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

Reproductive Biology and Endocrinology

Chawnshang Chang²  Chih-Rong Shyr²  Gonghui Li¹  Su Liu¹  Ning-Chun Liu¹  Yi-Fen Lee¹  Hsin-Chiu Ho¹  Shin-Jen Lin¹ 

[1] George Whipple Lab for Cancer Research, Departments of Pathology, Urology, Radiation Oncology, and The Wilmot Cancer center, University of Rochester Medical Center, Rochester, NY, 14642, USA;Sex Hormone Research Center, China Medical University/Hospital, Taichung, 404, Taiwan

关键词: Osteoporosis;    Bone;    Nuclear receptor;    TR4;   

Others  :  1150635
DOI  :  10.1186/1477-7827-10-43

 received in 2012-02-18, accepted in 2012-05-29,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Lin et al.; licensee BioMed Central Ltd.

【 图 表 】

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