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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 580 (1990), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 785 (1996), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 785 (1996), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 460 (1985), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using dual-laser sorted chromosomes and spot-blot analysis, we have previously assigned genomic DNA sequences coding for human α1(IV) procollagen to chromosome 13 (Pihlajaniemi et al. 1985). By in situ hybridization to normal chromosomes and chromosomes with 13q deletions, we now report the localization of this gene to the terminal end of the long arm of chromosome 13. In addition, Southern and slot blot hybridization analysis clearly show that these genomic sequences are present only once per haploid genome.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Modifications and adaptations of chemical techniques have been used to quantitate hydroxyproline (collagen), total protein and DNA in fixed, paraffin-embedded and sectioned tissue. The assays are rapid and sensitive to between 0.1 and 1.0 μg of each of the three components. Values for the ratio of collagen to total protein or collagen per DNA obtained from sectioned material are the same as values derived from fresh or fixed starting material. Depending on the tissue and the sample size, as little as three to five 5 μm thick sections can be analysed for these three components. The ratios of collagen per total protein or collagen per DNA are given for several murine tissues. The assay of these components in tissue sections will allow a more precise correlation between histological appearance and the quantitative biochemical changes that may accompany many pathological disorders. The techniques are particularly useful in retrospective studies where the experimental material may been bloc.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0730-2312
    Keywords: chondrocyte ; mRNA stability ; transient transfection ; promoter ; enhancer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Type II collagen is one of the predominant extracellular matrix macromolecules in cartilage responsible for maintenance of integrity of this specialized tissue. We showed previously that interleukin-1 (IL-1) and interferon-γ (IFN-γ) are capable of decreasing the levels of α1 (II) procollagen mRNA and suppressing the synthesis of type II collagen in cultured human chondrocytes. Data reported here show that these effects of IL-1 and IFN-γ on the expression of the human type II collagen gene (COL2A1) are mediated primarily at the transcriptional level. This conclusion is based on three types of experimental evidence: (1) in nuclear run-off assays, preincubation of chondrocytes with either IL-1 or IFN-γ decreased COL2A1 transcription; (2) experiments with the protein synthesis inhibitor cycloheximide and the transcriptional inhibitor 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB) indicated that the suppression of α1 (II) procollagen mRNA by IL-1 could not be ascribed to decreased mRNA stability; and (3) a plasmid (pCAT-B/4.0) containing 4.0 kb of 5′-flanking sequences of COL2A1 (-577/+3428), encompassing the promoter, exon 1 and the putative enhancer sequence in the first intron, linked to the chloramphenicol acetyltransferase (CAT) reporter gene, was transfected in human chondrocytes. A high level of expression of pCAT-B/4.0 was observed in human chondrocytes incubated with an insulin-containing serum substitute that is permissive for expression of the COL2A1 gene. Expression of pCAT-B/4.0 in these cells was inhibited by either IL-1 or IFN-γ. Furthermore, expression of pCAT-B/4.0 was not detected in human dermal fibroblasts. When the putative enhancer fragment in the first intron was removed, the expression in chondrocytes was greatly reduced. These studies demonstrate that expression of COL2A1 is tissue specific and that suppression by either IL-1 or IFN-γ is mediated primarily at the transcriptional level.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 28 (1994), S. 470-482 
    ISSN: 1059-910X
    Keywords: Cartilage ; Type II collagen ; Aggrecan ; Chondrogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cartilage is an important tissue in skeletogenesis, in the growth of long bones, and as a flexible component of the mature skeleton. The extracellular matrix proteins type II collagen and aggrecan comprise 90% of the matrix and are characteristic of cartilage. Type II collagen provides structural integrity to the tissue, while aggrecan confers resiliency. The quantity of type II procollagen is controlled at the level of transcription of mRNA from the COL2A1 gene. In addition, type II procollagen can be expressed in two isoforms by differential splicing of the primary gene transcript, a post-transcript, a post-transcriptional control mechanism. The two mRNAs either include exon 2 (type IIA) or exclude exon 2 (type IIB) which encodes the major portion of the amino (NH2)-propeptide [Ryan and Sandell (1990), J. Biol. Chem., 265:10334-10339]. The aggrecan gene also encodes alternative splice forms that may be developmentally expressed. The regulation of aggrecan splicing or transcription has not been studied in detail. To determine the spatial and temporal patterns of expression of extracellular matrix in the development of cartilage, we have examined the expression of type II collagen and aggrecan during chondrogenesis in the vertebral column and during elongation of a newborn growth plate. Our results indicate that there is a developmental sequence of type II collagen splice form expression during chondrogenesis with type IIA expressed in prechondrocytes and type IIB expressed in chondrocytes. During elongation of the growth plate, mature chondrocytes express type IIB procollagen and then differentiate into hypertrophic chondrocytes and initiate expression of type X collagen. In all cases, aggrecan was coordinately expressed with type IIB procollagen. As cartilage-like proteins have been observed in more primitive structures such as notochord, the expression of type II collagen mRNAs was also examined in the notochordal remnants of the vertebral column. In the notochord, the predominant collagen expressed was the type IIA collagen prechondrocyte isoform. Notochordal cells also expressed mRNAs more characteristic of fibroblasts such as versican and decorin: low expression of type I collagen, type IIB collagen, and aggrecan were observed. © 1994 Wiley-Liss, IncThis Article is a US Government work and, as such, is in the public domain in the United States of America..
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1058-8388
    Keywords: Collagen ; Chondrogenesis ; Joint development ; Apoptosis ; Alternative splicing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The developmental sequence of the embryonic joint has been well studied morphologically. There are, however, no definitive studies of cell function during joint development. In order to begin to understand the differentiation events that contribute to joint formation, we examined the expression of collagen mRNAs encoding types I, IIA, IIB, and XI. In situ hybridization was performed on chicken embryo hind limb buds and digits from day 7 to day 18 (Hamburger and Hamilton stages 31-44). In the day 7 (stage 31) limb bud, there was a condensation of mesenchyme forming the primitive tarsal and metatarsal bones that showed abundant expression of type IIA procollagen message, but no type IIB or type α1(XI) message. By day 8 (stage 33), co-expression of types IIA, and type XI procollagen mRNAs was observed in the condensations, with expression of IIB restricted to early chondrocytes with metachromatically staining matrix. At this stage, DNA fragmentation characteristic of apoptosis was observed in cells near the midline of the interzone region between the developing anlagen, and in areas between and around the individual digits of the paddle. The presumptive apoptotic cells were more numerous at day 9 (stage 35), and were not found in the developing joint at subsequent time points, including the initiation of spatial cavitation of the joint. From days 11-18, type IIA procollagen mRNA was expressed in flattened cells at the surface of the anlagen, and in the perichondrium and in the developing joint capsule; type IIB mRNA message was found only in chondrocytes. Type XI mRNA was expressed by all type II-expressing cells. Alpha 1(I) mRNA was expressed early by cells of th8e interzone and capsule, but as cavitation progressed, the type I expressing cells of the interzone merged with the superficial layer of the articular surface. Thus, at the time of joint cavitation, there was a distinct pattern of expression of procollagen messages at the articular surface, with type I being outermost, followed by morphologically similar cells expressing type IIA, then chondrocytes expressing type IIB. The progenitor cells expressing type IIA message define a new population of cells. These cell populations contribute to the molecular heterogeneity of the articular cartilage, and these same populations likely exist in the developing joints of other species. The transient transcription of type II and type XI collagen genes, characteristic of chondrocytes, by cells in the joint capsule demonstrates that these cells may have chondrogenic potential. ©1995 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1058-8388
    Keywords: Type II collagen ; Chondrogenesis ; Pattern formation ; Alternative splicing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Type II collagen, generally considered to be characteristic of cartilage, has been localized in specific non-cartilaginous structures during embryogenesis and development of the skeleton. Type II procollagen is synthesized in two different forms generated by alternative splicing of exon 2 in the precursor mRNA transcript. One form (type IIA procollagen) contains a large cysteine-rich domain in the NH2-terminal propeptide, while the second form (type IIB procollagen) does not. These two forms are spatially expressed during development and chondrogenesis with the type IIB procollagen mRNA primarily expressed by chondrocytes while the IIA form is expressed in chondroprogenitor cells (Sandell et al. [1991] J. Cell Biol. 114:1307-1319). The present study demonstrates that the early non-cartilage expression, by somites, mesenchymal and epithelial cells, is predominately the alternate splice form, type IIA procollagen mRNA. Later in development, the type IIB mRNA splice form is expressed by chondrocytes. During the development of intramembranous bones, such as the mandible, type IIA procollagen mRNA is also expressed. In this tissue, the splice form does not switch to type IIB mRNA and no cartilage is formed. These results show that expression of type IIA mRNA, whether by epithelial or mesenchymal cells, precedes formation of overt skeletal structures. © 1994 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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