TGF-β signaling is essential for joint morphogenesis

Despite its clinical significance, joint morphogenesis is still an obscure process. In this study, we determine the role of transforming growth factor β (TGF-β) signaling in mice lacking the TGF-β type II receptor gene (Tgfbr2) in their limbs (Tgfbr2PRX-1KO). In Tgfbr2PRX-1KO mice, the loss of TGF-β responsiveness resulted in the absence of interphalangeal joints. The Tgfbr2Prx1KO joint phenotype is similar to that in patients with symphalangism (SYM1-OMIM185800). By generating a Tgfbr2–green fluorescent protein–β–GEO–bacterial artificial chromosome β-galactosidase reporter transgenic mouse and by in situ hybridization and immunofluorescence, we determined that Tgfbr2 is highly and specifically expressed in developing joints. We demonstrated that in Tgfbr2PRX-1KO mice, the failure of joint interzone development resulted from an aberrant persistence of differentiated chondrocytes and failure of Jagged-1 expression. We found that TGF-β receptor II signaling regulates Noggin, Wnt9a, and growth and differentiation factor-5 joint morphogenic gene expressions. In Tgfbr2PRX-1KO growth plates adjacent to interphalangeal joints, Indian hedgehog expression is increased, whereas Collagen 10 expression decreased. We propose a model for joint development in which TGF-β signaling represents a means of entry to initiate the process.

[1]  Chenbei Chang,et al.  TGF-beta signaling in human skeletal and patterning disorders. , 2003, Birth defects research. Part C, Embryo today : reviews.

[2]  W. Reardon,et al.  Identification of mutations in the MSX2 homeobox gene in families affected with foramina parietalia permagna. , 2000, Human molecular genetics.

[3]  H. Moses,et al.  Conditional inactivation of the TGF-beta type II receptor using Cre:Lox. , 2002, Genesis.

[4]  J. Graff,et al.  Smad3 Mutant Mice Develop Metastatic Colorectal Cancer , 1998, Cell.

[5]  D. Mortlock,et al.  The molecular basis of hypodactyly (Hd): a deletion in Hoxa13 leads to arrest of digital arch formation , 1996, Nature Genetics.

[6]  H. Moses,et al.  Effect of IGF‐I in the Chondrogenesis of Bone Marrow Mesenchymal Stem Cells in the Presence or Absence of TGF‐β Signaling , 2005, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[7]  P. Chuang,et al.  Wnt/β-catenin signaling interacts differentially with Ihh signaling in controlling endochondral bone and synovial joint formation , 2006, Development.

[8]  A. Wilkie,et al.  Alx4 and Msx2 play phenotypically similar and additive roles in skull vault differentiation , 2004, Journal of anatomy.

[9]  G. Proetzel,et al.  Targeted disruption of the mouse transforming growth factor-β1 gene results in multifocal inflammatory disease , 1992, Nature.

[10]  C. Deng,et al.  Failure of egg cylinder elongation and mesoderm induction in mouse embryos lacking the tumor suppressor smad2. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[11]  D. Kingsley,et al.  Joint patterning defects caused by single and double mutations in members of the bone morphogenetic protein (BMP) family. , 1996, Development.

[12]  W. Wuyts,et al.  The ALX4 homeobox gene is mutated in patients with ossification defects of the skull (foramina parietalia permagna, OMIM 168500) , 2000, Journal of medical genetics.

[13]  H. Moses,et al.  Conditional deletion of the TGF-beta type II receptor in Col2a expressing cells results in defects in the axial skeleton without alterations in chondrocyte differentiation or embryonic development of long bones. , 2004, Developmental biology.

[14]  K. Krieglstein,et al.  Targeted mutations of transforming growth factor-beta genes reveal important roles in mouse development and adult homeostasis. , 2000, European journal of biochemistry.

[15]  A. McMahon,et al.  Noggin, cartilage morphogenesis, and joint formation in the mammalian skeleton. , 1998, Science.

[16]  V. Kaartinen,et al.  Abnormal lung development and cleft palate in mice lacking TGF–β3 indicates defects of epithelial–mesenchymal interaction , 1995, Nature Genetics.

[17]  E. Canalis,et al.  Bone morphogenetic proteins induce the expression of noggin, which limits their activity in cultured rat osteoblasts. , 1998, The Journal of clinical investigation.

[18]  The murine type II TGF-beta receptor has a coincident embryonic expression and binding preference for TGF-beta 1. , 1994, Development.

[19]  D. Livingston,et al.  Self-excising retroviral vectors encoding the Cre recombinase overcome Cre-mediated cellular toxicity. , 2001, Molecular cell.

[20]  G. Boivin,et al.  TGFbeta2 knockout mice have multiple developmental defects that are non-overlapping with other TGFbeta knockout phenotypes. , 1997, Development.

[21]  H. Moses,et al.  Immunohistochemical localization of TGF beta 1, TGF beta 2, and TGF beta 3 in the mouse embryo: expression patterns suggest multiple roles during embryonic development , 1991, The Journal of cell biology.

[22]  D. Kingsley,et al.  Efficient studies of long-distance Bmp5 gene regulation using bacterial artificial chromosomes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[23]  K. K. Deal,et al.  Distant regulatory elements in a Sox10‐βGEO BAC transgene are required for expression of Sox10 in the enteric nervous system and other neural crest‐derived tissues , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[24]  G. Nishimura,et al.  Mutations of the NOG gene in individuals with proximal symphalangism and multiple synostosis syndrome , 2001, Clinical genetics.

[25]  T. Okamoto,et al.  Induction of Notch signaling by tumor necrosis factor in rheumatoid synovial fibroblasts , 2003, Oncogene.

[26]  K. Schughart,et al.  Retinoic Acid Receptor α Function in Vertebrate Limb Skeletogenesis: a Modulator of Chondrogenesis , 1997, The Journal of cell biology.

[27]  Philippe Soriano.  Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[28]  Y. Gañán,et al.  Role of TGF beta s and BMPs as signals controlling the position of the digits and the areas of interdigital cell death in the developing chick limb autopod. , 1996, Development.

[29]  C. Tabin,et al.  Expression of Cre recombinase in the developing mouse limb bud driven by a Prxl enhancer , 2002, Genesis.

[30]  C. Guenther,et al.  A general approach for identifying distant regulatory elements applied to the Gdf6 gene. , 2003, Genome research.

[31]  D. Court,et al.  A highly efficient Escherichia coli-based chromosome engineering system adapted for recombinogenic targeting and subcloning of BAC DNA. , 2001, Genomics.

[32]  A. Broadus,et al.  Initial Characterization of PTH‐Related Protein Gene‐Driven lacZ Expression in the Mouse , 2005, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[33]  C. Hartmann,et al.  Wnt9a signaling is required for joint integrity and regulation of Ihh during chondrogenesis , 2006, Development.

[34]  Mahlon D. Johnson,et al.  Expression of Transforming Growth Factor-β1, -β2, and -β3 mRNA and Protein in the Murine Lung , 1991 .

[35]  E. Vuorio,et al.  Specific hybridization probes for mouse type I, II, III and IX collagen mRNAs. , 1991, Biochimica et biophysica acta.

[36]  M. Taketo,et al.  TGF-beta receptor type II deficiency results in defects of yolk sac hematopoiesis and vasculogenesis. , 1996, Developmental biology.

[37]  Nancy A. Jenkins,et al.  Simple and highly efficient BAC recombineering using galK selection , 2005, Nucleic acids research.

[38]  P. Kondaiah,et al.  Embryonic gene expression patterns of TGF beta 1, beta 2 and beta 3 suggest different developmental functions in vivo. , 1991, Development.

[39]  J. Nichols,et al.  Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[40]  M. Washington,et al.  TGF-ß Signaling in Fibroblasts Modulates the Oncogenic Potential of Adjacent Epithelia , 2004, Science.

[41]  Changshan Wu,et al.  Cellular and Molecular Mechanisms of Synovial Joint and Articular Cartilage Formation , 2006, Annals of the New York Academy of Sciences.

[42]  N. Copeland,et al.  Limb alterations in brachypodism mice due to mutations in a new member of the TGFβ-superfamily , 1994, Nature.

[43]  C. Archer,et al.  Development of synovial joints. , 2003, Birth defects research. Part C, Embryo today : reviews.

[44]  Christine Hartmann,et al.  Wnt-14 Plays a Pivotal Role in Inducing Synovial Joint Formation in the Developing Appendicular Skeleton , 2001, Cell.

[45]  C. Deng,et al.  TGF-β/Smad3 Signals Repress Chondrocyte Hypertrophic Differentiation and Are Required for Maintaining Articular Cartilage , 2001, The Journal of cell biology.

[46]  Mahlon D. Johnson,et al.  Expression of a Truncated, Kinase-Defective TGF-β Type II Receptor in Mouse Skeletal Tissue Promotes Terminal Chondrocyte Differentiation and Osteoarthritis , 1997, The Journal of cell biology.

[47]  Ilyas M. Khan,et al.  The surface of articular cartilage contains a progenitor cell population , 2004, Journal of Cell Science.

[48]  S. Dey,et al.  Inappropriate expression of human transforming growth factor (TGF)-alpha in the uterus of transgenic mouse causes downregulation of TGF-beta receptors and delays the blastocyst-attachment reaction. , 1997, Journal of molecular endocrinology.

[49]  U. Lendahl,et al.  Cross-talk between the Notch and TGF-β signaling pathways mediated by interaction of the Notch intracellular domain with Smad3 , 2003, The Journal of cell biology.

[50]  H. Moses,et al.  Expression of transforming growth factor-beta 1, -beta 2, and -beta 3 mRNA and protein in the murine lung. , 1991, American journal of respiratory cell and molecular biology.

[51]  C. Shuler,et al.  Conditional inactivation of Tgfbr2 in cranial neural crest causes cleft palate and calvaria defects , 2003, Development.

[52]  Stefan Mundlos,et al.  Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2. , 2005, The Journal of clinical investigation.

[53]  D. Rice,et al.  Msx2 and Twist cooperatively control the development of the neural crest-derived skeletogenic mesenchyme of the murine skull vault , 2003, Development.

[54]  J. Rossant,et al.  The tumor suppressor gene Smad4/Dpc4 is required for gastrulation and later for anterior development of the mouse embryo. , 1998, Genes & development.

[55]  Chenbei Chang,et al.  TGF‐β signaling in human skeletal and patterning disorders , 2003 .

[56]  D. Kingsley,et al.  GDF5 coordinates bone and joint formation during digit development. , 1999, Developmental biology.

[57]  C. Cremers,et al.  Heterozygous mutations in the gene encoding noggin affect human joint morphogenesis , 1999, Nature Genetics.

[58]  S. V. Webster,et al.  The distribution of Notch receptors and their ligands during articular cartilage development , 2003, Journal of anatomy.