title: Ameloblastin, an Extracellular Matrix Protein, Affects Long Bone Growth and Mineralization.
abstract: Matrix molecules such as the enamel-related calcium-binding phosphoprotein ameloblastin (AMBN) are expressed in multiple tissues, including teeth, bones, and cartilage. Here we have asked whether AMBN is of functional importance for timely long bone development and, if so, how it exerts its function related to osteogenesis. Adolescent AMBN-deficient mice (AMBN(Δ5-6) ) suffered from a 33% to 38% reduction in femur length and an 8.4% shorter trunk spinal column when compared with WT controls, whereas there was no difference between adult animals. On a cellular level, AMBN truncation resulted in a shortened growth plate and a 41% to 49% reduction in the number of proliferating tibia chondrocytes and osteoblasts. Bone marrow stromal cells (BMSCs) isolated from AMBN mutant mice displayed defects in proliferation and differentiation potential as well as cytoskeleton organization. Osteogenesis-related growth factors, such as insulin-like growth factor 1 (IGF1) and BMP7, were also significantly (46% to 73%) reduced in AMBN-deficient BMSCs. Addition of exogenous AMBN restored cytoskeleton structures in AMBN mutant BMSCs and resulted in a dramatic 400% to 600% increase in BMP2, BMP7, and Col1A expression. Block of RhoA diminished the effect of AMBN on osteogenic growth factor and matrix protein gene expression. Addition of exogenous BMP7 and IGF1 rescued the proliferation and differentiation potential of AMBN-deficient BMSCs. Confirming the effects of AMBN on long bone growth, back-crossing of mutant mice with full-length AMBN overexpressors resulted in a complete rescue of AMBN(Δ5-6) bone defects. Together, these data indicate that AMBN affects extracellular matrix production and cell adhesion properties in the long bone growth plate, resulting in altered cytoskeletal dynamics, increased osteogenesis-related gene expression, as well as osteoblast and chondrocyte proliferation. We propose that AMBN facilitates rapid long bone growth and an important growth spurt during the skeletogenesis of adolescent tooth-bearing vertebrates. © 2016 American Society for Bone and Mineral Research.
authors.affiliation: Department of Oral Biology, Brodie Laboratory for Craniofacial Genetics, University of Illinois College of Dentistry, Chicago, IL, USA.
authors.forename: Xuanyu
authors.initials: X
authors.lastname: Lu
authors.affiliation: Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Fukumoto
authors.affiliation: Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Yamada
authors.affiliation: Department of Orthodontics, Brodie Laboratory for Craniofacial Genetics, University of Illinois College of Dentistry, Chicago, IL, USA.
authors.forename: Carla A
authors.initials: CA
authors.lastname: Evans
authors.affiliation: Department of Oral Biology, Brodie Laboratory for Craniofacial Genetics, University of Illinois College of Dentistry, Chicago, IL, USA.
authors.forename: Thomas Gh
authors.initials: TG
authors.lastname: Diekwisch
authors.affiliation: Department of Oral Biology, Brodie Laboratory for Craniofacial Genetics, University of Illinois College of Dentistry, Chicago, IL, USA.|Department of Orthodontics, Brodie Laboratory for Craniofacial Genetics, University of Illinois College of Dentistry, Chicago, IL, USA.
authors.forename: Xianghong
authors.initials: X
authors.lastname: Luan
mesh_terms: D000818:Animals; D015519:Bone Density / Q000502:physiology*; D055419:Bone Morphogenetic Protein 7 / Q000235:genetics / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000235:genetics / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D005269:Femur / Q000254:growth & development*; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010012:Osteogenesis / Q000502:physiology*
chemical_list: C490890:Ambn protein, mouse; D055419:Bone Morphogenetic Protein 7; D003746:Dental Enamel Proteins; D016326:Extracellular Matrix Proteins; C525820:bmp7 protein, mouse; C519530:insulin-like growth factor-1, mouse; D007334:Insulin-Like Growth Factor I
keywords: AMELOBLASTIN; BMP7; BONE DEVELOPMENT; IGF1; MESENCHYMAL CELLS