Kidney International, Vol. 32 (1987), pp.

443—451

EDITORIAL REVIEW

Thyroid hormones and renal transport: Cellular

and biochemical aspects

The thyroid hormones tri-iodothyronine (T3) and thyroxine where, due to the close association between the glands, simul-
(T4) are found in all vertebrates, but it is only in the homeo- taneously parathyroidectomy and thyroidectomy have been
thermic animals that most of the effects are observed [I]. These performed surgically, one has to replace PTH or add CaCl2 to
include stimulation of 02 consumption, effects on growth and the drinking water in order to minimize changes in calcium and
maturation, regulations of sugar, and lipid metabolism. The phosphate metabolism. Also the induction of hypothyroidism
hormones furthermore influence mineral metabolism and the by using high doses (1 mCi) of 131j may damage the parathyroid
distribution of water and electrolytes between body fluid com- glands to a certain extent, as evidenced by the tendency of the
partments. This paper will review the action of thyroid hor- calcium blood levels to decline after 13hI treatment. It is
mones on the kidney with particular emphasis on the biochem- currently not possible to ignore the fact that after parathyroid-
ical and biophysical aspects of thyroid hormones action on ectomy the response of renal cells to thyroid hormones is
transport processes of well—defined tubular segments and cell modified. The same holds for the use of the antithyroid drugs,
membranes. Moreover, we will discuss the role of thyroid propyithiouracil or aminotriazole, which are known to exert
hormones in regulating renal cell growth and differentiation. non-specific side effects.
Finally we will outline a working hypothesis for the action of
The hypothyroid state of an animal can be documented by the
thyroid hormones on the kidney with specific indication of their
following criteria: failure to gain weight as rapidly as controls,
role in modulating trans-epithelial transport processes.
For an in depth analysis of the clinical aspects of the effect of decreased heart rate, decreased metabolic activity, low blood
thyroid hormones on kidney function and electrolyte metabo- levels of T3, T4, and high blood concentration of TSH. The
lism, the reader is referred to other reviews [2—4] which have integrity of parathyroid hormone system can be checked by the
been published recently. determination of the calcium and PTH concentration in the
plasma, although the latter is not easily performed in the rat.
General considerations With regard to the duration of experimental hypothyroidism
one has to take into account the turnover ofT3 in the animal. In
The understanding of the effect of thyroid hormones on renal rabbits and rats, two weeks are required to obtain T3 free
function is complicated by several factors. One is that primary
animals. The same time period suffices to reduce the T3 level
cellular events can be modified by systemic changes in hemo-
dynamic parameters that maintain excretory balance. The sec- significantly by antithyroid drugs. In the following, animals
ond point is the axial and longitudinal heterogeneity of the studied within this two week period will be referred to as
nephron which leads to intratubular compensation of hormonal short—term hypothyroid, while animals kept without T3 for a
alterations induced in specific tubular segments. Another prob- more extended time will be referred to as long—term hypothy-
lem in developing a unifying hypothesis for the action of thyroidroid. This distinction is important because, as will be shown
hormones on renal functions is that different investigators have below, the response of the kidney to thyroid hormones depends
used vastly different experimental conditions. In order to be on the duration of the hypothyroid state. In addition long—term
hypothyroidism is associated with increased levels of plasma
able to compare the findings present in the literature it is first
necessary to carefully define the state of the animals and the lipids, changes in hemodynamics and probably alterations in the
extent and mode of hormone application. This is attempted in plasma level of other hormones [5—6].
the next section. Hormone application should be aimed to obtain physiological
plasma levels of the hormone. In this regard the plasma level of
Experimental approaches T3 and TSH should be monitored. In the rat no more than 3 to
In preparing hypothyroid animals, ideally the thyroid glands 10/pg/kg body weight per day should be administered [71; in the
of the animal should be selectively removed. This is easily rabbit similar doses should be used. Most of the studies have
possible in the rabbit where parathyroid glands and thyroid been performed using a 50-fold higher dosage. The animals of
glands are anatomically separated; in the rat autotranspianta- the latter study should be regarded as undergoing transition
tion of the parathyroid glands into the neck muscles is possible from hypothyroidism to hyperthyroidism rather than from
which leaves the parathyroid system unaltered. If rats are used hypothyroidism to euthyroidism. Therefore, treatment with low
doses and with high doses will be distinguished. Since, as will
be shown below, T3 changes cellular functions in just 12 to 24
Received for publication May 15, 1986 hours after administration, the duration of hormone application
and in revised form November 24, 1986 and the time course of the response are also critical parameters
© 1987 by the International Society of Nephrology in comparing studies from various investigators.

443
444 Capasso et a!

Effects on whole kidney function Table 1. Effect of thyroid hormones on transport properties in
proximal tubules of longterm hypothyroid rats (TX)
Long—term hypothyroidism causes significant changes in Na transport Effect Refs.
renal hemodynamics and in the renal handling of salt and water.
No hormone substi- 28% decrease in sodium re- 9
In the hypothyroid experimental animal the changes in renal tution absorption (TF/P inulin)
function include: (a) increased urine flow rate and increased No hormone substi- 70% decrease in J (split 19
water intake [8], (b) reduced GFR with diminished effective renl tution droplet method)
plasma flow and increased filtration fraction [9, 10], (c) inability 50 pg/kg body wt T3
I day 83% increase 29
to produce maximally concentrated or diluted urine [11—13], (d)
2 days 86% compared to TX
reduced cortico-papillary tissue concentration gradients for 4 days 107% (split droplet method)
urea [14, 15], (e) impaired urinary acidification [16, 17], (1) 7 days 135%
aldosterone—independent natriuresis which is exaggerated by 7 days 65% restoration compared 19
water ingestion and mannitol or saline infusion [12, 18], and (g) to euthyroid
reduction in proximal tubule fluid and phosphate reabsorption 10 pg/kg body wt 13
1 day 37% increase 29
[9, 19, 20]. 2 days 43% compared to TX
Hypothyroid humans also exhibit numerous defects in renal 4 days 100% (split droplet method)
function that are similar to those found in experimental animals 4 days 60% restoration compared
[2]. In addition, humans develop myxedema which is accompa- to euthyroid
nied by a marked thickening of the basement membrane in the Histidine transport
No hormone substi- 20% decrease 19
glomeruli [21]. These changes contribute to the decrease in tution
renal plasma flow and glomerular filtration rate found in hypo- 50 zgIkg body wt T3 full restoration 19
thyroid patients [22, 23]. (7 days)
Although clinically a severe impairment of renal function in
humans is unusual, it should be stressed that most of the
described abnormalities improve after administration of thyroid
hormone. In uncomplicated thyrotoxicosis, on the other hand, Effects on development and growth
glomerular filtration rate, renal plasma flow, and tubular reab-
sorptive and secretory ability are usually increased [24, 25]. The Thyroxine has received wide attention as possible trigger of
clinically, most sigificant manifestation in the course of hyper- developmental changes in newborn rats because its circulating
thyroidism is hypercalcemia [261. It is usually moderate and concentration rises significantly during the second postnatal
produces little, if any, secondary renal pathology or dysfunc- week [30].
tion. Occasionally, however, serious renal damage occurs. This Thyroid deficiency is accompanied by a delay in kidney
damage is characterized by a substantial impairment of renal growth and in kidney maturation in young rats, and by a
concentrating ability and, occasionally, may result in decreased decrease in the kidney size in adult animals [31, 31. These
renal function, hypertension or renal tubular acidosis [27, 28]. effects are reversed by administration of physiological amounts
of T3 and T4. Overall renal growth is much more markedly
Effects on single nephrons affected than total body growth in young rats. On the other
hand, body growth is much more markedly accelerated by
At the present time only studies on rat proximal tubule are thyroid hormones in the hypothyroid than in euthyroid animal
available [9, 19, 29]. They have been performed using micro- [3]. In the long—term hypothyroid rat the decrease in kidney size
puncture techniques in a model of long—term hypothyroidism. and weight involves all nephron segments in the cortex and
The results are summarized in Table 1. Using TF/P inulin as a outer medulla and is especially pronounced in the medullary
marker of proximal tubule sodium reabsorption Michael et al [9] thick ascending limb. Morphometric analysis revealed a signif-
found a 28% decrase in JNa• These findings could not be icant decrease in the cross—sectional area of the medullary thick
explained by an adaptation phenomenon to a decrease in ascending limb (MAL) in both the inner and outer stripe of the
SNGFR, or by a decrease in the secretory rate of aldosterone outer medulla. No changes were observed in the surace density
secretion. Moreover, they were not related to chronic expan- of either the apical or basolateral plasma membrane. However,
sion of extracellular volume or to changes in peritubular Star- when calculated per millimeter tubular length there was a
ling forces. Therefore, the authors concluded that the decrease significant decrease in the surface area of both the apical and
in Na was directly related to the lack of thyroid hormones. This basolateral plasma membranes of MAL in long—term hypothy-
was confirmed by De Santo et al [19] who, by using the roid animals. Treatment with T4 prevented the reduction in
shrinking droplet method, could not only demonstrate the T3 tubule cross—sectional area and in the surface area of the plasma
dependence of the proximal fluid sodium reabsorption, but also membranes [32].
the presence of a defect in sodium coupled L-histidine transport Thyroid hormones have also been associated with impair-
in hypothyroid rats (Table 1). When time—course substitution ment of the renal response to unilateral nephrectomy. Renal
studies were performed, it was observed that the administration compensatory growth after unilateral nephrectomy does occur
of T3 induced a fast, dose and time—dependent increase in L. in hypothyroid rats, but to a lesser extent than in euthyroid
The increment was already detectable after 24 hours, but even animals [33]. In hypothyroid animals the compensatory re-
after six days of hormone substitution J, did not reach the value sponse involves an increase in cell size, since the DNA content
found in euthyroid animals [29]. of the kidney does not change but the protein/DNA ratio
 Thyroid hormones and the kidney 445

increases in the compensating kidney [34]. On the other hand hypothesis is hardly adequate to account for other thyroid
the administration of thyroxine enhances renal compensatory hormone effects such as the stimulation of growth, for example.
growth mainly through an increase in cell number, since the Thus the uncoupling theory has been generally discarded. Since
DNA content rises, but the protein/DNA ratio does not change under conditions of tight mitochondrial coupling between ATP
in the compensating kidney. It is not known, however, whether production and ATP utilization a sustained increase in respira-
the different parts of the nephron respond uniformly to the tion and metabolic rate can only be brought about by a
hormonal or compensatory stimulus. concomitant increase in ATP utilization, and since the active
Na transport uses a high proportion of the cellular energy
Although it is reasonable to suppose that kidney epithelial
cells have a requirement for thyroid hormone as a growth supply, it was proposed that a large part of the calorigenic effect
factor, it is also important to note that the long—term hypothy- of the thyroid hormones could be accounted for by a stimulation
roid state is associated with a reduction of glomerular filtration of the Na pump [47, 48]. The mechanism proposed envisioned
rate [9, 10, 35] and proximal tubular reabsorption of sodium [9, an increase in ATP hydrolysis linked to active extrusion of Na
19]. Since GFR seems to be the key determinant of the by the Na pump, which in turn would stimulate mitochondrial
compensatory response [36—38] it is also possible that the effect respiration. This hypothesis has been confirmed by data dem-
of thyroid hormone on kidney growth is secondary to the onstrating that acute administration of T3 to long—term hypo-
increased tubular load. However, in time course experiments it thyroid rats elicited similar time—dependent and dose—depend-
was found that in long—term hypothyroid rats after a single ent increases in total oxygen consumption, ouabain—dependent
injection of a large dose of T3, the increase in RNA/DNA and respiration, a-glycerophosphate dehydrogenase activity, and
protein/DNA ratios preceded the increase in PAH clearance Na,K-ATPase activity in liver and kidney [49]. The increase in
and renal sodium reabsorption, suggesting a direct effect of T3 Na,K-ATPase represented an increase in the number of Na,K-
on renal cortical growth [39]. ATPase units [50]. Also in rat skeletal muscle, small intestine,
The isolated microdissected rabbit tubules T3 also has been
salivary glands, and cardiac muscle, Na,K-ATPase activity
found to play an important role in the outgrowth, multiplication, increased significantly in the transition from the hypothyroid to
and differentiation of the tubular cells in primary culture. Thus,
the euthyroid and to the hyperthyroid state [5 1—54]. Under
in cortical collecting tubules, T3 at 10— M increases Na,
similar conditions the activities of succinate dehydrogenase and
K-ATPase activity and transepithelial voltage [40].
Another role of thyroid hormones during development is their cytochrome c increased in a variety of tissues including the
'permissive action" with regard to the effect of other hormones kidney [55].
on cellular functions. Thus T4 seems to play an important
permissive role for glucocorticoid hormones in controlling the Effects on plasma membrane composition
rate at which sucrase activity in the intestine rises to adult
levels [41]. T4 also plays a role in the ontogeny of serum
corticosteroid—binding globulin (CBG) and corticosterone. The Enzyme composition
postnatal increase in serum T4 concentration causes the devel-
opmental rise of both CBG and corticosterone in the rat [42]. As With respect to the activity of membrane enzymes and
a result of these interrelationships the ultimate control of many thyroid hormones great emphasis has been placed on alkaline
developmental events shown to be triggered by corticosterone phosphatase. In the kidney the enzyme is localized in the brush
in the rat might actually be related to the increasing concentra- border membrane of the proximal tubule. The administration of
tion of T4 that occurs during the early postnatal period. high doses of thyroid hormones to long—term hypothyroid rats
leads to a decrease in alkaline phosphatase activity in kidney
Effects on energy metabolism cortex homogenates [56], which was even more pronounced in
The activation of metabolism is the most conspicuous effect isolated brush border membranes [56, 57]. The physiological
of the thyroid hormone and is called its calorigenic effect. implication of this finding is unclear since the functional role of
Complete thyroidectomy leads to a decrease in basal metabolic the alkaline phosphatase has not yet been elucidated. The long
rate. All tissues except brain, testes and spleen appear to take maintained notion that the activity of this enzyme and the
part in this reduction of metabolism. The action of thyroid transport of inorganic phosphate across the brush border are
hormone on the kidney is of particular interest because, al-
correlated has been recently questioned by several investiga-
though the kidney contributes only approximately 0.5% of the
tors [58]. It remains to be established whether the observed
body weight in mammals, the process of reabsorption of sodium
decrease in enzyme activity is due to a reduction of the number
is energetically demanding, and as a result the kidney ordinarily
accounts for about 10% of whole body 02 consumption [43]. of enzyme molecules or a change in the turnover rate of the
Since the mitochondria have a crucial role in energy metabolism individual molecules. Changes in the latter can be induced by
they have been considered as possible subcellular locus of alterations in the lipid environment of the molecule. Since
thyroid hormone action. long—term thyroidectomy is associated with changes in the lipid
In earlier reports thyroid thermogenesis was attributed to metabolism of the body it is a tempting possibility that the
uncoupling of oxidative phosphorylation, which would result in fluidity of brush border membranes changes in parallel and
an elevated, mitochondrial oxygen consumption without a thereby alters the turnover rate of the alkaline phosphatase. A
corresponding increase in ATP synthesis [44—46]. This uncou- regulation by lipid fluidity of the activity of membrane—bound
pling effect might explain some toxic manifestations of exces- enzymes and membrane—bound transport systems has been
sive amounts of thyroid hormone; however, the uncoupling described in a variety of instances [59—6 11.
446 Capasso et a!

Table 2. Effect of thyroid hormones on Na,K-ATPase in microdissected tubular segments

1'CT PST MAL CCT MCT
Long—term Short—term Long—term Short—term Long—term Short—term Long—term Short—term Long—term Short—term
Treatment TX TX TX TX TX TX TX TX TX TX
No substitution
Rabbit 40% de- 40% de- decreased 70% de- decreased no change 65% de- 65% de- decreased 80% de-
creased creased activity creased activity creased creased activity creased
activity activity activity activity activity activity
Rat 60% de- 40% de- 60% de- no effect no effect ND.
creased creased creased
activity activity activity
High dosage
100-500 g/
kg/day
Rabbit complete complete no change complete complete
restora- restora- restora- restora-
tion after tion after tion after tion alter
48 hours 48 hours 48 hours 48 hours
Rat after 2
weeks
Low dosage
3—10 /.Lg/kg/
day
Rat 3 days no 3 days no
change change
Rat 7 days par- 7 days par-
tial res- tial res-
toration toration
(75% of (70% of
control) control)
Abbreviations are: PCT, proximal convoluted tubule; PST, proximal straight tubule; MAL, medullary thick ascending limb; CCT, cortical
collecting tubule; MCT, medullary collecting tubule; TX, hypothyroidism; Long—term TX, hypothyroidism lasting more than two weeks; Short—
term Tx, hypothyroidism lasting less than two weeks; N.D., not determined.
The data have been taken from references 64 to 66 and Note added in proof.

Effects on specific transport systems vented if the animals receive T3 during the period of
hypothyroidism. The time couse of restoration of enzyme
Na,K-ATPase capacity in rat proximal tubule has been studied in detail in
Most of the studies on the effect of thyroid hormones on parallel to the measurements of J,,, mentioned above [66]. At a
Na,K-ATPase have been performed on kidney homogenates or low dosage, which restores the physiological T3 level, Na,K-
membrane fractions prepared from them. Due to the heteroge- ATPase activity remained low for at last three days; after seven
neity of Na,K-ATPase distribution along thhe nephron [62—63], daily injections the enzyme activity reached about 75% of the
however, no definite conclusions on the response of the Na,K- control level. This time course differs markedly from the time
ATPase in single nephron segments could be drawn from these course of restoration of sodium transport in the same tubules.
experiments. The studies performed on the effect of thyroid
hormones on the Na,K-ATPase activity in microdissected Sodium—cotransport systems
tubular segments are summarized in Table 2. It is evident from In view of the well—established importance of sodium co-
this table that the response of the various tubular segments is transport in the function of the proximal tubule [67] the effect of
different and markedly affected by the duration of the hypothy- thyroid hormones on several sodium cotransport systems was
roid status of the animal. In short—term hypothyroid rabbits a investigated in isolated brush border membrane vesicles. The
decreased activity of Na,K-ATPase is found in the proximal results of these experiments are compiled in Table 3.
convoluted tubule, proximal straight tubule, cortical collecting Since it is known that excess of thyroid hormones is associ-
tubule, and medullary collecting tubule. In all other segments ated with hyperphosphatemia related to an increase in renal
no change in Na,K-ATPase activity was found [64]. Similarly, tubular phosphate reabsorption independent from PTH or cal-
in short—term hypothyroid rats the enzyme activity decreased citonin [20], several investigators focused on the Na phosphate
only in the proximal convoluted tubule [65]. In all these cotransport system [57, 68, 69]. It was found that brush border
segments the Na,K-ATPase activity could be restored within 48 membranes from hypothyroid rats had a lower capacity of Na
hours to normal values when T3 was administered to the gradient—dependent phosphate uptake than vesicles isolated
animals in high doses (100 to 500Ig/kg x day). In long—term from hyperthyroid animals [57]. Similarly, in short—term hypo-
hypothyroid animals all segments of the nephron exhibit re- thyroid rats an extremely high dosage of T3 (1 mg/kg body wt for
duced Na,K-ATPase activity in rabbit (Note added in proof) as 4 days) increased the Ymax of the sodium phosphate cotransport
do the rat kidney proximal convoluted tubule and proximal system by 25% and decreased the affinity of the transport
straight tubule [66]. The fall in enzyme activity can be pre- system by 15%. It is interesting to note that this change was
 Thyroid hormones and the kidney 447

Table 3. Effect of thyroid hormones on renal sodium—cotransport systems

Tubular segment
PCT PST
Treatment Long—term TX Short—term TX Long—term TX Short—term TX Refs.
Na/H exchanger
No hormone substitution decreaseda 68
high dosage (1 mg/kg 4 days 30% in- 69
body wt) crease
hyperthyroid no change Sabolic et al
hyperthyroid increased 68
low dosage (3—10 pg/kg 3 days no 71
body wt) change
Na/P1 cotransporter
high dosage (1 mg/kg no change 4 days 25% in- 57, 68
body wt) crease in
Vmax 15%
decrease in
P1 Vn,ax
hyperthyroid versus hy- increased 68
pothyroid
All the experiments were performed in rats. Abbreviations are: PCT, proximal convoluted tubule, mixture of S1 and S2 segments; PST, proximal
straight tubule, mixture of S2 and S3 segments.
a segment not separated, mainly PCT

only observed in brush border membranes isolated from prox- changer activity was determined in the presence of a proton
imal straight tubules [69]. The relevance of these data to the gradient (pH1 = 5.5, pH0 = 7.4). In the latter experiment [71] no
overall renal handling of phosphate was evident from the pH gradient was present. Moreover, when the Na/H ex-
decrease in phosphate clearance after hormone administration. change was measured in brush border membrane vesicles
In interpreting the above results one has to keep in mind that obtained from euthyroid or long—term hyperthyroid animals
the effects occurred after administration of high doses of under "short circuit" conditions, with high K concentrations
thyroid hormones. It remains to be determined whether similar at both sides of the vesicle membranes and in the presence of
responses can be evoked by physiological levels of thyroid valinomycin, no difference in electroneutral Na/H exchange
hormone or whether these effects are manifested only in long— was found (Sabolic et al, personal communication).
term hypothyroidism or hyperthyroidism. It should be mentioned that also the effect of thyroid hor-
Sodium/hydrogen ion exchange plays a major role in sodium mones on sodium—glucose, sodium—proline, and sodium—histi-
reabsorption as well as in proton secretion in the proximal dine cotransport has been investigated in isolated brush border
tubule [70]. In brush border membrane vesicles isolated from membrane vesicles. No change in transport activity was ob-
long—term hypothyroid rats the Na/H exchange activity, that is, served under any experimental condition [57, 68, 69, 71].
amiloride—sensitive Na and H fluxes, were decreased in hypo-
thyroid rats, while it increased in hyperthyroid rats relative to Conductive pathways in plasma membranes
euthyroid animals. A positive correlation between Na/H ex- About twenty years ago Adamson and Ingbar suggested that
change activity in isolated brush border vesicles and serum short—term treatment of cells or animals with low doses of T3
concentration of T4 and T3 was postulated, although even using can lead to modifications of membrane transport processes
a semilogarithmic plot the scatter of the data was quite high without involving de novo synthesis of membrane components
[68]. Yusufi et a! demonstrated that in short—term hypothyroid [72]. Further studies demonstrated that the effect was evident
rats a large dose of T4 given for four days increased Na/H within 20 minutes and was accompanied by an increase in
exchange in brush border membrane vesicles isolated from the intracellular calcium and an increase in intracellular cyclic AMP
outer cortical zone (proximal convoluted tubule) but not in levels [73—75]. More specifically, Haber and Loeb demon-
membrane vesicles prepared from the juxtamedullary cortex strated in rat diaphragm and in liver cells that single adminis-
[69] (Table 3). tration of a high dose of T3 to euthyroid rats increased the
In brush border membrane vesicles isolated from long—term potassium permeability of the cell membranes well before a rise
hypothyroid rats treated for three days with low doses of T3, in Na,K-ATPase activity was observed [76—77]. After adminis-
Capasso et al were unable to demonstrate any statistically tration of low doses ofT3 for two weeks to long—term hypothy-
significant changes in the amiloride—sensitive Na uptake via roid rats, rubidium efflux from liver cells also increased 55% in
the Na/H exchanger compared to untreated hypothyroid rats the absence of any change of the Na,K-ATPase activity [78].
[71]. Therefore, the effect of thyroid hormones on Na/H ex- With regard to the effect of thyroid hromones on ion perme-
change might be critically dependent on the hormone concen- abilities of renal cells, to date only indirect studies on rat
tration and the duration of administration. The different results proximal tubules are available. In micropuncture experiments
might also be due to differences in the experimental procedures. on long—term hypothyroid rats local application of the potas-
In the two former experimental setups [68, 69] Na/H ex- sium ionophore valinomycin to the luminal and contraluminal
448 Capasso et a!

surface enhanced isotonic fluid reabsorption from 1.81 X 10-2

ni/mm X sec to 2.23 as measured by the split droplet technique. Denovo
This stimulation is equivalent to about 40% of the effect exerted Modification synthesis General cell
of membrane :of specific growth
by T3 treatment on the tubule. In long—term hypothyroid proteins proteins
animals treated with low doses of T3, valinomycin had no effect Euthyroid
on isotonic fluid reabsorption [66. These data provide indirect
evidence that K permeability (probably in the contraluminal
membrane) is decreased after long—term hypothyroidism and
that it plays a critical role in the early effect of thyroid hormones V
on the proximal tubule. Hypothyroid
With regard to the sodium permeability, in brush border I I
1 2 3 4
vesicles isolated from long—term hypothyroid rats and long—
Duration of treatment with thyroid hormone, weeks
term hypothyroid rats treated with low doses ofT3 an identical
permeability for sodium was found independent of the accom-
Fig. 1. Postulated action of thyroid hormones on the kidney. Symbols
) sodium reabsorption; (- - -) ion permeability; (— • —)
panying anion. A minor role of changes in sodium permeability are: (
in stimulation of J by thyroid hormones was also suggested by Na,K-ATPase, Na cotransport systems; and (— 0 —) tubular mass.
the lack of amphotericin action on isotonic fluid reabsorption in
the proximal tubule of the same rats [711. In long—term hyper-
thyroid rats Sabolic et al observed an increase in sodium and course of hormone action and is most probably independent of
proton conductivity in isolated brush border vesicles. In these protein synthesis. We would like to propose as a possible
animals, however, already a large increase in Na,K-ATPase mechanism a modification of the properties of proteins already
had occurred (Sabolic, personal communication). inserted in the plasma membrane. The exact mechanism of this
modification is presently unknown; it is tempting to speculate,
Mechanism of action of thyroid hormones on renal function: a however, that the increase in intracelluar calcium found in
synopsis thymocytes [741 also occurs in proximal tubule cells and leads
to an activation of the Ca-dependent K channels found in a
General considerations
variety of epithelia.
In considering the above detailed observations one can try to In addition to the change in membrane conductivity a third
establish a working hypothesis for the action of thyroid hor- action of thyroid hormones can be noted. This action involves
mones on the kidney. In doing so three different actions can be the de novo synthesis of proteins, These proteins represent
postulated to occur, which differ in their underlying biochemi- enzymes of the oxidative metabolism (a-glycerophosphate de-
cal events, in their time course, and in their specficity of the hydrogenase, succinic dehydrogenase, cytochrome C) and tran-
target cell (Fig. 1). sport systems such as the Na,K-ATPase, and probably the
The first distinct action of thyroid hormones is their role in sodium—phosphate cotransport system and the Na/H ex-
supporting the growth and differentiation of renal cells in vitro changer. The stimulation of protein synthesis is strongly corre-
and in vivo during postnatal development, after partial nephrec- lated with the occupancy of the thyroid hormones receptors in
tomy and in adult rats. This effect includes hypertrophy and the cell nucleus; therefore, at high doses the increase in enzyme
hyperplasia of the cells and involves as one factor a permissive activity is high and occurs early. When a lower dose is used the
role of thyroid hormones for the action of growth factors and increase is smaller and becomes evident after a longer lag
other hormones such as glucocorticoids. This action is exerted phase.
on all renal cells and has the slowest time course of all actions,
both after removal of the hormone as well as after the repletion Action of thyroid hormones on the proximal tubule
of long—term hypothyroid animals with thyroid hormones. The above—mentioned events can at least qualitatively ex-
Therefore, short—term and long—term hypothyroid animals can plain the action of thyroid hormones on transport properties in
exhibit different functional defects because in the latter the the proximal tubule of long—term hypothyroid rats (Fig. 1). First
diameter of the tubules and the renal mass is already markedly let us consider the time course and extent of restoration of the
reduced. This reduction has to be taken into account when isotonic sodium reabsorption measured as L. There is an early
interpreting data obtained on whole kidneys. Thus, for example increase in transport which is not accompanied by an increase
the 25% decrease of TF/Pjnu:jn observed in micropuncture in the total Na,K-ATPase activity determined in vitro. The
studies in long—term hypothyroid rats [9] (Table I) can easily be increase in transport without an apparent change in the maxi-
explained by the decrease in tubular mass in the proximal mum Na,K-ATPase activity can be explained by the following
nephron. Recovery of cell mass and tubular diameter after assumptions. First, it has been shown that the Na,K-ATPase
long—term hypothyroidism seems, however, not to be a prereq- activity in the intact cell in vivo only approaches 30% of the
uisite for other actions of the thyroid hormones described activity found in vitro [79], that is the cell has reserve Na,K-
below. ATPase activity that under normal instances is not utilized [80].
After repletion of hypothyroid animals with thyroid hor- Second, the rate of Na,K-ATPase activity in vivo is not only
mones—independent of the length of the hypothyroid period— determined by the number of enzyme molecules but also by the
two other actions of the hormones can be distinguished. One is rate of potassium recycling across the contraluminal membrane
an increase in membrane permeability for potassium (and [81]. Increased sodium transport therefore can be brought about
perhaps Na and H). This effect appears very early in the time by an increase in potassium permeability of the contraluminal
 Thyroid hormones and the kidney 449

membrane, which in turn augments the activity of the Na,K- Acknowledgments

ATPase in the cell and transepithelial sodium transport. In a It is a pleasure to acknowledge the secretarial assistance of D.
second phase, the activity of Na,K-ATPase measured in vitro, Magdefessel and the artwork of F. Drager. The authors express their
and proportionally the activity in vivo, increases due de novo gratitude to all the members of the Max—Planck—Institut für System-
protein synthesis, and concomitantly sodium reabsorption reaches physiologie, Dortmund, in particular to Dr. Eva Kinne—Saifran, for
about 75% of the euthyroid level. The lack of complete resto- their enthusiastic and active support during the preparation of the
manuscript. We thank Drs. A. Doucet and I. Sabolic for personal
ration of Na,K-ATPase activity and J,., (both expressed per communications included in this manuscript.
millimeter tubular length) using low doses of T3 can be ex- This study was supported by N.I.H. Grant no. 27441 and by funds
plained by the decrease in tubular radius, that is, cell mass per from the Italian Research Council and Ministry of Education. Dr.
millimeter tubule, caused by the long duration of the hypothy- Capasso was the recipient of Fogarty International, NATO Senior, and
roidism. Complete restoration can be expected when in a third Fulibright Fellowships Awards.
phase, general cell growth achieves normalization of the mor- GIOVAMBATFISTA CApAsso, NATALE G. Dr SANTO,
phological appearance and structural organization of the tubule. and ROLF KINNE
In short—term hypothyroid animals where morphological Policlinico Nuovo, Naples, Italy, and Max—Planck--Insiitut für
changes are absent, complete restoration of the Na,K-ATPase Systemphysiologie
activity would be expected, as observed by Barlet et a! [64] and Dortmund 1, Federal Republic of Germany
Garg et al [65]. With regard to the reduced tubular histidine Reprint requests to Dr. Giovambattista Capasso, Chair Pediatric
transport a restoration of transport was found in a phase where Nephrology, 1st Faculty of Medicine, Padiglione 17, Policlinico Nuovo,
the Na,K-ATPase was already increased [19]. Since in isolated Via Pansini, 80100 Napoli, Italy,
brush border membranes no difference in histidine transport
was found, the change observed in the intact cells is most likely References
due to changes in driving forces for the histidine—sodium 1. HULBERT AJ: The thyroid hormones: A thesis concerning their
cotransport system caused by the altered Na,K-ATPase activ- action. I Theor Biol 73:81—100, 1978
ity. 2. KATZ Al, EMMANOUEL DS, LINDREIMER MD: Thyroid hormone
and the kidney. Nephron 15:223—249, 1975
3. BRADLEY SE, STEPHAN F, COELHO JF, REVILLE P: The thyroid
Conclusions and perspectives and the kidney. Kidney mt 6:346—365, 1974
4. VAAMONDE CA, MICHAEL UF: The kidney in thyroid dysfunction,
The studies reviewed above demonstrate that thyroid hor- in The Kidney in Systemic Diseases, edited by SUKI KN, EKNOYAN
mones exert a general action on the kidney and specific actions G, (2nd ed.) New York, John Wiley and Son, 1981, pp. 361—415
on defined tubular segments, on defined transport proteins and 5. DORY L, ROHEIM PS: Rat plasma lipoproteins and apoliproteins in
enzymes involved in oxidative metabolism and on perme- experimental hypothyroidism. J Lipid Res 22:287—296, 1981
6. ABRAMS JJ, GRUNDY SM: Cholesterol metabolism in hypothyroid-
abilities of defined membranes. It also has become clear that the ism and hyperthyroidism in man. J Lipid Res 22:323—338, 1981
action of thyroid hormone extends beyond its calorigenic action 7. DILLMANN WH, BEIUY S, ALEXANDER NH: A physiological dose
in being a modulator of transepithelial transport. To the known of triiodothyronine normalizes cardiac myosin adenosine triphos-
mechanism of action via de novo synthesis of proteins, post- phatase activity and changes myosin isoenzyme distribution in
semistarved rats. Endocrinology 112:2081—2087, 1983
translational modification of membrane proteins has to be 8. STEPHAN F, JAHN H, METZ B: Action de l'insuffisance thyroid-
added. This modification can account for the changes in mem- ienne sur l'élimination de l'eau, du sodium et du potassium chez le
brane permeability, the permissive action, and modulation of rat. C R Soc Biol 153:332—335, 1959
cell development and growth. 9. MICHAEL UF, BARENBERG RL, CHAVEZ R, VAAMONDE CA,
PAPPER S: Renal handling of sodium and water in the hypothyroid
From the review also some guidelines for future experiments rat. J Gun Invest 51:1405—1415, 1972
can be derived. The hormonal status of the animal has to be 10. Osoiuo JA, ZADUNAISKY JA: Clearance de inulina y driodrast y
clearly defined with regard to the duration of hypothyroidism peso renal et ratas tratadas con '31J y polvo de tiroides. Rev Soc
and with regard to the amount of hormone administered. In Arg Biol 32:195—203, 1956
addition, whenever possible, defined tubular segments should 11. EMMANOUEL DS, LINDHEIMAR MD, KATZ Al: Mechanism of
impaired water excretion in hypothyroid rat. J Gun Invest 54:
be studied physiologically and biochemically. Further investi- 926—934, 1974
gations are also needed to understand the mechanism of the 12. HOLMES EW, DI SCALA VA: Studies on the exaggerated natriuretic
rapid action of the hormones on cell membranes, and to define response to a saline infusion in the hypothyroid rat. J Clin Invest
the interplay of thyroid hormone and other hormones in regu- 49:1224—1236, 1970
13. MICHAEL UF, KELLEY J, ALPERT H, VAAMONDE CA: Role of
lating transport, development and growth. distal delivery of filtrate in impaired renal dilution on the hypothy-
With the availability of microdissected tubules, primary roid rat. Am J Physiol 230:699—705, 1976
cultures of renal cells, and established renal cell lines with 14. REVILLE P, URBAN M, STEPHAN F: Diminution du gradient intra-
retained differentiation, the tools are at hand to address these renal de concentration de l'uree par l'hypothyroidisme chez le rat.
questions. Thus, hopefully in the not too distant future it will be CR Soc Biol 159:2031—2033, 1965
15. STEPHAN F, JAHN H, REVILLE P, URBAN M: Action de l'insuf-
possible to describe in more detail the chain of events compris- fisance thyroidienne chronique sur le debit urinaire et Ic pouvoir de
ing the action of thyroid hormones on the kidney. concentration de rein chez le rat. Rev Franc Etudes Gun Biol
8:890—899, 1963
16. MICHAEL UF, CHAVEZ R, VAAMONDE CA: Impaired urinary
Note added in proof acidification in the hypothyroid rat. Pflugers Arch 361:215—220,
1976
Dr. Daucet has recently published his paper on Na,K- 17. MICHAEL UF, KELLEY J, VAAMONDE CA: Impaired renal bicar-
ATPase in Pflugers Arch 407:428—431, 1986. bonate reabsorption in the hypothyroid rat. Am I Physiol 236:
450 Capasso et
F536—F540, 1979
18, TAYLOR RE, FREGLY JM: Renal response to propyithiouracil—
treated rats to injected mineralcorticoids. Endocrinology 75:33—38,
1964
19. DE SANTO NG, CAPASSO G, CALELLA C, PADUANO C, GIORDANO
C: Tubular transport processes in proximal tubules of hypothyroid
rats. Micropuncture studies on isotonic fluid, aminoacid and buffer
reabsorption. PfliAgers Arch 384:117—122, 1980
20. BOMMER J, BONJOUR JP, RITZ E, FLEISCH H: Parathyroid—inde-
pendent change in renal handling of phosphate in hyperthyroid rats.
Kidney In! 15:325—334, 1979
21. SALOMON MI, Di SCALA V, GRISHAM E, BRENER J, CHURG J
Renal lesions in hypothyroidism: A study based on kidney biopsies.
Metabolism 16:846—852, 1967
22. GOLDBERG M, REIvIcH M: Studies on the mechanism of hypona-
tremia and impaired water excretion in myxedema. Ann In! Med
56:120—130, 1962
23. DE RIJBERTIS FR, MICHELIS MF, BLOOM ME, MINTZ DH, FIELD
JB, DAVIS BB: Impaired water excretion in myxedema. Am J Med
51:41—53, 1971
24. FORD RV, OWENS JC, CURD GW, MOYER JH, SPURR CL: Kidney
function in various thyroid states, J Clin Endocr Metab 21:548—553,
1961
25. HANDLEY CA: Effect of thyroxine and dinitrophenol on renal
functions. Proc Fed Am Soc Exp Biol 9:281—282, 1950
26. BAXTER JD, BONDY PK: Hypercalcemia of thyrotoxicosis. Ann Int
Med 65:429—442, 1966
27. EPSTEIN FH, FRIEDMAN LR, LEVITIN H: Hypercalcemia, nephro-
calcinosis and reversible renal insufficiency associated with hyper-
thyroidism. N Engl J Med 258:782—785, 1958
28. WIJDEVELD PGAB, JANSEN AP: Renal concentrating and water—
excreting capacity in hyperthyroidism. Clin Chim Acta 5:618—631,
1960
29. DE SANTO NG, CAPASSO G, KINNE R, MOEWES B, ANASTASIO P,
GIORDANO C: Tubular transport processes in proximal tubules of
hypothyroid rats. Lack of relationship between thyroidal depen-
dent rise of isotonic fluid reabsorption and Na-K-ATPase activity.
Pflugers Arch 394:294—301, 1982
30. HENNING SJ: Postnatal development: Coordination of feeding,
digestion and metabolism. Am J Physiol 24l:Gl99.-G214, 1981
31. KATZ Al, LINDHEIMER MD: Renal sodium and potassium activated
adenosine triphosphatase and sodium reabsorption in the hypothy-
roid rat. J Clin lnvest 52:796—804, 1973
32. BENTLEY AG, MADSEN KM, DAVIS RG, TISHER CC: Response of
the medullary thick ascending limb to hypothyroidism in the rat.
AmJPatho! 120:215—221, 1985
33. ZECHWER IT: Compensatory growth of the kidney after unilateral
nephrectomy in thyroidectomized rats. Am J Physiol 145:681—684,
1946
34. STEPHAN F, REVILLE P, DE LAHARPE F, KOELL—BAEK MM:
Impairment of renal compensatory hypertrophy by hypothyroid-
ism. Life Sd 30:623—631, 1982
35. BRADLEY SE, COELH0 JB, SEALEY JE, EDWARDS KDG, STEPHAN
F: Changes in glomerular ifitration rate and the renin—angiotensin
system in hypothyroid rat. Life Sd 30:633—639, 1982
36. LUBOWITZ H, PURKERSON ML, SUGITA M, BRICKER NS: GFR per
nephron and per kidney in the chronically diseased (pyelonephritic)
kidney of the rat. Am J Physiol 217:853—857, 1969
37. KNEPPER MA, BURG MB: Increased fluid reabsorption and cell
volume in isolated rabbit proximal straight tubules after in vivo
DOCA administration. Am J Physiol 241 :F502—F508, 1981
38. FINE L: The biology of renal hypertrophy. Kidney In! 29:619—634,
1986
39. Lo CS, GERENDASY D, Lo TN: Effect of triiodothyronine on renal
growth and renal sodium reabsorption in hypothyroid rats. Pflugers
Arch 390:186—190, 1981
40. HORSTER MF, SCHONOLKE M, STOP? M: Induction of differentiated
functions by hormones in cultured cortical collecting tubule epithe-
ha. (abstract) IX In!
Congress of Nephrology 4l8a, 1984
41. HENNING SJ: Permissive role of thyroxine in the ontogeny of
jejunal sucrose. Endocrinology 102:9—15, 1978
42. D'AGOSTINO J, HENNING S: Role of thyroxine in coordinate
control of corticosterone and CBC in postnatal development. Am J
a!
Physiol 242:E33—E39, 1982
43. EDELMAN IS: Thyroid thermogenesis. N Engi J Med 290:1303—
1308, 1974
44. MARTIUS C, HESS B: The mode of action of thyroxine. Arch
Biochem Biophys 33:486—487, 1951
45. HOCH FL, LIPMANN F: The uncoupling of respiration and phos-
phorylation by thyroid hormones. Proc Nat! Acad Sci USA 40:
909—921, 1954
46. TAPLEY DF, COOPER C, LEHNINGER AL: The action of thyroxine
on mitochondria and oxidative phosphorylation. Biochim Biophys
Acta 18:597—598, 1955
47. ISMAIL—BEIGI F, EDELMAN IS: The mechanism of thyroid calori-
genesis: Role of active sodium transport. Proc Nail Acad Sci USA
67: 1071—1078, 1970
48. IsMAIL—BEIGI F, EDELMAN IS: The Mechanism of the calorigenic
action of thyroid hormone: Stimulation of Na-K-activated ade-
nosinetriphosphatase activity. J Gen Physiol 57:710—722, 1971
49. SOMJEN D, ISMAIL-BEIGI F, EDELMAN IS: Nuclear binding of T3
and effects on Q02, Na-K-ATPase and -GPDH in liver and kidney.
Am J Physiol 240:E146—El54, 1981
50. LO CS, AUGUST TR, LIBERMAN UA, EDELMAN IS: Dependence of
renal Na-K-adenosine triphosphatase activity on thyroid status.
J Biol Chem 251:7826-7833, 1976
51. ASANO Y, LIBERMAN UA, EDELMAN IS: Thyroid thermogenesis:
Relationship between Nat-dependent respiration and Na-K-
adenosine triphosphatase activity in rat skeletal muscle. J Clin
Invest 57:368—379, 1976
52. LIBERMAN UA, ASANO Y, Lo CS, EDELMAN IS: Relationship
between Na-dependent respiration and Na1-K-adenosine tn-
phosphatase activity in the action of thyroid hormone or rat jejunal
mucosa. Biophys J 27:127—144, 1979
53. Lo CS, CHENG W, KLEIN LE: Effect of triiodothyronine on
(NatK) adenosine triphosphatase and (Na-Mg2)-dependent
phosphorylated intermediate in rat salivary glands. Pflugers Arch
392:134—138, 1981
54. PHILIPSON KD, EDELMAN IS: Thyroid hormone control of Nat-
K-ATPase and K-dependent phosphatase in rat heart. Am J
Physiol 232:C196—C201, 1977
55. KADENBACH B: Effect of thyroid hormones on mitochondnial
enzymes, in Regulation of Metabolic processes in Mitochondria,
edited by TAGER, PAPA, QUAGLIARIELLO, SLATER, Heidelburg,
Elsevier Publishing Company, 1966, pp 508—517
56. CAPASSO G, KINNE R, MOEWES B, ANASTASIO P, DE SANTO NG,
GIORDANO C: Thyroidal regulation of alkaline phosphatase activity
in kidney cortex brush border membranes. Renal Physiol 5:206—
208, 1982
57. ESPINOSA RE, KELLER MJ, YUSUFI ANK, DOUSA T: Effect of
thyroxine administration on phosphate transport across renal cor-
tical brush border membrane. Am J Physiol 246:F133—F139, 1984
58. DE SMEDT H, KINNE R: Temperature dependence of solute trans-
port and enzyme activities in hog renal brush border membrane
vesicles. Biochim Biophys Acta 648:247—253, 1981
59. WILEY JS, COOPER RA: Inhibition of cation cotransport by choles-
terol enrichment of human red cell membranes. Biochim Biophys
Acta 413:425—431, 1975
60. SINHA AK, SIIATTIL SJ, COLMAN RW: Cyclic AMP metabolism in
cholesterol—rich platelets. J Biol Chem 252:3310—3314, 1977
61. SOLOMONSON LP, LIEPKALNS VA, SPECTOR AA: Changes in
(Na-K)-ATPase activity of Ehrlich ascites tumor cells produced
by alteration of membrane fatty acid composition. Biochemistry
15:892—897, 1976
62. DOUCET A, KATZ Al, MOLEL F: Determination of Na-K-ATPase
activity in single segments of mammalian nephron. Am J Physiol
240:F105—F1l3, 1979
63. GARG LC, KNEPPER MA, BURG MB: Mineralcorticoid effects on
Na-K-ATPase in individual nephron segments. Am J Physiol 240:
F536—F544, 1981
64. BARLET C, ABDELKHALEK B, DOUCET A: Sites of thyroid hormone
action on Na-K-ATPase along the rabbit nephron. Pflugers Arch
405:52—57, 1985
65. GARG LC, TISHER CC: Effects of thyroid hormone on Na-K-
adenosine triphosphatase activity along the rat nephron. J Lab Clin
Med 106:568—572, 1985
 Thyroid hormones and the kidney
66. CAPASSO G, LIN J-T, DE SANTO NG, KINNE R: Short term effect of
low doses of tri-iodothyronine on proximal tubular membrane
Na-K-ATPase and potassium permeability in thyroidectomized
rats. Pflugers Arch 403:90—96, 1985
67. ULLRICI-I KJ: Sugar, amino—acid and Na cotransport in the
proximal tubule. Ann Rev Physiol 41:181—195, 1979
68. KINSELLA J, SACKTOR B: Thyroid hormones increase Na-H
exchange activity in renal brush border membranes. Proc Nat!
Acad Sci USA 82:3606—3610, 1985
69. YusuFI ANK, MURAYAMA N, KELLER MJ, DOUSA T: Modulatory
effect of thyroid hormones on uptake of phosphate and other
solutes across luminal brush border membrane of kidney cortex.
Endocrinology 116:2438—2449, 1985
70. KINSELLA JL, ARONSON PS: Properties of the Na-H exchanger
in renal microvillus membrane vesicles. Am J Physiol 238:F461—
F469, 1980
71. CAPASSO G, KINNE—SAFFRAN E, DE SANTO NO, KINNE R: Regu-
lation of volume reabsorption by thyroid hormones in the proximal
tubule of rat: Minor role of luminal sodium permeability. Pflugers
Arch 403:97—104, 1985
72. ADAMSON LF, INGBAR SH: Selective alteration by triiodothyronine
of aminoacid transport in embryonic bone. Endocrinology 81:1362—
1371, 1967
73. SEGAL J, INGBAR SH: Stimulation by triiodothyronine of the in
vitro uptake of sugars by rat thymocytes. J Clin Invest 63:507—515,
1979
451
74. SEGAL J, INGBAR SH: An immediate increase in calcium accumu-
lation by rat thymocytes induced by triiodothyronine: Its role in the
subsequent metabolic response. Endocrinology 115:160—166, 1985
75. SEGAL J, INGBAR SH: In vivo stimulation of sugar uptake in rat
thymocytes. An extranuclear action of 3,5,3'-triiodothyronine. J
Clin Invest 76:1575—1580, 1985
76. HABER RS, LOEB JN: Effect of 3,5,3'-triiodothyronine treatment on
potassium efflux from isolated rat diaphragm: Role of increased
permeability in the thermogenic response. Endocrinology 111:
1217—1223, 1982
77. HABER RS, LOEB JN: Early enhancement of passive potassium
efilux from rat liver by thyroid hormone: Relation to induction of
Na-K-ATPase. Endocrinology 115:291—297, 1984
78. HABER RS, LOEB JN: Stimulation of potassium efflux in rat liver by
a low dose of thyroid hormone: Evidence for enhanced cation
permeability in the absence of Na,K-ATPase induction. Endocri-
nology 118:207—211, 1986
79. HARRIS SL, BALABAN RS, BARRETT L, MANDEL L: Mitochondrial
respiratory capacity and Na-
and K-dependent adenosine tn-
phosphatase—mediated ion transport in the intact renal cell. J Biol
Chem 256:10319—10328, 1981
80. KATZ AL: Renal Na-K-ATPase: Its role in tubular sodium and
potassium transport. Am J Physiol 242:F207—F219, 1982
81. SCHULTZ SG: Homocellular regulatory mechanism in sodium—
transporting epithelia. Avoidance of extinction by "flush—through"
Am J Physiol 241:F579—F590, 1981