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Reproductive Biology and Endocrinology

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Open Access 01-12-2012 | Research

Reduced osteoblast activity in the mice lacking TR4 nuclear receptor leads to osteoporosis

Authors: Shin-Jen Lin, Hsin-Chiu Ho, Yi-Fen Lee, Ning-Chun Liu, Su Liu, Gonghui Li, Chih-Rong Shyr, Chawnshang Chang

Published in: Reproductive Biology and Endocrinology | Issue 1/2012

Abstract

Background

Early studies suggested that TR4 nuclear receptor might play important roles in the skeletal development, yet its detailed mechanism remains unclear.

Methods

We generated TR4 knockout mice and compared skeletal development with their wild type littermates. Primary bone marrow cells were cultured and we assayed bone differentiation by alkaline phosphatase and alizarin red staining. Primary calvaria were cultured and osteoblastic marker genes were detected by quantitative PCR. Luciferase reporter assays, chromatin immunoprecipitation (ChIP) assays, and electrophoretic mobility shift assays (EMSA) were performed to demonstrate TR4 can directly regulate bone differentiation marker osteocalcin.

Results

We first found mice lacking TR4 might develop osteoporosis. We then found that osteoblast progenitor cells isolated from bone marrow of TR4 knockout mice displayed reduced osteoblast differentiation capacity and calcification. Osteoblast primary cultures from TR4 knockout mice calvaria also showed higher proliferation rates indicating lower osteoblast differentiation ability in mice after loss of TR4. Mechanism dissection found the expression of osteoblast markers genes, such as ALP, type I collagen alpha 1, osteocalcin, PTH, and PTHR was dramatically reduced in osteoblasts from TR4 knockout mice as compared to those from TR4 wild type mice. In vitro cell line studies with luciferase reporter assay, ChIP assay, and EMSA further demonstrated TR4 could bind directly to the promoter region of osteocalcin gene and induce its gene expression at the transcriptional level in a dose dependent manner.

Conclusions

Together, these results demonstrate TR4 may function as a novel transcriptional factor to play pathophysiological roles in maintaining normal osteoblast activity during the bone development and remodeling, and disruption of TR4 function may result in multiple skeletal abnormalities.

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Lee YF, Lee HJ, Chang C: Recent advances in the TR2 and TR4 orphan receptors of the nuclear receptor superfamily. J Steroid Biochem Mol Biol. 2002, 81 (4–5): 291-308.PubMed

Chang C, Da Silva SL, Ideta R, Lee YF, Yeh S, Burbach JPH: Human and rat TR4 orphan receptors specify a subclass of the steroid receptor superfamily. Proceedings of the National Academy of Sciences USA. 1994, 1994: 6040-6044.CrossRef

Lee YF, Pan HJ, Burbach PH, Morkin E, Chang C: Identification of direct repeat 4 as a positive regulatory element for the human TR4 orphan receptor: a modulator for the thyroid hormone target genes. J Biol Chem. 1997, 272: 12215-12220. 10.1074/jbc.272.18.12215.CrossRefPubMed

Lee YF, Young WJ, Burbach JPH, Chang C: Negative feedback control of the retinoid-retinoic acid/retinoid X receptor pathway by the human TR4 orphan receptor, a member of the steroid receptor superfamily. J Biol Chem. 1998, 273: 13437-13443. 10.1074/jbc.273.22.13437.CrossRefPubMed

Lee YF, Young WJ, Lin WJ, Shyr CR, Chang C: Differential regulation of direct repeat 3 vitamin D3 and direct repeat 4 thyroid hormone signaling pathways by the human TR4 orphan receptor. J Biol Chem. 1999, 274: 16198-16205. 10.1074/jbc.274.23.16198.CrossRefPubMed

Lee Y-F, Shyr CR, Thin TH, Lin WJ, Chang C: Convergence of two repressors through heterodimer formation of androgen receptor and testicular orphan receptor-4: a unique signaling pathway in the steroid receptor superfamily. Proceedings of the National Academy of Sciences USA. 1999, 96: 14724-14729. 10.1073/pnas.96.26.14724.CrossRef

Collins LL, Lee YF, Heinlein CA, Liu NC, Chen YT, Shyr CR, Meshul CK, Uno H, Platt KA, Chang C: Growth retardation and abnormal maternal behavior in mice lacking testicular orphan nuclear receptor 4. Proc Natl Acad Sci USA. 2004, 101 (42): 15058-15063. 10.1073/pnas.0405700101.PubMedCentralCrossRefPubMed

Chen LM, Wang RS, Lee YF, Liu NC, Chang YJ, Wu CC, Xie S, Hung YC, Chang C: Subfertility with defective folliculogenesis in female mice lacking testicular orphan nuclear receptor 4. Mol Endocrinol. 2008, 22 (4): 858-867. 10.1210/me.2007-0181.PubMedCentralCrossRefPubMed

Chen YT, Collins LL, Uno H, Chang C: Deficits in motor coordination with aberrant cerebellar development in mice lacking testicular orphan nuclear receptor 4. Mol Cell Biol. 2005, 25 (7): 2722-2732. 10.1128/MCB.25.7.2722-2732.2005.PubMedCentralCrossRefPubMed

10.

Chen YT, Collins LL, Chang SS, Chang C: The roles of testicular orphan nuclear receptor 4 (TR4) in cerebellar development. Cerebellum. 2008, 7 (1): 9-17. 10.1007/s12311-008-0006-3.CrossRefPubMed

11.

Mu X, Lee Y, Liu N, Chen Y, Kim E, Shyr C, Chang C: Targeted inactivation of Testicular nuclear Orphan Receptor 4 delays and disrupts late meiotic prophase and subsequent meiotic divisions of spermatogenesis. Mol Cell Biochem. 2004, 24: 5887-5899. 10.1128/MCB.24.13.5887-5899.2004.CrossRef

12.

Zhang Y, Chen YT, Xie S, Wang L, Lee YF, Chang SS, Chang C: Loss of testicular orphan receptor 4 impairs normal myelination in mouse forebrain. Mol Endocrinol. 2007, 21 (4): 908-920.CrossRefPubMed

13.

Liu NC, Lin WJ, Kim E, Collins LL, Lin HY, Yu IC, Sparks JD, Chen LM, Lee YF, Chang C: Loss of TR4 orphan nuclear receptor reduces phosphoenolpyruvate carboxykinase-mediated gluconeogenesis. Diabetes. 2007, 56 (12): 2901-2909. 10.2337/db07-0359.CrossRefPubMed

14.

Kim E, Liu NC, Yu IC, Lin HY, Lee YF, Sparks JD, Chen LM, Chang C: Metformin inhibits nuclear receptor TR4-mediated hepatic stearoyl-coenzyme a desaturase 1 gene expression with altered insulin sensitivity. Diabetes. 2011, 60 (5): 1493-1503. 10.2337/db10-0393.PubMedCentralCrossRefPubMed

15.

Xie S, Lee YF, Kim E, Chen LM, Ni J, Fang LY, Liu S, Lin SJ, Abe J, Berk B, et al: TR4 nuclear receptor functions as a fatty acid sensor to modulate CD36 expression and foam cell formation. Proc Natl Acad Sci U S A. 2009, 106 (32): 13353-13358. 10.1073/pnas.0905724106.PubMedCentralCrossRefPubMed

16.

Collins LL, Lee YF, Heinlein CA, Liu NC, Chen YT, Shyr CR, Meshul CK, Uno H, Platt KA, Chang C: Growth retardation and abnormal maternal behavior in mice lacking testicular orphan nuclear receptor 4. Proc Natl Acad Sci U S A. 2004, 101 (42): 15058-15063. 10.1073/pnas.0405700101.PubMedCentralCrossRefPubMed

17.

Zhang X, Schwarz EM, Young DA, Puzas JE, Rosier RN, O'Keefe RJ: Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair. J Clin Invest. 2002, 109 (11): 1405-1415.PubMedCentralCrossRefPubMed

18.

Roy ME, Nishimoto SK, Rho JY, Bhattacharya SK, Lin JS, Pharr GM: Correlations between osteocalcin content, degree of mineralization, and mechanical properties of C. carpio rib bone. J Biomed Mater Res. 2001, 54 (4)): 547-553.CrossRefPubMed

19.

Lee NK, Sowa H, Hinoi E, Ferron M, Ahn JD, Confavreux C, Dacquin R, Mee PJ, McKee MD, Jung DY, et al: Endocrine regulation of energy metabolism by the skeleton. Cell. 2007, 130 (3): 456-469. 10.1016/j.cell.2007.05.047.PubMedCentralCrossRefPubMed

20.

Gutierrez S, Liu J, Javed A, Montecino M, Stein GS, Lian JB, Stein JL: The vitamin D response element in the distal osteocalcin promoter contributes to chromatin organization of the proximal regulatory domain. J Biol Chem. 2004, 279 (42): 43581-43588. 10.1074/jbc.M408335200.CrossRefPubMed

21.

Sierra J, Villagra A, Paredes R, Cruzat F, Gutierrez S, Javed A, Arriagada G, Olate J, Imschenetzky M, Van Wijnen AJ, et al: Regulation of the bone-specific osteocalcin gene by p300 requires Runx2/Cbfa1 and the vitamin D3 receptor but not p300 intrinsic histone acetyltransferase activity. Mol Cell Biol. 2003, 23 (9): 3339-3351. 10.1128/MCB.23.9.3339-3351.2003.PubMedCentralCrossRefPubMed

22.

Javed A, Gutierrez S, Montecino M, van Wijnen AJ, Stein JL, Stein GS, Lian JB: Multiple Cbfa/AML sites in the rat osteocalcin promoter are required for basal and vitamin D-responsive transcription and contribute to chromatin organization. Mol Cell Biol. 1999, 19 (11): 7491-7500.PubMedCentralCrossRefPubMed

23.

Shyr CR, Collins LL, Mu XM, Platt KA, Chang C: Spermatogenesis and testis development are normal in mice lacking testicular orphan nuclear receptor 2. Mol Cell Biol. 2002, 22 (13): 4661-4666. 10.1128/MCB.22.13.4661-4666.2002.PubMedCentralCrossRefPubMed

24.

Kawai M, Rosen CJ: PPARgamma: a circadian transcription factor in adipogenesis and osteogenesis. Nat Rev Endocrinol. 2010, 6 (11): 629-636. 10.1038/nrendo.2010.155.PubMedCentralCrossRefPubMed

Title: Reduced osteoblast activity in the mice lacking TR4 nuclear receptor leads to osteoporosis
Authors: Shin-Jen Lin
Hsin-Chiu Ho
Yi-Fen Lee
Ning-Chun Liu
Su Liu
Gonghui Li
Chih-Rong Shyr
Chawnshang Chang
Publication date: 01-12-2012
Publisher: BioMed Central
Published in: Reproductive Biology and Endocrinology / Issue 1/2012
Electronic ISSN: 1477-7827
DOI: https://doi.org/10.1186/1477-7827-10-43

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Reduced osteoblast activity in the mice lacking TR4 nuclear receptor leads to osteoporosis

Abstract

Background

Methods

Results

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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