6

PHYSIOLOGICAL REVIEWS
Vol. 80, No. 3, July 2000

Printed in U.S.A.
Molecular Analysis of the Sodium/Iodide Symporter:

Impact on Thyroid and Extrathyroid Pathophysiology
ANTONIO DE LA VIEJA, ORSOLYA DOHAN, ORLIE LEVY, AND NANCY CARRASCO
Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York
I. Introduction 1084
II. Characterization of the Sodium/Iodide Symporter Protein 1086
A. Generation of anti-NIS antibodies 1086
B. N-linked glycosylation of NIS: implications for the NIS secondary structure model 1087
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C. Extracellular orientation of the NH2 terminus of NIS 1087
D. Several hydroxyl-containing amino acid residues in the transmembrane segment IX (Ser-353,
Thr-354, Ser-356, and Thr-357) are important for NIS function 1088
E. Electrophysiological analysis of NIS: mechanism, stoichiometry, and specificity 1088
F. The NIS inhibitor perchlorate is not translocated by NIS into the cell 1089
III. Regulation of Sodium/Iodide Symporter Protein Expression 1090
IV. Transcriptional Regulation of the Sodium/Iodide Symporter 1090
A. Thyroid-specific transcription factors 1090
B. Transcriptional regulation of Tg and TPO 1091
C. Transcriptional regulation of TSHr 1091
D. Analysis of the human NIS promoter 1092
E. Analysis of the rat NIS promoter 1092
V. Expression of Sodium/Iodide Symporter in Nonthyroid Tissues 1093
A. NIS in salivary glands, stomach, mammary gland, and other tissues 1093
B. Identification of mammary gland NIS and gastric NIS 1094
C. Regulation of mammary gland NIS 1094
VI. The Sodium/Iodide Symporter in Autoimmune Thyroid Disease 1094
VII. The Sodium/Iodide Symporter and Cancer 1096
VIII. Congenital Iodide Transport Defect Due to Sodium/Iodide Symporter Mutations 1097
IX. Concluding Remarks 1101
De La Vieja, Antonio, Orsolya Dohan, Orlie Levy, and Nancy Carrasco. Molecular Analysis of the Sodium/
Iodide Symporter: Impact on Thyroid and Extrathyroid Pathophysiology. Physiol Rev 80: 1083–1105, 2000.—The
Na⫹/I⫺ symporter (NIS) is an intrinsic membrane protein that mediates the active transport of iodide into the thyroid
and other tissues, such as salivary glands, gastric mucosa, and lactating mammary gland. NIS plays key roles in
thyroid pathophysiology as the route by which iodide reaches the gland for thyroid hormone biosynthesis and as a
means for diagnostic scintigraphic imaging and for radioiodide therapy in hyperthyroidism and thyroid cancer. The
molecular characterization of NIS started with the 1996 isolation of a cDNA encoding rat NIS and has since
continued at a rapid pace. Anti-NIS antibodies have been prepared and used to study NIS topology and its secondary
structure. The biogenesis and posttranslational modifications of NIS have been examined, a thorough electrophys-
iological analysis of NIS has been conducted, the cDNA encoding human NIS (hNIS) has been isolated, the genomic
organization of hNIS has been elucidated, the regulation of NIS by thyrotropin and I⫺ has been analyzed, the
regulation of NIS transcription has been studied, spontaneous NIS mutations have been identified as causes of
congenital iodide transport defect resulting in hypothyroidism, the roles of NIS in thyroid cancer and thyroid
autoimmune disease have been examined, and the expression and regulation of NIS in extrathyroidal tissues have
been investigated. In gene therapy experiments, the rat NIS gene has been transduced into various types of human
cells, which then exhibited active iodide transport and became susceptible to destruction with radioiodide. The
continued molecular analysis of NIS clearly holds the potential of an even greater impact on a wide spectrum of
fields, ranging from structure/function of transport proteins to the diagnosis and treatment of cancer, both in the
thyroid and beyond.
www.physrev.physiology.org 0031-9333/00 $15.00 Copyright © 2000 the American Physiological Society 1083
1084 DE LA VIEJA, DOHAN, LEVY, AND CARRASCO Volume 80
I. INTRODUCTION by the Na⫹-K⫹-ATPase. The ability of the thyroid to ac-

cumulate I⫺ via NIS has long provided the basis for diag-
The thyroid is a master endocrine gland that plays a nostic scintigraphic imaging of the thyroid with radio-
central role in the intermediary metabolism of virtually all iodide and has served as an effective means for
tissues and is of fundamental importance for the devel- pharmacological doses of radioiodide to target and de-
opment of the central nervous system in the fetus and the stroy hyperfunctioning thyroid tissue, such as in Graves’
newborn. The widespread effects of the thyroid result disease or I⫺-transporting thyroid cancer and its metas-
from the biosynthesis and secretion of two rather unique tases (23, 124). Therefore, the study of NIS is of great
hormones, triiodothyronine (T3) and thyroxine (or tet- relevance to thyroid pathophysiology. Nevertheless, no
raiodothyronine) (T4), the only iodine-containing hor- molecular information on NIS was available until 1996,
mones in vertebrates. Iodide (I⫺) is an essential constit- when after a decades-long search by numerous investiga-
uent of T3 and T4 so that both thyroid function as a whole tors, a cDNA encoding rat NIS was finally isolated by
and its systemic ramifications depend on an adequate expression cloning in Xenopus laevis oocytes (21). This
supply of I⫺ to the gland. A remarkably efficient and development, a major breakthrough in the study of I⫺
specialized system has evolved in the thyroid that ensures transport processes and thyroid physiology, marked the
that most of the ingested dietary I⫺ (the only source of I⫺) beginning of the molecular characterization of NIS.
is accumulated in the gland and thus made available for T3 On the basis of the cloned cDNA, rat NIS was deter-

and T4 biosynthesis. The significance of this becomes mined to be a protein of 618 amino acids (relative molec-
more apparent when one considers that I⫺ is scarce in the ular mass 65,196) (Fig. 2). The hydropathic profile and
environment. Endemic goiter and cretinism caused pri- initial secondary structure predictions (20, 21) of the
marily by insufficient dietary supply of I⫺ remain a major protein suggested an intrinsic membrane protein with 12
health problem in many parts of the world, affecting putative transmembrane segments. The NH2 terminus
millions of people (24). I⫺ deficiency still often leads to was originally placed on the cytoplasmic side, given the
various degrees of impaired brain development, mostly in absence of a signal sequence. The COOH terminus, which
populations of children living in poor regions (114). These was also predicted to be on the cytoplasmic side, was
public health problems could conceivably be solved rela- found to contain a large hydrophilic region of ⬃70 amino
tively easily by ensuring that all table salt consumed in the acids within which the only potential canonical cAMP-
affected areas is iodized, as has been done in many coun- dependent protein kinase A (PKA) phosphorylation se-
tries. However, the sociopolitical realities of the affected quence of the molecule was located (positions 549 –552).
regions have prevented solutions like this from being Three potential Asn-glycosylation sites were identified in
implemented, at great human cost. Still, this situation the deduced amino acid sequence at positions 225, 485,
dramatizes not only the health value of I⫺ as a nutrient and 497. The first was located in a predicted intracellular
and the consequences to society of its environmental hydrophilic loop, while the last two were located in the
scarcity, but also underscores how difficult it would be last hydrophilic loop, a segment predicted to be on the
for people to stay euthyroid and healthy in the absence of extracellular face of the membrane. The length of the 12
a specialized I⫺ transport mechanism in the thyroid. transmembrane segments in this original model ranged
The ability of thyroid follicular cells to concentrate from 20 to 28 amino acid residues, except transmembrane
I⫺ was first reported as early as 1915 (68). The thyroid segment V, which contained 18 residues. Only three
gland was found to be capable of concentrating I⫺ by a charged residues were predicted to lie within transmem-
factor of 20 – 40 with respect to its concentration in the brane segments, namely, Asp-16 in transmembrane seg-
plasma under physiological conditions. Hence, the exis- ment I, Glu-79 in transmembrane segment II, and Arg-208
tence of a thyroid I⫺ transporter was inferred, and some in transmembrane segment VI. Out of a total of eight Trp
of its properties were elucidated over the years (see Refs. residues found in the membrane, six were located near
16, 42, 43, 61, 63, 98, 107 and 125 for reviews). Briefly, I⫺ the extremes of transmembrane segments. Four Leu res-
accumulation in the thyroid has long been shown to be an idues (positions 199, 206, 213, and 220) appeared to com-
active transport process that occurs against the I⫺ elec- prise a putative leucine zipper motif in transmembrane
trochemical gradient, stimulated by thyrotropin (TSH), segment VI. This motif, which has been proposed to play
and blocked by the well-known “classic” competitive in- a role in the oligomerization of subunits in the membrane,
hibitors, the anions thiocyanate and perchlorate. Eventu- has been conserved in all cloned neurotransmitter trans-
ally, it was determined that the thyroid I⫺ transporter is a porters (20, 21). NIS is often regarded to be a member of
Na⫹/I⫺ symporter (NIS) (6, 9, 45, 79, 122), i.e., an intrinsic the Na⫹/glucose cotransporter (SGLT1) family of trans-
plasma membrane transport protein that couples the in- porters. For a detailed review of this family of transport-
ward “downhill” translocation of Na⫹ to the inward “up- ers, see Reference 113a.
hill” translocation of I⫺ (Fig. 1). The driving force for the The molecular characterization of NIS has pro-
process is the inwardly directed Na⫹ gradient generated ceeded at an astounding pace with a wide variety of
July 2000 MOLECULAR ANALYSIS OF THE SODIUM/IODIDE SYMPORTER 1085

FIG. 1. Schematic representation of the biosynthetic pathway of thyroid hormones triiodothyronine (T3) and
thyroxine (T4) in the thyroid follicular cell. Thyroid follicles are comprised of a layer of epithelial cells surrounding the
colloid. The basolateral surface of the cell is shown on the left, and the apical surface on the right. Circle, active
accumulation of I⫺, mediated by the Na⫹/I⫺symporter (NIS); triangle, Na⫹-K⫹-ATPase; square, thyrotropin (TSH)
receptor; diamond, adenylate cyclase; ellipse, G protein; cylinder, I⫺ efflux toward the colloid; TPO, thyroid peroxidase
(TPO)-catalyzed organification of I⫺; arrows, endocytosis of iodinated thyroglobulin (Tg), followed by phagolysosomal
hydrolysis of endocytosed iodinated Tg and secretion of both thyroid hormones.
approaches and techniques, leading to numerous re- transcription has been studied; spontaneous NIS muta-
ports in just the last 3 years (61, 63, 98, 107). This tions have been identified as causes of congenital hy-
review provides an overview of the most recent devel- pothyroidism; and the roles of NIS in thyroid cancer
opments on the molecular analysis of NIS, among and thyroid autoimmune disease have been examined.
which are the following: anti-NIS antibodies have been In addition, it has been shown that I⫺ transport in
prepared and used to study the topology of NIS and to at least some extrathyroidal tissues, such as breast and
experimentally test the proposed NIS secondary model gastric mucosa, is also mediated by NIS expressed in
so that some aspects of the above-described model these tissues, in which NIS is differently regulated and
have been experimentally confirmed and others re- subjected to distinct posttranslational modifications.
vised. The most recent revised secondary structure This notion effectively invalidates the previously held
model for NIS proposes 13 (Fig. 2B) rather than 12 view that NIS was a major thyroid-specific protein, like
transmembrane segments (Fig. 2A). The biogenesis and thyroglobulin (Tg) and thyroid peroxidase (TPO), pre-
posttranslational modifications of NIS have been exam- sumably not expressed in any other tissues. Gene ther-
ined; a thorough electrophysiological analysis of NIS apy experiments have recently been reported in which
has been conducted, in which NIS specificity and stoi- the rat NIS gene was transduced into human melanoma,
chiometry were studied and a mechanistic model was murine liver, murine colon, and human carcinoma cells,
proposed; the cDNA encoding human NIS (hNIS) has all of which then exhibited active I⫺ transport and
been isolated; the genomic organization of hNIS has became susceptible to destruction with radioiodide.
been elucidated (Fig. 7) and the hNIS gene has been Therefore, it is now clearer than ever before that the
mapped to chromosome 19p; the regulation of NIS by continued molecular analysis of NIS holds the potential
TSH and I⫺ has been analyzed; the regulation of NIS of an even greater impact on a wide spectrum of fields,
FIG. 2. Original and current NIS secondary struc-

ture models. Top: original NIS secondary structure
model with 12 putative transmembrane segments
(21). Both the NH2 and COOH termini are proposed
to face intracellularly. Two putative N-linked glyco-
sylation consensus sequences are indicated with

branched symbols at positions 485 and 497. The third
N-linked glycosylation sequence at position 225 is
located in the third predicted intracellular loop. Bot-
tom: current NIS secondary structure mode with 13
putative transmembrane segments. Both the hydro-
philic loop containing N225 and the NH2 terminus
face extracellularly (64). All three N-linked glycosyl-
ation consensus sequences are indicated at positions
225, 485, and 497. Amino acid residues 389 – 410 form
a new transmembrane segment (X) between seg-
ments 9 and 10 of the original model (see top panel).
ranging from structure/function of transport proteins to COOH terminus of the protein. The Ab immunoreacts
the diagnosis and treatment of cancer, both in the with a mature ⬃87-kDa polypeptide (i.e., NIS) and a par-
thyroid and beyond. tially glycosylated (⬃56 kDa) polypeptide in a line of
highly functional thyroid rat cells (FRTL-5 cells). Immu-
noreactivity is also observed in X. laevis oocytes and COS
II. CHARACTERIZATION OF THE
cells expressing NIS, and it is competitively blocked by
SODIUM/IODIDE SYMPORTER PROTEIN
the presence of excess synthetic peptide. This anti-COOH-
terminal NIS Ab was the first available tool to experimen-
A. Generation of Anti-NIS Antibodies tally probe the NIS secondary structure model, and it was
used to confirm the model-predicted cytosolic-side loca-
A major development in the molecular characterization of the COOH terminus by indirect immunofluores-
tion of NIS has been the generation of anti-NIS antibodies cence experiments in permeabilized FRTL-5 cells (62).
(Ab). Levy et al. (62) generated a high-affinity [dissocia- Subsequently and independently, another site-directed Ab
tion constant (Kd) ⬃100 pM] site-directed polyclonal anti- against the same COOH-terminal segment of NIS was
NIS Ab against the last 16 amino acid residues of the generated by Paire et al. (81), which similarly immunore-
acts with an ⬃80- to 90-kDa glycosylated NIS protein from loop containing N225 are predicted to be on the extracel-
FRTL-5 cells. lular side, and the COOH terminus facing the cytosol, as
confirmed previously (Fig. 2B).
B. N-Linked Glycosylation of NIS: Implications for

the NIS Secondary Structure Model C. Extracellular Orientation of the NH2 Terminus
of NIS
Paire et al. (81) used their anti-NIS COOH Ab to
explore the regulation of NIS by TSH in FRTL-5 cells. Levy et al. (64) have recently demonstrated unequiv-
They observed that tunicamycin, an inhibitor of the syn- ocally that the NH2 terminus faces the external milieu, as
thesis of N-linked oligosaccharides, prevented both the proposed in the current model. This conclusion was
synthesis of mature NIS and the TSH-dependent reinduc- reached using two independent experimental approaches.
tion of NIS activity in FRTL-5 cells. On this basis, they First, these authors introduced a FLAG (MDYKDDDDK)
suggested that N-linked glycosylation of NIS was essential epitope into the NH2 terminus. COS cells transfected with
for NIS biosynthesis, correct folding, and stability. How- FLAG-containing NIS displayed undistinguishable I⫺ up-
ever, because tunicamycin inhibits the synthesis of N- take accumulation from COS cells transfected with wild-
linked oligosaccharides and thus prevents N-linked glyco- type NIS. Immunofluorescence experiments demon-

sylation of all proteins in the cell, it is clear that the strated positive immunoreactivity with anti-Flag Ab in
observed effect of tunicamycin on NIS activity is not nonpermeabilized COS cells transfected with FLAG-con-
necessarily due specifically to the lack of N-linked glyco- taining NIS. Positive immunoreactivity in nonpermeabil-
sylation of NIS. Levy et al. (64) have obtained conclusive ized cells indicates that the NH2 terminus faces exter-
evidence showing that, contrary to the conclusion of nally. In contrast, immunoreactivity using anti-COOH Ab
Paire et al., neither partial nor total lack of N-linked requires permeabilization because the COOH terminus
glycosylation impairs activity, stability, or targeting of faces the cytosol. The second technique took advantage
NIS. Using site-directed mutagenesis, Levy et al. (64) sub-
of a previous observation that unglycosylated NIS is ac-
stituted both separately and simultaneously the Asn resi-
tive. The N-linked glycosylation amino acid sequence
dues (amino acids 225, 485, and 497, see Fig. 2) in all three
NNSS was introduced into the NH2 terminus of unglyco-
putative N-linked glycosylation consensus sequences of
sylated NIS (25). They observed glycosylation of NIS at
NIS with Gln and assessed the effects of the mutations on
the NH2 terminus upon transfection of NNSS-containing
function, targeting, and stability of NIS in COS cells. All
NIS into COS cells, thus proving that the NH2 terminus
mutants were active and displayed 50 –100% of wild-type
faces the lumen of the endoplasmic reticulum during
NIS activity, including the completely nonglycosylated
biosynthesis and therefore faces the external milieu upon
triple mutant, which migrated as a ⬃50-kDa NIS polypep-
reaching the plasma membrane (25).
tide. They observed that the half-life of nonglycosylated
NIS was similar to wild-type NIS and that the Michaelis In addition, utilizing the same strategy of N-linked
constant (Km) value for I⫺ (⬃30 ␮M) in nonglycosylated glycosylation scanning mutagenesis, De la Vieja et al. (25)
NIS was virtually identical to wild-type NIS. These find- have demonstrated that the hydrophilic loop between
ings demonstrate that, to a considerable extent, function, putative transmembrane segments VIII and IX faces the
targeting, and stability of NIS are present even in the total external milieu (Fig. 2B). In a complementary approach to
absence of N-linked glycosylation (64). Therefore, a bac- study the topology of NIS in the plasma membrane, a Cys
terial expression system, in which no N-linked glycosyla- residue was placed at position 160 (in the hydrophilic
tion occurs, may be used to overproduce NIS for struc- loop between putative transmembrane segments IV and
tural studies. V) in an outside Cys-less background, a mutant that re-
In their report of N-linked glycosylation of NIS, Levy tains total activity. NIS activity was modified by mem-
et al. (64) demonstrated that the putative N-linked glyco- brane-impermeable sulfhydryl reagents such as sodium
sylation site at N225, which had originally been predicted (2-sulfonatoethyl) methanethiosulfonate (MTSES) and
to face intracellularly (Fig. 2A), is indeed glycosylated. 2-(trimethylamonium)ethyl methanethiosulfonate (MT-
Therefore, it is now clear that the hydrophilic loop that SET), indicating the external localization of this residue.
contains this sequence faces the extracellular milieu In summary, 5 NIS loops (NH2 terminus, loops between
rather than the cytosol. They have proposed a 13-trans- transmembrane segments IV and V, VI and VII, VIII and IX,
membrane segment model to be the most likely secondary and XII and XIII) out of a total of 7 have experimentally
structure for NIS. In contrast to the original model, in been confirmed to have the external disposition predicted
which the NH2 terminus was predicted to face the cytosol in the current 13-transmembrane segment secondary
based on the lack of a signal sequence in NIS, in the structure model. Additional experiments are being carried
current model both the NH2 terminus and the hydrophilic out to complete the topological analysis of NIS.
D. Several Hydroxyl-Containing Amino Acid

Residues in the Transmembrane Segment IX
(Ser-353, Thr-354, Ser-356, and Thr-357) Are
Important for NIS Function
Levy et al. (65) have demonstrated that a hydroxyl

group at the ␤-carbon at position 354 (in the transmem-
brane segment IX) is essential for NIS function. Such a
hydroxyl group is present in Thr-354. This discovery fol-
lowed reports that a spontaneous mutation consisting of
the single amino acid substitution of Pro instead of Thr at
position 354 (T354P) is the cause of congenital lack of I⫺
transport in several patients (39, 40, 55, 70). Patients with
this condition do not accumulate I⫺ in their thyroids,
often resulting in severe hypothyroidism. Seeking to char-
acterize this transport defect at the molecular level, Levy
et al. (65) set out to determine whether T354P NIS is a

nonfunctional but stable polypeptide properly targeted to
the plasma membrane, or a fully or partially functional
protein that is retained in intracellular organelles as a
result of the mutation. For this purpose, Levy et al. (65)
generated T354P NIS by site-directed mutagenesis and
used their high-affinity anti-NIS Ab to monitor T354P ex-
pression in COS cells transfected with NIS cDNA. COS FIG. 3. Amino acid sequence and ␣-helical wheel projection of the
cells transfected with T354P NIS cDNA were assayed for transmembrane segment IX of NIS. A: amino acid sequence. Hydroxyl-
I⫺ uptake activity and found to display no I⫺ accumula- containing residues are shown in bold. B: ␣-helical wheel projection.
Hydroxyl-containing residues are circled. Important residues for NIS
tion. After demonstrating that T354P was properly tar- activity are shaded.
geted to the plasma membrane, these authors carried out
several additional amino acid substitutions at position 354
and determined that the lack of I⫺ transport activity is not E. Electrophysiological Analysis of NIS:
due to a structural change induced by proline, as had been Mechanism, Stoichiometry, and Specificity
previously proposed (39), but rather to the absence of a
hydroxyl group at the ␤-carbon at position 354. Eskandari et al. (34) have examined the mechanism,
Significantly, the transmembrane segment IX, in stoichiometry, and specificity of NIS by means of electro-
which Thr-354 is located, is where the highest incidence physiological, tracer uptake, and electron microscopic
of hydroxyl-containing amino acids is found in NIS (Fig. methods in X. laevis oocytes expressing NIS. These in-
2B). Hence, De La Vieja et al. (unpublished data) assessed vestigators obtained electrophysiological recordings us-
the role played by these other hydroxyl groups in NIS ing the two-microelectrode voltage-clamp technique.
function by replacing the corresponding amino acid resi- They showed that an inward steady-state current (i.e., a
dues with Ala and Pro. Substituting Ser-349, Thr-351, Ser- net influx of positive charge) is generated in NIS-express-
358, and Thr-366 (Fig. 3) for similar amino acid residues ing oocytes upon addition of I⫺ to the bathing medium,
devoid of hydroxyl groups did not affect either NIS ex- leading to depolarization of the membrane. Because the
pression or activity. In contrast, the hydroxyl groups of recorded current is attributable to NIS activity, this ob-
Ser-353, Ser-356, and Thr-357 seem to be essential for NIS servation confirms that NIS activity is electrogenic. Simul-
activity, given that NIS functioned to a significant extent taneous measurements of tracer fluxes and currents re-
only when Ser or Thr was present at these positions. The vealed that two Na⫹ are transported with one anion,
lack of function when Ala or Pro was present instead at demonstrating unequivocally a 2:1 Na⫹/I⫺ stoichiometry.
these positions was not due to an effect on expression or Therefore, the observed inward steady-state current is
trafficking, given that the mutant NIS proteins were due to a net influx of Na⫹ (Fig. 4).
shown by confocal immunofluorescence to reach the Eskandari et al. (34) observed also that in response
plasma membrane properly. Additional experiments are to step voltage changes, NIS exhibited current transients
being carried out to determine the mechanistic role of that relaxed with a time constant of 8 –14 ms and that
these hydroxyl groups in NIS activity. Kinetic analyses are pre-steady-state charge movements (integral of the cur-
also being performed to differentiate between kinetically rent transients) versus voltage relations obeyed a Boltz-
impaired and uncoupled NIS molecules. mann distribution. These charge movements are attrib-
gesting that it is not transported (34). Yoshida et al. (130)

have reported, similarly, that perchlorate did not induce
an inward current in Chinese hamster ovary (CHO) cells
stably expressing NIS, as measured using the whole cell
patch-clamp technique. The most likely interpretation of
these observations is that perchlorate is not transported
by NIS, although the unlikely possibility that perchlorate
is translocated by NIS on a 1:1 Na⫹/ClO⫺ 4 stoichiometry
cannot be ruled out. Therefore, perchlorate is a potent
inhibitor of NIS most probably acting as a blocker, not as
a substrate.
FIG. 4. Schematic representation of a NIS mechanistic model. As
These results raise the question of whether or not
shown in this scheme, first 1 Na⫹ binds to NIS, which in the absence of perchlorate is indeed translocated into thyroid cells ex-
substrate is able to cross the membrane via NIS in a Na⫹ uniport mode pressing endogenous NIS or into cells other than oocytes
(CNa⬘3 CNa⬙; C, carrier). Release of Na⫹ into the cytoplasm is followed
by the return of the empty binding site to complete the pathway (C⬙ 3 expressing transfected NIS, and this question has been
C⬘). The kinetic data suggest that Na⫹ binds to NIS before I⫺. In the the subject of some controversy. In the acknowledgments
presence of I⫺, the complex CNa2I⬘ is formed which undergoes a con- section of his recent extensive review on perchlorate and
formational change to expose the bound I⫺ and 2 Na⫹ to the interior of

the cell (CNa2I⬘ 3 CNa2I⬙). Both Na⫹ and I⫺ are released into the the thyroid gland, Wolff (127) asserts that “perchlorate
cytoplasmic compartment, and the empty carrier undergoes another accumulation in thyroid tissue has been repeatedly dem-
conformational change to expose the binding sites to the external solu- onstrated” and suggests that the absence of perchlorate
tion again. Charge movement data suggest that the Na⫹ binding disso-
ciation does not contribute greatly to the total observed charge. Thus it transport by NIS in oocytes reported by Eskandari et al.
is proposed that NIS charge movements arise primarily from conforma- (34) may be due to differences between the oocyte system
tional changes of the empty carrier (C⬘3 C⬙). For more details, refer to and thyroid tissue. However, Yoshida et al. (131) thereaf-
Reference 34.
ter showed that perchlorate elicits no change in the mem-
brane current in the highly functional rat thyroid cell line
FRTL-5, as revealed by the whole cell patch-clamp tech-
uted to the conformational changes of the empty
nique, thus strongly suggesting that perchlorate is not
transporter within the membrane electric field. These au-
transported into FRLT-5 cells and supporting both their
thors determined that the turnover rate of NIS is ⬃36 s⫺1
previous observations in CHO cells (130) and the results
and reported that expression of NIS in oocytes led to a
of Eskandari et al. (34) in oocytes.
⬃2.5-fold increase in the density of plasma membrane
With the consideration that the properties of NIS
protoplasmic face intramembrane particles, as ascer-
expressed in oocytes are virtually indistinguishable from
tained by freeze-fracture electron microscopy. This is the
those of endogenous NIS in thyroid cells, including
first direct electron microscopy visualization of ostensible
FRTL-5 cells, it appears that earlier experiments (42, 43,
NIS molecules present in the oocyte plasma membrane.
Moreover, on the basis of their kinetic results, these in-
vestigators proposed an ordered simultaneous transport
mechanism in which Na⫹ binds to NIS before I⫺, i.e.,
whereas transport of both ions is simultaneous and bind-
ing is ordered and sequential. The combined data from
electrophysiological measurements and freeze-fracture
electron microscopy (9-nm-diameter NIS particles) pro-
vided in this report suggest that NIS may be multimeric in
its functional form (34).
F. The NIS Inhibitor Perchlorate Is Not

Translocated by NIS Into the Cell
Similar steady-state inward currents were generated

by a wide variety of anions in addition to I⫺ (including FIG. 5. Substrate selectivity of NIS. Inward currents induced by
various anions (500 ␮M) were recorded at a membrane potential of ⫺50
ClO⫺ ⫺ ⫺ ⫺ ⫺ ⫺ ⫺
3 , SCN , SeCN , NO3 , Br , BF4 , IO4 , and BrO3 ),
⫺
mV. Currents were normalized with respect to the current generated by
indicating that these anions are also transported by NIS. I⫺. I⫺-induced current in the absence of Cl⫺ did not differ from that in
However, perchlorate (ClO⫺ the presence of 100 mM Cl⫺. Other anions tested that did not induce a
4 ) (Fig. 5), the most widely
detectable inward current were as follows: NO⫺ ⫺ 2⫺ 2⫺
2 , HCO3 , SO3 , CO3 ,
characterized inhibitor of thyroidal I⫺ uptake, was sur- 3 , P2O4 . Data are reported as means ⫾ SE (n ⫽ 3). [Adapted from
S2O2⫺ 4⫺
prisingly found not to generate a current, strongly sug- Eskandari et al. (34).]
125) ostensibly showing that 36Cl-labeled perchlorate en- et al. (75) reported that the biosynthesis of thyroid hor-
ters the cell may have been misinterpreted. Because mones by sheep thyroid slices was inhibited by high doses
[36Cl]chlorate (ClO⫺3 ) is a
36
Cl-labeled by-product of the of I⫺. Wolff and Chaikoff reported in 1948 (128) that
reaction employed to chemically synthesize [36Cl]per- organic binding of iodide in the rat thyroid was blocked
chlorate (ClO⫺4 ) for these uptake studies, it seems likely when I⫺ thyroid levels reached a critical high threshold, a
that [36Cl]chlorate, rather than perchlorate, accounts for phenomenon known as the acute Wolff-Chaikoff effect.
the presence of label in the cytosol of thyrocytes, given These researchers observed further that ⬃2 days later, in
that chlorate is readily translocated via NIS into the cell the presence of continued high plasma I⫺ concentrations,
(34). Current electrophysiological data, in conclusion, an “escape” or adaptation from the acute effect is ob-
strongly indicate that perchlorate is not translocated via served so that the level of organification of I⫺ is restored
NIS into the cell. and normal hormone biosynthesis resumes (129).
Whereas the mechanism responsible for the acute Wolff-
Chaikoff effect has yet to be fully elucidated, it has been
III. REGULATION OF SODIUM/IODIDE proposed to be the result of organic iodocompounds act-
SYMPORTER PROTEIN EXPRESSION ing as mediators. The iodolipid ␣-iodohexadecanal has
been suggested to be one such mediator, on account of its
TSH is the primary hormonal regulator of thyroid ability to inhibit NADPH oxidase, TPO, and TSH-induced

function overall and has long been known to stimulate cAMP formation in the thyroid (28). The less studied
thyroidal I⫺ accumulation. TSH is a glycoprotein of mechanism for the escape from the acute Wolff-Chaikoff
⬃30,000 Da, biosynthesized in the adenohypophysis by effect has been proposed by Braverman and Ingbar (12) to
basophilic cells known as thyrotropes. The release of TSH be due to a decrease in I⫺ transport, which would pre-
from the pituitary is stimulated by thyrotropin-releasing sumably lead to sufficiently low intracellular I⫺ concen-
hormone (TRH) from the hypothalamus and inhibited trations to remove inhibition of I⫺ organification.
through a negative-feedback mechanism by the thyroid As in the case of NIS regulation by TSH, the regula-
hormones T3 and T4. Most actions of TSH take place tory role played by I⫺ on NIS function was explored at the
through activation of adenylate cyclase via the GTP bind- molecular level only after the cDNA that encodes NIS was
ing protein G␣ (117). This cascade of events is initiated by isolated. In vivo studies have shown that I⫺ inhibits the
the interaction of TSH with its receptor [i.e., TSH receptor expression of both TPO and NIS mRNA in dog thyroid
(TSHr)] on the basolateral membrane of the follicular (115), a finding consistent with the Wolff-Chaikoff effect.
cells (Fig. 1). Early observations made before the isola- More recently, Eng et al. (33) measured the levels of NIS
tion of the NIS cDNA suggested that TSH stimulation of I⫺ mRNA and NIS protein in response to both chronic and
accumulation results, at least in part, from the cAMP- acute I⫺ excess in rats in vivo, thus showing, specifically,
mediated increased biosynthesis of NIS (51, 122). Using that the decrease of I⫺ transport observed during the
high affinity anti-NIS Ab, Levy et al. (62) demonstrated in escape from the Wolff-Chaikoff effect is due to a decrease
rats that NIS protein expression is upregulated by TSH in in NIS expression by a mechanism that is at least in part
vivo. They prepared thyroid membrane fractions from transcriptional.
control, propylthiouracil (PTU)-treated, iodine-deficient,
and hypophysectomized rats, the latter with or without
IV. TRANSCRIPTIONAL REGULATION OF THE
subsequent injection of TSH. Membrane fractions were
SODIUM/IODIDE SYMPORTER
then subjected to immunoblot analysis with anti-NIS Ab.
They observed NIS upregulation in rats with increased
TSH circulating levels caused either by PTU treatment A. Thyroid-Specific Transcription Factors
(which inhibits I⫺ organification) or an I⫺-deficient diet.
Conversely, NIS protein expression was decreased in hy- The transcriptional regulation of three other genes of
pophysectomized rats, which exhibit markedly lower TSH major significance in thyroid physiology had been ana-
levels. A single injection of TSH to hypophysectomized lyzed, namely, the Tg, TPO, and TSHr genes. Initially, all
rats reinduced the expression of NIS protein back to basal three gene products were proposed to be thyroid specific,
levels. Consistent with these findings is a later observa- i.e., to be expressed exclusively in the thyroid. However,
tion by Uyttersprot et al. (115) that the expression of NIS TSHr expression and activity have recently been demon-
mRNA in dog thyroid (⬃3.9 kb) is dramatically upregu- strated in adipocytes (100), leaving Tg and TPO as the
lated by goitrogenic treatment (i.e., PTU treatment, which only thyroid-specific proteins.
leads to elevated TSH circulating levels in vivo). Three different transcription factors have been impli-
The main factor regulating the accumulation of I⫺ in cated in thyroid-specific gene transcription (22, 73): 1)
the thyroid (i.e., NIS activity) other than TSH has long thyroid transcription factor (TTF)-1, a homeodomain
been considered to be I⫺ itself. As early as 1944, Morton (HD)-containing protein present in the developing thy-
roid, lung, forebrain, and pituitary and in the adult thyroid (95) have recently shown that both TTF-2 and HNF-3␤,
and lung; 2) TTF-2, a forkhead protein detected in devel- another forkhead transcription factor also present in the
oping thyroid and anterior pituitary and in the adult thy- thyroid, bind to the same site and thus participate in the
roid; and 3) Pax8, a nuclear protein member of the murine regulation of the TPO but not the Tg promoter. In addi-
family of paired-domain (PD) containing genes, present in tion, upstream enhancers have also been identified in
the developing thyroid, kidney, and midbrain boundary both promoters. Three TTF-1 binding sites for the bovine
and in the adult thyroid and kidney. Specific combinations Tg promoter are present 2 kb upstream. Two TTF-1 and
of these factors have been proposed to regulate transcrip- one Pax8 binding site, recently reported (35), are found in
tion of the thyroid-specific proteins Tg and TPO and also a 230-bp enhancer located 5.5 kb upstream of the TPO
of the TSHr. initiation codon (22).
B. Transcriptional Regulation of Tg and TPO C. Transcriptional Regulation of the TSHr
The Tg and TPO minimal promoter structures are The TSHr promoter is very different from the Tg and
very similar to each other but different from the TSHr TPO promoters (Fig. 6). The TSHr minimal promoter
promoter (22) (Fig. 6). The Tg and TPO promoters from region is localized in the 5⬘-upstream region between

different species exhibit three binding sites for TTF-1 (A, ⫺220 and ⫺39 bp and shows thyroid-specific expression
B, and C), one binding site for TTF-2, and one for Pax8. All and TSH/cAMP regulation. Several binding regulatory el-
of these sites are localized 5⬘-upstream between ⫺170 and ements have been proposed to play a role in the regula-
⫹1 bp (A in the ATG initiation codon) and both contain a tion of TSHr transcription. A canonical cAMP responsive
TATA box. The TTF-1 site C and Pax8 site overlap, sug- element (CRE) (⫺139 to ⫺131 bp) site that is autoregu-
gesting that this overlapping site area may play an impor- lated by TSH via cAMP has been shown to first increase
tant role in thyroid-specific transcription. TTF-2 plays an and then decrease the expression of TSHr as a function of
important role in the modulation of the Tg and TPO genes time (93). One TTF-1 binding site is present at ⫺189 to
by insulin and insulin-like growth factor I, but this action ⫺175 bp; binding of phosphorylated TTF-1 to this site
is more evident for the TPO than for the Tg promoter (8, produces a modest effect on the activation of transcrip-
80, 94). Both promoters have been shown to have binding tion (77, 101). Single-strand DNA-binding proteins
sites for different ubiquitous transcription factors, (SSBP), whose binding sites overlap with the 5⬘-end of the
namely, ubiquitous factor A (UFA) in the Tg promoter and TTF-1 site, have been implicated in the regulation of the
ubiquitous factor B (UFB) in the TPO promoter. Sato et al. TSHr (102). In contrast to the Tg and TPO promoters,
FIG. 6. Schematic representation of the structures of the Tg, TPO, TSH receptor (TSHr), and NIS promoters. Symbols
refer to binding sites mapped by DNase I footprinting. Numbers indicate the distances from the corresponding
transcriptional start sites. For specific details, see Reference 22.
neither TTF-2 nor Pax8 binding sites have been identified could result from the presence of an upstream tissue-
in the TSHr promoter. The TSHr transcript has also been specific enhancer or from the influence of chromatin
found in retro-orbital fibroblasts and in adipose tissue (32, structure and/or methylation, as there are numerous CpG
60). Rat adipose tissue expresses TSHr mRNA levels sim- dinucleotides.
ilar to those found in the thyroid (100). The promoter
regulation is different in adipocytes from thyroid cells,
but both display multiple start sites and important regu- E. Analysis of the Rat NIS Promoter
lation by CREB (99).
TPO and Tg expression are both upregulated by TSH Tong et al. (112) isolated a 16.4-kb fragment of
(41, 116) but by different regulatory mechanisms. TPO genomic rNIS DNA. They localized the start site at ⫺98 bp
and TSHr regulation takes place faster than Tg regulation. and one TATA box between ⫺124 and ⫺118 (AATAAAT),
Although Tg transcriptional activation kinetics depend on with a minimal promoter localized between ⫺199 and
the experimental model used, i.e., it is rapid (⬃1 h) in ⫺110 bp. Surprisingly, however, they concluded that 8 kb
thyroid slides and slow (⬃8 h) in primary cultures, TPO of the 5⬘-flanking region of the NIS gene is not sufficient to
induction is rapid in both experimental models (41). All confer thyroid-specific transcription. Neither TTF-2 nor
three genes are controlled in the thyroid at the transcrip- Pax8 putative binding sites were localized within 2 kb
tional level mainly by a cAMP-dependent mechanism, upstream of the initiation codon. However, the conclu-

whose intracellular level is elevated by TSH/forskolin. sion of Tong et al. (112) proved incorrect, because Endo
However, only the TSHr gene has been shown to be et al. (30) localized a TTF-1 binding site between ⫺245
regulated by cAMP through a CRE sequence. The exact and ⫺230 bp, in the proximal rNIS promoter (2 kb), that
mechanism by which cAMP regulates Tg and TPO is still confers thyroid-specific transcription but only exerts a
an open question. modest effect.
The same group (76) subsequently identified, in the
5⬘-flanking region between ⫺1,968 and ⫹1 bp, a novel TSH-
D. Analysis of the Human NIS Promoter responsive element (TRE) between ⫺420 and ⫺385 bp up-
stream of the TTF-1 site in the rNIS promoter, that upregu-
It has long been established that TSH stimulates NIS lated two- to threefold NIS expression. The TSH effect is
activity via the cAMP pathway (62, 81, 122) and, more cAMP mediated and thyroid specific. They showed that the
recently, that it upregulates NIS mRNA levels (53). How- protein that binds this site is different from TTF-1, TTF-2,
ever, to fully understand these mechanisms, it is neces- Pax8, or other known transcription factors, and the re-
sary to study the transcriptional regulation of NIS. The searchers named the putative binding protein in the TRE site
human NIS promoter has been sequenced by three differ- NIS TSH-responsive factor 1 (NTF-1). However, because the
ent groups (10, 90, 118). Venkataraman et al. (118) iso- TSH upregulation of NIS via this TRE site is lower than the
lated a 1.2-kb fragment of the 5⬘-flanking region of the regulation that the same group reported previously (⬃6-
hNIS gene. Significantly, they characterized the promoter fold) (53), they have suggested that other transcription bind-
in a human thyroid cell line, KAT-50, and found thyroid- ing sequences may be present upstream from the 1,968-bp
specific expression between ⫺1,044 and ⫺336 bp. Some region that they studied.
putative transcription factor binding sites, both thyroid A thorough characterization of the upstream en-
and nonthyroid, are localized in this region (1.2 kb) but hancer of the rNIS gene has recently been reported by
were not analyzed. It is noteworthy that this list includes Ohno et al. (78). These investigators showed that the rNIS
the putative TTF-2 and Pax8 binding sites, both of which regulatory region contains a nonthyroid-specific promoter
are found within the 2-kb portion of the 5⬘-flanking region between ⫺564 and ⫺2 bp and an enhancer located be-
in the rat promoter (see below). Ryu et al. (90) isolated 2 tween ⫺2,264 and ⫺2,495 bp that recapitulates the most
kb of the 5⬘-flanking region from the hNIS promoter. They relevant aspects of NIS regulation. This rNIS enhancer
mapped the start site to ⫺375 bp and delimited the min- mediates thyroid-specific gene expression by the interac-
imal promoter between ⫺478 and ⫺389 bp. A 90-bp frag- tion of Pax8 with a novel cAMP-dependent pathway. The
ment exhibited a high identity (73%) with the minimal NIS upstream enhancer (NUE) stimulates transcription in
rNIS promoter. These investigators reported some mis- a thyroid-specific and cAMP-dependent manner. NUE
matches with the previously reported sequence. However, contains the following: two Pax8 binding sites (PA and
this group did not find thyroid-specific regulatory ele- PB); two TTF-1 binding sites (TA and TB) that have no
ments. Finally, Behr et al. (10) isolated 1.6 kb of the effect on rNIS transcription; and a degenerate CRE se-
5⬘-flanking region of the hNIS gene. Like Ryu et al. (90), quence (5⬘-TGACGCA-3⬘), which is important for NUE
Behr et al. (10) also found strong promoter activity in transcriptional activity. Interestingly, the same degener-
thyroid and nonthyroid cells within the minimal pro- ate CRE sequence has been implicated in tissue-specific
moter. Behr et al. (10) suggested that the nonspecificity cAMP response of other promoters, such as the dopamine
␤-hydroxylase (111), prohormone convertase 1 (47), and ability to organify accumulated I⫺; therefore, they behave
proenkephalin genes (19). In NUE, both Pax8 and the like PTU-treated thyroid tissue; 2) TSH exerts no regula-
unidentified CRE-like binding factor act synergistically to tory influence on nonthyroid I⫺ accumulation; and 3) at
obtain full TSH/cAMP-dependent transcription. However, least salivary glands and gastric mucosa concentrate thio-
this enhancer is also able to mediate cAMP-dependent cyanate, unlike the thyroid where it is metabolized (oxi-
transcription by a novel PKA-independent mechanism dized). Despite these differences, several reports of pa-
(78). tients suffering the simultaneous genetic absence of I⫺
The present picture of the NIS promoter shows two transport in the thyroid, the salivary glands, and the gas-
important regions (Fig. 6): 1) a proximal promoter, still tric mucosa strongly hinted at a “genetic link” among
questionably thyroid specific, reported to be TSH/cAMP these I⫺ transport systems, suggesting that extrathyroidal
regulated by a TRE binding sequence, mediated by a new I⫺ transport is catalyzed by plasma membrane proteins
NTF-1 protein (76). This upregulation, however, is lower that are very similar, if not identical, to thyroid NIS (13,
than the TSH-induced mRNA expression reported by the 16, 42, 125). Moreover, thyroidal and extrathyroidal I⫺
same group (53). 2) The NIS promoter also has an up- accumulation generate I⫺ concentration gradients of sim-
stream enhancer that is thyroid specific, with Pax8 and ilar magnitude (⬃20- to 40-fold under steady-state condi-
CRE-like binding factors. It seems likely that both regions tions). Hence, the isolation and characterization of the
in the promoter act synergistically to achieve full high- NIS cDNA from rat thyroid (21) made it possible to ex-

level transcription. amine NIS expression in nonthyroid tissues, leading to the
Transcriptional regulation of the Tg, TPO, and TSHr conclusion that I⫺ transport in most (and probably all)
genes by TSH/forskolin is mediated by cAMP. However, extrathyroid tissues in which it is present is also mediated
no CRE sequences have been identified for any of these, by NIS itself, like in the thyroid. However, NIS is clearly
except the TSHr gene. Kambe et al. (49) have reported regulated and processed differently in each tissue.
that redox regulation of TTF-1 and Pax8 was involved in
the TSH-increased DNA-binding activities of these two
factors. However, no clear evidence has been provided on A. NIS in Salivary Glands, Stomach, Mammary
the control exerted by TSH/cAMP. Thus the novel PKA- Gland, and Other Tissues
independent mechanism reported by Ohno et al. (78) is
highly significant, because it establishes a direct relation- Spitzweg et al. (108) subjected several extrathyroidal
ship between thyroid-specific transcription factors and human tissues to Northern blot and RT-PCR analysis with
the TSH/cAMP regulation in rNIS. This picture clearly human thyroid NIS cDNA and detected the NIS transcript
separates rNIS gene regulation from that of true thyroid- by Northern blot predominatly in the parotid salivary
restricted genes (Tg and TPO) and also from TSHr, a gland but not in the other tissues. With the use of RT-PCR,
notion consistent with the fact that NIS is not exclusively hNIS gene expression was detectable in salivary glands,
expressed in the thyroid gland. It is clearly of much pituitary gland, pancreas, testis, mammary gland, gastric
interest to investigate the transcriptional regulation of mucosa, prostate, and ovary but not in orbital fibroblasts,
NIS in extrathyroidal tissues. These studies will reveal colon, and nasopharyngeal mucosa. Kotani et al. (56),
mechanisms of different tissue-specific regulation of the who carried out Northern analyses with rat thyroid NIS
same gene and may provide insights into possible appli- cDNA in extrathyroidal rat tissues, detected the NIS tran-
cations of NIS in the diagnosis and treatment of both script primarily in the stomach. Subsequently, Spitzweg et
thyroid and nonthyroid cancer. al. (108) reported the cloning of human NIS cDNA from
gastric mucosa, parotid, and mammary glands, all of
which exhibited full identity to thyroid hNIS cDNA. Con-
V. EXPRESSION OF SODIUM/IODIDE sistent with these findings, immunohistochemical studies
SYMPORTER IN NONTHYROID TISSUES by Jhiang et al. (48) and by Kotani et al. (56) detected NIS
protein in the acinar cells of human salivary glands and in
The topic of I⫺ transport systems outside the thyroid surface epithelial and parietal cells of the rat stomach,
was extensively reviewed in 1961 by Brown-Grant (13). respectively. Ajjan et al. (4) have reported variable
The main vertebrate nonthyroid tissues reported to ac- amounts of hNIS amplified product, as detected by RT-
tively accumulate I⫺ are salivary glands, gastric mucosa, PCR, in human stomach, salivary gland, and mammary
lactating mammary gland, choroid plexus, and the ciliary tissue, but not in the ovary, esophagus, colon, extraocular
body of the eye. Many of these transport systems exhibit fat, or skin. Surprisingly, they failed to detect NIS expres-
functional similarities with their thyroid counterpart, no- sion using Northern blot analysis in stomach, salivary
tably a susceptibility to inhibition by thiocyanate and gland, intestinal fat, and multinodular goiter tissue sam-
perchlorate, but they also display important differences: ples.
1) nonthyroid I⫺ transporting tissues do not have the It must be pointed out that the RT-PCR technique
yields a large number of false positives due to the high Ref. 62), a finding consistent with the identity among
sensitivity of the RT-PCR technique (38). Therefore, the human thyroid NIS, mg-NIS, and g-NIS predicted by the
detection of the NIS amplified product by RT-PCR in a cloning of human NIS cDNA from gastric mucosa and
given tissue cannot be regarded as sufficient evidence that mammary glands by Spitzweg et al. (108).
NIS is functionally expressed in that tissue. A thorough
characterization of NIS protein expression is necessary to
C. Regulation of Mammary Gland NIS
properly evaluate the significance or results obtained by
RT-PCR and even Northern analyses, and once NIS pro-
Subsequently, Tazebay et al. (111a) investigated
tein expression has been demonstrated, a correlation with
whether a correlation existed between I⫺ accumulation
Na⫹-dependent, perchlorate-sensitive, active I⫺ accumu-
activity and mg-NIS expression in the various physiolog-
lation in that tissue must be established. By these criteria,
ical stages of the mammary gland. They found that mg-
and with the consideration of the above results, NIS ap-
NIS was absent in nubile and pregnant mammary gland,
pears to be expressed and active in extrathyroidal tissues
but clearly present in lactating mammary gland. To ascer-
previously known to exhibit NIS activity, such as salivary
tain whether weaning had an effect on mg-NIS expres-
glands and gastric mucosa. The significance of the detec-
sion, mother rats were removed from their litters. Twenty-
tion of the NIS-amplified product by RT-PCR in other
four hours after weaning, mg-NIS expression was
human and rat tissues still awaits further characteriza-
significantly decreased, and after 48 h, it was barely de-

tion.
tectable. Furthermore, mg-NIS expression was reversible
upon reestablishment of nursing. These results indicate
B. Identification of Mammary Gland NIS and that mg-NIS expression is clearly upregulated during ac-
Gastric NIS tive nursing, rapidly downregulated upon cessation of it,
and exquisitely regulated in a reversible manner by suck-
ling.
Levy et al. (62) performed immunoblot analyses to
To assess the issue of tissue distribution of I⫺ trans-
assess whether a high-affinity antithyroid NIS Ab would
port activity, Tazebay et al. (111a) carried out in vivo
react with a mammary gland membrane protein (111a).
scintigraphic imaging of 131I⫺ (or the substitute isotope
They observed immunoreactivity against a single broad 99m
TcO⫺4 ) tissue distribution in rats. In nonlactating rats,
⬃75-kDa polypeptide in rat lactating mammary gland
the isotope was concentrated in the stomach and later in
membranes, but not in membranes from nonlactating
the thyroid, whereas in lactating rats it was evident in the
mammary gland or from lung, muscle, or heart, all tissues
stomach and soon thereafter in all pairs of mammary
that do not transport I⫺. This immunoreactive polypep-
glands. Concentration of the isotope in all these tissues
tide is mammary gland NIS (mg-NIS). These authors then
was inhibited by perchlorate.
investigated the difference in electrophoretic mobilities
between mg-NIS (⬃75 kDa) and thyroid NIS (⬃90 kDa)
and found that it is due to differences in their posttrans- VI. THE SODIUM/IODIDE SYMPORTER
lational modifications. They treated membrane proteins IN AUTOIMMUNE THYROID DISEASE
from thyroid and lactating mammary gland with N-glyco-
sidase F, an enzyme that removes N-linked carbohy- An important aspect of the wide pathophysiological
drates, and probed membranes with anti-NIS Ab. Under impact of Tg, TPO, and TSHr is the fact that autoAb
these conditions, anti-NIS Ab recognized a ⬃50-kDa against all three proteins have been demonstrated in pa-
polypeptide in membranes from both thyroid and lactat- tients suffering from autoimmune thyroid disease (AITD).
ing mammary gland. Significantly, both nonglycosylated AITD is significantly more prevalent in women; it affects
NIS in FRTL-5 cells and NIS expressed in Escherichia coli ⬃2% of the female population but only ⬃0.2% of males
exhibit an identical electrophoretic mobility (i.e., ⬃50 (121). The pathogenesis of AITD across the spectrum
kDa). These results demonstrate that the ⬃75- and ⬃50- from Graves’ disease to Hashimoto’s thyroiditis has been
kDa immunoreactive polypeptides detected in lactating suggested to involve a deficiency in the number and/or
mammary gland correspond to glycosylated and nongly- function of antigen-specific and nonspecific population of
cosylated mg-NIS, respectively. Moreover, Tazebay et al. T lymphocytes (37). As is the case in all autoimmune
(111a) also observed immunoreactivity of anti-NIS Ab disorders, multiple factors are believed to play a role in
with a ⬃100-kDa gastric polypeptide, which upon degly- the development of AITD, including genetic predisposi-
cosylation migrated, too, at ⬃50 kDa. In all likelihood, tion, environmental factors, and a state of dysregulation
these polypeptides correspond, respectively, to glyco- of the immune system (37). In the presence of AITD, the
sylated and nonglycosylated gastric NIS (g-NIS). CNBr thyroid is infiltrated with mononuclear cells, including
treatment of rat thyroid NIS, mg-NIS, and g-NIS proteins activated T and B plasma cells and macrophages, thus
yielded the same peptide map in each case (Levy et al., creating a local cytokine milieu (120). A distinctive fea-
ture of AITD is the expression of HLA class II molecules, CHO cells stably expressing recombinant rat NIS. The
induced by the T cell secreted interferon (IFN)-␥, on the investigators observed also that some normal sera and
follicular epithelial cells, which are normally class II neg- patients’ sera that did not immunoreact on immunoblots
ative. Another characteristic feature of AITD is the pres- nevertheless caused ⬃90% inhibition of NIS activity. Be-
ence in patients’ sera of autoAb that are directed against cause this inhibitory effect was lost after dialyzing these
specific thyroid antigens (Tg, TPO, and TSHr) (121). Io- sera, the authors concluded that some sera contained a
dine is an important susceptibility and modulating factor dialyzable inhibitor, independent of the presence or ab-
in the development of thyroid autoimmunity. High levels sence of anti-NIS autoAb, and thus suggested that purified
of iodine can cause thyroid damage and thyroiditis, par- IgG be used to accurately evaluate the putative autoAb
ticularly against the background of a previously low in- activity. The investigators also concluded that the pres-
take of iodine. Significantly, the incidence of AITD is ence of anti-NIS autoAb may play a role in the pathogen-
higher in areas of iodine sufficiency than in areas of iodine esis of Hashimoto’s thyroiditis and thus may modulate
deficiency (37). thyroid function. All of these preliminary observations
Given that thyroid-associated proteins such as Tg, call for further study and confirmation.
TPO, and TSHr are known targets for autoAb in AITD, the Morris et al. (74) synthesized 21 peptides correspond-
molecular identification of NIS has made it possible for ing to putative extracellular segments of rat NIS, based on
the first time to experimentally assess whether NIS is also the first 12 transmembrane segment secondary structure

a target autoantigen, and whether any anti-NIS autoAb (if model proposed for rat NIS (21). Samples were analyzed
present) have an effect on NIS function. In addition, it is by ELISA using the synthetically made peptides. Because
also possible that NIS is one of the modulators of the local of the high variability of background binding, the normal
autoimmune processes, conceivably regulating the indi- range of binding was determined in the case of each
vidual I⫺ supply to the follicular cell. peptide by a pool of normal IgG, and thereafter the num-
Even before the isolation of the NIS cDNA, Raspe et ber of standard deviation units from the mean of the
al. (88) screened sera from 147 patients with AITD by control was calculated. Eighty-eight percent of Graves’ (n
measuring the effect of the sera on I⫺ transport activity in ⫽ 27) and 63% of Hashimoto (n ⫽ 27) serum samples
primary cultures of dog thyrocytes. They observed that recognized at least one synthetic peptide. The most highly
the serum from a 58-yr-old female patient suffering from recognized eight peptides were those corresponding to
Hashimoto’s thyroiditis, autoimmune gastritis, and rheu- the fourth, fifth, and sixth extracellular loops and the
matoid arthritis, with high autoAb titers against Tg, TPO, intracellular COOH-terminal tail of the first secondary
and gastric parietal cells, completely blocked TSH-in- structure model (Fig. 2A), which correspond, respec-
duced I⫺ uptake. Inhibition was specific, given that the tively, to the fourth and sixth intracellular and sixth ex-
serum was still active at 1:1,000 dilution and did not tracellular loops, and the intracellular COOH-terminal tail
inhibit the activity of the Na⫹-K⫹-ATPase. Unfortunately, of the current 13 transmembrane segments model (Fig.
no further experiments were performed because only a 2B). In contrast, none of the control sera displayed any
limited amount of serum was available. Nevertheless, the immunoreactivity. The occurrence of Ab that recognized
investigators still concluded that the serum contained intracellular epitopes was explained by the investigators
NIS-inhibiting autoAb. The low prevalence of the autoAb as a result of exposure of these internal sequences to the
positivity (1 of 147 sera) may be due to the purely func- thyroiditis-induced follicular cell damage. Unfortunately,
tional nature of the assay, as only Ab against external there were no data regarding recognition of the entire NIS
epitopes and with an I⫺ transport inhibitory effect would molecule by these antisera.
be detected, as well as to the use of dog thyrocytes to Ajjan et al. (3) established a CHO cell line stably
screen human sera. expressing human NIS devoid of the last 31 amino acids,
Endo et al. (31) screened sera from patients with thus generating a valuable system (CHO-NIS9 cells) for
AITD by recombinant NIS protein slot blotted onto nitro- the evaluation of anti-NIS Abs, on account of the absence
cellulose sheets. Twenty-two of 26 Graves’ disease sera of other thyroid-specific antigens. Eighty-eight sera from
and 3 of 20 Hashimoto’s thyroiditis sera seemed to detect patients with Graves’ disease were tested for their effect
the recombinant NIS protein, but the effect of these pos- on I⫺ uptake. Twenty-seven of 88 (30.7%) of the Graves’
itive sera on I⫺ uptake was not tested. In a subsequent disease sera (and also their corresponding purified IgG)
study, the same group (29) concentrated exclusively on but none of the controls inhibited I⫺ uptake. The autoAb
Hashimoto’s thyroiditis samples. Four of 34 sera from were not immunoreactive in immunoblot experiments us-
patients with Hashimoto’s thyroiditis immunoreacted ing extracts from the same cells, an observation that may
with a ⬃80-kDa polypeptide, as seen upon immunoblot relate to antigen concentration and/or the absence of
analysis of FRTL-5 cell membranes (which contain not linear epitopes in NIS.
only NIS but also other thyroid antigens), and these same These experimental data suggest that NIS might be
four sera caused 14 – 62% inhibition of I⫺ accumulation in an autoantigen in AITD and that autoAb are generated
against it. It is important to continue the analysis of this noma, either follicular or papillary, is one of the most
topic utilizing experimental systems free of other thyroid curable cancers; the overall survival rate at 10 years for
autoantigens, and to investigate the presence of anti-NIS middle-aged adults with thyroid carcinoma is ⬃80 –90%
autoAb against both linear and conformational epitopes, (97). The reduced I⫺ accumulation detected in the major-
that are no longer present after SDS-PAGE electrophore- ity of thyroid cancers suggests that malignant transforma-
sis. Clearly, a wide range of experimental approaches will tion of these cells may have an effect on the NIS molecule.
be necessary to unequivocally determine the existence, Thus the continued characterization of NIS may shed light
real prevalence, functional effects, and pathological sig- on I⫺ transport changes observed in thyroid cancer.
nificance of autoAb against NIS. Furthermore, another The treatment of differentiated thyroid carcinoma is
interesting aspect to consider is the role of certain cyto- total or near-total thyroidectomy followed by 131I abla-
kines in NIS function, because recent reports indicate that tion. Radioiodide ablation destroys occult microscopic
tumor necrosis factor (TNF)-␣ and, to lesser extent, in- carcinomas and also any remaining normal thyroid tissue,
terleukin (IL)-1␣ inhibit both basal and TSH-induced NIS permitting postablative 131I total body scanning search for
expression (5). High concentrations of IFN-␥ also down- possible persistent carcinoma (119). The tools that allow
regulate TSH-stimulated NIS mRNA expression. Conse- the sensitive and specific detection and subsequent treat-
quently, IL-1␣, TNF-␣, and IFN-␥ all inhibit I⫺ uptake in ment of differentiated thyroid carcinoma are the uptake
FRTL-5 cells. Caturegli et al. (17) have recently generated (mediated by NIS) and organification (mediated by TPO)

transgenic mice that produce IFN-␥ in the thyroid. The of I⫺, both TSH-regulated processes. The tumorigenic role
presence of this cytokine in the thyroid had no effect on of radioidine in the thyroid, transported also by NIS into
Tg expression. In contrast, the expression of both TPO the follicular cell, is debated, but the increased incidence
and TSHr was slightly increased, whereas NIS expression of papillary thyroid carcinomas in children (100 times
was significantly decreased. Consistently, I⫺ accumula- higher than in nonexposed children) after nuclear testing
tion was severely reduced in these animals (17). Further and nuclear accidents suggests that radioactive isotopes
investigation on the effect of cytokines on I⫺ uptake may of iodine have a direct tumorigenic effect on the thyroid
reveal some correlations between I⫺ supply and the (97).
pathogenesis of the autoimmune process. To understand the involvement of NIS in the ob-
served patterns of I⫺ transport in thyroid carcinomas with
respect to healthy thyroid tissue, several groups have
VII. THE SODIUM/IODIDE SYMPORTER looked for a possible correlation between the lower or
AND CANCER absent radioiodine uptake activity in thyroid carcinomas
and the expression of NIS mRNA or protein, using North-
Although cancer of the thyroid is a relatively infre- ern blot analysis, RT-PCR, or immunological analyses.
quent condition in the United States (0.6% of all cancers in Smanik et al. (106) observed, by Northern blot analysis,
men and 1.6% in women, Ref. 104), it has a considerably lower expression of hNIS in two papillary, one follicular,
higher impact on endocrinological practice than would be and one anaplastic carcinoma. Upon further analysis of
expected solely on the basis of its incidence. This is so additional papillary carcinomas by RT-PCR, a more sen-
because the possible existence of thyroid cancer must be sitive technique, they found that different papillary can-
ruled out whenever a thyroid nodule is detected. There is cers expressed hNIS at variable levels.
a high estimated incidence of solitary palpable nodules in Considering the limitations of RT-PCR as a quantita-
United States adults (⬃2– 4%) (89). The degree of accu- tive method, Arturi et al. (7) were able to evaluate only the
mulation of I⫺, as revealed by scans of the gland, is used presence or absence of the NIS transcript in a series of
as an aid in the differential diagnosis of thyroid nodules. malignant and benign thyroid tumors. They first analyzed
Thyroid nodules that accumulate I⫺ equally or more effi- by RT-PCR 26 primary thyroid carcinomas (19 papillary, 5
ciently than the normal surrounding tissue are generally follicular, and 2 anaplastic), and subsequently a new se-
benign, whereas most thyroid cancers display markedly ries of 15 follicular adenomas (11 cold and 4 hot as
reduced I⫺ accumulation relative to healthy tissue. On the classified by thyroid scintigraphy), and found loss of ex-
other hand, carcinomas behaving as hyperfunctioning hot pression of NIS in 5 of 19 papillary cancers, 1 of 5 follic-
nodules are very rare. Thus radioactive I⫺ plays a major ular cancers, and none detected in anaplastic tissues. All
diagnostic and therapeutic role in the management of the (benign) follicular adenomas except one expressed
differentiated thyroid carcinoma. The main morphologi- NIS. According to these authors, the fact that even one
cal types of differentiated thyroid cancer are the papillary benign follicular adenoma failed to express NIS means
carcinoma (75– 85% of all thyroid cancers) and the follic- that the absence of the NIS mRNA signal may not be
ular carcinoma (10 –20%). In most instances, both types restricted to the malignant phenotype. In all the studied
are well-differentiated tumors that originate in the thyroid tumors, except anaplastic, both Tg and TPO transcripts
follicular cells (89). Well-differentiated thyroid carci- were expressed.
Caillou et al. (14) conducted an immunohistochemi- have recently introduced rNIS into melanoma, ovarian,
cal analysis of NIS protein expression in thyroid patho- liver, and colon carcinoma cells. The resulting rNIS trans-
logical tissues, including six benign adenomas, five follic- duced tumor cells exhibited I⫺ uptake activity. In vitro
ular, and nine papillary carcinomas. No immunostaining experiments showed that these transduced cells could be
for hNIS was detected in any of the six benign adenomas destroyed by accumulation of 131I. If positive results were
that appeared to be hypofunctioning on thyroid scintigra- obtained in subsequent in vivo experiments, this gene
phy. hNIS expression was lower or absent in the papillary therapy approach would undoubtedly be one of the most
and follicular well-differentiated carcinomas, but more promising developments concerning the possible uses of
NIS was present in these than in the poorly differentiated the molecular characterization of NIS in the diagnosis and
follicular carcinomas. Consistent with these results, treatment of cancer.
Jhiang et al. (48) detected no hNIS by immunohistochem-
ical staining in any of the three papillary or the single
follicular carcinoma that they studied. In contrast, Saito et VIII. CONGENITAL IODIDE TRANSPORT
al. (92) found increased NIS expression in papillary car- DEFECT DUE TO SODIUM/IODIDE
cinomas. They investigated 31 papillary carcinomas by SYMPORTER MUTATIONS
either Northern blot or immunological analysis. Based on
their observations of higher NIS expression in ⬃50% of Congenital lack of I⫺ transport is a rather uncommon

the analyzed carcinomas compared with nonmalignant thyroid condition that has been defined as an I⫺ transport
thyroid tissue, the investigators concluded that despite defect (ITD). The general clinical picture consists of hy-
the high amounts of NIS protein present, well-differenti- pothyroidism (which can be normalized in some cases
ated thyroid carcinomas do not accumulate radioiodide, with high I⫺ supplementation or L-T4 substitutive ther-
suggesting that the activity of NIS is impaired or absent as apy), goiter, low thyroid I⫺ uptake (as determined by
a result of different posttranslational modifications, traf- scintigraphy), and low saliva-to-plasma I⫺ ratio (109, 126).
ficking alterations, or other unknown modulatory factors. Even though congenital hypothyroidism is an infrequent
In cancerous follicular cells positive for hNIS, the local- disease (incidence 1:3,000 –1:4,000 in neonates) (113), it
ization of the protein was described as basolateral and has an irreversible deleterious effect on the development
intracellular (92). of the newborn, finally resulting in cretinism if untreated.
In light of the data summarized above, one may con- Mutations in thyroid-specific molecules, such as TPO (2,
clude that hNIS expression may be either increased, de- 11), Tg (60, 72), and TSHr (1, 110), have been identified
creased, or absent in well-differentiated thyroid cancer, among causes of congenital hypothyroidism. Most re-
even though I⫺ transport activity is consistently de- cently, NIS mutations have also been demonstrated to
creased in the majority of tumors. Therefore, the expla- cause this disease. In the absence of a functional NIS
nation for the decreased I⫺ uptake in well-differentiated molecule, I⫺ has no access to the thyroid epithelial cells,
thyroid carcinomas lies not simply in lower NIS expres- resulting in decreased thyroid hormone biosynthesis (see
sion, but more likely involves a more complex combina- sect. I) and higher circulating levels of TSH, which in turn
tion of regulatory changes ultimately affecting NIS ex- stimulate the morphological and biochemical changes in
pression, targeting, and/or activation. To date, there are the thyroid that lead to the development of goiter.
still no data concerning the regulation of expression, Ever since the first case of congenital hypothyroid-
posttranslational modifications, targeting to the plasma ism due to an I⫺ transport defect was described by Fed-
membrane, or other factors regulating NIS in cancerous ermann et al. (36), several explanations have been pro-
follicular cells. Thus there clearly is a pressing need to posed to better define the nature of the defect. However,
investigate a much larger number of thyroid cancer spec- the molecular basis of this condition has only begun to be
imens by more quantitative techniques to better under- examined after the cloning of NIS cDNA (21, 105) and the
stand the expression of hNIS both at the transcript and elucidation of the exon-intron organization of the NIS
protein levels, to elucidate the cellular localization of NIS, gene (106) (Fig. 7). About 48 cases of ITD, belonging to 33
and to be able to compare such findings to the clinical families, have been reported worldwide to date. Seven-
behavior of the carcinomas. teen cases from 13 families have been studied at the
Some in vitro experiments concerning NIS-based molecular level and shown to have a mutation in NIS (see
gene therapy for both diagnostic and therapeutic pur- Table 1).
poses have been reported, in which NIS-mediated radio- Shortly after isolation of the cDNA that encodes NIS,
iodide uptake was used to visualize and destroy malignant two groups reported almost simultaneously a homozy-
tumor cells. Schimura et al. (96) transfected the hNIS gous missense mutation in patients that had previously
gene into malignant rat thyroid cells that did not concen- been diagnosed with congenital hypothyroidism caused
trate I⫺, resulting in increased I⫺ accumulation in these by ITD (39, 70) (Table 1, cases 1 and 6, respectively). Both
cells in vitro. Using a retroviral vector, Mandell et al. (67) groups found a DNA substitution of adenine with cytosine

FIG. 7. Correlation of the structural organization of the human NIS (hNIS) gene with the NIS protein. Exons in hNIS
gene are represented with hatched boxes; the 5⬘- and 3⬘-untranslated regions are represented by open boxes. The
putative 13 transmembrane segments in the protein are represented with cylinders. Exons are connected to the
corresponding amino acid regions of the protein by dotted lines (64, 106). Localization of the I⫺ transport defect
mutations is indicated under the schematic representation of the hNIS protein. [Modified from Smanik et al. (106) with
data from Levy et al. (64).]
in nucleotide 1060 in exon 9, resulting in a Pro instead of Subsequently, Levy et al. (65) studied the T354P mu-
Thr at position 354 (T354P) in NIS. The T354P mutation is tation by site-directed mutagenesis and transfection of
located in the putative transmembrane segment IX of the COS cells. They determined that the T354P NIS was not
NIS protein (Figs. 2B, 3, and 7). active, but they observed by immunofluorescence analy-
The patient in the report of Fujiwara et al. (39) was sis that it was properly targeted to the plasma membrane.
homozygous for the T354P mutation and had consanguin- Then various other amino acids substitutions (Ala, Pro,
eous parents; her two siblings were heterozygous for Cys, Tyr, Ser) at position 354 were explored to determine
T354P and were unaffected, suggesting the recessive na- the structural change in putative transmembrane segment
ture of the disease. Extremely high doses of potassium IX, leading to the conclusion that the hydroxyl group at
iodide treatment maintained this patient euthyroid. HEK- the ␤-carbon of the amino acid residue at position 354 is
293 cells (human embryonic kidney cells) transfected essential for NIS function (see sect. II).
with the mutant T354P NIS cDNA did not exhibit any I⫺ Fujiwara et al. (40) have reported four new cases of
uptake activity. Based on these results, the authors pro- patients with the T354P mutation from two families (Ta-
posed that T354P probably disrupts (due to the presence ble 1, cases 2–5). Case 2 was diagnosed at age 5, treated
of proline), through a structural change, the putative with thyroid powder, and thus maintained euthyroid;
transmembrane segment IX of NIS, where the mutation is however, this patient’s thyroid gland enlarged gradually
located. and developed multinodular goiter at the age of 14. The
The second case (70) was diagnosed 23 years ago patient’s parents were heterozygous for the T354P muta-
with an I⫺ transport defect (44) (Table 1, case 6). The tion without thyroid disorders. The other three patients
patient was euthyroid apparently on account of a very reported by Fujiwara et al. (40) are siblings (Table 1, cases
high iodide dietary intake. However, diffuse goiter with 3–5). These patients and their mother were heterozygous
slightly elevated TSH was noted at 18 yr of age. The for T354P, but their father was negative for this mutation.
patient exhibited greatly increased NIS mRNA levels The patients presented a small goiter that developed late
(⬎100-fold) in his thyroid. Hence, these investigators pro- in life with only slight symptoms of hypothyroidism. Even
posed that the observed increase in NIS transcription may though the patients were heterozygous for the T354 mu-
reflect a compensating mechanism for low NIS activity, tation, they were previously diagnosed as having ITD but
possibly related to transcriptional regulation of NIS by I⫺ were not screened for other NIS mutations.
itself. This patient was born from a consanguineous mar- Kosugi et al. (55) have reported more cases of T354P
riage, and his daughter was heterozygous for the mutation mutations, including three new and four reevaluated
but had no abnormal phenotype, as would be expected in cases with known ITD from five families (Table 1, cases
a recessive condition. 6 –12). All these cases were homozygous for the T354P
July 2000
TABLE 1. Summary of clinical and laboratory parameters of ITD patients and correlation to the NIS molecular defect
Year Age of Detection TSH,

of Reference T4, ␮g/l T3, ␮g/l mU/l TRIU, % S/P ratio DNA Protein
Case Birth Sex Country Hypothyroidism Goiter Development Cons No. (50–110) (0.94–1.54) (0.3–4.0) (10–40) (⬎20) Treatment Exon Mutation Zygosity Mutation Localization
1 1986 F Japan 3 mo 8 yr (FA) Normal ⫹ 39, 40 2 0.40 730 4.5 1.61 L-T4 9 A1060C Homozygous T354P TM IX
2 1958 M Japan 5 yr 13 yr (FA) Retarded ⫹ 40 25 1.0 0.94 Thyroid powder 9 A1060C Homozygous T354P TM IX
3 1968 M Japan 3 yr 8 yr Retarded ⫺ 40 27 0.41 71 1.2 1.1 Thyroid powder 9 A1060C/? Heterozygous T354P/? TM IX
4 1966 F Japan 10 yr 10 yr Retarded ⫺ 40 36 1.42 110 4.8 0.93 Thyroid powder 9 A1060C/? Heterozygous T354P/? TM IX
5 1963 F Japan 13 yr 13 yr Normal ⫺ 40 12 0.46 300 1.2 1.2 9 A1060C/? Heterozygous T354P/? TM IX
6 1937 M Japan Euthyroid 18 yr Normal ⫹ 44, 55, 70 69 1.06 3.3 2.5 1.5 L-T4 9 A1060C Homozygous T354P TM IX
7 1942 F Japan Euthyroid 20 yr Normal ⫹ 55 54 1.4 1.3 2.0 1.1 L-T4 9 A1060C Homozygous T354P TM IX
8 1975 F Japan 6 yr 6 yr Retarded ⫹ 46, 55 91 2.35 13.3 2.1 1.0 12.6 mg I/day 9 A1060C Homozygous T354P TM IX
9 1978 M Japan 1 yr None Retarded ⫹ 46, 55 ⬍10 0.61 148 1.3 1.0 12.6 mg I/day 9 A1060C Homozygous T354P TM IX
10 1971 M Japan 8 mo 21 yr Retarded ⫺ 55 11 1.06 37.5 3.0 1.5 1 mg I/day 9 A1060C Homozygous T354P TM IX
11 1989 M Japan 1.5 yr 1.5 yr Retarded ⫹ 55 1.2 1.25 ⬎100 1.4 2.6 L-T4 9 A1060C Homozygous T354P TM IX
12 1974 F Japan 4 mo None Retarded ⫺ 55, 71 5 0.46 430 2.4 1.8 L-T4 9 A1060C Homozygous T354P TM IX
13 1947 M Japan 22 yr 16 yr Normal ⫺ 54, 91, 126 10 0.39 217 0.9 0.7 189 mg I/day 1 G277C Heterozygous G93R TM III
9 A1060C Heterozygous T354P TM IX
14 1984 F Japan 3 yr 10 yr Retarded ⫺ 54 9 0.8 1,044 4.0 1.0 L-T4 13 G1628A Homozygous G543E TM XIII
15 1985 F Japan 2 yr 9 yr Retarded ⫺ 54 4 0.31 484 3.6 1.0 L-T4 13 G1628A Homozygous G543E TM XIII
16 1958 M Brazil 3 mo 3 mo Normal ⫺ 15, 85 37 104 1.0 3.0 12 mg I/day 6 C1163A Homozygous C272X Loop ÷ TM
VII–VIII
6 C1146G Heterozygous Q267E Loop ÷ TM
VII–VIII
17 1958 F USA Neonatal 12 yr (FA) Normal ⫺ 86 67 142 3.0 3 L-T4 13 C1940G Heterozygous Y531X TM XIII
and/or Alt. Spliced 515X Loop ÷ TM
XII–XIII
MOLECULAR ANALYSIS OF THE SODIUM/IODIDE SYMPORTER
S/P, salivary to plasma ratio of I; FA, follicular adenoma; empty box, test not done or unknown; Cons, parental consanguinity; TRIU, thyroid radioiodide uptake (%); T4, thyroxine; T3, triiodothyronine; TSH,
thyrotropin; TM, transmembrane segment. Patients 3–5 only have been screened for the T354P mutation.
1099

mutation. Whenever the patients’ parents were analyzed, segments VII and VIII (Fig. 6), and no I⫺ uptake activity
they were heterozygous for the T354P mutation and had was detected after transfecting mutant NIS cDNA into
no obvious thyroid disorders. The first case (case 6) was COS-7 cells. The patient was homozygous for the muta-
previously described by the same group (see above) (70). tion. His parents were not consanguineous; his mother,
Cases 6 and 7, siblings, were euthyroid due to an ex- son, and a paternal aunt were heterozygous for C272X,
tremely high I⫺ diet. They presented large diffuse goiters and his father was not investigated. All of the carriers
at ages 18 and 20, respectively. In cases 9 and 12, hypo- were euthyroid with normal or slightly enlarged thyroid
thyroidism was diagnosed early so that L-T4 treatment was glands. The patient’s goiter was noted at 3 mo of age and
started and goiter development was prevented. The rest treated with L-T4, but examination after several years
of the cases had different degrees of diffuse goiter and revealed low T4 and high TSH levels. A nodule developed
hypothyroidism. NIS protein determinations were carried in each thyroid lobe. Subsequent treatment with high I⫺
out in cases 6 and 10 and were found to be increased restored the patient to the euthyroid state.
⬃10-fold with respect to normal tissue. Immunohisto- Case 17 in Table 1 was reported by Pohlenz et al.
chemical staining of NIS in these two patients indicated (86). This was a patient of Hispanic ancestry who carried
that NIS was overexpressed and properly localized in the a compound heterozygous mutation. From the paternal
basolateral plasma membrane of the thyroid cells (55). allele she presented a substitution of cytosine for guani-
The same group has subsequently reported three pa- dine in nucleotide 1146 in exon 6 that resulted in a Gln for

tients with ITD from two families, presenting new muta- Glu amino acid substitution (Q267E) in NIS. This muta-
tions of the NIS gene (54) (Table 1, cases 13–15). Case 13 tion is located between the putative transmembrane seg-
had already been reported (91) and reviewed earlier by ments VII and VIII of NIS (Fig. 7). COS cells transfected
Wolff (126). This patient carried compound heterozygous with the mutated cDNA did not exhibit I⫺ transport ac-
mutations. From the paternal allele he presented a DNA tivity. The patient’s brother, father, and a paternal uncle
substitution of guanidine with cytosine in the 277 nucle- and aunt were heterozygous for the mutation, and they
otide in exon 1. From the maternal allele he presented a were clinically unaffected. From the maternal allele she
substitution of adenine with cytosine in nucleotide 1060 presented a substitution of cytosine with guanidine in
in exon 9. These resulted in the compound G93R/T354P nucleotide 1940 in exon 13. This point mutation has a
mutation, affecting the putative transmembrane segments double effect. First, it creates a stop codon at amino acid
III and IX of NIS (Fig. 7). The patient’s father was het- 531 (Y531X) with the consequent deletion of the entire
erozygous for the G93R mutation and his mother for the hydrophilic COOH terminus (Fig. 7). Second, it generates
T354P mutation, both without thyroid disorders. They a new 3⬘-acceptor splice site for intron 12 that produces a
were not consanguineous. The patient had developed dif- 67-nucleotide deletion in exon 13, rendering a protein
fuse goiter at age 15, and he had I⫺ responsive hypothy- with only 515 amino acids that has a deleted part of the
roidism. putative transmembrane segment XIII and of the COOH
Cases 14 and 15 are siblings. They carried the ho- terminus of NIS. The in vitro activity of these truncated
mozygous substitution of guanidine for adenine in the proteins was not evaluated. None of the heterozygous
1,628 nucleotide in exon 13. The resulting G543E muta- maternal members was affected. This patient was diag-
tion is located in the putative transmembrane segment nosed with hypothyroidism during neonatal screening
XIII of NIS (Fig. 7). The patients’ parents, who were not and treated with L-T4. At 12 yr of age she was diagnosed as
consanguineous, were heterozygous for G543E, without having ITD. She presented a nodule that had not been
thyroid disorders. The patients’ younger sister did not observed 1 yr before. Histological investigation revealed a
carry the mutation. Neither G93R, T354P, G93R/T354P, follicular adenoma.
nor G543E mutant cDNA transiently transfected into In vitro experiments in cells transfected with the
COS-7 displayed any I⫺ transport activity (54). Neverthe- various NIS mutants found in ITD patients have shown
less, these mutations were not characterized at the pro- that the mutant NIS proteins do not elicit I⫺ transport.
tein level so that the possibility that these mutations However, the precise structural change brought about by
resulted in a nonfunctional protein or one not properly a NIS mutation has only been determined for T354P.
targeted to the plasma membrane remains open. Therefore, it remains to be determined whether the other
Only two ITD cases characterized at the molecular NIS mutations give rise to proteins that are nonfunctional
level are not from Japan (85, 86) (Table 1, cases 16 and but properly targeted to the plasma membrane, proteins
17). The first one (Table 1, case 16), from Brazil, was also that are rapidly degraded, or retained in intracellular or-
reviewed by Camargo et al. (15). The patient presented a ganelles. It is unknown how I⫺ enters the thyroid follic-
substitution of cytosine 1163 with adenine in exon 6, ular cells in these patients. However, a high I⫺ diet has
resulting in a premature stop codon at position 272 been shown to be able to maintain some of these patients
(C272X). The mutant NIS protein is truncated in the hy- in a euthyroid state. It is possible that thyroidal I⫺ trans-
drophilic segment between the putative transmembrane port in these patients occurs by “simple diffusion” or by
nonspecific channels, as suggested by Wolff (126). More- cases (cases 6 and 7) remained euthyroid. Most probably,
over, Cl⫺ channels have been shown to be able to trans- this variability reflects the roles played by multiple factors
port other halides with different affinities, and in some in the onset and evolution of ITD due to NIS mutations,
cases, these channels display a considerable affinity for I⫺ including the type of mutation, time of detection of the
(26, 27, 52). Administration of high doses of potassium condition, time and type of administered therapy, and I⫺
iodide (KI) (1–200 mg/day) has been reported to maintain content in the diet. In any case, the considerable strides
these patients euthyroid, without apparent long-term side made in this area of research in recent years illustrate the
effects. powerful impact that detailed function/structure studies
The inheritance pattern of ITD is autosomal recessive of this key molecule can have on health and disease.
in all cases. In the investigated families, the heterozygous
carriers were always clinically unaffected, and only a few
of them presented slight goiter. Nevertheless, cases 3–5 in IX. CONCLUDING REMARKS
Table 1, reported by Fujiwara et al. (40), were heterozy-
gous for T354P. However, other mutations could have The landscape of NIS research has been dramatically
been present in these patients and missed in the study transformed in a very short time. Just six years ago, in
because only T354P was screened. 1993, the following was written in a review on “Iodide
Most of the patients suffering from this condition Transport in the Thyroid Gland” (16): “The chemical na-

have been from Japan. The high incidence of ITD in the ture of the putative I⫺ carrier has been the subject of
Japanese population could be related to the high I⫺ diet some debate in the past. At one time, phospholipids were
prevalent in Japan, which provides an adequate I⫺ supply considered potential I⫺ carriers as much as proteins, but
even for ITD patients, and allows them to maintain ac- it has long been clear that transport of ions and small
ceptable thyroid hormone synthesis. In a few ITD cases, molecules (such as carbohydrates and amino acids)
expression and activity of the other TSH-regulated thy- across cell membranes is mediated by intrinsic membrane
roid-specific proteins (TPO and Tg) were also investi- proteins, not phospholipids.” Later in the same article, it
gated. Low or normal levels of Tg protein were observed was stated that “the Na⫹/I⫺ symporter molecule has not
in the cases that were studied [Table 1, cases 13 and 16; been fully characterized yet, as its size, number of sub-
and 5 more cases reported by Wolff (126)]. In case 16, Tg units, and amino acid sequence remain unknown,” and
was poorly iodinated, but the patient was euthyroid as a with respect to NIS the article concludes by saying that
result of a high I⫺ diet. This indicates that under certain “there is no question that our understanding of I⫺ trans-
circumstances “non-NIS mediated transport” is able to location processes, both in the thyroid and in other tis-
supply enough I⫺ into the follicle to ensure hormonogen- sues, will be greatly enhanced when this molecule is fully
esis. Furthermore, Lamas et al. (58) have shown that the identified and characterized. Once this is achieved, insight
efficiency of hormonogenesis is determined by the avail- may subsequently be gained into various important re-
able amount of I⫺. Lamas et al. (57) also reported that lated research fields, given the relevance of I⫺ transport
under conditions of low I⫺ intake the synthesis of thyroid to Na⫹-dependent symport mechanisms, thyroid physiol-
hormones on the Tg molecule occurred at the main hor- ogy and pathophysiology, hormone action mechanisms,
monogenic site, and Passler et al. (83) showed that this and cell differentiation.” The present review, in fact, fo-
utilization was regulated by TSH. cuses on reports that started to appear shortly after pub-
In two of the patients reported by Wolff (126), TPO lication of the quoted article, covering the isolation of a
activity was normal or slightly increased. In case 16 (15), cDNA encoding rat NIS, the preparation of anti-NIS anti-
the activity and immunostaining of TPO were threefold bodies to study NIS topology and its secondary structure,
with respect to normal thyroid tissue, probably due to the the examination of the biogenesis and posttranslational
high TSH levels. These findings suggest that only NIS is modifications of NIS, its electrophysiological analysis, the
affected in ITD patients, whereas Tg and TPO expression isolation of the cDNA encoding hNIS, the elucidation of
and activity remain intact despite the upregulated TSH the genomic organization of hNIS, the analysis of NIS
levels. Anti-Tg, anti-TPO, and anti-TSHr autoantibodies regulation by TSH and I⫺, and the regulation of NIS
were negative when tested in all these patients. transcription, among several other topics. It also dis-
Thyroid morphological findings are fairly heteroge- cusses spontaneous NIS mutations that have been identi-
neous, even in patients carrying the same NIS mutation. fied as causes of congenital ITD resulting in hypothyroid-
Among the patients in which NIS mutations have been ism, the roles of NIS in thyroid cancer and thyroid
characterized at the molecular levels (as summarized in autoimmune disease, and the expression and regulation
Table 1), we find 2 cases without goiter (cases 9 and 12), of NIS in extrathyroidal tissues. Perhaps most signifi-
11 cases with small or large diffuse goiter (cases 3– 8, 10, cantly, the above accomplishments have made it possible
11, and 13–15), 1 case with nodular goiter (case 16), and to carry out gene therapy experiments in which the rat
3 cases with follicular adenoma (cases 1, 2, and 17). Two NIS gene has been transduced into various types of hu-
man cells, which then exhibited active iodide transport 14. CAILLOU B, TROALEN F, BAUDIN E, TALBOT M, FILETTI S,
SCHLUMBERGER M, AND BIDART JM. Na⫹/I⫺ symporter distribu-
and became susceptible to destruction with radioiodide. tion in human thyroid tissues: an immunohistochemical study.
Hence, the impact of the continued molecular analysis of J Clin Endocrinol Metab 83: 4102– 4106, 1998.
NIS has already reached farther than predicted a few 15. CAMARGO RYA, GROSS JL, SILVEIRO SP, KNOBEL M, AND ME-
DEIROS-NETO G. Pathological findings in dyshormonogenetic goi-
years ago, given that it extends not only to structure/ ter with defective iodide transport. Endocr Pathol 9: 225–233, 1998.
function of transport proteins, thyroid pathophysiology, 16. CARRASCO N. Iodide transport in the thyroid gland. Biochim
hormone action mechanisms, and cell differentiation, but Biophys Acta 1154: 65– 82, 1993.
17. CATUREGLI, P, HEJAZI M, SUZUKI K, DOHAN O, CARRASCO N,
also, quite possibly, to the diagnosis and treatment of KOHN LK, AND ROSE NR. Hypothyroidism in transgenic mice ex-
cancer, both in the thyroid and beyond. pressing IFN-gamma in the thyroid. Proc Natl Acad Sci USA 97:
1719 –1724, 2000.
We are grateful to Drs. U. Tazebay, C. Riedel, and C. Ginter 18. CHENG SH, GREGORY RJ, MARSHALL J, SUCHARITA P, SOUZA
for critical reading of the manuscript. DW, WHITE GA, O’RIORDAN CR, AND SMITH A. Defective intra-
A. De La Vieja was supported in part by Ministry of Edu- cellular transport and processing of CFTR is the molecular basis of
most cystic fibrosis. Cell 63: 827– 834, 1990.
cation and Science of Spain Grant PF 97 52094152. This project 19. COMB M, MERMOD N, HYMAN SE, PEARLBERG J, ROSS ME, AND
was supported by National Institute of Diabetes and Digestive GOODMAN HM. Proteins bound at adjacent DNA elements act
and Kidney Diseases Grant DK-41544 (to N. Carrasco). synergistically to regulate human proenkephalin cAMP inducible
Address for reprint requests and other correspondence: N. transcription. EMBO J 7: 3793–3805, 1988.
20. DAI G, LEVY O, AMZEL LM, AND CARRASCO N. The mediator of
Carrasco, Dept. of Molecular Pharmacology, Albert Einstein
thyroidal iodide accumulation: the sodium/iodide symporter. In:

College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461 Handbook of Biological Physics. Transport Processes in Eukary-
(E-mail: carrasco@aecom.yu.edu). otic and Prokaryotic Organisms, edited by W. N. Konings, H. R.
Kaback, and J. S. Lolkema. Amsterdam: Elsevier, 1996, vol. II, p.
343–367.
REFERENCES 21. DAI G, LEVY O, AND CARRASCO N. Cloning and characterization of
the thyroid iodide transporter. Nature 379: 458 – 460, 1996.
1. ABRAMOWICZ MJ, DUPREZ L, PARM J, VASSART G, AND HEIN- 22. DAMANTE G AND DI LAURO R. Thyroid-specific gene expression.
RICHS C. Familial congenital hypothyroidism due to inactivating Biochim Biophys Acta 1218: 255–266, 1994.
mutation of the thyrotropin receptor causing profound hypoplasia 23. DEGROOT LJ (Editor). Endocrinology. Orlando, FL.: Grune &
of the thyroid gland. J Clin Invest 99: 3018 –3024, 1997. Stratton, 1995.
2. ABRAMOWICZ MJ, TARGONIK HM, VARELA V, COCHAUX P, 24. DELANGE F. The disorders induced by iodine deficiency. Thyroid
KRAWIEC L, AND PISAREV MA. Identification of a mutation in the 4: 107–128, 1994.
coding sequence of the human thyroid peroxidase gene causing 25. DE LA VIEJA A, GINTER CS, AND CARRASCO N. Topology of the
congenital goiter. J Clin Invest 90: 1200 –1204, 1992. sodium/iodide symporter (Abstract). In: Proceedings of the AAPS
3. AJJAN RA, FINDLAY C, METCALFE RA, WATSON PF, CRISP M, Frontier Symposium: Membrane Transporters and Drug Ther-
LUDGATE M, AND WEETMAN AP. The modulation of the human apy. Bethesda, MD: NIH, 1999. p. 49.
sodium iodide symporter activity by Graves sera. J Clin Endocri- 26. DEVIDAS S AND GUGGINO WB. CFTR: domains, structure, and
nol Metab 83: 1217–1221, 1998. function. J Bioenerg Biomembr 29: 443– 451, 1997.
4. AJJAN RA, KAMARUDDIN NA, CRISP M, WATSON PF, LUDGATE 27. DEVIDAS S, YUE H, AND GUGGINO WB. The second half of the
M, AND WEETMAN AP. Regulation and tissue distribution of the cystic fibrosis transmembrane concductance regulator forms a
human sodium iodide symporter gene. Clin Endocrinol 49: 517– functional chloride channel. J Biol Chem 273: 29373–29380, 1998.
523, 1998. 28. DUGRILLON, A. Iodolactones and iodoaldehydes: mediators of
5. AJJAN RA, WATSON PF, FINDLAY C, METCALFE RA, CRISP M, iodine in thyroid autoregulation. Exp Clin Endocrinol Diabetes
LUDGATE M, AND WEETMAN AP. The sodium iodide symporter 104, Suppl 4: 41– 45, 1996.
gene and its regulation by cytokines found in autoimmunity. J 29. ENDO T, KANESHIGE M, NAKAZATO M, KOGAI T, SAITO T, AND
Endocrinol 158: 351–358, 1998. ONAYA T. Autoantibody against thyroid iodide transporter in the
6. ALEXANDER WD AND WOLFF I. Cation requirements for iodide sera from patients with Hashimoto’s thyroiditis possesses iodide
transport Arch Biochem Biophys 106: 525–528, 1964. transport inhibitory activity. Biochem Biophys Res Commun 228:
7. ARTURI F, RUSSO D, SCHLUMBERGER M, DUVILLARD JA, AND 199 –202, 1996.
CAILLOU B. Iodide symporter gene expression in human thyroid 30. ENDO T, KANESHIGE M, NAKAZATO M, OHMORI M, HARRI N,
tumors. J Clin Endocrinol Metab 83: 2493–2496, 1998. AND ONAYA T. Thyroid transcription factor-1 activated the pro-
8. AZA-BLANZ P, DI LAURO R, AND SANTISTEBAN P. Identification moter activity of rat Na⫹/I⫺ symporter gene. Mol Endocrinol 11:
of a cis-regulatory element and a thyroid-specific factor mediating 1747–1755, 1997.
the hormonal regulation of rat thyroid peroxidase promoter activ- 31. ENDO T, KOGAI T, NAKAZATO M, SAITO T, KANESHIGE M, AND
ity. Mol Endocrinol 7: 1297–1306, 1993. ONAYA T. Autoantibody against Na⫹/I⫺ symporter in the sera of
9. BAGCHI N AND FAWCETT DM. Role of sodium ion in active trans- patients with autoimmune thyroid disease. Biochem Biophys Res
port of iodide by cultured thyroid cells. Biochim Biophys Acta 318: Commun 224: 92–95, 1996.
235–251, 1973. 32. ENDO T, OHNO M, KOTANI S, GUNJI K, AND ONAYA T. Thyro-
10. BEHR M, SCHMITT TL, ESPINOZA CR, AND LOOS U. Cloning of a tropin receptor in nonthyroid tissues. Biochem Biophys Res Com-
functional promoter of the human sodium/iodide symporter gene. mun 190: 774 –779, 1993.
Biochem J 331: 359 –363, 1998. 33. ENG PHK, CARDONA GR, FANG SL, ALEX S, CARRASCO N, CHIN
11. BIKKER H, VULSMA T, BAAS F, AND DE VIJLDER JJM. Identifica- WW, AND BRAVERMAN LE. “Escape from the acute Wolff-Chaikoff
tion of five novel inactivating mutations in the human thyroid effect is associated with a decrease in thyroid sodium/iodide sym-
peroxidase gene by denaturing gradient gel electrophoresis. Hum porter (NIS) expression.” Endocrinology. In press.
Mutat 6: 9 –16, 1995. 34. ESKANDARI S, LOO DDF, DAI G, LEVY O, WRIGHT EW, AND
12. BRAVERMAN, LE AND INGBAR SH. Changes in thyroidal function CARRASCO N. Thyroid Na⫹/I⫺ symporter: mechanism, stoichiom-
during adaptation to large doses of iodide. J Clin Invest 42: 1216 – etry, and specificity. J Biol Chem 272: 27230 –27238, 1997.
1231, 1963. 35. ESPOSITO C, MICCADEI S, SAIARDI A, AND CIVITAREALE D.
13. BROWN-GRANT K. Extra thyroidal iodide concentrating mecha- Pax8 activates the enhancer of the human thyroperoxidase gene.
nisms. Physiol Rev 41: 189 –213, 1961. Biochem J 331: 37– 40, 1998.
36. FEDERMAN D, ROBBINS J, AND RALL JE. Some observations on 57. LAMAS L, ANDERSON PC, FOX JW, AND DUNN JT. Consensus
cretinism and its treatment. N Engl J Med 259: 610 – 616, 1958. sequences for early iodination and hormonogenesis in human thy-
37. FISFALEN ME AND DEGROOT LJ. Graves disease and autoimmune roglobulin. J Biol Chem 264: 13541–13545, 1989.
thyroiditis. In: Molecular Endocrinology: Basic Concepts and Clin- 58. LAMAS L, SANTISTEBAN P, TURMO C, AND SEGUIDO AM. Evi-
ical Correlations, edited by B. D. Weintraub. New York : Raven, dence, by in vitro enzymatic iodination of thyroglobulin, that the
1995, p. 319 –369. efficiency of coupling is determined by the initial iodide concen-
38. FOLEY KP, LEONARD MW, AND ENGEL JD. Quantitation of RNA tration. Endocrinology 118: 2131–2136, 1986.
using the polymerase chain reaction. Trends Genet 9: 380 –385, 59. LAZZARO D, PRICE M, FELICE D, AND DI LAURO R. The transcrip-
1993. tion factor TTF-1 is expressed at the onset of thyroid and lung
39. FUJIWARA H, TATSUMI K, MIKI K, HARADA T, MIYAI K, TAKAI S, morphogenesis and in restricted regions of the fetal brain. Devel-
AND AMINO N. Congenital hypothyroidism caused by a mutation in opment 113: 1093–1104, 1991.
the Na⫹/I⫺ symporter. Nature Genet 16: 124 –125, 1997. 60. LEIRI T, COCHAUX P, TARGOVNIK HM, SUZUKI M, SHIMODA SI,
40. FUJIWARA H, TATSUMI K, MIKI K, HARADA T, OKADA S, NOSE AND PERRET J. A 3⬘ splice site mutation in the thyroglobulin gene
O, KODAMA S, AND AMINO N. Recurrent T354P mutation of the responsible for congenital goiter with hypothyroidism. J Clin In-
Na⫹/I symporter in patients with iodide transport defect. J Clin vest 88: 1901–1905, 1991.
Endocrinol Metab 83: 2940 –2943, 1998. 61. LEVY O AND CARRASCO N. Structure and function of the thyroid
41. GERARD CM, LEFORT A, LIBERT F, CHRISTOPHE D, DUMONT iodide transporter and its implications for thyroid disease. Curr
JE, AND VASSART G. Transcriptional regulation of the thyroperoxi- Opin Endocr Diabetes 4: 364 –370, 1997.
dase gene by thyrotropin and forskolin. Mol Cell Endocrinol 60: 62. LEVY O, DAI G, RIEDEL C, GINTER CS, PAUL EM, LEBOWITZ AN,
⫹ ⫺
239 –242, 1988. AND CARRASCO N. Characterization of the thyroid Na /I sym-
42. HALMI NS. Thyroidal iodide transport. Vitam Horm 19: 133–163, porter with an anti-COOH terminus antibody. Proc Natl Acad Sci
1961. USA 94: 5568 –5573, 1997.
43. HALMI NS, NISSEN WM, AND SCRANTON JR. The kinetics of the 63. LEVY O, DE LA VIEJA A, AND CARRASCO N. The Na⫹/I⫺ symporter

enhancement of thyroid iodide accumulation after actinomysin D (NIS): recent advances. J Bioenerg Biomembr 30: 195–206, 1998.
administration. Endocrinology 84: 943–945, 1969. 64. LEVY O, DE LA VIEJA A, GINTER CS, DAI G, RIEDEL C, AND
44. HAMADA S, MATSUMURA T, AND YAWATA M. A case of iodide CARRASCO N. N-linked glycosylation of the thyroid Na⫹/I⫺ sym-
concentration disorder thyroid disease accompanied by citrullin- porter (NIS): implications for its secondary structure model. J Biol
emia. Nippon Riosho 32: 2439 –2442, 1974. Chem 273: 22657–22663, 1998.
45. IFF HW AND WILLBRANDT W. Die Abhengigkeit der Jodakkumu- 65. LEVY O, GINTER CS, DE LA VIEJA A, LEVY D, AND CARRASCO N.
lation in Schilddrüsenschnitten von der ionalen Zusammensetzung Identification of a structural requirement for thyroid Na⫹/I⫺ sym-
des Inkubationsmediums und ihre Beeinflussung durch Herzglyco- porter (NIS) function from analysis of a mutation that causes
side. Biochim Biophys Acta 78: 711–752, 1963. human congenital hypothyroidism. FEBS Lett 429: 36 – 40, 1998.
46. INOMATA H, TAMRU K, SATO H, SASAKI N, NIIMI H, AND NAKA- 66. LUDGATE M, CRISP M, LANE C, COSTAGLIOLA S, VASSART G,
JIMA H. Two sibbigs of absence of iodide-concentrating mecha- WEETMAN A, DAUNERIE C, AND MANY MC. The thyrotropin
nism. Nippon Shonika Gakkai Zasshi 92: 2383–2388, 1988. receptor in thyroid eye disease. Thyroid 8: 411– 413, 1998.
47. JANSEN E, AYOUBI TAY, MEULEMAN SMP, AND VAN DE VEN. 67. MANDELL RB, MANDEL LZ, AND LINK CJ. Radioisotope concen-
Cell type-specific protein-DNA interactions at the cAMP response trator gene therapy using the sodium/iodide symporter gene. Can-
elements of the prohormone convertase 1 promoter. J Biol Chem cer Res 59: 661– 668, 1999.
272: 2500 –2508, 1997. 68. MARINE D AND FEISS HO. The absorption of potassium iodide by
48. JHIANG SM, CHO J, RYU KY, DEYOUNG BR, SMANIK PA, MC- perfused thyroid glands and some of the factors modifying it.
GAUGHY VR, FISCHER AH, AND MAZZAFERRI EL. An immunohis- J Pharmacol Exp Ther 7: 557–576, 1915.
tochemical study of Na⫹/I⫺ symporter in human thyroid tissues 69. MARTIN, GM, TURK E, LOSTAO MP, KERNER C, AND WRIGHT
and salivary gland tissues. Endocrinology 139: 4416 – 4419, 1998. EM. Defects in Na⫹/glucose cotransporter (SGLT1) trafficking and
49. KAMBE F, NOMURA Y, OKAMOTO T, AND SEO H. Redox regula- function caused glucose galactose malabsorption. Nature Genet 12:
tion for thyroid-transcription factors, Pax-8 and TTF-1, is involved 216 –220, 1996.
in their increased DNA-binding activities by thyrotropin in rat 70. MATSUDA A AND KOSUGI S. A homozygous missense mutation of
thyroid FRTL-5 cells. Mol Endocrinol 10: 801– 812, 1996. the sodium/iodide symporter gene causing iodide transport defect.
50. KAMINSKY SM, LEVY O, GARRY MT, AND CARRASCO N. Inhibition J Clin Endocrinol Metab 82: 3966 –3971, 1997.
of the Na⫹/I⫺symporter by harmaline and TRP-P2 in thyroid cells 71. MATSURA M, NISHIHATA N, KONDO M, ZENSAK N, AND SUWA S.
and membrane vesicles. Eur J Biochem 200: 203–207, 1991. An infant case of goitrous hypothyroidism due to iodide concen-
51. KAMINSKY SM, LEVY O, SALVADOR C, DAI G, AND CARRASCO N. tration disorders. Clin Endocrinol 23: 2383–2388, 1975.
Na⫹/I⫺ symport activity is present in membrane vesicles from 72. MEDEIROS-NETO G, TARGOVNIK HM, AND VASSART G. Defective
TSH-deprived non-I⫺ transporting cultured thyroid cells. Proc Natl thyroglobulin synthesis and secretion causing goiter and hypothy-
Acad Sci USA 91: 3789 –3793, 1994. roidism. Endocr Rev 14: 165–183, 1993.
52. KATAYAMA Y, AND WIDDICOMBE JH. Halide transport in Xeno- 73. MISSERO C, COBELLIS G, DE FELICE M, AND DI LAURO R.
pus oocytes. J Physiol (Lond) 443: 587–599, 1991. Molecular events involved in differentiation of thyroid follicular
53. KOGAI T, ENDO T, SAITO T, MIYAZAKI A, KAWAGUCHI A, AND cells. Mol Cell Endocrinol 141: 37– 43, 1998.
ONAYA T. Regulation by thyroid-stimulating hormone of sodium/ 74. MORRIS JC, BERGERT ER, AND BRYANT WP. Binding of immu-
iodide symporter gene expression and protein levels in FRTL-5 noglobulin G from patients with autoimmune thyroid disease to rat
cells. Endocrinology 138: 2227–2232, 1997. sodium-iodide symporter peptides: evidence for the iodide trans-
54. KOSUGI S, INOUE S, MATSUDA A, AND JHIANG S. Novel, missense porter as an autoantigen. Thyroid 4: 527–534, 1997.
and loss-of-function mutations in the sodium/iodide symporter 75. MORTON ME, CHAIKOFF IL, AND ROSENFELD S. Inhibiting effect
gene causing iodide transport defect in three Japanese patients. of inorganic iodide on the formation in vitro of thyroxine and
J Clin Endocrinol Metab 83: 3373–3376, 1998. diiodotyrosine by surviving thyroid tissue. J Biol Chem 154: 381,
55. KOSUGI S, SATO Y, MATSUDA A, OHYAMA Y, FUJIEDA K, INO- 1944.
MATA H, KAMEYA T, ISOZAKI O, AND JHIANG S. High prevalence 76. OHMORI M, ENDO T, HARII N, AND ONAYA T. A novel thyroid
of T354P sodium/iodide symporter gene mutation in Japanese pa- transcription factor is essential for thyrotropin-induced up-regula-
tients with iodide transport defect who have heterogeneous clinical tion of Na⫹/I⫺ symporter gene expression. Mol Endocrinol 12:
pictures. J Clin Endocrinol Metab 83: 4123– 4129, 1998. 727–736, 1998.
56. KOTANI T, OGATA Y, YAMAMOTO I, ARATAKE Y, KAWANO JI, 77. OHMORI M, SHIMURA H, SHIMURA Y, IKUYAMA S, AND KOHN
SUGANUMA T, AND OHTAKI S. Characterization of gastric Na⫹/I⫺ LD. Characterization of an up-stream thyroid transcription factor-
symporter of the rat Clin. Immunol Immunopathol 89: 271–278, 1-binding site in the thyrotropin receptor promoter. Endocrinology
1998. 136: 269 –282, 1995.
78. OHNO M, ZANNINI M, LEVY O, CARRASCO N, AND DI LAURO R. 97. SCHLUMBERGER MJ. Papillary and follicular thyroid carcinoma.
The paired-domain transcription factor Pax8 binds to the upstream N Engl J Med 338: 297–306, 1998.
enhancer of the rat sodium/iodide symporter gene and participates 98. SCHMUTZLER C AND KÖHRLE J. Implications of the molecular
in both thyroid-specific and cyclic-AMP-dependent transcription. characterizations on the sodium-iodide symporter (NIS). Exp Clin
Mol Cell Biol 19: 2051–2060, 1999. Endocrinol Diabetes 106 Suppl 3: S1–S10, 1998.
79. O’NEILL B, MAGNOLATO D, AND SEMENZA G. The electrogenic, 99. SHIMURA H, MIYAZAKI A, HARAGUCHI K, ENDO T, AND ONAYA
Na⫹-dependent I⫺ transport system in plasma membrane vesicles T. Analysis of differentiation-induced expression mechanisms of
from thyroid glands. Biochim Biophys Acta 896: 263–274, 1987. thyrotropin receptor gene in adipocytes. Mol Endocrinol 12: 1473–
80. ORTIZ L, ZANNINI M, DI LAURO R, AND SANTISTEBAN P. Tran- 1486, 1998.
scriptional control of the forkhead transcription factor TTF-2 by 100. SHIMURA H, HARAGUCHI K, ENDO T, AND ONAYA T. Regulation
thyrotropin, insulin, and insulin-like factor 1. J Biol Chem 272: of thyrotropin receptor gene expression in 3T3–L1 adipose cells is
23334 –23339, 1997. distinct from its regulation in FRTL-5 thyroid cells. Endocrinology
81. PAIRE A, BERNIER-VALENTIN F, SELMI-RUBY S, AND ROUSSET 138: 1483–1490, 1997.
B. Characterization of the rat thyroid iodide transporter using 101. SHIMURA H, OKAJIMA F, IKUYAMA S, SHIMURA Y, KIMURA S,
anti-peptide antibodies. Relationship between its expression and SAJI M, AND KOHN LD. Thyroid-specific expression and cyclic
activity. J Biol Chem 272: 18245–18249, 1997. adenosine 3⬘,5⬘-monophosphate autoregulation of the thyrotropin
82. PANG XP, YOSHIMURE M, AND HERSHMEN JM. Suppression of receptor gene involves thyroid transcription factor-1. Mol Endocri-
rat thyrotroph and thyroid cell function by tumor necrosis factor ␣. nol 8: 1049 –1069, 1994.
Thyroid 3: 325–330, 1993. 102. SHIMURA H, SHIMURA Y, OHMORI M, IKUYAMA S, AND KOHN
83. PASSLER CA, DUNN JT, ANDERSON PC, FOX JW, DUNN AD, LD. Single strand DNA-binding proteins and thyroid transcription
HITE LA, MOORE RC, AND KIM PS. Thyrotropin alters utilization of factor-1 conjointly regulate thyrotropin receptor gene expression.
thyroglobulin’s hormonogenic sites. J Biol Chem 263: 17366, 1988. Mol Endocrinol 9: 527–539, 1995.
103. SHIMURA Y, SHIMURA H, OHMORI M, IKUYAMA S, AND KOHN

84. PIACHOW D, CHOWDHORY R, WALTHER C, SIMON D, GROOT
JL, AND CRUSE P. Pax8 a murine paired box gene expressed in the LD. Identification of a novel insulin-responsive element in the rat
developing excretory system an thyroid gland. Development 110: thyrotropin receptor promoter. J Biol Chem 269: 31908 –31914,
643– 651, 1990. 1994.
85. POHLENZ J, MEDEIROS-NETO G, GROSS JL, SILVEIRO SP, KNO- 104. SILVERBERG E AND LUBERA JA. Cancer statistics. CA cancer.
BEL M, AND REFETOFF S. Hypothyroidism in a Brazilian kindred J Clin Cancer Statistics 39: 3–20, 1989.
due to iodide trapping defect caused by a homozygous mutation in 105. SMANIK PA, LIU Q, FURMINGER TL, RYU K, XING S, MAZZA-
the sodium/iodide symporter gene. Biochem Biophys Res Commun FERRI EL, AND JHIANG SM. Cloning of the human sodium iodide
240: 488 – 491, 1997. symporter. Biochem Biophys Res Commun 226: 339 –345, 1996.
86. POHLENZ J, ROSENTHAL IM, WEISS RE, JHIANG SM, BURANT 106. SMANIK PA, RYU K, THEIL KS, MAZZAFERRI EL, AND JHIANG
SM. Expression, exon-intron organization and chromosome map-
C, AND REFETOFF S. Congenital hypothyroidism due to mutations
ping of the human sodium iodide symporter. Endocrinology 138:
in the sodium/iodide symporter. Identification of a nonsense mu-
3555–3558, 1997.
tation producing a downstream cryptic 3⬘ splice site. J Clin Invest
107. SPITZWEG C AND HEUFELDER AE. The sodium iodide symporter:
101: 1028 –1035, 1998.
its emerging relevance to clinical thyroidology. Eur J Endocrinol
87. RASMUSSEN AK, KAYSER L, RASMUSSEN UF, AND BENDTZEN
138: 374 –375, 1998.
K. Influence of tumor necrosis factor-␤ and interferon-␥, separately
108. SPITZWEG C, JOBA W, EISENMENGER W, AND HEUFELDER AE.
and added together with interleukin-1␤, on the function of cultured
Analysis of human sodium iodide symporter gene expression in
human thyroid cells. J Endocrinol 143: 359 –365, 1994.
extrathyroidal tissues and cloning of its complementary DNA from
88. RASPE E, COSTAGLIOLA S, RUF J, MARIOTTI S, DUMONT JE,
⫹ ⫺ salivary gland, mammary gland, and gastric mucosa J. Clin Endo-
AND LUDGATE M. Identification of the Na /I cotransporter as a
crinol Metab 83: 1746 –1751, 1998.
potential autoantigen in thyroid autoimmune disease. Eur J Endo- 109. STANBURY JB AND DUMONT JE. Familial goiter and related dis-
crinol 132: 399 – 405, 1995. orders. In: The Metabolic Basis of Inherited Disease, edited by J. B.
89. ROBBINS. Pathological Basis of Disease. Philadelphia, PA: Saun- Stanbury, J. B. Wyngaarden, D. S. Fredrickson, J. L. Goldstein, and
ders, 1994, p. 1133–1142. M. S. Brown. New York: McGraw-Hill, 1983, p. 231–269.
90. RYU KY, TONG Q, AND JHIANG SM. Promoter characterization of 110. SUNTHORNTHEPVARUKUI T, GOTTSCHALK ME, HAYASHI Y,
the human Na⫹/I⫺ symporter. J Clin Endocrinol Metab 83: 3247– AND REFETTOF S. Brief report: resistance to thyrotropin caused by
3251, 1998. mutations in the thyrotropin receptor gene. N Engl J Med 332:
91. SAITO K, YAMAMOTO K, YOSHIDA S, MANABE S, SUZUKI M, 155–160, 1995.
TAKAI T, SAITO T, KUZUYA T, AND MORIYAMA S. Goitrous hypo- 111. SWANSON DJ, ZELLMER E, AND LEWIS EJ. The homeodomain
thyroidism due to iodide-trapping defect. J Clin Endocrinol Metab protein Arix interacts synergistically with cAMP to regulate expres-
53: 1267–1272, 1981. sion of neurotransmitter biosynthetic genes. J Biol Chem 272:
92. SAITO T, ENDO T, KAWAGUCHI A, IKEDA M, KATOH R, KAAOI A 27382–27392, 1997.
MURAMATSU A, AND ONAYA T. Increased expression of the so- 111a.TAZEBAY UH, WAPNIR L, LEVY O, ZUCKIER L, PESTELL RG, AND
dium/iodide symporter in papillary thyroid carcinomas J Clin In- CARRASCO N. Hormonal regulation of the mammary gland Na⫹/I⫺
vest 101: 1296 –1300, 1998. symporter (mgNIS) (Abstract). In: Proceedings of the AAPS Fron-
93. SAJI M, AKAMIZU T, SANCHEZ M, OBICI S, AVVEDIMENTO E, tier Symposium: Membrane Transporters and Drug Therapy. Be-
GOTTESMAN ME, AND KOHN LD. Regulation of thyrotropin recep- thesda, MD: NIH, 1999, p. 45.
tor gene expression in rat FRTL-5 thyroid cells. Endocrinology 130: 112. TONG Q, RYU KY, AND JHIANG SM. Promoter characterization of
520 –533, 1992. the rat Na⫹/I⫺ symporter gene. Biochem Biophys Res Commun
94. SANTISTEBAN P, ACEBRON A, POLYCARPOU-SCHWARZ M, AND 239: 34 – 41, 1997.
DI LAURO R. Insulin and insulin-like factor regulated a thyroid- 113. TOUBLANC J. Comparison of epidemiological data on congenital
specific nuclear protein that binds to the thyroglobulin promoter. hypothyroidism in Europe with those of other parts of the world.
Mol Cell Biol 5: 1310 –1317, 1992. Horm Res 38: 230 –235, 1992.
95. SATO K AND DI LAURO R. Hepatocyte nuclear factor 3␤ partici- 113a.TURK E AND WRIGHT EM. Membrane topology motifs in the SGLT
pated in the transcriptional regulation of the thyroperoxidase pro- cotransporter family. J Membr Biol 159: 1–20, 1997.
moter. Biochem Biophys Res Commun 220: 86 –93, 1996. 114. TYLER P. China confronts retardation of millions who lack iodine.
96. SCHIMURA H, HARAGUCHI K, MIYAZAKI A, ENDO T, AND ONAYA New York Times June 4, 1996.
T. Iodide uptake and experimental 131iodide therapy in trans- 115. UYTTERSPROT N, PELGRIMS N, CARRASCO N, GERVY C,
planted undifferentiated thyroid cancer cells expressing the Na⫹/I⫺ MAENHAUT C, DUMONT JE, AND MIOT F. Moderate doses of
symporter gene. Endocrinology 138: 4493– 4496, 1997. iodide in vivo inhibit cell proliferation and the expression of thy-
roid peroxidase and the Na⫹/I⫺ symporter mRNAs in dog thyroid. Clinical Text, edited by L. E. Braverman and R. D. Utiger. Phila-
Mol Cell Endocrinol 131: 195–203, 1997. delphia, PA: Lippincott, 1991, p. 1–1362.
116. VAN HEUVERSWYN B, LERICHE A, VAN SANDE J, DUMONT J, 125. WOLFF J. Transport of iodide and other anions in the thyroid
AND VASSART G. Transcriptional control of thyroglobulin gene gland. Physiol Rev 44: 45–90, 1964.
expression by cyclic AMP. FEBS Lett 188: 192–196, 1985. 126. WOLFF J. Congenital goiter with defective iodide transport. En-
117. VASSART G, DESARNAUD F, DUPREZ L, EGGERICKX D, docr Rev 4: 240 –254, 1983.
LABBE F LIBERT O, MOLLEREAU C, PARMA J, PASCHKE R, 127. WOLFF J. Perchlorate and the thyroid gland. Pharmacol Rev 50:
AND TONACCHERA M. The G protein-coupled receptor family 89 –105, 1998.
and one of its members, the TSH receptor. Ann NY Acad Sci 766: 128. WOLFF J AND CHAIKOFF IL. Plasma inorganic iodide as a homeo-
23–30, 1995. static regulator of thyroid function. J Biol Chem 174: 555–560, 1948.
118. VENKATARAMAN MG, YATIN M, AND AIN KB. Cloning of the 129. WOLFF J, CHAIKOFF IL, GOLDBERG RC, AND MEIER JR. The
human sodium-iodide symporter promoter and characterization in temporary nature of the inhibitory action of excess iodide on
a differentiated human thyroid cell line, KAT-50. Thyroid 8: 63– 69, organic synthesis in the normal thyroid. Endocrinology 45: 504 –
1998. 513, 1949.
119. WARTOFSKY L, SHERMAN SI, GOPAL J, SCHLUMBERGER M, 130. YOSHIDA A, SASAKI N, MORI A, TANIGUCHI S, MITANI Y, UETA
AND HAY ID. The use of radioactive iodine in patients with papillary Y, HATTORI K, SATO R, HISATOME I, MORI T, SHIGEMASA C,
⫺
and follicular cancer. J Clin Endocrinol Metab 83: 4195– 4203, 1998. AND KOSUGI S. Different electrophysiological character of I ,
120. WEETMAN AP. The Na⫹/I⫺ cotransporter: a thyroid autoantigen? ClO⫺4 , and SCN
⫺
in the transport by Na⫹/I⫺ symporter. Biochem
Eur J Endocrinol 132: 397–398, 1995. Biophys Res Commun 231: 731–734, 1997.
121. WEETMAN AP. New aspects of thyroid immunity. Horm Res 48: 131. YOSHIDA A, SASAKI N, MORI A, TANIGUCHI S, UETA Y, HAT-
51–54, 1997. TORI K, TANAKA Y, IGAWA O, TSUBOI M, SUGAWA H, SATO R,
122. WEISS SJ, PHILIP NJ, AMBESI-IMPIOMBATO FS, AND GROLLMAN HISATOMEM I, SHIGEMASA C, GROLLMAN EF, AND KOSUGI S.
EF. Thyrotropin-stimulated iodide transport mediated by adeno- Differences in the electrophysiological response to I⫺ and the

sine 3⬘,5⬘-monophosphate and dependent on protein synthesis. En- inhibitory anions SCN⫺ and ClO⫺ 4 , studied in FRTL-5 cells. Bio-
docrinology 114: 1099 –1107, 1984. chim Biophys Acta 11: 231–237, 1998.
123. WEISS SJ, PHILIP NJ, AND GROLLMANN EF. Iodide transport in a 132. ZANINI M, AZA-BLANZ P, DI LAURO R, AND SANTISTEBAN P.
continuous line of cultured cells from rat thyroid. Endocrinology Identification of a cis-regulatory element and a thyroid-specific
114: 1090 –1098, 1984. factor mediating the hormonal regulation of rat thyroid peroxidase
124. WERNER SC AND INGBAR S. The Thyroid: A Fundamental and promoter activity. Mol Endocrinol 7: 1297–1306, 1993.
Molecular Analysis of the Sodium/Iodide Symporter:
Impact on Thyroid and Extrathyroid Pathophysiology
Antonio De la Vieja, Orsolya Dohan, Orlie Levy and Nancy Carrasco
Physiol Rev 80:1083-1105, 2000.
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PHYSIOLOGICAL REVIEWS

Vol. 80, No. 3, July 2000

Molecular Analysis of the Sodium/Iodide Symporter:

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I. INTRODUCTION by the Na⫹-K⫹-ATPase. The ability of the thyroid to ac-

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FIG. 2. Original and current NIS secondary struc-

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B. N-Linked Glycosylation of NIS: Implications for

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D. Several Hydroxyl-Containing Amino Acid

Levy et al. (65) have demonstrated that a hydroxyl

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gesting that it is not transported (34). Yoshida et al. (130)

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F. The NIS Inhibitor Perchlorate Is Not

Similar steady-state inward currents were generated

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B. Transcriptional Regulation of Tg and TPO C. Transcriptional Regulation of the TSHr

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Year Age of Detection TSH,

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You might find this additional info useful...

Influence of Vitamin C on Salivary Absorbed Dose of 131I in Thyroid Cancer Patients: A

Downloaded from physrev.physiology.org on March 26, 2011

Human Sodium/Iodide Symporter−Mediated Radioiodine Gene Therapy Enhances the

Subclinical Hypothyroidism in Korean Preterm Infants Associated with High Levels of

Targeted Radioiodine Therapy of Neuroblastoma Tumors following Systemic Nonviral

This infomation is current as of March 26, 2011.

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