in specific segments of rat inner medullary thin limbs of Henle’s
loops. FASEB J 23: 970.3, 2009.
30. Dantzler WH, Kim YK, Abbott DE, Serrano OK, Brokl OH. Intracellu-
lar pH in isolated rat renal papillary thin limbs of Henle’s loop. Pflugers
Arch 440: 140-148, 2000.
31. Dantzler WH, Silbernagl S. Amino acid transport by juxtamedullary
nephrons: Distal reabsorption and recycling. Am J Physiol 255: F397-
F407, 1988.
32. Dantzler WH, Silbernagl S. Amino acid transport: Microinfusion and
micropuncture of Henle’s loops and vasa recta. Am J Physiol 258:
F504-F513, 1990.
33. Estevez R, Bottger T, Stein V, Birkenhager R, Otto E, Hildebrandt F,
Jentsch TJ. Barttin is a Cl- channel beta-subunit crucial for renal Cl-
reabsorption and inner ear K +secretion. Nature 414: 558-561, 2001.
34. Fenton RA, Brond L, Nielsen S, Praetorius J. Cellular and subcellular
distribution of the type-2 vasopressin receptor in the kidney. Am J
Physiol Renal Physiol 293: F748-F760, 2007.
35. Fenton RA, Chou C-L, Stewart GS, Smith CP, Knepper MA. Uri-
nary concentrating defect in mice with selective deletion of phloretin-
sensitive urea transporters in the renal collecting duct. Proc Natl Acad
Sci U S A 101: 7469-7474, 2004.
36. Fenton RA, Knepper MA. Mouse models and the urinary concentrating
mechanism in the new millennium. Physiol Rev 87: 1083-1112, 2007.
37. Fenton RA, Stewart GS, Carpenter B, Howorth A, Potter EA, Cooper
GJ, Smith CP. Characterization of mouse urea transporters UT-A1 and
UT-A2. Am J Physiol Renal Physiol 283: F817-F825, 2002.
38. Fischer M, Janssen AGH, Fahlke C. Barttin activates CIC-K channel
function by modulating gating. J Am Soc Nephrol 21: 1281-1289, 2010.
39. Fujiwara I, Kondo Y, Igarashi Y, Inoue CN, Takahashi N, Tada K, Abe
K. Amiloride-sensitive Na+ /H+ antiporter in basolateral membrane of
hamster ascending thin limb of Henle’s loop. Am J Physiol 268: F410-
F415, 1995.
40. Furuse M, Tsukita S. Claudins in occluding junctions of humans and
flies. Trends Cell Biol 16: 181-188, 2006.
41. Gonzalez-Mariscal L, Namorado MC, Martin D, Luna J, Alarcon L,
Islas S, Valencia L, Muriel P, Ponce L, Reyes JL. Tight junction proteins
ZO-1, ZO-2, and occludin along isolated renal tubules. Kidney Int 57:
2386-2402, 2000.
42. Gottschalk CW, Mylle M. Micropuncture study of the mammalian
urinary concentrating mechanism: Evidence for the countercurrent hy-
pothesis. Am J Physiol 196: 927-936, 1959.
43. Han JS, Thompson KA, Chou C-L, Knepper MA. Experimental tests
of three-dimensional model of urinary concentrating mechanism. J Am
Soc Nephrol 2(12): 1677-1688, 1992.
44. Hanner F, Schnichels M, Zheng-Fischhofer Q, Yang LE, Toma I, Wil-
lecke K, McDonough AA, Peti-Peterdi J. Connexin 30.3 is expressed
in the kidney but not regulated by dietary salt or high blood pressure.
Cell Commun Adhes 15: 219-230, 2008.
45. Hoffert JD, Chou CL, Fenton RA, Knepper MA. Calmodulin is required
for vasopressin-stimulated increase in cyclic AMP production in inner
medullary collecting duct. J Biol Chem 280: 13624-13630, 2005.
46. Humbert F, Pricam C, Perrelet A, Orci L. Freeze-fracture differences
between plasma-membranes of descending and ascending branches of
rat Henles thin loop. Lab Invest 33: 407-411, 1975.
47. Imai M. Function of the thin ascending limb of Henle of rats and
hamsters perfused in vitro. Am J Physiol 232: F201-F209, 1977.
48. Imai M. Functional heterogeneity of the descending limbs of Henle’s
loop. ii. Interspecies differences among rabbits, rats, and hamsters.
Pflugers Arch 402: 393-401, 1984.
49. Imai M, Hayashi M, Araki M. Functional heterogeneity of the descend-
ing limbs of Henle’s loop. I. Internephron heterogeneity in the hamster
kidney. Pflugers Arch 402: 385-392, 1984.
50. Imai M, Kokko JP. Sodium chloride, urea, and water transport in the thin
ascending limb of Henle. Generation of osmotic gradients by passive
diffusion of solutes. J Clin Invest 53: 393-402, 1974.
51. Imai M, Taniguchi J, Tabei K. Function of thin loops of Henle. Kidney
Int 31: 565-579, 1987.
52. Imai M, Taniguchi J, Yoshitomi K. Transition of permeability properties
along the descending limb of long-loop nephron. Am J Physiol 254:
F323-F328, 1988.
53. Imai M, Yasoshima K, Yoshitomi K. Mechanism of water transport
across the upper portion of the descending thin limb of long-looped
nephron of hamsters. Pflugers Arch 415: 630-637, 1990.
54. Imai M, Yoshitomi K. Heterogeneity of the descending thin limb of
Henle’s loop. Kidney Int 38: 687-694, 1990.
55. Iwaki T, Kume-Iwaki A, Goldman JE. Cellular distribution of αB-
crystallin in non-lenticular tissues. J Histochem Cytochem 38: 31-39,
1990.
56. Jamison RL. Short and long loop nephrons. Kidney Int 31: 597-605,
1987.
57. Jamison RL, Kriz W. Urinary Concentrating Mechanism. New York:
Oxford University Press, 1982.
2084
Comprehensive Physiology
58. Jenq W, Mathieson IM, Ihara W, Ramirez G. Aquaporin-1: An osmoin-
ducible water channel in cultured mIMCD-3 cells. Biochem Biophys
Res Comm 245: 804-809, 1998.
59. Jentsch TJ, Stein V, Weinreich F, Zdebik AA. Molecular structure and
physiological function of chloride channels. Physiol Rev 82: 503-568,
2002.
60. Johnston PA, Battilana CA, Lacy FB, Jamison RL. Evidence for a
concentration gradient favoring outward movement of sodium from the
thin loop of Henle. J Clin Invest 59: 234-240, 1977.
61. Jung JY, Madsen KM, Han KH, Yang CW, Knepper MA, Sands JM,
Kim J. Expression of urea transporters in potassium-depleted mouse
kidney. Am J Physiol Renal Physiol 285: F1210-F1224, 2003.
62. Kaissling B, Kriz W. Structural analysis of the rabbit kidney. Adv Anat
Embryol Cell Biol 56: 1-123, 1979.
63. Kaissling B, Kriz W. Morphology of the loop of Henle, distal tubule,
and collecting duct. In: Windhager EE, editor. Handbook of Physiology.
New York: Oxford University Press, 1992, Sec. 8, pp. 109-167.
64. Kashgarian M, Biemesderfer D, Caplan M, Forbush B III. Mono-
clonal antibody to Na,K-ATPase: Immunocytochemical localization
along nephron segments. Kidney Int 28: 899-913, 1985.
65. Kersting U, Dantzler WH, Oberleithner H, Silbernagl S. Evidence for an
acid pH in rat renal inner medulla: Paired measurements with liquid ion-
exchange microelectrodes on collecting ducts and vasa recta. Pflugers
Arch 426: 354-356, 1994.
66. Kieferle S, Fong PY, Bens M, Vandewalle A, Jentsch TJ. Two highly
homologous members of the ClC chloride channel family in both
rat and human kidney. Proc Natl Acad Sci U S A 91: 6943-6947,
1994.
67. Kim J, Pannabecker TL. Two-compartment model of inner medullary
vasculature supports dual modes of vasopressin-regulated inner
medullary blood flow. Am J Physiol Renal Physiol, 299: F273-F279,
2010.
68. Kim YH, Kim DU, Han KH, Jung JY, Sands JM, Knepper MA, Madsen
KM, Kim J. Expression of urea transporters in the developing rat kidney.
Am J Physiol Renal Physiol 282: F530-F540, 2002.
69. Kiuchi-Saishin Y, Gotoh S, Furuse M, Takasuga A, Tano Y, Tsukita S.
Differential expression patterns of claudins, tight junction membrane
proteins, in mouse nephron segments. J Am Soc Nephrol 13: 875-886,
2002.
70. Klein JD, Le Quach D, Cole JM, Disher K, Mongiu AK, Wang XD,
Bernstein KE, Sands JM. Impaired urine concentration and absence of
tissue ACE: Involvement of medullary transport proteins. Am J Physiol
Renal Physiol 283: F517-F524, 2002.
71. Knepper MA, Danielson RA, Saidel GM, Post RS. Quantitative analysis
of renal medullary anatomy in rats and rabbits. Kidney Int 12: 313-323,
1977.
72. Knepper MA, Roch-Ramel F. Pathways of urea transport in the mam-
malian kidney. Kidney Int 31: 629-633, 1987.
73. Knepper MA, Saidel GM, Hascall VC, Dwyer T. Concentration of
solutes in the renal inner medulla: Interstitial hyaluronan as a mechano-
osmotic transducer. Am J Physiol Renal Physiol 284: F433-F446, 2003.
74. Kokko JP. Sodium chloride and water transport in the descending limb
of Henle. J Clin Invest 49: 1838-1846, 1970.
75. Kokko JP. Urea transport in the proximal tubule and the descending
limb of Henle. J Clin Invest 51: 1999-2008, 1972.
76. Kokko JP, Rector FC. Countercurrent multiplication system without
active transport in inner medulla. Kidney Int 2: 214-223, 1972.
77. Kondo Y, Yoshitomi K, Imai M. Effect of pH on Cl- transport in TAL
of Henle’s loop. Am J Physiol 253: F1216-F1222, 1987.
78. Kondo Y, Yoshitomi K, Imai M. Effect of Ca2+ on Cl- transport in thin
ascending limb of Henle’s loop. Am J Physiol 254: F232-F239, 1988.
79. Koyama S, Yoshitomi K, Imai M. Effect of protamine on ion con-
ductance of ascending thin limb of Henle’s loop from hamsters. Am J
Physiol 261: F593-F599, 1991a.
80. Koyama S, Yoshitomi K, Imai M. Effect of protamine on ion conduc-
tance of upper portion of descending limb of long-looped nephron from
hamsters. Am J Physiol 260: F839-F847, 1991b.
81. Kriz W. Structural organization of the renal medulla: Comparative and
functional aspects. Am J Physiol 241: R3-R16, 1981.
82. Kriz W, Bankir L. A standard nomenclature for structures of the kidney.
Am J Physiol 254: F1-F8, 1988.
83. Kriz W, Kaissling B. Structural organization of the mammalian kid-
ney. In: Seldin DW, Giebisch G, editors. The Kidney: Physiology and
Pathophysiology. New York: Raven Press Ltd., 1992, pp. 707-777.
84. Kriz W, Schnermann J, Koepsell H. The position of short and long
loops of Henle in the rat kidney. Z Anat Entwickl-Gesch 138: 301-319,
1972.
85. Kurtz I. Apical and basolateral Na+ /H+ exchange in the rabbit outer
medullary thin descending limb of Henle: Role of intracellular pH
regulation. J Membr Biol 106: 253-260, 1988.
86. Kwon O, Myers BD, Sibley R, Dafoe D, Alfrey E, Nelson WJ. Distri-
bution of cell membrane-associated proteins along the human nephron.
J Histochem Cytochem 46: 1423-1434, 1998.
Volume 2, July 2012
Comprehensive Physiology
87. Lam AKM, Ko BCB, Tam S, Morris R, Yang JY, Chung SK, Chung
SSM. Osmotic response element-binding protein (OREBP) is an essen-
tial regulator of the urine concentrating mechanism. J Biol Chem 279:
48048-48054, 2004.
88. Lassiter WE, Gottschalk CW, Mylle M. Micropuncture study of net
transtubular movement of water and urea in nondiuretic mammalian
kidney. Am J Physiol 200: 1139-1147, 1961.
89. Layton AT, Layton HE. A region-based mathematical model of the
urine concentrating mechanism in the rat outer medulla. I. Formulation
and base-case results. Am J Physiol 289: F1346-F1366, 2005.
90. Layton AT, Layton HE, Dantzler WH, Pannabecker TL. The mam-
malian urine concentrating mechanism: Hypotheses and uncertainties.
Physiology 24: 250-256, 2009.
91. Layton AT, Pannabecker TL, Dantzler WH, Layton HE. Two modes for
concentrating urine in rat inner medulla. Am J Physiol Renal Physiol
287: F816-F839, 2004.
92. Layton AT, Pannabecker TL, Dantzler WH, Layton HE. Functional
implications of the three-dimensional architecture of the rat renal inner
medulla. Am J Physiol Renal Physiol 298: F973-F987, 2010.
93. Layton HE. Distribution of Henle’s loops may enhance urine concen-
trating capability. Biophys J 49: 1033-1040, 1986.
94. Layton HE. Concentrating urine in the inner medulla of the kidney.
Comments Theor Biol 1: 179-196, 1989.
95. Layton HE, Davies JM. Distributed solute and water reabsorption in
a central core model of the renal medulla. Math Biosci 116: 169-196,
1993.
96. Layton HE, Knepper MA, Chou CL. Permeability criteria for effective
function of passive countercurrent multiplier. Am J Physiol 270: F9-
F20, 1996.
97. Lemley KV, Kriz W. Cycles and separations: The histotopography of
the urinary concentrating process. Kidney Int 31: 538-548, 1987.
98. Leroy C, Basset G, Gruel G, Ripoche P, Trinh-Trang-Tan MM, Rous-
selet G. Hyperosmotic NaCl and urea synergistically regulate the ex-
pression of the UT-A2 urea transporter in vitro and in vivo. Biochem
Biophys Res Comm 271: 368-373, 2000.
99. Li WY, Huey CL, Yu AS. Expression of claudin-7 and -8 along the
mouse nephron. Am J Physiol Renal Physiol 286: F1063-F1071, 2004.
100. Liantonio A, Picollo A, Carbonara G, Fracchiolla G, Tortorella P,
Loiodice F, Laghezza A, Babini E, Zifarelli G, Pusch M, Camerino
DC. Molecular switch for CLC-K Cl- channel block/activation: Opti-
mal pharmacophoric requirements towards high-affinity ligands. Proc
Natl Acad Sci U S A 105: 1369-1373, 2008.
101. Lim S-W, Han K-H, Jung J-Y, Kim W-Y, Yang C-W, Sands JM, Knep-
per MA, Madsen KM, Kim J. Ultrastructural localization of UT-A,
UT-B in rat kidneys with different hydration status. Am J Physiol Regul
Integr Comp Physiol 290: R479-R492, 2006.
102. Liu W, Morimoto T, Kondo Y, Iinuma K, Uchida S, Imai M. “Avian-
type” renal medullary tubule organization causes immaturity of urine-
concentrating ability in neonates. Kidney Int 60: 680-693, 2001.
103. Liu W, Morimoto T, Kondo Y, Iinuma K, Uchida S, Sasaki S, Marumo
F, Imai M. Analysis of NaCl transport in thin ascending limb of Henle’s
loop in CLC-K1 null mice. Am J Physiol Renal Physiol 282: F451-
F457, 2002.
104. Lopes AG, Amzel LM, Markakis D, Guggino WB. Cell-volume regu-
lation by the thin descending-limb of Henles loop. Proc Natl Acad Sci
U S A 85: 2873-2877, 1988.
105. Maeda Y, Smith BL, Agre P, Knepper MA. Quantification of aquaporin-
CHIP water channel protein in microdissected renal tubules by
fluorescence-based ELISA. J Clin Invest 95: 422-428, 1995.
106. Marin-Castano ME, Schanstra JP, Neau E, Praddaude F, Pecher C,
Ader JL, Girolami JP, Bascands JL. Induction of functional bradykinin
B-1-receptors in normotensive rats and mice under chronic angiotensin-
converting enzyme inhibitor treatment. Circ 105: 627-632, 2002.
107. Marsh DJ. Solute and water flows in thin limbs of Henle’s loop in the
hamster kidney. Am J Physiol 218: 824-831, 1970.
108. Marsh DJ, Azen SP. Mechanism of NaCl reabsorption by hamster thin
ascending limbs of Henle’s loop. Am J Physiol 228: 71-79, 1975.
109. Marsh DJ, Solomon S. Analysis of electrolyte movement in thin Henle’s
loops of hamster papilla. Am J Physiol 208: 1119-1128, 1965.
110. Matsumura Y, Uchida S, Kondo Y, Miyazaki H, Ko SBH, Hayama A,
Morimoto T, Liu W, Arisawa M, Sasaki S, Marumo F. Overt nephro-
genic diabetes insipidus in mice lacking the CLC-K1 chloride channel.
Nat Genet 21: 95-98, 1999.
111. Mejia R, Wade JB. Immunomorphometric study of rat renal inner
medulla. Am J Physiol Renal Physiol 282: F553-F557, 2002.
112. Michl M, Ouyang N, Fraek ML, Beck FX, Neuhofer W. Expression
and regulation of alpha B-crystallin in the kidney in vivo and in vitro.
Pflugers Archiv 452: 387-395, 2006.
113. Moffat DB, Fourman J. The vascular pattern of the rat kidney. J Anat
97: 543-553, 1963.
114. Morel F, Imbert-Teboul M, Chabardes D. Receptors to vasopressin and
other hormones in the mammalian kidney. Kidney Int 31: 512-520,
1987.
Volume 2, July 2012
Structure and Function of the Thin Limbs of the Loop of Henle
115. Morgan T. A microperfusion study in the rat of the permeability of the
papillary segments of the nephron to Na24 . Clin Exp Pharmacol Physiol
1: 23-30, 1974.
116. Morgan T, Berliner RW. Permeability of the loop of Henle, vasa recta,
and collecting duct to water, urea, and sodium. Am J Physiol 215:
108-115, 1968.
117. Moridaira K, Nodera M, Sato G, Yanagisawa H. Detection of Prepro-
ET-1 mRNA in normal rat kidney by in situ RT-PCR. Nephron Exp
Nephrol 95: E55-E61, 2003.
118. Nielsen S, Frokiaer J, Marples D, Kwon TH, Agre P, Knepper MA.
Aquaporins in the kidney: From molecules to medicine. Physiol Rev
82: 205-244, 2002.
119. Nielsen S, Smith BL, Christensen EI, Knepper MA, Agre P. Chip28 wa-
ter channels are localized in constitutively water-permeable segments
of the nephron. J Cell Biol 120: 371-383, 1993.
120. Nielsen S, Terris J, Smith CP, Hediger MA, Ecelbarger CA, Knepper
MA. Cellular and subcellular localization of the vasopressin- regulated
urea transporter in rat kidney. Proc Natl Acad Sci U S A 93: 5495-5500,
1996.
121. Nishimura H. Urine concentration and avian aquaporin water channels.
Pflugers Archiv 456: 755-768, 2008.
122. Ong ACM, Jowett TP, Firth JD, Burton S, Karet FE, Fine LG. An
endothelin-1 mediated autocrine growth loop involved in human renal
tubular regeneration. Kidney Int 48: 390-401, 1995.
123. Pallone TL, Turner MR, Edwards A, Jamison RL. Countercurrent ex-
change in the renal medulla. Am J Physiol Regul Integr Comp Physiol
284: R1153-R1175, 2003.
124. Pannabecker TL. Loop of Henle interaction with interstitial nodal
spaces in the renal inner medulla. Am J Physiol Renal Physiol 295:
F1744-F1751, 2008.
125. Pannabecker TL, Abbott DE, Dantzler WH. Three-dimensional func-
tional reconstruction of inner medullary thin limbs of Henle’s loop. Am
J Physiol Renal Physiol 286: F38-F45, 2004.
126. Pannabecker TL, Brokl OH, Kim YK, Abbott DE, Dantzler WH. Reg-
ulation of intracellular pH in rat renal inner medullary thin limbs of
Henle’s loop. Pflugers Arch 443: 446-457, 2002.
127. Pannabecker TL, Dahlmann A, Brokl OH, Dantzler WH. Mixed
descending- and ascending-type thin limbs of Henle’s loop in mam-
malian renal inner medulla. Am J Physiol Renal Physiol 278: F202-
F208, 2000.
128. Pannabecker TL, Dantzler WH. Three-dimensional lateral and vertical
relationships of inner medullary loops of Henle and collecting ducts.
Am J Physiol Renal Physiol 287: F767-F774, 2004.
129. Pannabecker TL, Dantzler WH. Three-dimensional architecture of in-
ner medullary vasa recta. Am J Physiol Renal Physiol 290: F1355-
F1366, 2006.
130. Pannabecker TL, Dantzler WH. Three-dimensional architecture of col-
lecting ducts, loops of Henle, and blood vessels in the renal papilla. Am
J Physiol Renal Physiol 293: F696-F704, 2007.
131. Pannabecker TL, Dantzler WH, Layton HE, Layton AT. Role of three-
dimensional architecture in the urine concentrating mechanism of the
rat renal inner medulla. Am J Physiol Renal Physiol 295: F1271-F1285,
2008.
132. Pannabecker TL, Henderson C, Dantzler WH. Quantitative analysis of
functional reconstructions reveals lateral and axial zonation in the renal
inner medulla. Am J Physiol Renal Physiol 294: 1306-1314, 2008.
133. Pannabecker TL, Völker K, Silbernagl S, Dantzler WH. Cycloleucine
fluxes during rat vasa recta and loop microinfusions in vivo and loop
microperfusions in vitro. Pflugers Arch 439: 517-523, 2000.
134. Pennell JP, Lacy FB, Jamison RL. An in vivo study of the concentrating
process in the descending limb of Henle’s loop. Kidney Int 5: 337-347,
1974.
135. Pennell JP, Sanjana V, Frey NR, Jamison RL. The effect of urea infusion
on the urinary concentrating mechanism in protein-depleted rats. J Clin
Invest 55: 399-409, 1975.
136. Piepenhagen PA, Peters LL, Lux SE, Nelson WJ. Differential expres-
sion of Na+ -K+ -ATPase, ankyrin, fodrin, and E-cadherin along the
kidney nephron. Am J Physiol Cell Physiol 269: C1417-C1432, 1995.
137. Pihakaski-Maunsbach K, Vorum H, Honore B, Tokonabe S, Frokiaer J,
Garty H, Karlish SJD, Maunsbach AB. Locations, abundances, and pos-
sible functions of FXYD ion transport regulators in rat renal medulla.
Am J Physiol Renal Physiol 291: F1033-F1044, 2006.
138. Preston GM, Carroll TP, Guggino WB, Agre P. Appearance of water
channels in Xenopus oocytes expressing red-cell Chip28 protein. Sci
256: 385-387, 1992.
139. Promeneur D, Bankir L, Hu MC, Trinh-Trang-Tan M-M. Renal tubular
and vascular urea transporters: Influence of antidiuretic hormone on
messenger RNA expression in Brattleboro rats. J Am Soc Nephrol 9:
1359-1366, 1998.
140. Pupilli C., Brunori M, Misciglia N, Selli C, Ianni L, Yanagisawa M,
Mannelli M, Serio M. Presence and distribution of endothelin-1 gene
expression in human kidney. Am J Physiol Renal Physiol 267: F679-
F687, 1994.
2085
Structure and Function of the Thin Limbs of the Loop of Henle
141. Reinking LN, Schmidt-Nielsen B. Peristaltic flow of urine in the renal
papillary collecting ducts of hamsters. Kidney Int 20: 55-60, 1981.
142. Rollhauser H, Kriz W, Heinke W. Das gefass-system der rattenniere. Z
Zellforsch 64: 381-403, 1964.
143. Sabolic I, Herak-Kramberger CM, Breton S, Brown D. Na/K-ATPase
in intercalated cells along the rat nephron revealed by antigen retrieval.
J Am Soc Nephrol 10: 913-922, 1999.
144. Sands JM, Kokko JP, Jacobson HR. Intrarenal heterogeneity: Vascular
and tubular. In: Seldin DW, Giebisch G, editors. The Kidney: Physiol-
ogy and Pathophysiology. New York: Raven, 1992, pp. 1087-1155.
145. Sands JM, Layton HE. The urine concentrating mechanism and urea
transporters. In: Alpern RJ, Hebert SC, editors. The Kidney: Physiology
and Pathophysiology. Philadelphia: Elsevier, 2007, pp. 1143-1177.
146. Sands JM, Nonoguchi H, Knepper MA. Vasopressin effects on urea
and H2 O transport in inner medullary collecting duct subsegments. Am
J Physiol 253: F823-F832, 1987.
147. Sands JM, Nonoguchi H, Knepper MA. Hormone effects on NaCl per-
meability of rat inner medullary collecting duct. Am J Physiol 255(3):
F421-F428, 1988.
148. Sands JM, Terada Y, Bernard LM, Knepper MA. Aldose reductase
activities in microdissected rat renal tubule segments. Am J Physiol
256(4): F563-F569, 1989.
149. Schmidt-Nielsen B, Graves B. Changes in fluid compartments in ham-
ster renal papilla due to peristalsis in the pelvic wall. Kidney Int 22:
613-625, 1982.
150. Schneeberger EE, Lynch RD. The tight junction: A multifunctional
complex. Am J Physiol Cell Physiol 286: C1213-C1228, 2004.
151. Scholl U, Hebeisen S, Janssen AGH, Mueller-Newen G, Alekov A,
Fahlke C. Barttin modulates trafficking and function of ClC-K channels.
Proc Natl Acad Sci U S A 103: 11411-11416, 2006.
152. Schwartz MM, Karnovsky MJ, Venkatachalam MA. Regional mem-
brane specialization in the thin limbs of Henle loops as seen by freeze-
fracture electron-microscopy. Kidney Int 16: 577-589, 1979.
153. Schwartz MM, Venkatachalam MA. Structural differences in thin limbs
of Henle: Physiological implications. Kidney Int 6: 193-208, 1974.
154. Shayakul C, Knepper MA, Smith CP, DiGiovanni SR, Hediger MA.
Segmental localization of urea transporter mRNAs in rat kidney. Am J
Physiol Renal Physiol 272: F654-F660, 1997.
155. Smith CP, Lee WS, Martial S, Knepper MA, You GF, Sands JM,
Hediger MA. Cloning and regulation of expression of the rat-kidney
urea transporter (Rut2). J Clin Invest 96: 1556-1563, 1995.
156. Stephenson JL. Concentration of urine in a central core model of the
renal counterflow system. Kidney Int 2: 85-94, 1972.
157. Takada T, Yamamoto A, Omori K, Tashiro Y. Quantitative immunogold
localization of Na, K-ATPase along rat nephron. Histochem 98: 183-
197, 1992.
158. Takahashi N, Kondo Y, Fujiwara I, Ito O, Igarashi Y, Abe K. Charac-
terization of Na+ transport across the cell membranes of the ascending
thin limb of Henle’s loop. Kidney Int 47: 789-794, 1995.
159. Takahashi N, Kondo Y, Ito O, Igarashi Y, Omata K, Abe K. Vasopressin
stimulates Cl− transport in ascending thin limb of Henle’s loop in
hamster. J Clin Invest 95: 1623-1627, 1995.
160. Terada Y, Tomita K, Nonoguchi H, Marumo F. Polymerase chain re-
action localization of constitutive nitric oxide synthase and soluble
guanylate cyclase messenger RNAs in microdissected rat nephron seg-
ments. J Clin Invest 90: 659-665, 1992.
161. Terada Y, Tomita K, Nonoguchi H, Yang TX, Marumo F. Expression of
endothelin-3 messenger-RNA along rat nephron segments using poly-
merase chain-reaction. Kidney Int 44: 1273-1280, 1993.
162. Thomson RB, Igarashi P, Biemesderfer D, Kim R, Abualfa A,
Soleimani M, Aronson PS. Isolation and cDNA cloning of Ksp-
cadherin, a novel kidney-specific member of the cadherin multigene
family. J Biol Chem 270: 17594-17601, 1995.
163. Thorens B. Facilitated glucose transporters in epithelial-cells. Annu Rev
Physiol 55: 591-608, 1993.
164. Thorens B, Lodish HF, Brown D. Differential localization of two glu-
cose transporter isoforms in rat kidney. Am J Physiol Cell Physiol 259:
C286-C294, 1990.
2086
Comprehensive Physiology
165. Tom B, Dendorfer A, Danser AHJ. Bradykinin, angiotensin-(1-7), and
ACE inhibitors: how do they interact? Int J Biochem Cell Biol 35:
792-801, 2003.
166. Tsukita S, Furuse M, Itoh M. Multifunctional strands in tight junctions.
Nat Rev Mol Cell Biol 2: 285-293, 2001.
167. Uchida S, Endou H. Substrate specificity to maintain cellular ATP along
the mouse nephron. Am J Physiol 255: F977-F983, 1988.
168. Uchida S, Sasaki S, Furakawa T, Hiraoka M, Imai T, Hirata Y, Marumo
F. Molecular cloning of a chloride channel that is regulated by dehydra-
tion and expressed predominantly in the kidney medulla. J Biol Chem
268: 3821-3824, 1993.
169. Uchida S, Sasaki S, Nitta K, Uchida K, Horita S, Nihei H, Marumo
F. Localization and functional characterization of rat kidney-specific
chloride channel. J Clin Invest 95: 104-113, 1995.
170. Uchida S, Sohara E, Rai T, Ikawa M, Okabe M, Sasaki M. Impaired urea
accumulation in the inner medulla of mice lacking the urea transporter
UT-A2. Mol Cell Biol 25: 7357-7363, 2005.
171. Umenishi F, Schrier RW. Hypertonicity-induced aquaporin-1 (AQP1)
expression is mediated by the activation of MAPK pathways and
hypertonicity-responsive element in the AQP1 gene. J Biol Chem 278:
15765-15770, 2010.
172. Van Itallie CM, Fanning AS, Bridges A, Anderson JM. ZO-1 stabilizes
the tight junction solute barrier through coupling to the perijunctional
cytoskeleton. Mol Biol Cell 20: 3930-3940, 2009.
173. Van Itallie CM, Rogan S, Yu A, Vidal LS, Holmes J, Anderson JM.
Two splice variants of claudin-10 in the kidney create paracellular
pores with different ion selectivities. Am J Physiol Renal Physiol 291:
F1288-F1299, 2006.
174. Vandewalle A, Cluzeaud F, Bens M, Kieferle S, Steinmeyer K, Jentsch
TJ. Localization and induction by dehydration of ClC-K chloride chan-
nels in the rat kidney. Am J Physiol 272: F678-F688, 1997.
175. Verbavatz JM, Brown D, Sabolic I, Valenti G, Ausiello DA, Vanhoek
AN, Ma T, Verkman AS. Tetrameric assembly of Chip28 water channels
in liposomes and cell-membranes - a freeze-fracture study. J Cell Biol
123: 605-618, 1993.
176. Wade JB, Lee AJ, Liu C, Ecelbarger C, Mitchell C, Bradford AD, Terris
J, Kim G-H, Knepper MA. UT-A2: A 55-kDa urea transporter in thin
descending limb whose abundance is regulated by vasopressin. Am J
Physiol 278: F52-F62, 2000.
177. Waldegger S, Busch AE, Kern C, Capasso G, Murer H, Lang F. Func-
tion and dysfunction of renal transport molecules: Lessons from elec-
trophysiology. Renal Physiol Biochem 19: 155-159, 1996.
178. Wetzel RK, Sweadner KJ. Immunocytochemical localization of Na-K-
ATPase alpha- and gamma-subunits in rat kidney. Am J Physiol Renal
Physiol 281: F531-F545, 2001.
179. Wilkes BM, Susin M, Mento PF, Macica CM, Girardi EP, Boss E, Nord
EP. Localization of endothelin-like immunoreactivity in rat kidneys. Am
J Physiol 260: F913-F920, 1991.
180. Wolf K, Meier-Meitinger M, Bergler T, Castrop H, Vitzthum H, Rieg-
ger GAJ, Kurtz A, Kramer BK. Parallel down-regulation of chloride
channel ClC-K1 and barttin mRNA in the thin ascending limb of the
rat nephron by furosemide. Pflugers Arch 446: 665-671, 2003.
181. Wu F, Park F, Cowley AW, Mattson DL. Quantification of nitric oxide
synthase activity in microdissected segments of the rat kidney. Am J
Physiol Renal Physiol 276: F874-F881, 1999.
182. Yool AJ, Brokl OH, Pannabecker TL, Dantzler WH, Stamer WD.
Tetraethylammonium block of water flux in aquaporin-1 channels ex-
pressed in kidney thin limbs of Henle’s loop and a kidney-derived cell
line. BMC Physiol 2: 4, 2002.
183. You G, Smith CP, Kanai Y, Lee W-S, Stelzner M, Hediger MA. Cloning
and characterization of the vasopressin-regulated urea transporter. Na-
ture 365: 844-847, 1993.
184. Zhai X-Y, Thomsen JS, Birn H, Kristoffersen IB, Andreasen A, Chris-
tensen EI. Three-dimensional reconstruction of the mouse nephron. J
Am Soc Nephrol 17: 77-88, 2006.
185. Zhai XY, Fenton RA, Andreasen A, Thomsen JS, Christensen EI.
Aquaporin-1 is not expressed in descending thin limbs of short-loop
nephrons. J Am Soc Nephrol 18: 2937-2944, 2007.
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