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. 2001 May;2(5):446-52.
doi: 10.1093/embo-reports/kve094.

The collagen receptor DDR2 regulates proliferation and its elimination leads to dwarfism

J P Labrador  1 , V Azcoitia, J Tuckermann, C Lin, E Olaso, S Mañes, K Brückner, J L Goergen, G Lemke, G Yancopoulos, P Angel, C Martínez, R Klein
Affiliations

The collagen receptor DDR2 regulates proliferation and its elimination leads to dwarfism

J P Labrador et al. EMBO Rep. 2001 May.

Abstract

The discoidin domain receptor 2 (DDR2) is a member of a subfamily of receptor tyrosine kinases whose ligands are fibrillar collagens, and is widely expressed in postnatal tissues. We have generated DDR2-deficient mice to establish the in vivo functions of this receptor, which have remained obscure. These mice exhibit dwarfism and shortening of long bones. This phenotype appears to be caused by reduced chondrocyte proliferation, rather than aberrant differentiation or function. In a skin wound healing model, DDR2-/- mice exhibit a reduced proliferative response compared with wild-type littermates. In vitro, fibroblasts derived from DDR2-/- mutants proliferate more slowly than wild-type fibroblasts, a defect that is rescued by introduction of wild-type but not kinase-dead DDR2 receptor. Together our results suggest that DDR2 acts as an extracellular matrix sensor to modulate cell proliferation.

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Figures

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Fig. 1. Generation of DDR2 null mice. (A) Genomic organization of the DDR2 wild-type allele, targeting vector and targeted allele after homologous recombination. (B, BamHI; E, EcoRI). (B) Southern blot analysis of genomic DNA from ES cell clones. EcoRI fragments corresponding to wild-type (3 kb) and targeted (5 kb) alleles are detected. (C) Detection of DDR2 protein in muscle extracts by immunoprecipitation (IP) and western blot analysis (WB) using DDR2-specific antiserum. (D) Growth curve of DDR2–/– mutant versus heterozygous littermates (129/Sv × C57BL/6J background). Each point represents the mean (± SEM) of at least nine individuals. (E) Representative images of Alizarin red-stained bones from wild-type (+/+) and homozygous mutant (–/–) mice. Scale bar = 2 mm.
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Fig. 2. DDR2 expression in bone. In situ hybridization analysis of sections from 1-week-old tibia from wild-type mice, hybridized with DDR2 antisense (A, B) or sense (C) cRNA probes. (B) High power bright field image of DDR2 mRNA expression (black silver grains) in a hematoxylin/eosin-stained section from proliferating chondrocytes. The area shown corresponds to the white square in (A). Scale bar = 0.5 mm.
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Fig. 3. Differentiation markers in DDR2–/– growth plates. In situ hybridization analysis of sections from 2-week-old wild-type (A, C, E, G) and homozygous mutant (B, D, F, H) tibias. Antisense cRNA probes for type II collagen (A and B), type X collagen (C and D), MMP-9 (E and F), and MMP-13 (G and H). Lack of differences between +/+ and DDR2–/– material suggests normal differentiation in mutant bones. Scale bar = 0.5 mm.
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Fig. 4. Reduced chondrocyte proliferation in DDR2–/– mice. (A) In vivo BrdU labelling (brown) in metatarsals of wild-type (+/+) and DDR2–/– mice (–/–) at 2 weeks of age. (B) Quantification of BrdU incorporation. Data represent mean ± SEM (** P <0.001). (C) Apoptotic cells in metatarsals from 2-week-old wild-type (+/+) and homozygous mutant (–/–) mice, stained by TUNEL. Arrowheads indicate apoptotic hypertrophic chondrocytes. There is no apparent difference in the number of TUNEL-positive cells in DDR2–/– or wild-type bones. Scale bar = 0.5 mm.
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Fig. 5. Reduced BrdU incorporation in skin. (A) Broad tissue distribution of DDR2 (top panel) shown by immunoprecipitation (IP) and western blotting (WB) with anti-DDR2 antibodies. WB with anti-phosphotyrosine specific antibodies (bottom panel) reveals high levels of phosphorylated DDR2 protein in the skin. (B, C, D, E) In vivo BrdU labelling in skin wounds of wild-type (B and D) and DDR2–/– mice (C and E ), 2.5 days (B and C) and 4.5 days after wounding (D and E). Skin sections were labelled with PO-conjugated anti-BrdU antibody (brown nuclei) and counterstained with hematoxylin. Abbreviations: B, brain; C, cerebellum; H, heart; I, intestine; K, kidney; Li, liver; Lu, lung; M, skeletal muscle; O, ovary; Sk, skin; Sp, spleen; St, stomach; T, thymus.
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Fig. 6. In vitro rescue of DNA synthesis and proliferation defects of DDR2–/– fibroblasts. Proliferation rate of adult skin fibroblasts derived from wild-type (+/+) or DDR2–/– mice. Mutant cells were infected with recombinant retroviruses expressing GFP alone, wild-type, myc-tagged DDR2 or a kinase-dead version of DDR2 (K608E). Wild-type, but not kinase-dead DDR2 rescues the proliferation defect of DDR2–/– fibroblasts.
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References

    1. Alves F., Vogel, W., Mossie, K., Millauer, B., Hofler H. and Ullrich, A. (1995) Distinct structural characteristics of discoidin I subfamily receptor tyrosine kinases and complementary expression in human cancer. Oncogene, 10, 609–618. - PubMed
    1. Ankoma-Sey V., Matli, M., Chang, K.B., Lalazar, A., Donner, D.B., Wong, L., Warren, R.S. and Friedman, S.L. (1998) Coordinated induction of VEGF receptors in mesenchymal cell types during rat hepatic wound healing. Oncogene, 17, 115–121. - PubMed
    1. Barker K.T., Martindale, J.E., Mitchell, P.J., Kamalati, T., Page, M.J., Phippard, D.J., Dale, T.C., Gusterson, B.A. and Crompton, M.R. (1995) Expression patterns of the novel receptor-like tyrosine kinase, DDR, in human breast tumours. Oncogene, 10, 569–575. - PubMed
    1. Bhatt R.S., Tomoda, T., Fang, Y. and Hatten, M.E. (2000). Discoidin domain receptor 1 functions in axon extension of cerebellar granule neurons. Genes Dev., 14, 2216–2228. - PMC - PubMed
    1. Brückner K., Labrador, J.P., Scheiffele, P., Herb, A., Seeburg, P.H. and Klein, R. (1999) EphrinB ligands recruit GRIP family PDZ adaptor proteins into raft membrane microdomains. Neuron, 22, 511–524. - PubMed

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