Search PubMed⌕ Search

Biomedical subjects

B R Olsen

Publications and source records attributed to B R Olsen.

At least 73 records · Page 4Linked to original sources
PubMed1997

Abnormal compartmentalization of cartilage matrix components in mice lacking collagen X: implications for function.

There are conflicting views on whether collagen X is a purely structural molecule, or regulates bone mineralization during endochondral ossification. Mutations in the human collagen alpha1 (X) gene (COL10A1) in Schmid metaphyseal chondrodysplasia (SMCD) suggest a supportive role. But mouse collagen alpha1 (X) gene (Col10a1) null mutants were previously reported to show no obvious phenotypic change. We have generated collagen X deficient mice, which shows that deficiency does have phenotypic consequences which partly resemble SMCD, such as abnormal trabecular bone architecture. In particular, the mutant mice develop coxa vara, a phenotypic change common in human SMCD. Other consequences of the mutation are reduction in thickness of growth plate resting zone and articular cartilage, altered bone content, and atypical distribution of matrix components within growth plate cartilage. We propose that collagen X plays a role in the normal distribution of matrix vesicles and proteoglycans within the growth plate matrix. Collagen X deficiency impacts on the supporting properties of the growth plate and the mineralization process, resulting in abnormal trabecular bone. This hypothesis would accommodate the previously conflicting views of the function of collagen X and of the molecular pathogenesis of SMCD.

Animals↗
PubMed1997

Endostatin: an endogenous inhibitor of angiogenesis and tumor growth.

We previously identified the angiogenesis inhibitor angiostatin. Using a similar strategy, we have identified endostatin, an angiogenesis inhibitor produced by hemangioendothelioma. Endostatin is a 20 kDa C-terminal fragment of collagen XVIII. Endostatin specifically inhibits endothelial proliferation and potently inhibits angiogenesis and tumor growth. By a novel method of sustained release, E. coli-derived endostatin was administered as a nonrefolded suspension. Primary tumors were regressed to dormant microscopic lesions. Immunohistochemistry revealed blocked angiogenesis accompanied by high proliferation balanced by apoptosis in tumor cells. There was no toxicity. Together with angiostatin data, these findings validate a strategy for identifying endogenous angiogenesis inhibitors, suggest a theme of fragments of proteins as angiogenesis inhibitors, and demonstrate dormancy therapy.

Amino Acid Sequence↗
PubMed1997

Disproportionate micromelia (Dmm) in mice caused by a mutation in the C-propeptide coding region of Col2a1.

Mice that are homozygous for the autosomal semidominant disproportionate micromelia (Dmm) mutation are characterized by disproportionate micromelia, thoracic dysplasia, and cleft palate. Chondrocytes of the epiphyseal growth plates are not organized into columns, and ultrastructural analysis reveals excessive dilation of the endoplasmic reticulum and a paucity of collagen fibrils in the extracellular matrix. To map the Dmm locus, Dmm mice were crossed with the multiple ecotropic viral (MEV) linkage testing stock. Significant linkage of Dmm to the fourteen MEV linkage markers was not observed, thereby excluding approximately 50% of the genome as candidate regions encoding Dmm. Subsequently, microsatellite markers were used to assess linkage to the nonexcluded regions of the genome, revealing tight linkage to the locus of Col2a1, the gene encoding the alpha-chains of type II collagen. alpha 1(II) collagen cDNA, synthesized with RNA from homozygotes, was cloned and sequenced, revealing a three-nucleotide deletion in the region encoding the C-propeptide globular domain. The deletion leads to the substitution of one amino acid, Asn, in the mutant for two amino acids, Lys and Thr, in the wild type. Several human chondrodysplasias with similar phenotypes to that of Dmm are associated with defects in type II collagen. Thus, mice bearing the Dmm mutation serve as a model for studying the pathogenesis of these disorders while revealing novel insights into normal skeletal morphogenesis.

Amino Acid Sequence↗
PubMed1997

Craniofacial abnormalities in mice carrying a dominant interference mutation in type X collagen.

Type X collagen is a short, non-fibril forming collagen restricted to hypertrophic cartilage, and has been hypothesized to play a role in endochondral ossification. The purpose of the study was to investigate the consequences resulting from the interference of type X collagen function on the growth and development of the craniofacial skeleton through analysis of transgenic mice with a dominant interference mutation for type X collagen. The craniofacial tissues of 21-day-old transgenic mice were examined by: cephalometric and radiographic densitometry analyses, conventional histology, and immunohistochemistry using antibodies specific for either endogenous mouse type X collagen or the transgene product. Genotypically positive mutant mice showed moderate but statistically significant craniofacial skeletal abnormalities, including the underdevelopment of the chondrocranium and mandible, but no cleft palate. Mean radiographic optical densities of the mutant condylar cartilage and the subchondylar areas were 32% less than the corresponding areas of normal mandibles, while mean radiographic optical density measured at the incisor tooth point remained constant. Histologically, transgene-positive mice revealed compressed hypertrophic cartilage zones and reduced trabeculae in both the mandibular condyle and the synchondroses of the chondrocranium. In the normal condyle, mouse type X collagen was localized by the monospecific antibody against a synthetic rat type X collagen NC1 peptide throughout the hypertrophic cartilage layer; in the mutant condyle, immunoreactivity to endogenous type X collagen was only seen sporadically. The truncated type X collagen transgene product, identified with the monoclonal antibody against an epitope within the chick type X collagen NC2 domain, persisted in the lower hypertrophic cartilage layer and the primary spongiosa, rather than being removed by subsequent endochondral ossification. The data suggested that the expression of the chick type X collagen transgene product was strongly associated with the craniofacial skeletal abnormalities that were distinct from other cartilage-related phenotypes.

Animals↗
PubMed1997

Mouse clavicular development: analysis of wild-type and cleidocranial dysplasia mutant mice.

Cleidocranial dysplasia (CCD) is an autosomal dominant disease characterized by hypoplasia or aplasia of clavicles, open fontanelles, and other skeletal anomalies. A mouse mutant, shown by clinical and radiographic analysis to be strikingly similar to the human disorder and designated Ccd, was used as a model for the human disorder. Since malformation of the clavicle is the hallmark of CCD, we studied clavicular development in wild-type and Ccd mice. Histology and in situ hybridization experiments were performed to compare the temporal and spatial expression of several genes in wild-type and Ccd mutant mouse embryos. Bone and cartilage specific markers--type I, II, and X collagens, Sox9, aggrecan, and osteopontin were used as probes. The analyses covered the development of the clavicle from the initial mesenchymal condensation at embryonic day 13 (E13) to the late mineralization stage at embryonic day 15.5. At day 13.5, cells in the center of the condensation differentiate into characteristic precursor cells that were not observed in other bone anlagen. In the medial part of the anlage these cells express markers of the early cartilage lineage (type II collagen and Sox9), whereas cells of the lateral part express markers of the osteoblast lineage (type I collagen). With further development the medial cells differentiate into chondrocytes and start to express chondrocyte-specific markers such as aggrecan. Cells of the lateral part differentiate into osteoblasts as indicated by the production of bone matrix and the expression of osteopontin. At day 14.5 a regular growth plate has developed between the two parts where type X collagen expression can be demonstrated in hypertrophic chondrocytes. The data indicate that the medial part of the clavicle develops by endochondral bone formation while the lateral part ossifies as a membranous bone. The clavicle of Ccd mice showed a smaller band of mesenchymal cell condensation than in wild-type mice. Cells of the condensation failed to express type I and type II collagen at E13.5. In the lateral part of the clavicle type I collagen expression was not detected until E14.5 and osteopontin expression only appeared at E15.5. At E15.5, a small ossification center appears in the lateral part which is, in contrast to the wild-type clavicular bone, solid and without primary spongiosa as well as bone marrow. In the medial portion, type II collagen expression and endochondral ossification never occurs in Ccd mice; this portion of the clavicle is therefore missing in Ccd.

Animals↗
PubMed1997

Murine models of human genetic skeletal disorders.

Genetic and molecular biological studies have recently resulted in the identification of gene mutations responsible for a large number of human osteochondrodysplasias, skeletal disorders associated with abnormalities of cartilage and/or bone development and growth. Mouse strains carrying mutations in homologous genes are useful, not only because they provide sources of abnormal tissue, but also because specific hypotheses concerning pathogenetic mechanisms can be readily tested. Many of the strains with abnormalities in cartilage and bone have been studied by morphological and biochemical methods for years without successful identification of the genes involved, but recent use of linkage mapping, followed by positional candidate gene cloning, has greatly improved the situation. In a number of instances, the mutations are now known. This is also true for the three "classical" mouse mutants chondrodysplasia (cho), disproportionate micromelia (Dmm) and cartilage matrix deficiency (cmd). In the three strains, mutations in alpha 1(XI) collagen, alpha 1(II) collagen or aggrecan lead to severe defects in the structure and function of cartilage with significant negative effects on longitudinal bone growth.

Animals↗
PubMed1997

The role of sonic hedgehog in vertebrate development.

Members of the hedgehog family are important signalling molecules during embryonic development. One member, Sonic hedgehog, is expressed in embryonic structures such as the zone of polarizing activity in the posterior limb bud, the notochord, and the floor plate of the neural tube, where it plays a role in patterning of the embryo. Sonic hedgehog is synthesized as an inactive precursor which must be proteolytically cleaved and modified by the addition of a cholesterol moiety to become active as a signalling molecule. In this processing, the C-terminal region of Sonic hedgehog serves as both the endoprotease and a cholesterol transferase. The importance of cholesterol for Sonic hedgehog function may explain many of the profound developmental defects caused by perturbations of cholesterol metabolism. The receptor for Sonic hedgehog is Patched, a multi-pass transmembrane protein which forms a complex with Smoothened Mutations in Patched are associated with basal cell naevus syndrome, while mutations in Sonic hedgehog cause holoprosencephaly. Downstream targets of Sonic hedgehog signalling are transcription factors like Gli3, responsible for Greigs polycephalosyndactyly in humans and Hoxd13, responsible for polysyndactyly.

Animals↗
PubMed1997

Collagen IX.

Collagen IX was identified as a distinct component of cartilage about 10 years ago. Composed of three polypeptide chains, its heterotrimeric molecules are located on the surface of type II collagen fibrils in cartilage. The interaction between collagens II and IX is stabilized by covalent crosslinks. The location of collagen IX molecules on fibril surfaces suggests that they represent macromolecular bridges between fibrils and other matrix components in cartilage, and that collagen IX is important for the cohesive and compressive properties of cartilage. Transgenic mice with mutations in a type IX gene develop normally, but show degenerative changes in articular cartilage after birth. Additional evidence for the importance of collagen IX in human articular cartilage comes from the recent finding that a mutation in one of the collagen IX genes causes multiple epiphyseal dysplasia. The mild complaints in affected individuals with the mutation suggest that mutations in type IX collagen genes may represent genetic risk factors for late onset osteoarthritis.

Animals↗
PubMed1997

Heritable diseases of the skeleton. Part I: Molecular insights into skeletal development-transcription factors and signaling pathways.

The recent identification of the genetic basis of hereditary skeletal disorders is providing important insights into the intricate processes of skeletal formation, growth, and homeostasis. These processes include patterning events during condensation and differentiation of mesenchymal cells to form cartilage precursors of the future bones, the replacement of cartilage by bones through endochondral ossification, the growth of long bones through proliferation and differentiation of chondrocytes in growth plates, and bone formation through differentiation of osteoblasts from mesenchymal cells in areas of intramembranous ossification. Defects in any of these processes can give rise to skeletal abnormalities. Mutations in transcription factors such as HOX and PAX and members of the transforming growth factor-beta superfamily cause disorders associated with abnormal mesenchymal condensation, whereas defects in the transcription factor SOX-9 lead to abnormalities in chondrocyte differentiation. Abnormal growth plate function, resulting in dwarfism, is the consequence of mutations in receptors for fibroblast growth factors and parathyroid hormone-related peptide. Premature closure of cranial sutures in intramembranous ossification is a feature of syndromes due to mutations in fibroblast growth factor receptors.

Animals↗
PubMed1997

Tissue inhibitor of metalloproteinases-3 is a component of Bruch's membrane of the eye.

Mutations in tissue inhibitor of metalloproteinases (TIMP)-3 are found in some patients with Sorsby's fundus dystrophy, a retinal degeneration characterized by abnormal deposits in Bruch's membrane and choroidal neovascularization. The purpose of this study was to localize TIMP-3 in the retina/choroid of normal human and animal eyes. Immunolabeling was performed on unfixed and fixed sections of human eyes aged 24 to 85 years and unfixed sections of baboon, chicken, cow, pig, and rat eyes using a monoclonal antibody against a human TIMP-3 synthetic peptide. The antibody produced strong immunolabeling of Bruch's membrane and drusen and weak labeling of retina blood vessels in unfixed human and baboon eyes. Unfixed chicken, cow, pig, and rat tissues showed no reactivity. After antigen retrieval, all fixed human eyes showed specific labeling of Bruch's membrane and drusen, which was strongest in eyes from elderly donors. The results indicate that TIMP-3 is an extracellular matrix component of Bruch's membrane. Thus, abnormal local function of TIMP-3 may lead to the characteristic Bruch's membrane deposits and choroidal neovascularization found in Sorsby's fundus dystrophy. Specific labeling of drusen raises the possibility that altered TIMP-3-mediated matrix remodeling may contribute to age-related degenerative changes in Bruch's membrane.

Adult↗
PubMed1997

Heritable diseases of the skeleton. Part II: Molecular insights into skeletal development-matrix components and their homeostasis.

A range of osteochondrodysplasias is caused by mutations in components of the extracellular matrix in cartilage and bone and in molecules that are important for posttranslational processing of such components. Mutations in the genes encoding the two polypeptide subunits of collagen I cause defects in the structure of bone matrix while mutations in genes encoding cartilage-specific collagens are responsible for several chondrodysplasias. Abnormalities in cartilage structure and function can also be due to mutations in structural noncollagenous components such as aggrecan and cartilage oligomeric matrix protein. Finally, several cartilage and bone disorders are due to abnormalities in sulfate transport and regulation of bone matrix homeostasis.

Animals↗
PubMed1996

Vascular dysmorphogenesis caused by an activating mutation in the receptor tyrosine kinase TIE2.

Venous malformations (VMs), the most common errors of vascular morphogenesis in humans, are composed of dilated, serpiginous channels. The walls of the channels have a variable thickness of smooth muscle; some mural regions lack smooth muscle altogether. A missense mutation resulting in an arginine-to-tryptophan substitution at position 849 in the kinase domain of the receptor tyrosine kinase TIE2 segregates with dominantly inherited VM in two unrelated families. Using proteins expressed in insect cells, we demonstrate that the mutation results in increased activity of TIE2. We conclude that an activating mutation in TIE2 causes inherited VMs in the two families and that the TIE2 signaling pathway is critical for endothelial cell-smooth muscle cell communication in venous morphogenesis.

Amino Acid Sequence↗
PubMed1996

Morphogenesis: collagen it takes and bone it makes.

The same enzyme is responsible for the activation of transforming growth factor-beta-like signalling molecules and the processing of fibrillar procollagens. This finding implicates a metalloendoprotease in the control of cell behavior and matrix assembly during morphogenesis.

Animals↗
PubMed1996

Altered growth and branching patterns in synpolydactyly caused by mutations in HOXD13.

Hox genes regulate patterning during limb development. It is believed that they function in the determination of the timing and extent of local growth rates. Here, it is demonstrated that synpolydactyly, an inherited human abnormality of the hands and feet, is caused by expansions of a polyalanine stretch in the amino-terminal region of HOXD13. The homozygous phenotype includes the transformation of metacarpal and metatarsal bones to short carpal- and tarsal-like bones. The mutations identify the polyalanine stretch outside of the DNA binding domain of HOXD13 as a region necessary for proper protein function.

Amino Acid Sequence↗