title: TOPGAL mice show that the canonical Wnt signaling pathway is active during bone development and growth and is activated by mechanical loading in vitro.
abstract: We identified cellular targets of canonical Wnt signaling within the skeleton, which included chondrocytes, osteoblasts, and osteocytes in growing bone, but only osteocytes and chondrocytes in the mature skeleton. Mechanical deformation induced Wnt signaling in osteoblasts in vitro.
  
  
  BACKGROUND
  Genetic evidence in mice and humans has implicated the canonical Wnt signaling pathway in the control of skeletal development and bone mass. However, little is known of the details of Wnt signaling in the skeleton in vivo. We used Wnt indicator TOPGAL mice to identify which cells activated this pathway during bone development and in the mature skeleton.
  
  
  METHODS
  We examined canonical Wnt signaling during embryonic and neonatal bone development in TOPGAL mice. The TOPGAL transgene consists of a beta-galactosidase gene driven by a T cell factor (TCF)beta-catenin responsive promoter so that canonical Wnt activity can be detected by X-gal staining. Expression of Wnt signaling components was examined in primary calvarial cell cultures by RT-PCR. The effect of mechanical deformation on Wnt signaling was examined in primary calvarial cells grown on collagen I and stretched using Flexercell Tension Plus System FX-4000T. Immunohistochemistry was used to examine the localization of beta-catenin in cartilage, bone, and cultured calvarial cells exposed to physical deformation.
  
  
  CONCLUSIONS
  Canonical Wnt signaling was active in several cell types in the fetal and neonatal skeleton, including chondrocytes, osteoblasts, and osteocytes. With age, activation of Wnt signaling became less prominent but persisted in chondrocytes and osteocytes. Although osteoblasts in culture expressed many different individual Wnt's and Wnt receptors, the TOPGAL transgene was not active in these cells at baseline. However, Wnt signaling was activated in these cells by physical deformation. Together with the activation of canonical Wnt signaling in osteocytes seen in vivo, these data suggest that Wnt signaling may be involved in the coupling of mechanical force to anabolic activity in the skeleton.
authors.affiliation: Section of Endocrinology and Metabolism, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520-8020, USA.
authors.forename: Julie R
authors.initials: JR
authors.lastname: Hens
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Wilson
authors.forename: Pamela
authors.initials: P
authors.lastname: Dann
authors.forename: Xuesong
authors.initials: X
authors.lastname: Chen
authors.forename: Mark C
authors.initials: MC
authors.lastname: Horowitz
authors.forename: John J
authors.initials: JJ
authors.lastname: Wysolmerski
mesh_terms: D000818:Animals; D001846:Bone Development; D001842:Bone and Bones / Q000166:cytology / Q000196:embryology*; D003598:Cytoskeletal Proteins / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D015398:Signal Transduction; D012886:Skull / Q000737:chemistry / Q000166:cytology; D013314:Stress, Mechanical; D013601:T-Lymphocytes / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D051153:Wnt Proteins; D051176:beta Catenin; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
chemical_list: C495267:CTNNB1 protein, mouse; D003598:Cytoskeletal Proteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D051153:Wnt Proteins; D051176:beta Catenin; D001616:beta-Galactosidase