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. 2014 Oct;24(10):1266-9.
doi: 10.1038/cr.2014.111. Epub 2014 Aug 22.

Osteogenic fate of hypertrophic chondrocytes

Guan Yang  1 , Liang Zhu  1 , Ning Hou  1 , Yu Lan  1 , Xi-Mei Wu  2 , Bin Zhou  3 , Yan Teng  1 , Xiao Yang  1
Affiliations

Osteogenic fate of hypertrophic chondrocytes

Guan Yang et al. Cell Res. 2014 Oct.
No abstract available

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Figures

Figure 1
Figure 1
Multipotential fates of hypertrophic chondrocyte. (A) The transgenic vector contained an 8.2 kb promoter of mouse type X collagen (Col10a1), Cre cDNA, human growth hormone (hGH) polyadenylation signal and a 3.2 kb 2nd intron of Col10a1. Isotopic in situ hybridization assay was used to detect the transgenic Cre mRNA in tibia proximal growth plate from Col10a1int2-Cre transgenic mice at E16.5, P1, P5 and P10. (B) Double staining of YFP (blue) by immunohistochemistry (IHC) and Cre mRNA (fuchsia) by in situ hybridization in 10-day-old Col10a1int2-Cre;ROSAEYFP tibia. The right panel shows the high-magnification image of the area boxed in red on the left. (C) After 1 and 5 days following tamoxifen administration (50 mg/kg) at P5, Col2a1-CreERT;ROSAEYFP tibia was stained by YFP (blue) and Col1a1 (fuchsia) antibodies. The clonal columns of YFP+ cells were detectable in Col2a1-CreERT;ROSAEYFP chondrocyte zone. (D) After 10 days following tamoxifen administration at P5, Col2a1-CreERT;ROSAEYFP tibia was stained by YFP (blue) antibody, showing a column of YFP+ cells entering into the metaphysis. IHC double staining of YFP (blue) and type I collagen (Col1a1) (fuchsia) showed that some YFP+ cells underneath the growth plate were labeled with Col1a1 (red frames, high-magnification images of areas boxed on the left). (E) After 10 days following tamoxifen administration (50 mg/kg) at P5, Col2a1-CreERT;ROSAConfetti tibia was stained by RFP (fuchsia) antibody. Double staining of RFP (fuchsia) by IHC and Col1a1 (purple grey) mRNA by in situ hybridization showed that some RFP+ cells in metaphysis were Col1a1-positive (red frames, high-magnification images of areas boxed on the left). (F) Double staining of YFP (fuchsia) by IHC and the transcripts of Col1a1 or osteocalcin (Ocn) (blue) by in situ hybridization in metaphyseal, trabecular and cortical bones from 20-day-old Col10a1int2-Cre;ROSAEYFP tibia. The percentages of YFP+Col1a1+ and YFP+Ocn+osteogenic cells in total Col1a1+and Ocn+ cells from 3 images per mouse (n = 3) are shown in the top-right corner. Von Kossa staining showed that some YFP+cells (fuchsia) within the mineralized bone were osteocytes. (G) In 20-day-old Col10a1int2-Cre;ROSAEYFP bone marrow (BM), double immunostaining (left) of YFP (fuchsia) and endomucin (blue, marker for endothelial cells) as well as double immunofluorescence analysis (middle) using YFP (red) and PDGFR-β (green, marker for pericytes) antibodies indicated that some YFP+ cells were adjacent to the endothelial cells and vascular pericytes. Double immunostaining (right) of YFP (fuchsia) and perilipin (blue, marker for adipocytes) showed that some YFP+ cells were adipocytes (red arrow). (H) Triple staining of YFP (blue) and BrdU (brown) by IHC along with Col1a1 mRNA (fuchsia) by in situ hybridization in 20-day-old Col10a1int2-Cre;ROSAEYFP metaphyseal bones (MB). PZ, proliferation zone; HZ, hypertrophic zone; MB, metaphyseal bone; TB, trabecular bone; BM, bone marrow. Scale bars are 200 μm (A-C), 50 μm (D-F) and 25 μm (G, H).
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