title: Aggrecan is required for growth plate cytoarchitecture and differentiation.
abstract: The proteoglycan aggrecan is a prominent component of the extracellular matrix in growth plate cartilage. A naturally occurring, recessive, perinatally lethal mutation in the aggrecan core protein gene, cmd(bc) (Acan(cmd-Bc)), that deletes the entire protein-coding sequence provided a model in which to characterize the phenotypic and morphologic effects of aggrecan deletion on skeletal development. We also generated a novel transgenic mouse, Tg(COL2A1-ACAN), that has the chick ACAN coding sequence driven by the mouse COL2A1 promoter to enable the production of cmd(bc)/cmd(bc); Tg(COL2A1-ACAN) rescue embryos. These were used to assess the impact of aggrecan on growth plate organization, chondrocyte survival and proliferation, and the expression of mRNAs encoding chondrocyte differentiation markers and growth factors. Homozygous mutant (cmd(bc)/cmd(bc)) embryos exhibited severe defects in all skeletal elements with deformed and shortened (50%) limb elements. Expression of aggrecan in rescue embryos reversed the skeletal defects to varying degrees with a 20% increase in limb element length and near-full reversal (80%) of size and diameter of the ribcage and vertebrae. Aggrecan-null growth plates were devoid of matrix and lacked chondrocyte organization and differentiation, while those of the rescue embryos exhibited matrix production concomitant with partial zonation of chondrocytes having proliferative and hypertrophic morphologies. Deformation of the trachea, likely the cause of the mutation's lethality, was reduced in the rescue embryos. Aggrecan-null embryos also had abnormal patterns of COL10A1, SOX9, IHH, PTCH1, and FGFR3 mRNA expression in the growth plate. Expression of chick aggrecan in the rescue embryos notably increased COLX expression, accompanied by the reappearance of a hypertrophic zone and IHH expression. Significantly, in transgenic rescue embryos, the cell death and decreased proliferation phenotypes exhibited by the mutants were reversed; both were restored to wild-type levels. These findings suggest that aggrecan has a major role in regulating the expression of key growth factors and signaling molecules during development of cartilaginous tissue and is essential for proper chondrocyte organization, morphology, and survival during embryonic limb development.
authors.affiliation: Department of Pediatrics, The University of Chicago, Chicago IL 60637, USA.
authors.forename: Kristen L
authors.initials: KL
authors.lastname: Lauing
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Cortes
authors.affiliation: Department of Pediatrics, The University of Chicago, Chicago IL 60637, USA.
authors.forename: Miriam S
authors.initials: MS
authors.lastname: Domowicz
authors.affiliation: Department of Pediatrics, The University of Chicago, Chicago IL 60637, USA.
authors.forename: Judith G
authors.initials: JG
authors.lastname: Henry
authors.affiliation: Department of Pediatrics, The University of Chicago, Chicago IL 60637, USA.
authors.forename: Alexis T
authors.initials: AT
authors.lastname: Baria
authors.affiliation: Department of Pediatrics, The University of Chicago, Chicago IL 60637, USA; Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA. Electronic address: n-schwartz@uchicago.edu.
authors.forename: Nancy B
authors.initials: NB
authors.lastname: Schwartz
mesh_terms: D053674:Aggrecans / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D015139:Blotting, Southern; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002645:Chickens; D019902:Chondrocytes / Q000378:metabolism / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D005121:Extremities / Q000196:embryology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006132:Growth Plate / Q000166:cytology / Q000196:embryology*; D053823:Hedgehog Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D020287:In Situ Nick-End Labeling; D051379:Mice; D008822:Mice, Transgenic; D055758:SOX9 Transcription Factor / Q000378:metabolism
chemical_list: C508320:Acan protein, mouse; D053674:Aggrecans; D017931:DNA Primers; D053823:Hedgehog Proteins; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; C514061:ihh protein, mouse
keywords: Aggrecan; Cartilage matrix deficiency; Chondrocyte morphology; Extracellular matrix; Growth plate; Proteoglycan