The skeletal phenotypes of TRα and TRβ mutant mice

in Journal of Molecular Endocrinology
View More View Less
  • Molecular Endocrinology Group, Division of Medicine, Imperial College London, MRC Clinical Sciences Centre, Hammersmith Hospital, 5th Floor, Clinical Research Building, Du Cane Road, London W12 0NN, UK

Analysis of mice harbouring deletions or mutations of T3 receptor α (TRα) and β (TRβ) have clarified the complex relationship between central and peripheral thyroid status and emphasised the essential but contrasting roles of T3 in skeletal development and adult bone. These studies indicate that TRα1 is the predominant TR expressed in bone and that T3 exerts anabolic actions during growth but catabolic actions in the adult skeleton. Examination of key skeletal regulatory pathways in TR mutant mice has identified GH, IGF-1 and fibroblast growth factor signalling and the Indian hedgehog/parathyroid hormone-related peptide feedback loop as major targets of T3 action in chondrocytes and osteoblasts. Nevertheless, although increased osteoclastic resorption is a major feature of thyrotoxic bone loss and altered osteoclast activity is central to the skeletal phenotype of TR mutant mice, it remains unclear whether T3 has direct actions in osteoclasts. Detailed future analysis of the molecular mechanisms of T3 action in bone will enhance our understanding of this emerging field and has the potential to identify novel strategies for the prevention and treatment of osteoporosis.

Abstract

Analysis of mice harbouring deletions or mutations of T3 receptor α (TRα) and β (TRβ) have clarified the complex relationship between central and peripheral thyroid status and emphasised the essential but contrasting roles of T3 in skeletal development and adult bone. These studies indicate that TRα1 is the predominant TR expressed in bone and that T3 exerts anabolic actions during growth but catabolic actions in the adult skeleton. Examination of key skeletal regulatory pathways in TR mutant mice has identified GH, IGF-1 and fibroblast growth factor signalling and the Indian hedgehog/parathyroid hormone-related peptide feedback loop as major targets of T3 action in chondrocytes and osteoblasts. Nevertheless, although increased osteoclastic resorption is a major feature of thyrotoxic bone loss and altered osteoclast activity is central to the skeletal phenotype of TR mutant mice, it remains unclear whether T3 has direct actions in osteoclasts. Detailed future analysis of the molecular mechanisms of T3 action in bone will enhance our understanding of this emerging field and has the potential to identify novel strategies for the prevention and treatment of osteoporosis.

Introduction

Thyroid hormone is essential for normal skeletal development and the maintenance of adult bone mass. Recent studies of T3 receptor (TR) mutant mice have advanced our understanding and demonstrated the relevance of this field to osteoporosis, a major public health burden that affects half of all women and one-fifth of men over the age of 50, costing the European community €31 billion per annum (Kanis & Johnell 2005).

Systemic thyroid hormone levels are maintained by the hypothalamus–pituitary–thyroid (HPT) feedback axis. The cellular actions of 3,5,3′-l-triiodothyronine (T3) are mediated by TRs, which act as hormone-inducible transcription factors. Unliganded TRs bind T3 response elements (TREs) in T3-target genes and mediate transcriptional repression. T3 binding results in derepression and activation of gene transcription (Yen 2001). The THRA and THRB genes encode three functional receptors TRα1, TRβ1 and TRβ2 as well as a non-T3-binding isoform of unknown function TRα2 (Fig. 1). TRα1 and TRβ1 are expressed widely and the ratio of TRα1 to TRβ1 is spatio-temporally regulated. Thus, T3-target tissues may predominantly display either TRα1 or TRβ1 responsiveness or show no TR-isoform specificity. TRβ2 has a more restricted pattern of expression and regulates sensory organ development (Jones et al. 2007) as well as the HPT axis. In the skeleton, chondrocyte and osteoblast lineages express TRα and TRβ mRNAs, but in osteoclasts the position is less clear as studies have been restricted to precursor cells or in situ hybridisation analysis of osteoclastomas (Williams et al. 1994, Abu et al. 2000, Stevens et al. 2000, Kanatani et al. 2004). Several studies have demonstrated apparent expression of TR proteins in all bone cell lineages, but it is well recognised within the field that available TR antibodies are of low affinity, thus compromising the detection of endogenous TRs (Abu et al. 1997, Robson et al. 2000). For these reasons, a comprehensive understanding of TR expression in bone is lacking and a detailed analysis of cell-specific and temporal expression of TR isoforms is required.

Figure 1
Figure 1

Thyroid hormone receptor isoforms. (A) Schematic of the four functional domains of the thyroid hormone receptor. (B) Products of the Thra and Thrb genes in the mouse. Thra has two promoters, TRα1 and TRα2 are derived from the 5′ promoter whereas TRΔα1 and TRΔα2 are transcribed from an internal promoter in intron 7. TRα2 and TRΔα2 result from alternative splicing. TRα1 is a functional receptor whereas TRΔα1, TRα2 and TRΔα2 do not bind T3 and act as antagonists. Thrb also has two promoters and both N-terminal variants TRβ1 and TRβ2 bind T3 and act as functional receptors.

Citation: Journal of Molecular Endocrinology 42, 4; 10.1677/JME-08-0142

Long bones are formed by endochondral ossification and the skull by intramembranous ossification. During endochondral ossification, mesenchyme-derived chondrocytes form a cartilage model, undergo hypertrophic differentiation and then apoptose. The surrounding collagen X-rich cartilage matrix calcifies and forms a scaffold for bone formation by osteoblasts. Organised columns of proliferating and differentiating chondrocytes persist in the growth plate until adolescence and mediate linear growth and the acquisition of peak bone mass. By contrast, in intramembranous ossification, osteoblasts differentiate from mesenchyme to form bone directly. Adult bone structure and mechanical strength are preserved by a continuous process of skeletal remodelling during which precise coupling of osteoclastic bone resorption and subsequent osteoblastic bone formation is maintained.

The established view that skeletal responses to abnormal thyroid status result exclusively from altered T3 action in bone has been challenged recently by studies proposing a direct role for TSH in bone. We recently discussed this issue elsewhere (Bassett & Williams 2008) and the present review will therefore focus on the analysis of T3 and TR action in bone.

Thyroid hormone receptor mutant mice

Several TRα knockout mice have been generated and this led to the identification of additional TRα isoforms expressed from a promoter within intron 7 of the Thra gene (Chassande et al. 1997). As a result only TRα0/0 mice lack all TRα isoforms whereas other TRα mutants retain truncated isoforms with dominant-negative activity (Fig. 1 and Table 1). This review will focus on mice lacking all TRα (TRα0/0) or all TRβ (TRβ−/−) isoforms and those harbouring dominant-negative mutations of either TRα (TRα1PV/+, TRα1R384C/+) or TRβ (TRβPV/PV).

Table 1

Skeletal phenotypes of TH receptor mutant mice

GenotypeEndocrine statusJuvenile skeletonAdult skeletonSkeletal thyroid statusReferences
Model
TRα mutants
 TRα−/−No TRα1/α2 TRΔα1 and Δα2 preservedHypothyroid (T4 0.1×T3 0.4×, TSH 2×) GH normalSevere growth delay; delayed endochondral ossification; impaired chondrocyte differentiation; reduced mineralisationDie by weaning unless T3 treatedNRChassande et al. (1997), Fraichard et al. (1997) and Gauthier et al. (1999)
 TRα1−/−No TRα1/Δα1 TRα2/Δα2 preservedMild hypothyroidism (T4 0.7×T3 1×, TSH 0.8×)No growth retardation of skeletal phenotypeNRNRWikstrom et al. (1998)
 TRα2−/−No TRα2/Δα2 TRα1/Δα1 preserved; TRα1 over-expressionMild hypothyroidism (T4 0.8×T3 0.7×, TSH 1×) GH normal IGF1 lowNo growth retardation; normal endochondral ossificationReduced bone mineral density; reduced cortical boneNRSalto et al. (2001)
 TRα0/0No TRα transcripts TRβ unaffectedEuthyroid; normal GH and IGF1Transient growth delay; delayed endochondral ossification; impaired chondrocyte differentiation; reduced mineralisationOsteosclerosis; increased trabecular bone volume; reduced osteoclastic bone resorptionHypothyroidGauthier et al. (2001) and Bassett et al. (2007b)
 TRα1PV/+Heterozygous dominant-negative TRα receptorMild thyroid failure (T4 1× T3 1.2×, TSH 1.7×) GH normalSevere persistent growth retardation; delayed intramembranous and endochondral ossification; impaired chondrocyte differentiation; reduced mineralisationNRHypothyroidKaneshige et al. (2001) and O'Shea et al. (2005)
 TRα1R384C/+Heterozygous dominant-negative TRα receptor (10-fold reduced affinity for T3)Euthyroid adults; mild hypothyroidism P10-35 (T4 0.8× T3 0.7×, TSH 0.7×) (GH reduced in juveniles)Transient growth delay; delayed intramembranous and endochondral ossification; impaired chondrocyte differentiationOsteosclerosis; increased trabecular bone volume; increased mineralisation; reduced osteoclastic bone resorptionHypothyroidTinnikov et al. (2002) and Bassett et al. (2007a)
 TRα1R398H/+Heterozygous dominant-negative TRα receptorEuthyroid (T4 1.1× T3 1.1×, TSH 3.4×)NRNRNRLiu et al. (2003)
 TRα1AMI/+ SYCP1-CreCre-inducible dominant-negative receptor TRα1L400R (early global expression)Euthyroid (T4 1× T3 0.9×, TSH 0.3×) Reduced GH (0.4×)Severe persistent growth retardation; delayed endochondral ossificationNRNRQuignodon et al. (2007)
TRβ mutants
 TRβ−/−No TRβ transcripts TRα unaffectedRTH and goitre (T4 4× T3 4×, TSH 8×)1 (T4 3× T3 3×, TSH 2.6×)2Persistent short stature; advanced endochondral and intramembranous ossification; increased mineralisationOsteoporosis; reduced mineralisation; increased osteoclastic bone resorptionThyrotoxicForrest et al. (1996), 1Gauthier et al. (2001), 2Ng et al. (2001) and Bassett et al. (2007a,b)
 TRβ2−/−No TRβ2 TRβ1 preservedMild RTH (T4 3× T3 1.3×, TSH 2.5×)1 Small decrease in GH (T4 1× T3 1.5×, TSH 1.2×)2No reported growth abnormalityNRNR1Abel et al. (1999) and 2Ng et al. (2001)
 TRβPV/PVHomozygous dominant-negative TRβ receptorSevere RTH and goitre (T4 15× T3 9×, TSH >400×); reduced GHAccelerated prenatal growth; persistent post-natal growth retardation; advanced intramembranous and endochondral ossification; increased mineralisationNRThyrotoxicKaneshige et al. (2000), O'Shea et al. (2003, 2005) and Bassett et al. (2007a)
 TRβΔ377T/Δ377THomozygous dominant-negative TRβ receptorSevere RTH and goitre (T4 15× T3 10×, TSH >50×)No growth phenotypeNRNRHashimoto et al. (2001)
TRα and TRβ compound mutants
 TRα−/−β−/−No TRα1/α2 or TRβ TRΔα1/Δα2 preservedRTH and small goitre (T4 10× T3 10×, TSH >100×)Growth delay similar to TRα−/−; delayed endochondral ossification; impaired chondrocyte differentiation; reduced mineralisationDie at or near weaningNRGauthier et al. (1999) and Bassett et al. (2007b)
 TRα1−/−β−/−No TRα1/Δα1 or TRβ TRα2/Δα2 preservedRTH and large goitre (T4 60× T3 60×, TSH >160×); reduced GH/IGF1Persistent growth retardation; delayed endochondral ossification; reduced mineralisationReduced trabecular and cortical bone mineral density. GH treatment corrects growth but not ossification defectNRGothe et al. (1999), Kindblom et al. (2001), O'Shea et al. (2003, 2005) and Kindblom et al. (2005)
 TRα2−/−β−/−No TRα2/Δα2 or TRβ TRα1/Δα1 preserved, TRα1 over-expressionMild hypothyroidism (T4 0.7× T3 0.8×, TSH 1×)Transient growth delayNRNRNg et al. (2001)
 TRα0/0β−/−No TRα/ TRβ transcriptsRTH and goitre (T4 13× T3 14×, TSH >200×); reduced GH/IGF1More severe phenotype than TRα0/0; growth delay; delayed endochondral ossification; impaired chondrocyte differentiation; reduced mineralisationNRHypothyroidGauthier et al. (2001) and Flamant et al. (2002)
TRα, TRβ and Pax8 compound mutants
 Pax8−/−Apo TRα and TRβ receptorsNo thyroid (No T4 or T3, TSH 1900×)More severe growth defect than TRα0/0β−/−; severe persistent growth retardation; severely delayed endochondral ossification; impaired chondrocyte differentiationMajority die by weaning; coarse plate-like trabeculae with impaired trabecular remodelling; reduced cortical thickness; reduced mineralisationHypothyroidMansouri et al. (1998), Flamant et al. (2002) and Bassett et al. (2008)
 Pax8−/−TRα1−/−No T3 or TRα1/Δα1 TRα2/Δα2 preserved Apo TRβ receptorsNo thyroid (No T4 or T3, TSH NR)Severe growth retardation similar to Pax8−/−; skeletal phenotype not reportedDie by weaningNRMittag et al. (2005)
 Pax8−/−TRα0/0No T3 or TRα Apo TRβ receptorsNo thyroid (No T4 or T3, TSH >400×)Growth retardation less than Pax8−/− and similar to TRα0/0β−/−; delayed endochondral ossification; mice survive to adulthoodNRNRFlamant et al. (2002)
 Pax8−/−TRβ−/−No T3 or TRβ Apo TRα receptorsNo thyroid (No T4 or T3, TSH >400×)Severe growth retardation similar to Pax8−/−; severely delayed endochondral ossificationDie by weaningNRFlamant et al. (2002)

TR, T3 receptor; T4, thyroxine; T3, triiodothyronine; NR, not reported; IGF1, insulin-like growth factor 1; RTH, resistance to thyroid hormone; P10-35, postnatal day 10 to 35.

Thyroid hormone actions during skeletal development are anabolic

The developing skeleton is exquisitely sensitive to thyroid status and childhood hypothyroidism is characterised by growth retardation, delayed bone age and short stature, whereas juvenile thyrotoxicosis accelerates growth and advances bone age but results in short stature due to premature fusion of the epiphyses (Rivkees et al. 1988, Boersma et al. 1996, Segni & Gorman 2001).

Although TRα0/0 mice are systemically euthyroid, juveniles display post-natal growth retardation with delayed endochondral ossification characterised by impaired chondrocyte differentiation and decreased mineral deposition (Bassett et al. 2007b). TRα1R384C/+ mice, which harbour a dominant-negative mutation of TRα, are also euthyroid. They display a similar period of growth retardation and delayed endochondral ossification, but additionally have reduced cortical bone thickness, abnormal cortical bone remodelling and impairment of intramembranous ossification (Fig. 2; Bassett et al. 2007a). TRα1PV/+ mice, which express a potent dominant-negative TRα mutant, display a more severe skeletal phenotype. TRα1PV/+ mice have persistent post-natal growth retardation, markedly delayed endochondral ossification, decreased mineralisation, reduced cortical bone thickness and impaired intramembranous ossification (O'Shea et al. 2005). Similar findings have been reported in mice expressing the potent dominant-negative receptor TRα1L400R (Quignodon et al. 2007). Consistent with the delayed bone development in juvenile TRα0/0, TRα1R384C/+ and TRα1PV/+ mice, skeletal expression of the T3-target genes, fibroblast growth factor receptors 1 and 3 (FGFR1/3; Stevens et al. 2003, Barnard et al. 2005, O'Shea et al. 2007) were reduced (Barnard et al. 2005, O'Shea et al. 2005, Bassett et al. 2007a,b). Deletion or mutation of TRα does not affect systemic thyroid status but causes local skeletal hypothyroidism while the presence of a dominant-negative TRα leads to a more severe skeletal phenotype than receptor deficiency alone.

Figure 2
Figure 2

Skeletal development and growth in TR mutant mice. (A) Skull vaults from P1 mutant mice and littermate controls stained with alizarin red (bone) and alcian blue (cartilage) showing sutures and anterior and posterior fontanelles. Arrows indicate delayed intramembranous ossification in TRα1PV/+ and TRα1R384C/+ mice and advanced ossification in TRβ−/− and TRβPV/PV mice. Note that two TRβ−/− strains in different genetic backgrounds were analysed TRβ−/−(a) (Gauthier et al. 2001, Bassett et al. 2007b) and TRβ−/−(b) (Forrest et al. 1996, Ng et al. 2001, Bassett et al. 2007a). Scale bar, 1mm. (B) Graphs of longitudinal growth in mutant mice and littermate controls; *P<0.05; **P<0.01; ***P<0.001 mean tibia/ulna length in mutant versus control; two-tailed Students's t-test.

Citation: Journal of Molecular Endocrinology 42, 4; 10.1677/JME-08-0142

Two TRβ−/− strains have been generated in different genetic backgrounds and both show similar skeletal phenotypes (Table 1; Forrest et al. 1996, Gauthier et al. 1999). Deletion of TRβ results in elevated circulating TSH, T4 and T3 levels consistent with resistance to thyroid hormone (RTH). In contrast to TRα mutants, juvenile TRβ−/− mice display advanced endochondral ossification, accelerated chondrocyte differentiation, increased mineral deposition and persistent short stature due to premature growth plate quiescence. Furthermore, cortical thickness is increased and intramembranous ossification advanced in TRβ−/− mice (Fig. 2; Bassett et al. 2007b). TRβPV/PV mice express a potent dominant-negative TRβ and display severe RTH with a 400-fold elevation of TSH and 15-fold elevation of T4. TRβPV/PV animals exhibit a more severe phenotype than TRβ−/− mice with accelerated intrauterine growth characterised by advanced endochondral and intramembranous ossification. Premature ossification results in persistent post-natal growth retardation, premature growth plate quiescence, increased mineral deposition and craniosynostosis (O'Shea et al. 2003). Consistent with advanced skeletal development in TRβ−/− and TRβPV/PV mice, expression of the T3-target genes Fgfr1 and Fgfr3 was increased (O'Shea et al. 2003, Barnard et al. 2005, Bassett et al. 2007a,b). Thus, deletion or mutation of TRβ disrupts the HPT axis resulting in increased circulating thyroid hormone levels and skeletal thyrotoxicosis. The presence of a dominant-negative TRβ leads to a greater elevation of systemic thyroid hormone concentration and a more severe skeletal phenotype than receptor deficiency alone.

In the developing skeleton, reduced T3 action in TRα mutant mice results in delayed ossification and reduced mineralisation whereas increased T3 action in TRβ mutant mice leads to advanced ossification and increased mineralisation. Thus, during growth, T3 actions in bone are anabolic.

Thyroid hormone actions in adult bone are catabolic

Adult thyrotoxicosis results in both increased bone resorption and formation, but uncoupling of these processes favours osteoclastic resorption and leads to a 10% net bone loss per remodelling cycle (Mosekilde et al. 1990). Consequently, thyrotoxicosis is an important cause of secondary osteoporotic fracture (Mosekilde et al. 1990, Cummings et al. 1995, Franklyn et al. 1998, Vestergaard et al. 2000, Vestergaard & Mosekilde 2002, Murphy & Williams 2004) and even subclinical hyperthyroidism has been associated with decreased bone mineral density and increased fracture risk in postmenopausal women (Bauer et al. 2001, Quan et al. 2002, Murphy & Williams 2004, Heemstra et al. 2006, Kim et al. 2006, Morris 2007).

Remarkably, delayed ossification and reduced mineralisation in juvenile TRα0/0 mice were accompanied by greatly increased trabecular bone mass in adults (Fig. 3; Bassett et al. 2007b). Moreover, the robust and plate-like trabeculae contained highly mineralised calcified cartilage indicating a trabecular bone remodelling defect. Consistent with such a defect, TRα0/0 mice displayed reduced osteoclast numbers and activity (Bassett et al. 2007b). Trabecular bone mass increased progressively with age in TRα1R384C/+ mice with adults showing osteosclerosis (Bassett et al. 2007a). Consistent with a remodelling defect, trabeculae were of increased thickness and connectivity, showed increased mineralisation with extensive retention of calcified cartilage and reduced osteoclast numbers and activity (Fig. 4). Remarkably, brief T3 supplementation during growth, sufficient to overcome transcriptional repression by TRα1R384C, ameliorated the adult skeletal phenotype (Bassett et al. 2007a; Table 1). These data indicate that during development even transient relief from the transcriptional repression mediated by unliganded TRα1 (apo-TRα1) has long-term consequences for adult bone structure and mineralisation. Thus, in the adult skeleton, deletion or mutation of TRα results in persistently impaired bone remodelling. Similarly, the presence of a dominant-negative TRα leads to a more severe skeletal phenotype than receptor deficiency alone.

Figure 3
Figure 3

Trabecular bone micro-architecture in adult TR mutant mice. (A) Backscattered electron scanning electron microscopy (BSE-SEM) views showing trabecular bone in thyrotoxic wild-type (WT) mice treated with daily sc injections of T3 (30 ng T3/gram body weight) and congenitally hypothyroid Pax8−/− mice. (B) Animals lacking either TRα or TRβ and (C) mice harbouring a dominant-negative mutation of TRα (R384C). Scale bar, 200 μm.

Citation: Journal of Molecular Endocrinology 42, 4; 10.1677/JME-08-0142

Figure 4
Figure 4

Trabecular bone micro-mineralisation in adult TR mutant mice. Quantitative backscattered electron scanning electron microscopy (qBSE-SEM) images showing mineralisation densities of trabecular bone from mice lacking TRβ and mice harbouring a dominant-negative mutation of TRα (R384C) (A). Mineralisation densities were derived from halogenated standards and images pseudo-coloured according to the palette shown in which high mineralisation density is grey. (B) Relative and cumulative frequency histograms of bone micro-mineralisation densities are shown. Black arrow indicates the increased relative frequency of high mineralisation densities that correspond to retained calcified cartilage in TRα1R384C/+ mice. (C) Higher power views of trabecular bone are shown. Dashed white arrows indicate normal bone cement lines. Solid white arrows indicate retained calcified cartilage within which the outlines of chondrocyte lacunae remain evident. Scale bar 200 μm in all panels. ***P<0.001 micro-mineralisation density in mutant versus WT; Kolmogorov–Smirnov test.

Citation: Journal of Molecular Endocrinology 42, 4; 10.1677/JME-08-0142

Despite a juvenile phenotype of accelerated growth and increased ossification, adult TRβ−/− mice became progressively osteoporotic (Fig. 3; Bassett et al. 2007a). Two TRβ−/− strains in different genetic backgrounds were analysed (Gauthier et al. 2001, Bassett et al. 2007b). In one strain, cortical and trabecular bone mineralisation were both reduced and osteoclast numbers and activity increased (Bassett et al. 2007a,b). However, in the other strain, only trabecular bone mineralisation was affected but this was also accompanied by an increase in osteoclast numbers and activity (Bassett et al. 2007a; Fig. 4). Thus, in the adult skeleton, deletion of TRβ results in accelerated remodelling and bone loss although the severity may depend on genetic background.

In the adult skeleton reduced T3 action in TRα mutant mice results in osteosclerosis whereas increased T3 action in TRβ mutant mice leads to osteoporosis. Thus, the actions of T3 are catabolic in adult bone.

Mechanism of T3 action

The opposing skeletal phenotypes in TRα and TRβ mutant mice provide compelling evidence for the complex interaction between central and peripheral thyroid status (Fig. 5). Thus, delayed ossification and impaired bone remodelling in TRα mutant mice are secondary to the disruption of T3 action in bone, whereas advanced skeletal development and osteoporosis in TRβ mutant mice are due to disruption of the HPT axis, elevated systemic thyroid hormone levels and local supraphysiological stimulation of TRα in bone (O'Shea et al. 2006). This model is supported by T3-target gene expression in skeletal cells of TR mutant mice and by the demonstration of higher levels of TRα mRNA expression in bone compared with TRβ (O'Shea et al. 2003, 2005, Barnard et al. 2005, Bookout et al. 2006, Bassett et al. 2007a,b). Nevertheless, it is apparent that TRα0/0 TRβ−/− mice have a more severe skeletal phenotype than TRα0/0 mice, while TRα0/0 mice also remain sensitive to T4 treatment, thus suggesting a residual role for TRβ in skeletal cells. In this context, quantitative RT-PCR analysis has shown that TRα expression is 10- to 100-fold greater than TRβ expression in adult whole bone (O'Shea et al. 2003, Bookout et al. 2006). However, since the temporo-spatial patterns of TRα and TRβ expression in skeletal cells are unknown, a role for TRβ is possible. Furthermore, it is unclear whether individual skeletal cell co-express both TR isoforms or whether their patterns of expression are cell type specific.

Figure 5
Figure 5

Proposed molecular mechanism for TRα and TRβ mutant mice. The pituitary expresses predominantly TRβ T3 and TRβ act via a negative TRE to repress TSH transcription. The skeleton expresses predominantly TRα1 and T3 acts via TRα1 to activate target gene expression. Since the levels of TRα in the pituitary are low, its absence in TRα0/0 mice does not disrupt pituitary negative feedback and systemic TSH, T4 and T3 concentrations are normal. By contrast, because TRα predominates in bone, its absence in TRα0/0 mice results in impaired T3 responsiveness, skeletal hypothyroidism and reduced expression of T3-target genes. Similarly, in TRα1PV/+ and TRα1R384C/+ mice, the levels of the dominant-negative TRα receptor in the pituitary are insufficient to interfere with TRβ and disrupt TSH repression. However, in bone, the higher concentrations of mutant receptor interfere with the actions of wild-type TRα1. Thus, T3 responses are severely impaired and the skeleton is hypothyroid. In TRβ−/− mice, by contrast, the absence of TRβ disrupts pituitary T3 responses and impairs TSH repression leading to high circulating TSH, T4 and T3 concentrations. Since TRβ levels are low in bone, T3 responses are unaffected in TRβ−/− mice and high circulating levels of TH act via wild-type TRα1 to induce skeletal thyrotoxicosis. Similarly in TRβPV/PV mice, the dominant-negative TRβ disrupts TSH repression in the pituitary and results in grossly elevated TSH, T4 and T3 concentrations. The low levels of the mutant receptor in bone are insufficient to interfere with TRα1 and impair T3 responses and thus high circulating levels of TH increase expression of T3-target genes resulting in severe skeletal thyrotoxicosis.

Citation: Journal of Molecular Endocrinology 42, 4; 10.1677/JME-08-0142

Present understanding is that TRα1 is functionally predominant in bone.

T3 action in the developing skeleton

In vitro T3 inhibits chondrocyte proliferation and promotes differentiation (Robson et al. 2000, Shao et al. 2006; Fig. 6). Since growth plate architecture and linear growth are frequently disrupted in TR mutant mice, key regulators of endochondral ossification have been investigated as targets of thyroid hormone action.

Figure 6
Figure 6

Role of TH in endochondral ossification and bone remodelling. (A) The role of T3 in the growth plate is illustrated and the pattern of TR expression is shown. Reserve cells undergo clonal expansion to form columns of proliferating chondrocytes that secrete cartilage matrix stained blue in this section. Committed prehypertrophic chondrocytes undergo hypertrophic differentiation, enlarge and subsequently apoptose, the residual mineralised cartilage matrix forming the scaffold for trabecular bone formation. In chondrocyte cultures, T3 inhibits proliferation and increases the expression of cyclin-dependent kinase inhibitors P21cip-1 P27kip-1. T3 stimulates matrix synthesis and expression of the differentiation markers collagen X, alkaline phosphatase (ALP), matrix metalloproteinase 13 (MMP13) and aggrecanase 2. GH/insulin-like growth factor 1 (GH/IGF1), Indian hedgehog/parathyroid hormone-related peptide (Ihh/PTHrP) and fibroblast growth factor/fibroblast growth factor receptor (FGF/FGFR) pathways are T3 targets. Expression of heparan sulphate proteoglycans (HSPGs), which are essential for FGF and Ihh signalling, is negatively regulated by T3. (B) The role of T3 in the bone remodelling cycle, which is characterised by osteoclastic bone resorption followed by osteoblast migration, osteoid matrix deposition and mineralisation, is shown. Osteoclast differentiation requires direct osteoblast–osteoclast interaction mediated by receptor activator of nuclear factor-κB (RANK) and its ligand RANKL. Osteoprotegerin (OPG) is a decoy receptor secreted by osteoblasts that antagonises this interaction. Osteoclast differentiation also requires co-stimulation by osteoblast-derived cytokines. Thyroid hormone stimulates osteoblast proliferation, differentiation and apoptosis directly and also indirectly by regulating IGF1 and FGF signalling. It remains uncertain whether osteoclast differentiation is regulated directly by T3 or indirectly via T3 actions in osteoblasts.

Citation: Journal of Molecular Endocrinology 42, 4; 10.1677/JME-08-0142

GH and insulin-like growth factor 1 signalling pathways are thought to act as a point of convergence for many growth promoting factors since more than 80% of somatic growth can be attributable to the actions of the GH/IGF1 axis (Lupu et al. 2001). IGF1 is the major determinant of post-natal growth, it mediates both GH-dependent and -independent effects and is implicated in chondrocyte recruitment, proliferation and hypertrophic differentiation (van der Eerden et al. 2003). Examination of the GH/IGF1 pathway revealed that GH receptor (GHR) and Igf1 receptor (IGF1R) mRNA expression was reduced in growth plate chondrocytes in TRα0/0 and TRα1PV/+ mice. Furthermore, phosphorylation of their secondary messengers signal tranducer and activator of transcription 5 (STAT5) and protein kinase B (AKT) was also impaired (O'Shea et al. 2005, Bassett et al. 2007b). By contrast, GHR and IFG1R expression was increased in TRβ−/− and TRβPV/PV mice (O'Shea et al. 2005, Bassett et al. 2007b). Thus, GH/IGF1 signalling is also a local downstream mediator of T3 action in the growth plate (Fig. 6).

FGFs and their receptors have key roles in skeletal development with activating mutations of FGFR3 causing achondroplasia, the commonest form of genetic dwarfism. In the developing growth plate, FGFR3 is expressed in reserve and proliferating chondrocytes and negatively regulates their proliferation and differentiation (Murakami et al. 2004). By contrast, FGFR1 is expressed in prehypertrophic and hypertrophic chondrocytes and its location suggests a role in differentiation, matrix synthesis and apoptosis (Ornitz 2005). FGFR1 is also expressed in the osteoblast lineage with activating mutations result in Pfeiffer craniosynostosis. Investigation of the FGF/FGFR signalling pathway in TR mutant mice demonstrated that Fgfr3 and Fgfr1 expression was reduced in growth plates of TRα0/0, TRα1R384C/+ and TRα1PV/+ mice and Fgfr1 expression was reduced in osteoblasts from TRα0/0 and Pax8−/− mice (Stevens et al. 2003, Barnard et al. 2005, O'Shea et al. 2005, Bassett et al. 2007a,b, 2008). By contrast, Fgfr3 and Fgfr1 expression was increased in growth plates of TRβ−/− and TRβPV/PV mice and Fgfr1 expression was increased in osteoblasts from TRβPV/PV mice (O'Shea et al. 2003, Barnard et al. 2005, Bassett et al. 2007a,b). Thus, FGF/FGFR signalling is a downstream mediator of T3 action in chondrocytes and osteoblasts (Fig. 6).

The pace of chondrocyte differentiation is precisely regulated by the Indian hedgehog/parathyroid hormone-related peptide paracrine (Ihh/PTHrP) negative feedback loop. Prehypertrophic chondrocytes secrete Ihh which diffuses to periarticular cells to induce synthesis of PTHrP. PTHrP, acting via its receptor PTHR1, then completes the loop by stimulating chondrocyte proliferation and inhibiting further hypertrophic differentiation (Vortkamp et al. 1996, Dentice et al. 2005). Although this pathway has not been studied in TR mutant mice, previous experiments in thyroid-manipulated rats demonstrated increased PTHrP mRNA expression in hypothyroid animals and decreased PTHrP receptor mRNA expression in growth plates of thyrotoxic animals (Stevens et al. 2000). Furthermore, recent studies in chicken tibia explants have shown that Ihh stimulates degradation of the type 2 deiodinase enzyme resulting in an induction of PTHrP expression (Dentice et al. 2005). Together, these findings suggesting that thyroid hormone can inhibit chondrocyte proliferation and promote differentiation by local regulation of the Ihh/PTHrP negative feedback loop (Fig. 6).

T3 is essential for normal cartilage matrix synthesis. Heparan sulphate proteoglycans (HSPGs) are a key matrix component and are essential for functional FGF/FGFR signalling and extracellular diffusion of Ihh. Studies in thyroid manipulated rats and TRα0/0β−/− and Pax8−/− mice revealed reduced HSPG expression in thyrotoxic animals, increased expression in TRα0/0β−/−mice and more markedly increased expression in hypothyroid rats and congenitally hypothyroid Pax8−/− mice (Bassett et al. 2008).

These studies suggest T3 coordinately regulates FGF/FGFR and Ihh/PTHrP signalling within the growth plate.

T3 actions in the bone remodelling cycle

In vitro studies suggest that thyroid hormone stimulates osteoblast proliferation, differentiation and apoptosis by direct and indirect actions. Thus, T3 increases synthesis of osteocalcin, type 1 collagen, alkaline phosphatase and matrix metalloproteinases 9 and 13 (Pereira et al. 1999, Huang et al. 2000, Gouveia et al. 2001, Bassett & Williams 2003) and also regulates IGF1, parathyroid hormone (PTH) and FGF signalling (Huang et al. 2000, Gu et al. 2001, Pepene et al. 2001, Bassett & Williams 2003, Stevens et al. 2003, O'Shea et al. 2007; Fig. 6). In vivo, activation of FGFR1 stimulates osteoblast proliferation and differentiation (Zhou et al. 2000). Consistent with this, Fgfr1 expression in osteoblasts is reduced in the hypothyroid skeleton and increased in thyrotoxic bone (O'Shea et al. 2003, Stevens et al. 2003, Bassett et al. 2007a). Although osteoclasts have been reported to express TRs (Allain et al. 1992, Abu et al. 1997, 2000, Kanatani et al. 2004), it remains uncertain whether T3 regulates osteoclast differentiation directly or indirectly (Siddiqi et al. 1998, Miura et al. 2002, Varga et al. 2004). Previous studies are contradictory; some demonstrating that T3 acts directly in osteoclasts while others report indirect effects mediated via osteoblasts.

Future directions

Analyses of TR mutant and congenitally hypothyroid Pax8−/− mice have identified a key role for thyroid hormone and TRα1 in both the developing skeleton and adult bone. However, it remains unclear how the skeletal effects of apo-TRα1 differ from those of TRα1 deficiency and to what extent the skeletal effects of thyroid hormones result from central, systemic and local actions of T3. The more severe skeletal phenotype observed in congenitally hypothyroid Pax8−/− mice as compared with TRα0/0β−/− mice suggests that unliganded TRs may be more detrimental to skeletal development than TR deficiency (Mansouri et al. 1998, Flamant et al. 2002, Bassett et al. 2008; Table 1). In support of this, amelioration of the Pax8−/− skeletal phenotype in Pax8−/−TRα0/0 mice but not in Pax8−/−TRβ−/− mice suggests that some of the detrimental effects of hypothyroidism are mediated by unliganded TRα1 in bone (Flamant et al. 2002). Despite this, it is important to note from an additional study (Mittag et al. 2005) that deletion of TRα1 alone in Pax8−/−TRα1−/− mice did not prevent weight loss, early mortality and pituitary abnormalities although the skeletal consequences were not investigated. Thus, it remains possible that TRα2 has an additional and essential role in the manifestation of the Pax8−/− phenotype. This importance of apo-TRα1 is further supported by the more severe skeletal phenotype present in mice harbouring dominant-negative mutations of TRα1 (TRα1R384C/+ and TRα1PV/+) as compared with mice lacking all TRα isoforms (TRα0/0; O'Shea et al. 2005, Bassett et al. 2007a,b) and by the amelioration of the adult skeletal phenotype in TRα1R384C/+ mice following a transient reversal of TRα1R384C apo-receptor activity during development (Bassett et al. 2007a). Nevertheless, a complete understanding of the molecular mechanism of thyroid hormone action in bone will require at least two experimental approaches. First, it is clear that phenotyping of the existing mouse models is incomplete and more detailed studies including quantitative histomorphometry, mechanical testing and analysis of primary bone cell cultures will help to clarify the picture. However, such an approach cannot identify the cellular targets of T3 action in the skeleton in vivo and this will require the use of cell-specific gene-targeting strategies.

Analysis of TRα and TRβ mutant mice has demonstrated the complex relationship between central and peripheral thyroid status and established the predominant role of TRα1 in bone. These studies also highlight contrasting responses of the skeleton to T3 during developing and in adulthood. Although understanding of the molecular mechanism of T3 action in bone is still limited, coordinate regulation of key signalling pathways has now been identified in chondrocytes and osteoblasts. By contrast, the molecular mechanism of T3 action in the osteoclast lineage remains unclear. A more detailed understanding of the molecular basis of T3 action in bone will provide the rational for the development of novel strategies for the prevention and treatment of osteoporosis.

Declaration of interest

There is no conflict of interest.

Funding

This work was supported by the Medical Research Council (grant numbers G108/502 and G0501486) and the Wellcome Trust (grant number GR076584).

Acknowledgements

We wish to thank Prof. Alan Boyde, Queen Mary University of London, for BSE-SEM imaging, valuable advice and continuing collaboration.

References

  • Abel ED, Boers ME, Pazos-Moura C, Moura E, Kaulbach H, Zakaria M, Lowell B, Radovick S, Liberman MC & Wondisford F 1999 Divergent roles for thyroid hormone receptor beta isoforms in the endocrine axis and auditory system. Journal of Clinical Investigation 104 291300.

    • Search Google Scholar
    • Export Citation
  • Abu EO, Bord S, Horner A, Chatterjee VK & Compston JE 1997 The expression of thyroid hormone receptors in human bone. Bone 21 137142.

  • Abu EO, Horner A, Teti A, Chatterjee VK & Compston JE 2000 The localization of thyroid hormone receptor mRNAs in human bone. Thyroid 10 287293.

  • Allain TJ, Chambers TJ, Flanagan AM & McGregor AM 1992 Tri-iodothyronine stimulates rat osteoclastic bone resorption by an indirect effect. Journal of Endocrinology 133 327331.

    • Search Google Scholar
    • Export Citation
  • Barnard JC, Williams AJ, Rabier B, Chassande O, Samarut J, Cheng SY, Bassett JH & Williams GR 2005 Thyroid hormones regulate fibroblast growth factor receptor signaling during chondrogenesis. Endocrinology 146 55685580.

    • Search Google Scholar
    • Export Citation
  • Bassett JH & Williams GR 2003 The molecular actions of thyroid hormone in bone. Trends in Endocrinology and Metabolism 14 356364.

  • Bassett JH & Williams GR 2008 Critical role of the hypothalamic–pituitary–thyroid axis in bone. Bone 43 418426.

  • Bassett JH, Nordstrom K, Boyde A, Howell PG, Kelly S, Vennstrom B & Williams GR 2007a Thyroid status during skeletal development determines adult bone structure and mineralization. Molecular Endocrinology 21 18931904.

    • Search Google Scholar
    • Export Citation
  • Bassett JH, O'Shea PJ, Sriskantharajah S, Rabier B, Boyde A, Howell PG, Weiss RE, Roux JP, Malaval L & Clement-Lacroix P 2007b Thyroid hormone excess rather than thyrotropin deficiency induces osteoporosis in hyperthyroidism. Molecular Endocrinology 21 10951107.

    • Search Google Scholar
    • Export Citation
  • Bassett JH, Williams AJ, Murphy E, Boyde A, Howell PG, Swinhoe R, Archanco M, Flamant F, Samarut J & Costagliola S 2008 A lack of thyroid hormones rather than excess thyrotropin causes abnormal skeletal development in hypothyroidism. Molecular Endocrinology 22 501512.

    • Search Google Scholar
    • Export Citation
  • Bauer DC, Ettinger B, Nevitt MC & Stone LS 2001 Risk for fracture in women with low serum levels of thyroid-stimulating hormone. Annals of Internal Medicine 134 561568.

    • Search Google Scholar
    • Export Citation
  • Boersma B, Otten BJ, Stoelinga GB & Wit JM 1996 Catch-up growth after prolonged hypothyroidism. European Journal of Pediatrics 155 362367.

  • Bookout AL, Jeong Y, Downes M, Yu RT, Evans RM & Mangelsdorf DJ 2006 Anatomical profiling of nuclear receptor expression reveals a hierarchical transcriptional network. Cell 126 789799.

    • Search Google Scholar
    • Export Citation
  • Chassande O, Fraichard A, Gauthier K, Flamant F, Legrand C, Savatier P, Laudet V & Samarut J 1997 Identification of transcripts initiated from an internal promoter in the c-erbA alpha locus that encode inhibitors of retinoic acid receptor-alpha and triiodothyronine receptor activities. Molecular Endocrinology 11 12781290.

    • Search Google Scholar
    • Export Citation
  • Cummings SR, Nevitt MC, Browner WS, Stone K, Fox KM, Ensrud KE, Cauley J, Black D & Vogt TM 1995 Risk factors for hip fracture in white women. Study of Osteoporotic Fractures Research Group. New England Journal of Medicine 332 767773.

    • Search Google Scholar
    • Export Citation
  • Dentice M, Bandyopadhyay A, Gereben B, Callebaut I, Christoffolete MA, Kim BW, Nissim S, Mornon JP, Zavacki AM & Zeold A 2005 The Hedgehog-inducible ubiquitin ligase subunit WSB-1 modulates thyroid hormone activation and PTHrP secretion in the developing growth plate. Nature Cell Biology 7 698705.

    • Search Google Scholar
    • Export Citation
  • van der Eerden BC, Karperien M & Wit JM 2003 Systemic and local regulation of the growth plate. Endocrine Reviews 24 782801.

  • Flamant F, Poguet AL, Plateroti M, Chassande O, Gauthier K, Streichenberger N, Mansouri A & Samarut J 2002 Congenital hypothyroid Pax8(−/−) mutant mice can be rescued by inactivating the TRalpha gene. Molecular Endocrinology 16 2432.

    • Search Google Scholar
    • Export Citation
  • Forrest D, Hanebuth E, Smeyne RJ, Everds N, Stewart CL, Wehner JM & Curran T 1996 Recessive resistance to thyroid hormone in mice lacking thyroid hormone receptor beta: evidence for tissue-specific modulation of receptor function. EMBO Journal 15 30063015.

    • Search Google Scholar
    • Export Citation
  • Fraichard A, Chassande O, Plateroti M, Roux JP, Trouillas J, Dehay C, Legrand C, Gauthier K, Kedinger M & Malaval L 1997 The T3R alpha gene encoding a thyroid hormone receptor is essential for post-natal development and thyroid hormone production. EMBO Journal 16 44124420.

    • Search Google Scholar
    • Export Citation
  • Franklyn JA, Maisonneuve P, Sheppard MC, Betteridge J & Boyle P 1998 Mortality after the treatment of hyperthyroidism with radioactive iodine. New England Journal of Medicine 338 712718.

    • Search Google Scholar
    • Export Citation
  • Gauthier K, Chassande O, Plateroti M, Roux JP, Legrand C, Pain B, Rousset B, Weiss R, Trouillas J & Samarut J 1999 Different functions for the thyroid hormone receptors TRalpha and TRbeta in the control of thyroid hormone production and post-natal development. EMBO Journal 18 623631.

    • Search Google Scholar
    • Export Citation
  • Gauthier K, Plateroti M, Harvey CB, Williams GR, Weiss RE, Refetoff S, Willott JF, Sundin V, Roux JP & Malaval L 2001 Genetic analysis reveals different functions for the products of the thyroid hormone receptor alpha locus. Molecular and Cellular Biology 21 47484760.

    • Search Google Scholar
    • Export Citation
  • Gothe S, Wang Z, Ng L, Kindblom JM, Barros AC, Ohlsson C, Vennstrom B & Forrest D 1999 Mice devoid of all known thyroid hormone receptors are viable but exhibit disorders of the pituitary–thyroid axis, growth, and bone maturation. Genes and Development 13 13291341.

    • Search Google Scholar
    • Export Citation
  • Gouveia CH, Schultz JJ, Bianco AC & Brent GA 2001 Thyroid hormone stimulation of osteocalcin gene expression in ROS 17/2.8 cells is mediated by transcriptional and post-transcriptional mechanisms. Journal of Endocrinology 170 667675.

    • Search Google Scholar
    • Export Citation
  • Gu WX, Stern PH, Madison LD & Du GG 2001 Mutual up-regulation of thyroid hormone and parathyroid hormone receptors in rat osteoblastic osteosarcoma 17/2.8 cells. Endocrinology 142 157164.

    • Search Google Scholar
    • Export Citation
  • Hashimoto K, Curty FH, Borges PP, Lee CE, Abel ED, Elmquist JK, Cohen RN & Wondisford FE 2001 An unliganded thyroid hormone receptor causes severe neurological dysfunction. PNAS 98 39984003.

    • Search Google Scholar
    • Export Citation
  • Heemstra KA, Hamdy NA, Romijn JA & Smit JW 2006 The effects of thyrotropin-suppressive therapy on bone metabolism in patients with well-differentiated thyroid carcinoma. Thyroid 16 583591.

    • Search Google Scholar
    • Export Citation
  • Huang BK, Golden LA, Tarjan G, Madison LD & Stern PH 2000 Insulin-like growth factor I production is essential for anabolic effects of thyroid hormone in osteoblasts. Journal of Bone and Mineral Research 15 188197.

    • Search Google Scholar
    • Export Citation
  • Jones I, Ng L, Liu H & Forrest D 2007 An intron control region differentially regulates expression of thyroid hormone receptor beta2 in the cochlea, pituitary, and cone photoreceptors. Molecular Endocrinology 21 11081119.

    • Search Google Scholar
    • Export Citation
  • Kanatani M, Sugimoto T, Sowa H, Kobayashi T, Kanzawa M & Chihara K 2004 Thyroid hormone stimulates osteoclast differentiation by a mechanism independent of RANKL–RANK interaction. Journal of Cellular Physiology 201 1725.

    • Search Google Scholar
    • Export Citation
  • Kaneshige M, Kaneshige K, Zhu X, Dace A, Garrett L, Carter TA, Kazlauskaite R, Pankratz DG, Wynshaw-Boris A & Refetoff S 2000 Mice with a targeted mutation in the thyroid hormone beta receptor gene exhibit impaired growth and resistance to thyroid hormone. PNAS 97 1320913214.

    • Search Google Scholar
    • Export Citation
  • Kaneshige M, Suzuki H, Kaneshige K, Cheng J, Wimbrow H, Barlow C, Willingham MC & Cheng S 2001 A targeted dominant negative mutation of the thyroid hormone alpha 1 receptor causes increased mortality, infertility, and dwarfism in mice. PNAS 98 1509515100.

    • Search Google Scholar
    • Export Citation
  • Kanis JA & Johnell O 2005 Requirements for DXA for the management of osteoporosis in Europe. Osteoporosis International 16 229238.

  • Kim DJ, Khang YH, Koh JM, Shong YK & Kim GS 2006 Low normal TSH levels are associated with low bone mineral density in healthy postmenopausal women. Clinical Endocrinology 64 8690.

    • Search Google Scholar
    • Export Citation
  • Kindblom JM, Gothe S, Forrest D, Tornell J, Vennstrom B & Ohlsson CGH 2001 GH substitution reverses the growth phenotype but not the defective ossification in thyroid hormone receptor alpha 1−/−beta−/− mice. Journal of Endocrinology 171 1522.

    • Search Google Scholar
    • Export Citation
  • Kindblom JM, Gevers EF, Skrtic SM, Lindberg MK, Gothe S, Tornell J, Vennstrom B & Ohlsson C 2005 Increased adipogenesis in bone marrow but decreased bone mineral density in mice devoid of thyroid hormone receptors. Bone 36 607616.

    • Search Google Scholar
    • Export Citation
  • Liu YY, Schultz JJ & Brent GA 2003 A thyroid hormone receptor alpha gene mutation (P398H) is associated with visceral adiposity and impaired catecholamine-stimulated lipolysis in mice. Journal of Biological Chemistry 278 3891338920.

    • Search Google Scholar
    • Export Citation
  • Lupu F, Terwilliger JD, Lee K, Segre GV & Efstratiadis A 2001 Roles of growth hormone and insulin-like growth factor 1 in mouse postnatal growth. Developmental Biology 229 141162.

    • Search Google Scholar
    • Export Citation
  • Mansouri A, Chowdhury K & Gruss P 1998 Follicular cells of the thyroid gland require Pax8 gene function. Nature Genetics 19 8790.

  • Mittag J, Friedrichsen S, Heuer H, Polsfuss S, Visser TJ & Bauer K 2005 Athyroid Pax8−/− mice cannot be rescued by the inactivation of thyroid hormone receptor alpha1. Endocrinology 146 31793184.

    • Search Google Scholar
    • Export Citation
  • Miura M, Tanaka K, Komatsu Y, Suda M, Yasoda A, Sakuma Y, Ozasa A & Nakao K 2002 A novel interaction between thyroid hormones and 1,25(OH)(2)D(3) in osteoclast formation. Biochemical and Biophysical Research Communications 291 987994.

    • Search Google Scholar
    • Export Citation
  • Morris MS 2007 The association between serum thyroid-stimulating hormone in its reference range and bone status in postmenopausal American women. Bone 40 11281134.

    • Search Google Scholar
    • Export Citation
  • Mosekilde L, Eriksen EF & Charles P 1990 Effects of thyroid hormones on bone and mineral metabolism. Endocrinology and Metabolism Clinics of North America 19 3563.

    • Search Google Scholar
    • Export Citation
  • Murakami S, Balmes G, McKinney S, Zhang Z, Givol D & de Crombrugghe B 2004 Constitutive activation of MEK1 in chondrocytes causes Stat1-independent achondroplasia-like dwarfism and rescues the Fgfr3-deficient mouse phenotype. Genes and Development 18 290305.

    • Search Google Scholar
    • Export Citation
  • Murphy E & Williams GR 2004 The thyroid and the skeleton. Clinical Endocrinology 61 285298.

  • Ng L, Hurley JB, Dierks B, Srinivas M, Salto C, Vennstrom B, Reh TA & Forrest D 2001 A thyroid hormone receptor that is required for the development of green cone photoreceptors. Nature Genetics 27 9498.

    • Search Google Scholar
    • Export Citation
  • Ornitz DM 2005 FGF signaling in the developing endochondral skeleton. Cytokine and Growth Factor Reviews 16 205213.

  • O'Shea PJ, Harvey CB, Suzuki H, Kaneshige M, Kaneshige K, Cheng SY & Williams GR 2003 A thyrotoxic skeletal phenotype of advanced bone formation in mice with resistance to thyroid hormone. Molecular Endocrinology 17 14101424.

    • Search Google Scholar
    • Export Citation
  • O'Shea PJ, Bassett JH, Sriskantharajah S, Ying H, Cheng SY & Williams GR 2005 Contrasting skeletal phenotypes in mice with an identical mutation targeted to thyroid hormone receptor alpha1 or beta. Molecular Endocrinology 19 30453059.

    • Search Google Scholar
    • Export Citation
  • O'Shea PJ, Bassett JH, Cheng SY & Williams GR 2006 Characterization of skeletal phenotypes of TRalpha1 and TRbeta mutant mice: implications for tissue thyroid status and T3 target gene expression. Nuclear Receptor Signaling 4 e011

    • Search Google Scholar
    • Export Citation
  • O'Shea PJ, Guigon CJ, Williams GR & Cheng SY 2007 Regulation of fibroblast growth factor receptor-1 by thyroid hormone: identification of a thyroid hormone response element in the murine Fgfr1 promoter. Endocrinology

    • Search Google Scholar
    • Export Citation
  • Pepene CE, Kasperk CH, Pfeilschifter J, Borcsok I, Gozariu L, Ziegler R & Seck T 2001 Effects of triiodothyronine on the insulin-like growth factor system in primary human osteoblastic cells in vitro. Bone 29 540546.

    • Search Google Scholar
    • Export Citation
  • Pereira RC, Jorgetti V & Canalis E 1999 Triiodothyronine induces collagenase-3 and gelatinase B expression in murine osteoblasts. American Journal of Physiology 277 E496E504.

    • Search Google Scholar
    • Export Citation
  • Quan ML, Pasieka JL & Rorstad O 2002 Bone mineral density in well-differentiated thyroid cancer patients treated with suppressive thyroxine: a systematic overview of the literature. Journal of Surgical Oncology 79 6269.(discussion 69–70)

    • Search Google Scholar
    • Export Citation
  • Quignodon L, Vincent S, Winter H, Samarut J & Flamant F 2007 A point mutation in the activation function 2 domain of thyroid hormone receptor alpha1 expressed after CRE-mediated recombination partially recapitulates hypothyroidism. Molecular Endocrinology 21 23502360.

    • Search Google Scholar
    • Export Citation
  • Rivkees SA, Bode HH & Crawford JD 1988 Long-term growth in juvenile acquired hypothyroidism: the failure to achieve normal adult stature. New England Journal of Medicine 318 599602.

    • Search Google Scholar
    • Export Citation
  • Robson H, Siebler T, Stevens DA, Shalet SM & Williams GR 2000 Thyroid hormone acts directly on growth plate chondrocytes to promote hypertrophic differentiation and inhibit clonal expansion and cell proliferation. Endocrinology 141 38873897.

    • Search Google Scholar
    • Export Citation
  • Salto C, Kindblom JM, Johansson C, Wang Z, Gullberg H, Nordstrom K, Mansen A, Ohlsson C, Thoren P & Forrest D 2001 Ablation of TRalpha2 and a concomitant overexpression of alpha1 yields a mixed hypo- and hyperthyroid phenotype in mice. Molecular Endocrinology 15 21152128.

    • Search Google Scholar
    • Export Citation
  • Segni M & Gorman CA 2001 The aftermath of childhood hyperthyroidism. Journal of Pediatric Endocrinology and Metabolism 14 12771282.(discussion 1297–1298)

    • Search Google Scholar
    • Export Citation
  • Shao YY, Wang L & Ballock RT 2006 Thyroid hormone and the growth plate. Reviews in Endocrine and Metabolic Disorders 7 265271.

  • Siddiqi A, Burrin JM, Wood DF & Monson JP 1998 Tri-iodothyronine regulates the production of interleukin-6 and interleukin-8 in human bone marrow stromal and osteoblast-like cells. Journal of Endocrinology 157 453461.

    • Search Google Scholar
    • Export Citation
  • Stevens DA, Hasserjian RP, Robson H, Siebler T, Shalet SM & Williams GR 2000 Thyroid hormones regulate hypertrophic chondrocyte differentiation and expression of parathyroid hormone-related peptide and its receptor during endochondral bone formation. Journal of Bone and Mineral Research 15 24312442.

    • Search Google Scholar
    • Export Citation
  • Stevens DA, Harvey CB, Scott AJ, O'Shea PJ, Barnard JC, Williams AJ, Brady G, Samarut J, Chassande O & Williams GR 2003 Thyroid hormone activates fibroblast growth factor receptor-1 in bone. Molecular Endocrinology 17 17511766.

    • Search Google Scholar
    • Export Citation
  • Tinnikov A, Nordstrom K, Thoren P, Kindblom JM, Malin S, Rozell B, Adams M, Rajanayagam O, Pettersson S & Ohlsson C 2002 Retardation of post-natal development caused by a negatively acting thyroid hormone receptor alpha1. EMBO Journal 21 50795087.

    • Search Google Scholar
    • Export Citation
  • Varga F, Spitzer S & Klaushofer K 2004 Triiodothyronine (T3) and 1,25-dihydroxyvitamin D3 (1,25D3) inversely regulate OPG gene expression in dependence of the osteoblastic phenotype. Calcified Tissue International 74 382387.

    • Search Google Scholar
    • Export Citation
  • Vestergaard P & Mosekilde L 2002 Fractures in patients with hyperthyroidism and hypothyroidism: a nationwide follow-up study in 16 249 patients. Thyroid 12 411419.

    • Search Google Scholar
    • Export Citation
  • Vestergaard P, Rejnmark L, Weeke J & Mosekilde L 2000 Fracture risk in patients treated for hyperthyroidism. Thyroid 10 341348.

  • Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM & Tabin CJ 1996 Regulation of rate of cartilage differentiation by Indian hedgehog and PTH-related protein. Science 273 613622.

    • Search Google Scholar
    • Export Citation
  • Wikstrom L, Johansson C, Salto C, Barlow C, Campos Barros A, Baas F, Forrest D, Thoren P & Vennstrom B 1998 Abnormal heart rate and body temperature in mice lacking thyroid hormone receptor alpha 1. EMBO Journal 17 455461.

    • Search Google Scholar
    • Export Citation
  • Williams GR, Bland R & Sheppard MC 1994 Characterization of thyroid hormone (T3) receptors in three osteosarcoma cell lines of distinct osteoblast phenotype: interactions among T3, vitamin D3, and retinoid signaling. Endocrinology 135 23752385.

    • Search Google Scholar
    • Export Citation
  • Yen PM 2001 Physiological and molecular basis of thyroid hormone action. Physiological Reviews 81 10971142.

  • Zhou YX, Xu X, Chen L, Li C, Brodie SG & Deng CX 2000 A Pro250Arg substitution in mouse Fgfr1 causes increased expression of Cbfa1 and premature fusion of calvarial sutures. Human Molecular Genetics 9 20012008.

    • Search Google Scholar
    • Export Citation

If the inline PDF is not rendering correctly, you can download the PDF file here.

 

      Society for Endocrinology

Sept 2018 onwards Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 927 79 7
PDF Downloads 269 66 9
  • View in gallery

    Thyroid hormone receptor isoforms. (A) Schematic of the four functional domains of the thyroid hormone receptor. (B) Products of the Thra and Thrb genes in the mouse. Thra has two promoters, TRα1 and TRα2 are derived from the 5′ promoter whereas TRΔα1 and TRΔα2 are transcribed from an internal promoter in intron 7. TRα2 and TRΔα2 result from alternative splicing. TRα1 is a functional receptor whereas TRΔα1, TRα2 and TRΔα2 do not bind T3 and act as antagonists. Thrb also has two promoters and both N-terminal variants TRβ1 and TRβ2 bind T3 and act as functional receptors.

  • View in gallery

    Skeletal development and growth in TR mutant mice. (A) Skull vaults from P1 mutant mice and littermate controls stained with alizarin red (bone) and alcian blue (cartilage) showing sutures and anterior and posterior fontanelles. Arrows indicate delayed intramembranous ossification in TRα1PV/+ and TRα1R384C/+ mice and advanced ossification in TRβ−/− and TRβPV/PV mice. Note that two TRβ−/− strains in different genetic backgrounds were analysed TRβ−/−(a) (Gauthier et al. 2001, Bassett et al. 2007b) and TRβ−/−(b) (Forrest et al. 1996, Ng et al. 2001, Bassett et al. 2007a). Scale bar, 1mm. (B) Graphs of longitudinal growth in mutant mice and littermate controls; *P<0.05; **P<0.01; ***P<0.001 mean tibia/ulna length in mutant versus control; two-tailed Students's t-test.

  • View in gallery

    Trabecular bone micro-architecture in adult TR mutant mice. (A) Backscattered electron scanning electron microscopy (BSE-SEM) views showing trabecular bone in thyrotoxic wild-type (WT) mice treated with daily sc injections of T3 (30 ng T3/gram body weight) and congenitally hypothyroid Pax8−/− mice. (B) Animals lacking either TRα or TRβ and (C) mice harbouring a dominant-negative mutation of TRα (R384C). Scale bar, 200 μm.

  • View in gallery

    Trabecular bone micro-mineralisation in adult TR mutant mice. Quantitative backscattered electron scanning electron microscopy (qBSE-SEM) images showing mineralisation densities of trabecular bone from mice lacking TRβ and mice harbouring a dominant-negative mutation of TRα (R384C) (A). Mineralisation densities were derived from halogenated standards and images pseudo-coloured according to the palette shown in which high mineralisation density is grey. (B) Relative and cumulative frequency histograms of bone micro-mineralisation densities are shown. Black arrow indicates the increased relative frequency of high mineralisation densities that correspond to retained calcified cartilage in TRα1R384C/+ mice. (C) Higher power views of trabecular bone are shown. Dashed white arrows indicate normal bone cement lines. Solid white arrows indicate retained calcified cartilage within which the outlines of chondrocyte lacunae remain evident. Scale bar 200 μm in all panels. ***P<0.001 micro-mineralisation density in mutant versus WT; Kolmogorov–Smirnov test.

  • View in gallery

    Proposed molecular mechanism for TRα and TRβ mutant mice. The pituitary expresses predominantly TRβ T3 and TRβ act via a negative TRE to repress TSH transcription. The skeleton expresses predominantly TRα1 and T3 acts via TRα1 to activate target gene expression. Since the levels of TRα in the pituitary are low, its absence in TRα0/0 mice does not disrupt pituitary negative feedback and systemic TSH, T4 and T3 concentrations are normal. By contrast, because TRα predominates in bone, its absence in TRα0/0 mice results in impaired T3 responsiveness, skeletal hypothyroidism and reduced expression of T3-target genes. Similarly, in TRα1PV/+ and TRα1R384C/+ mice, the levels of the dominant-negative TRα receptor in the pituitary are insufficient to interfere with TRβ and disrupt TSH repression. However, in bone, the higher concentrations of mutant receptor interfere with the actions of wild-type TRα1. Thus, T3 responses are severely impaired and the skeleton is hypothyroid. In TRβ−/− mice, by contrast, the absence of TRβ disrupts pituitary T3 responses and impairs TSH repression leading to high circulating TSH, T4 and T3 concentrations. Since TRβ levels are low in bone, T3 responses are unaffected in TRβ−/− mice and high circulating levels of TH act via wild-type TRα1 to induce skeletal thyrotoxicosis. Similarly in TRβPV/PV mice, the dominant-negative TRβ disrupts TSH repression in the pituitary and results in grossly elevated TSH, T4 and T3 concentrations. The low levels of the mutant receptor in bone are insufficient to interfere with TRα1 and impair T3 responses and thus high circulating levels of TH increase expression of T3-target genes resulting in severe skeletal thyrotoxicosis.

  • View in gallery

    Role of TH in endochondral ossification and bone remodelling. (A) The role of T3 in the growth plate is illustrated and the pattern of TR expression is shown. Reserve cells undergo clonal expansion to form columns of proliferating chondrocytes that secrete cartilage matrix stained blue in this section. Committed prehypertrophic chondrocytes undergo hypertrophic differentiation, enlarge and subsequently apoptose, the residual mineralised cartilage matrix forming the scaffold for trabecular bone formation. In chondrocyte cultures, T3 inhibits proliferation and increases the expression of cyclin-dependent kinase inhibitors P21cip-1 P27kip-1. T3 stimulates matrix synthesis and expression of the differentiation markers collagen X, alkaline phosphatase (ALP), matrix metalloproteinase 13 (MMP13) and aggrecanase 2. GH/insulin-like growth factor 1 (GH/IGF1), Indian hedgehog/parathyroid hormone-related peptide (Ihh/PTHrP) and fibroblast growth factor/fibroblast growth factor receptor (FGF/FGFR) pathways are T3 targets. Expression of heparan sulphate proteoglycans (HSPGs), which are essential for FGF and Ihh signalling, is negatively regulated by T3. (B) The role of T3 in the bone remodelling cycle, which is characterised by osteoclastic bone resorption followed by osteoblast migration, osteoid matrix deposition and mineralisation, is shown. Osteoclast differentiation requires direct osteoblast–osteoclast interaction mediated by receptor activator of nuclear factor-κB (RANK) and its ligand RANKL. Osteoprotegerin (OPG) is a decoy receptor secreted by osteoblasts that antagonises this interaction. Osteoclast differentiation also requires co-stimulation by osteoblast-derived cytokines. Thyroid hormone stimulates osteoblast proliferation, differentiation and apoptosis directly and also indirectly by regulating IGF1 and FGF signalling. It remains uncertain whether osteoclast differentiation is regulated directly by T3 or indirectly via T3 actions in osteoblasts.

  • Abel ED, Boers ME, Pazos-Moura C, Moura E, Kaulbach H, Zakaria M, Lowell B, Radovick S, Liberman MC & Wondisford F 1999 Divergent roles for thyroid hormone receptor beta isoforms in the endocrine axis and auditory system. Journal of Clinical Investigation 104 291300.

    • Search Google Scholar
    • Export Citation
  • Abu EO, Bord S, Horner A, Chatterjee VK & Compston JE 1997 The expression of thyroid hormone receptors in human bone. Bone 21 137142.

  • Abu EO, Horner A, Teti A, Chatterjee VK & Compston JE 2000 The localization of thyroid hormone receptor mRNAs in human bone. Thyroid 10 287293.

  • Allain TJ, Chambers TJ, Flanagan AM & McGregor AM 1992 Tri-iodothyronine stimulates rat osteoclastic bone resorption by an indirect effect. Journal of Endocrinology 133 327331.

    • Search Google Scholar
    • Export Citation
  • Barnard JC, Williams AJ, Rabier B, Chassande O, Samarut J, Cheng SY, Bassett JH & Williams GR 2005 Thyroid hormones regulate fibroblast growth factor receptor signaling during chondrogenesis. Endocrinology 146 55685580.

    • Search Google Scholar
    • Export Citation
  • Bassett JH & Williams GR 2003 The molecular actions of thyroid hormone in bone. Trends in Endocrinology and Metabolism 14 356364.

  • Bassett JH & Williams GR 2008 Critical role of the hypothalamic–pituitary–thyroid axis in bone. Bone 43 418426.

  • Bassett JH, Nordstrom K, Boyde A, Howell PG, Kelly S, Vennstrom B & Williams GR 2007a Thyroid status during skeletal development determines adult bone structure and mineralization. Molecular Endocrinology 21 18931904.

    • Search Google Scholar
    • Export Citation
  • Bassett JH, O'Shea PJ, Sriskantharajah S, Rabier B, Boyde A, Howell PG, Weiss RE, Roux JP, Malaval L & Clement-Lacroix P 2007b Thyroid hormone excess rather than thyrotropin deficiency induces osteoporosis in hyperthyroidism. Molecular Endocrinology 21 10951107.

    • Search Google Scholar
    • Export Citation
  • Bassett JH, Williams AJ, Murphy E, Boyde A, Howell PG, Swinhoe R, Archanco M, Flamant F, Samarut J & Costagliola S 2008 A lack of thyroid hormones rather than excess thyrotropin causes abnormal skeletal development in hypothyroidism. Molecular Endocrinology 22 501512.

    • Search Google Scholar
    • Export Citation
  • Bauer DC, Ettinger B, Nevitt MC & Stone LS 2001 Risk for fracture in women with low serum levels of thyroid-stimulating hormone. Annals of Internal Medicine 134 561568.

    • Search Google Scholar
    • Export Citation
  • Boersma B, Otten BJ, Stoelinga GB & Wit JM 1996 Catch-up growth after prolonged hypothyroidism. European Journal of Pediatrics 155 362367.

  • Bookout AL, Jeong Y, Downes M, Yu RT, Evans RM & Mangelsdorf DJ 2006 Anatomical profiling of nuclear receptor expression reveals a hierarchical transcriptional network. Cell 126 789799.

    • Search Google Scholar
    • Export Citation
  • Chassande O, Fraichard A, Gauthier K, Flamant F, Legrand C, Savatier P, Laudet V & Samarut J 1997 Identification of transcripts initiated from an internal promoter in the c-erbA alpha locus that encode inhibitors of retinoic acid receptor-alpha and triiodothyronine receptor activities. Molecular Endocrinology 11 12781290.

    • Search Google Scholar
    • Export Citation
  • Cummings SR, Nevitt MC, Browner WS, Stone K, Fox KM, Ensrud KE, Cauley J, Black D & Vogt TM 1995 Risk factors for hip fracture in white women. Study of Osteoporotic Fractures Research Group. New England Journal of Medicine 332 767773.

    • Search Google Scholar
    • Export Citation
  • Dentice M, Bandyopadhyay A, Gereben B, Callebaut I, Christoffolete MA, Kim BW, Nissim S, Mornon JP, Zavacki AM & Zeold A 2005 The Hedgehog-inducible ubiquitin ligase subunit WSB-1 modulates thyroid hormone activation and PTHrP secretion in the developing growth plate. Nature Cell Biology 7 698705.

    • Search Google Scholar
    • Export Citation
  • van der Eerden BC, Karperien M & Wit JM 2003 Systemic and local regulation of the growth plate. Endocrine Reviews 24 782801.

  • Flamant F, Poguet AL, Plateroti M, Chassande O, Gauthier K, Streichenberger N, Mansouri A & Samarut J 2002 Congenital hypothyroid Pax8(−/−) mutant mice can be rescued by inactivating the TRalpha gene. Molecular Endocrinology 16 2432.

    • Search Google Scholar
    • Export Citation
  • Forrest D, Hanebuth E, Smeyne RJ, Everds N, Stewart CL, Wehner JM & Curran T 1996 Recessive resistance to thyroid hormone in mice lacking thyroid hormone receptor beta: evidence for tissue-specific modulation of receptor function. EMBO Journal 15 30063015.

    • Search Google Scholar
    • Export Citation
  • Fraichard A, Chassande O, Plateroti M, Roux JP, Trouillas J, Dehay C, Legrand C, Gauthier K, Kedinger M & Malaval L 1997 The T3R alpha gene encoding a thyroid hormone receptor is essential for post-natal development and thyroid hormone production. EMBO Journal 16 44124420.

    • Search Google Scholar
    • Export Citation
  • Franklyn JA, Maisonneuve P, Sheppard MC, Betteridge J & Boyle P 1998 Mortality after the treatment of hyperthyroidism with radioactive iodine. New England Journal of Medicine 338 712718.

    • Search Google Scholar
    • Export Citation
  • Gauthier K, Chassande O, Plateroti M, Roux JP, Legrand C, Pain B, Rousset B, Weiss R, Trouillas J & Samarut J 1999 Different functions for the thyroid hormone receptors TRalpha and TRbeta in the control of thyroid hormone production and post-natal development. EMBO Journal 18 623631.

    • Search Google Scholar
    • Export Citation
  • Gauthier K, Plateroti M, Harvey CB, Williams GR, Weiss RE, Refetoff S, Willott JF, Sundin V, Roux JP & Malaval L 2001 Genetic analysis reveals different functions for the products of the thyroid hormone receptor alpha locus. Molecular and Cellular Biology 21 47484760.

    • Search Google Scholar
    • Export Citation
  • Gothe S, Wang Z, Ng L, Kindblom JM, Barros AC, Ohlsson C, Vennstrom B & Forrest D 1999 Mice devoid of all known thyroid hormone receptors are viable but exhibit disorders of the pituitary–thyroid axis, growth, and bone maturation. Genes and Development 13 13291341.

    • Search Google Scholar
    • Export Citation
  • Gouveia CH, Schultz JJ, Bianco AC & Brent GA 2001 Thyroid hormone stimulation of osteocalcin gene expression in ROS 17/2.8 cells is mediated by transcriptional and post-transcriptional mechanisms. Journal of Endocrinology 170 667675.

    • Search Google Scholar
    • Export Citation
  • Gu WX, Stern PH, Madison LD & Du GG 2001 Mutual up-regulation of thyroid hormone and parathyroid hormone receptors in rat osteoblastic osteosarcoma 17/2.8 cells. Endocrinology 142 157164.

    • Search Google Scholar
    • Export Citation
  • Hashimoto K, Curty FH, Borges PP, Lee CE, Abel ED, Elmquist JK, Cohen RN & Wondisford FE 2001 An unliganded thyroid hormone receptor causes severe neurological dysfunction. PNAS 98 39984003.

    • Search Google Scholar
    • Export Citation
  • Heemstra KA, Hamdy NA, Romijn JA & Smit JW 2006 The effects of thyrotropin-suppressive therapy on bone metabolism in patients with well-differentiated thyroid carcinoma. Thyroid 16 583591.

    • Search Google Scholar
    • Export Citation
  • Huang BK, Golden LA, Tarjan G, Madison LD & Stern PH 2000 Insulin-like growth factor I production is essential for anabolic effects of thyroid hormone in osteoblasts. Journal of Bone and Mineral Research 15 188197.

    • Search Google Scholar
    • Export Citation
  • Jones I, Ng L, Liu H & Forrest D 2007 An intron control region differentially regulates expression of thyroid hormone receptor beta2 in the cochlea, pituitary, and cone photoreceptors. Molecular Endocrinology 21 11081119.

    • Search Google Scholar
    • Export Citation
  • Kanatani M, Sugimoto T, Sowa H, Kobayashi T, Kanzawa M & Chihara K 2004 Thyroid hormone stimulates osteoclast differentiation by a mechanism independent of RANKL–RANK interaction. Journal of Cellular Physiology 201 1725.

    • Search Google Scholar
    • Export Citation
  • Kaneshige M, Kaneshige K, Zhu X, Dace A, Garrett L, Carter TA, Kazlauskaite R, Pankratz DG, Wynshaw-Boris A & Refetoff S 2000 Mice with a targeted mutation in the thyroid hormone beta receptor gene exhibit impaired growth and resistance to thyroid hormone. PNAS 97 1320913214.

    • Search Google Scholar
    • Export Citation
  • Kaneshige M, Suzuki H, Kaneshige K, Cheng J, Wimbrow H, Barlow C, Willingham MC & Cheng S 2001 A targeted dominant negative mutation of the thyroid hormone alpha 1 receptor causes increased mortality, infertility, and dwarfism in mice. PNAS 98 1509515100.

    • Search Google Scholar
    • Export Citation
  • Kanis JA & Johnell O 2005 Requirements for DXA for the management of osteoporosis in Europe. Osteoporosis International 16 229238.

  • Kim DJ, Khang YH, Koh JM, Shong YK & Kim GS 2006 Low normal TSH levels are associated with low bone mineral density in healthy postmenopausal women. Clinical Endocrinology 64 8690.

    • Search Google Scholar
    • Export Citation
  • Kindblom JM, Gothe S, Forrest D, Tornell J, Vennstrom B & Ohlsson CGH 2001 GH substitution reverses the growth phenotype but not the defective ossification in thyroid hormone receptor alpha 1−/−beta−/− mice. Journal of Endocrinology 171 1522.

    • Search Google Scholar
    • Export Citation
  • Kindblom JM, Gevers EF, Skrtic SM, Lindberg MK, Gothe S, Tornell J, Vennstrom B & Ohlsson C 2005 Increased adipogenesis in bone marrow but decreased bone mineral density in mice devoid of thyroid hormone receptors. Bone 36 607616.

    • Search Google Scholar
    • Export Citation
  • Liu YY, Schultz JJ & Brent GA 2003 A thyroid hormone receptor alpha gene mutation (P398H) is associated with visceral adiposity and impaired catecholamine-stimulated lipolysis in mice. Journal of Biological Chemistry 278 3891338920.

    • Search Google Scholar
    • Export Citation
  • Lupu F, Terwilliger JD, Lee K, Segre GV & Efstratiadis A 2001 Roles of growth hormone and insulin-like growth factor 1 in mouse postnatal growth. Developmental Biology 229 141162.

    • Search Google Scholar
    • Export Citation
  • Mansouri A, Chowdhury K & Gruss P 1998 Follicular cells of the thyroid gland require Pax8 gene function. Nature Genetics 19 8790.

  • Mittag J, Friedrichsen S, Heuer H, Polsfuss S, Visser TJ & Bauer K 2005 Athyroid Pax8−/− mice cannot be rescued by the inactivation of thyroid hormone receptor alpha1. Endocrinology 146 31793184.

    • Search Google Scholar
    • Export Citation
  • Miura M, Tanaka K, Komatsu Y, Suda M, Yasoda A, Sakuma Y, Ozasa A & Nakao K 2002 A novel interaction between thyroid hormones and 1,25(OH)(2)D(3) in osteoclast formation. Biochemical and Biophysical Research Communications 291 987994.

    • Search Google Scholar
    • Export Citation
  • Morris MS 2007 The association between serum thyroid-stimulating hormone in its reference range and bone status in postmenopausal American women. Bone 40 11281134.

    • Search Google Scholar
    • Export Citation
  • Mosekilde L, Eriksen EF & Charles P 1990 Effects of thyroid hormones on bone and mineral metabolism. Endocrinology and Metabolism Clinics of North America 19 3563.

    • Search Google Scholar
    • Export Citation
  • Murakami S, Balmes G, McKinney S, Zhang Z, Givol D & de Crombrugghe B 2004 Constitutive activation of MEK1 in chondrocytes causes Stat1-independent achondroplasia-like dwarfism and rescues the Fgfr3-deficient mouse phenotype. Genes and Development 18 290305.

    • Search Google Scholar
    • Export Citation
  • Murphy E & Williams GR 2004 The thyroid and the skeleton. Clinical Endocrinology 61 285298.

  • Ng L, Hurley JB, Dierks B, Srinivas M, Salto C, Vennstrom B, Reh TA & Forrest D 2001 A thyroid hormone receptor that is required for the development of green cone photoreceptors. Nature Genetics 27 9498.

    • Search Google Scholar
    • Export Citation
  • Ornitz DM 2005 FGF signaling in the developing endochondral skeleton. Cytokine and Growth Factor Reviews 16 205213.

  • O'Shea PJ, Harvey CB, Suzuki H, Kaneshige M, Kaneshige K, Cheng SY & Williams GR 2003 A thyrotoxic skeletal phenotype of advanced bone formation in mice with resistance to thyroid hormone. Molecular Endocrinology 17 14101424.

    • Search Google Scholar
    • Export Citation
  • O'Shea PJ, Bassett JH, Sriskantharajah S, Ying H, Cheng SY & Williams GR 2005 Contrasting skeletal phenotypes in mice with an identical mutation targeted to thyroid hormone receptor alpha1 or beta. Molecular Endocrinology 19 30453059.

    • Search Google Scholar
    • Export Citation
  • O'Shea PJ, Bassett JH, Cheng SY & Williams GR 2006 Characterization of skeletal phenotypes of TRalpha1 and TRbeta mutant mice: implications for tissue thyroid status and T3 target gene expression. Nuclear Receptor Signaling 4 e011

    • Search Google Scholar
    • Export Citation
  • O'Shea PJ, Guigon CJ, Williams GR & Cheng SY 2007 Regulation of fibroblast growth factor receptor-1 by thyroid hormone: identification of a thyroid hormone response element in the murine Fgfr1 promoter. Endocrinology

    • Search Google Scholar
    • Export Citation
  • Pepene CE, Kasperk CH, Pfeilschifter J, Borcsok I, Gozariu L, Ziegler R & Seck T 2001 Effects of triiodothyronine on the insulin-like growth factor system in primary human osteoblastic cells in vitro. Bone 29 540546.

    • Search Google Scholar
    • Export Citation
  • Pereira RC, Jorgetti V & Canalis E 1999 Triiodothyronine induces collagenase-3 and gelatinase B expression in murine osteoblasts. American Journal of Physiology 277 E496E504.

    • Search Google Scholar
    • Export Citation
  • Quan ML, Pasieka JL & Rorstad O 2002 Bone mineral density in well-differentiated thyroid cancer patients treated with suppressive thyroxine: a systematic overview of the literature. Journal of Surgical Oncology 79 6269.(discussion 69–70)

    • Search Google Scholar
    • Export Citation
  • Quignodon L, Vincent S, Winter H, Samarut J & Flamant F 2007 A point mutation in the activation function 2 domain of thyroid hormone receptor alpha1 expressed after CRE-mediated recombination partially recapitulates hypothyroidism. Molecular Endocrinology 21 23502360.

    • Search Google Scholar
    • Export Citation
  • Rivkees SA, Bode HH & Crawford JD 1988 Long-term growth in juvenile acquired hypothyroidism: the failure to achieve normal adult stature. New England Journal of Medicine 318 599602.

    • Search Google Scholar
    • Export Citation
  • Robson H, Siebler T, Stevens DA, Shalet SM & Williams GR 2000 Thyroid hormone acts directly on growth plate chondrocytes to promote hypertrophic differentiation and inhibit clonal expansion and cell proliferation. Endocrinology 141 38873897.

    • Search Google Scholar
    • Export Citation
  • Salto C, Kindblom JM, Johansson C, Wang Z, Gullberg H, Nordstrom K, Mansen A, Ohlsson C, Thoren P & Forrest D 2001 Ablation of TRalpha2 and a concomitant overexpression of alpha1 yields a mixed hypo- and hyperthyroid phenotype in mice. Molecular Endocrinology 15 21152128.

    • Search Google Scholar
    • Export Citation
  • Segni M & Gorman CA 2001 The aftermath of childhood hyperthyroidism. Journal of Pediatric Endocrinology and Metabolism 14 12771282.(discussion 1297–1298)

    • Search Google Scholar
    • Export Citation
  • Shao YY, Wang L & Ballock RT 2006 Thyroid hormone and the growth plate. Reviews in Endocrine and Metabolic Disorders 7 265271.

  • Siddiqi A, Burrin JM, Wood DF & Monson JP 1998 Tri-iodothyronine regulates the production of interleukin-6 and interleukin-8 in human bone marrow stromal and osteoblast-like cells. Journal of Endocrinology 157 453461.

    • Search Google Scholar
    • Export Citation
  • Stevens DA, Hasserjian RP, Robson H, Siebler T, Shalet SM & Williams GR 2000 Thyroid hormones regulate hypertrophic chondrocyte differentiation and expression of parathyroid hormone-related peptide and its receptor during endochondral bone formation. Journal of Bone and Mineral Research 15 24312442.

    • Search Google Scholar
    • Export Citation
  • Stevens DA, Harvey CB, Scott AJ, O'Shea PJ, Barnard JC, Williams AJ, Brady G, Samarut J, Chassande O & Williams GR 2003 Thyroid hormone activates fibroblast growth factor receptor-1 in bone. Molecular Endocrinology 17 17511766.

    • Search Google Scholar
    • Export Citation
  • Tinnikov A, Nordstrom K, Thoren P, Kindblom JM, Malin S, Rozell B, Adams M, Rajanayagam O, Pettersson S & Ohlsson C 2002 Retardation of post-natal development caused by a negatively acting thyroid hormone receptor alpha1. EMBO Journal 21 50795087.

    • Search Google Scholar
    • Export Citation
  • Varga F, Spitzer S & Klaushofer K 2004 Triiodothyronine (T3) and 1,25-dihydroxyvitamin D3 (1,25D3) inversely regulate OPG gene expression in dependence of the osteoblastic phenotype. Calcified Tissue International 74 382387.

    • Search Google Scholar
    • Export Citation
  • Vestergaard P & Mosekilde L 2002 Fractures in patients with hyperthyroidism and hypothyroidism: a nationwide follow-up study in 16 249 patients. Thyroid 12 411419.

    • Search Google Scholar
    • Export Citation
  • Vestergaard P, Rejnmark L, Weeke J & Mosekilde L 2000 Fracture risk in patients treated for hyperthyroidism. Thyroid 10 341348.

  • Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM & Tabin CJ 1996 Regulation of rate of cartilage differentiation by Indian hedgehog and PTH-related protein. Science 273 613622.

    • Search Google Scholar
    • Export Citation
  • Wikstrom L, Johansson C, Salto C, Barlow C, Campos Barros A, Baas F, Forrest D, Thoren P & Vennstrom B 1998 Abnormal heart rate and body temperature in mice lacking thyroid hormone receptor alpha 1. EMBO Journal 17 455461.

    • Search Google Scholar
    • Export Citation
  • Williams GR, Bland R & Sheppard MC 1994 Characterization of thyroid hormone (T3) receptors in three osteosarcoma cell lines of distinct osteoblast phenotype: interactions among T3, vitamin D3, and retinoid signaling. Endocrinology 135 23752385.

    • Search Google Scholar
    • Export Citation
  • Yen PM 2001 Physiological and molecular basis of thyroid hormone action. Physiological Reviews 81 10971142.

  • Zhou YX, Xu X, Chen L, Li C, Brodie SG & Deng CX 2000 A Pro250Arg substitution in mouse Fgfr1 causes increased expression of Cbfa1 and premature fusion of calvarial sutures. Human Molecular Genetics 9 20012008.

    • Search Google Scholar
    • Export Citation