W. Wang, T. Hayami, S. Kapila Osteoarthritis and Cartilage

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Presentation transcript:

Female hormone receptors are differentially expressed in mouse fibrocartilages W. Wang, T. Hayami, S. Kapila Osteoarthritis and Cartilage Volume 17, Issue 5, Pages 646-654 (May 2009) DOI: 10.1016/j.joca.2008.09.015 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Localization of ERs ER-α and ER-β, relaxin receptors LGR7 and LGR8, and PR in mouse TMJ disc, KM, and pubic symphysis (PS) fibrochondrocytes. ER-α, ER-β, LGR7 and PR were highly expressed in the TMJ and pubic symphysis fibrochondrocytes while LGR8 was detected at higher levels in KM and PS than in TMJ cells. Negative controls (NC) show pubic symphysis cells subjected to this assay in the absence of primary antibody. The nuclei were stained with DAPI. (Magnification 40×; bar=50μm.) Osteoarthritis and Cartilage 2009 17, 646-654DOI: (10.1016/j.joca.2008.09.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 In situ immunolocalization of ERs ER-α and ER-β, relaxin receptors LGR7 and LGR8 and PR in mouse TMJ, KM, and pubic symphysis (PS) fibrocartilages. ER-α, ER-β, LGR7 and PR were more highly expressed in the TMJ and pubic symphysis fibrocartilages but only a few positively stained cells were found in the KM fibrocartilage. In contrast, LGR8 was mainly expressed in the cells of KM and pubic symphysis. Negative controls (NC) show pubic symphysis tissues subjected to this assay in the absence of primary antibody. Arrows point to positively stained cells. (Magnification 20×; bar=50μm.) Osteoarthritis and Cartilage 2009 17, 646-654DOI: (10.1016/j.joca.2008.09.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Quantitative differential gene expression for ERs, ER-α (A), ER-β (B), relaxin receptors, LGR7 (C) and LGR8 (D), and PR (E) in female mouse TMJ disc, KM and pubic symphysis (PS) fibrochondrocytes. ER-α, ER-β, LGR7 and PR were highly expressed in the TMJ and pubic symphysis cells, while LGR8 was highly expressed in KM cells. Data represent means±SD fold levels of respective receptors relative to those in the KM from three independent experiments, each performed in triplicate. (n=3; *P<0.01, **P<0.05.) Osteoarthritis and Cartilage 2009 17, 646-654DOI: (10.1016/j.joca.2008.09.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Differential expression of ERs ER-α and ER-β, relaxin receptors LGR7 and LGR8, and PR in mouse TMJ, KM, and pubic symphysis (PS) fibrochondrocytes. (A). Total RNA, standardized for RNA concentration, was subjected to semi-quantitative RT-PCR for hormone receptors. GAPDH was used as an internal control. (B) Alternatively, cell lysates, standardized for total protein, were subjected to Western blot assays for each of the hormone receptors. Actin was used as an internal control for the Western blots. Both the genes and protein for ER-α, ER-β, LGR-7, and PR were highly expressed in the TMJ and pubic symphysis fibrochondrocytes and were weakly expressed in the KM cells. In contrast, higher levels of LGR8 were detected in KM and pubic symphysis cells as compared to that in TMJ cells. Osteoarthritis and Cartilage 2009 17, 646-654DOI: (10.1016/j.joca.2008.09.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 TMJ, KM and pubic symphysis (PS) fibrochondrocytes show distinctly different ratios of gene levels for ERs ER-α:ER-β (A), and relaxin receptors LGR7:LGR8 (B). A significantly higher expression ratio of ERs ER-α:ER-β and relaxin receptors LGR7:LGR8 was found in the TMJ and pubic symphysis fibrochondrocytes than in KM cells. Data represent means±SD from three independent experiments, each performed in triplicate. (n=3; *P<0.01, **P<0.05.) Osteoarthritis and Cartilage 2009 17, 646-654DOI: (10.1016/j.joca.2008.09.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Gene expression levels for ERs but not relaxin receptors or PR demonstrate sexual dimorphism in mouse TMJ fibrochondrocytes. Hormone receptor gene expression levels in male cells were standardized to that in female cells. Male TMJ cells had a significantly lower gene expression levels for ER-α (67.3%) and ER-β (24.9%), but not for LGR7 (88.7%), LGR8 (75.9%) and PR (116%) relative to that in female TMJ cells. Data represent means±SD from three independent experiments, each performed in triplicate. (n=3; *P<0.01, **P<0.05.) Osteoarthritis and Cartilage 2009 17, 646-654DOI: (10.1016/j.joca.2008.09.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions