Thyroid Hormones Promote Chondrocyte Differentiation in Mouse ATDC5 Cells and Stimulate Endochondral Ossification in Fetal Mouse Tibias Through Iodothyronine Deiodinases in the Growth Plate

Abstract

Thyroid hormones (THs), 3,3,5-triiodo-L-thyronine (T3) and L-thyroxine (T4), are important for the normal development of the growth plate (GP); congenital TH deficiency leads to severe dwarfism. In mouse chondrogenic cell line, ATDC5, T3 enhanced differentiation and increased Alizarin red staining, but did not affect Alcian blue staining. In organ-cultured mouse tibias, THs stimulated the cartilage growth, especially in the hypertrophic zone. Interestingly, T4 was as equally potent as T3 in organ-cultured tibias, which suggests that T4 is metabolized locally to T3, because T4 is a prohormone and must be converted to T3 for its activity. Two enzymes catalyze the conversion; type I deiodinase (D1) and type II deiodinase (D2). D1 has a ubiquitous distribution and D2, with a high affinity for T4, is present where the maintenance of intracellular T3 concentration is critical. Messenger RNAs (mRNAs) for D1 and D2 were detected in neonatal mouse tibias and ATDC5 cells. The enzyme activity was unaffected by the D1 inhibitor 6-propyl-2-thiouracil, suggesting that D2 mainly catalyzes the reaction. D2 mRNA was detected in differentiated ATDC5 cells. In organ-cultured mouse tibias, D2 activity was greater at later stages. In contrast, thyroid hormone receptors (TRs) were expressed in neonatal mouse tibias and ATDC5 cells, but their expression levels in ATDC5 cells were stable throughout the culture periods. Therefore, increased T3 production at later stages by D2 is likely to contribute to the preferential effects of THs in the terminal differentiation of GP. This article is the first to show that T4 is activated locally in GP and enhances the understanding of TH effects in GP. (J Bone Miner Res 2002;17: 443-454)

American Thyroid Association Guide to Investigating Thyroid Hormone Economy and Action in Rodent and Cell Models

“…BSA with fatty acids may have T 3 , thus fatty acid-free BSA should be used (282). It should also be noted that most commercially available forms of T 4 have at least trace T 3 contamination that must be considered.…”

Molecular basis of imprinting disorders affecting chromosome 14: lessons from murine models

“…T 3 promotes chondrocyte differentiation in ATDC5 cells (Miura et al 2002) as well as mesenchymal stem cells (Karl et al 2014) and in cultured epiphyseal chondrocytes (Wang et al 2007). Dio3 is expressed in developing cartilage and the direct action of D3 in cartilage may thus result in a reduction in chondrogenesis.…”

O efeito molecular e estrutural do hormônio tiroideano no esqueleto

“…Demonstrou-se que o T 3 r ecruta e ativa osteoclastos em culturas de ossos de roedores por um mecanismo modulado pelos osteoblastos (40). Recentemente, demonstrou-se que o T 3 a u m e n t a a expressão do mRNA do ligante do receptor activator of nuclear factor-kB (RANKL) em células pré-osteoblás-ticas (42). O RANKL é uma molécula chave para a osteoclastogenese e ativação osteoclástica.…”

“…Esse achado levou à identificação da expressão do mRNA da desiodase das iodotironinas do tipo 2 (D2), uma das enzimas que convertem T 4 a T 3 , em células pré-osteoblásticas. Demonstrou-se, ainda, que a VD aumenta a expressão da D2 nessas células de maneira dependente da dose e do tempo, o que indica um novo mecanismo de sinergismo entre a VD e o T 3 na osteoclastogenese (42).…”