Immunohistochemical Localization of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the Forming Rat Incisor

Goldberg, Michel; Septier, D.; Bourd, Katia; Hall, Richard L.; George, Anne; Goldberg, Harvey E.; Ménashi, Suzanne

doi:10.1080/713713680

Cited by 44 publications

(65 citation statements)

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“…Three dentin proteases were evident on zymograms, which corresponded to enzymes previously shown to be expressed by porcine odontoblasts: MMP-2 (27), MMP-20 (28), and KLK4 (29). We did not observe MMP-9, which corresponds to a 102-kDa gelatinasepositive band in rat dentin extracts (30). Instead, we observed a 98-kDa protease-positive artifact band caused by DPP in the extract, which does not stain with CBB and therefore leaves an unstained band on the zymogram.…”

Section: Resultsmentioning

confidence: 54%

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Dentin Sialophosphoprotein Is Processed by MMP-2 and MMP-20 in Vitro and in Vivo

Yamakoshi

Iwata

et al. 2006

Journal of Biological Chemistry

116

135

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Dentin sialophosphoprotein (DSPP) is a major secretory product of odontoblasts and is critical for proper tooth dentin formation. During dentinogenesis, DSPP is proteolytically cleaved into smaller subunits. These cleavages are proposed activation steps, and failure to make these cleavages is a potential cause of developmental tooth defects. We tested the hypothesis that dentin-resident matrix metalloproteinases catalyze the cleavages that process DSPP. We defined the exact DSPP cleavages that are catalyzed by proteases during crown formation by isolating DSPP-derived proteins from developing porcine molars and characterizing their N-terminal sequences and apparent size on SDS-PAGE and Western blots. The in vivo DSPP cleavage sites were on the N-terminal sides of Thr Dentin sialophosphoprotein (DSPP)2 is a multidomain extracellular matrix protein that is critical for proper dentin formation. The major DSPP domains are dentin sialoprotein (DSP), dentin glycoprotein (DGP), and dentin phosphoprotein (DPP) (1-3). In humans, nine disease-causing mutations in the DSPP gene on chromosome 4q21.3 have been reported in kindreds with isolated inherited dentin defects (4 -10). No other genes have been implicated in their etiology by mutation analyses. DSPP mutations result in a variety of dental phenotypes, including dentin dysplasia type II and dentinogenesis imperfecta types II and III. In mice, targeted knock-out of the Dspp gene (Dspp Ϫ/Ϫ ) resulted in tooth defects resembling dentinogenesis imperfecta type III (11). DSPP is expressed in other tissues besides dentin, such as in bone (12), but the protein levels in the secondary locations are hundreds of times lower than in dentin. Outside of a poorly understood association with progressive high frequency sensorineural hearing loss, DSPP mutations result in defects limited to the teeth. Perhaps because DSPP functions are so specialized, relatively little research has been focused on it, and many of its basic structural features are still unknown. DSPP itself has never been isolated or detected in dentin extracts. DSPP was discovered through its proteolytic cleavage products, starting with DPP (13). DPP is a highly acidic phosphoprotein with an isoelectric pH of 1.1 (14). Besides its distinctive amino acid composition, dominated by serine and aspartic acid (15), DPP has a conserved N-terminal sequence of AspAsp-Pro-Asn (1). DPP is difficult to study at the protein level, because it is resistant to digestion by proteases, such as trypsin (16). Cloning and characterization of the first DPP cDNA transcript revealed the extreme redundancy of its DNA and deduced amino acid sequences (17). Surprisingly, the cloned DPP transcript also encoded DSP, another major noncollagenous protein in dentin (18,19). DSP is a highly glycosylated proteoglycan (19 -21). Demonstration of a continuous open reading frame between the DSP and DPP coding regions disclosed that DSP and DPP are cleavage products of a larger protein, called DSPP (22). DGP is the most recent and smallest DSPP-derived pro...

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Section: Resultsmentioning

confidence: 54%

“…An unusual feature of DPP is that it does not stain with CBB. Because of this, DPP made a clear band on gelatin zymograms that we originally mistook for being MMP-9, a protease reportedly in rat dentin that migrates at a similar position (30). We did not detect MMP-9 in porcine dentin extracts.…”

Section: Discussionmentioning

confidence: 69%

Dentin Sialophosphoprotein Is Processed by MMP-2 and MMP-20 in Vitro and in Vivo

Yamakoshi

Iwata

et al. 2006

Journal of Biological Chemistry

116

135

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“…Several proteolytic activities are present in the predentin and dentin 1,19) , including kallikrein 4 (Klk4), matrix metalloproteinase 2(Mmp-2), Mmp-9, a disintegrin and metalloproteinase with thrombospondin motifs 4 (Adamts4), and Mmp-20 [20][21][22][23][24] . These activated proteinases degrade the coexisting proteoglycans following the addition of calcium in vitro 2) .…”

Section: Discussionmentioning

confidence: 99%

A Large Chondroitin Sulfate Proteoglycan, Versican, in Porcine Predentin

Okahata

Yamamoto

Yamakoshi

et al. 2011

Journal of Oral Biosciences

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Proteoglycans and their constituent glycosaminoglycan (GAG) have been proposed to be involved in the inhibition of mineralization in unmineralized tissue, predentin. Among the proteoglycans secreted by odontoblasts, we focused on the large chondroitin sulfate proteoglycan, versican, for its large binding capacity for calcium ions. The aims of this study were the determination of the full-length sequence and splicing variants of the porcine versican, and the detection of versican in the porcine predentin. The complete coding sequence of the porcine versican mRNA was cloned to be 11,775 nucleotides long and encode 3,924 amino acids, and four splicing variants, V0, V1, V2 and V3, were characterized in the isolated porcine cartilage cells. The number of potential GAG attachment sites was 15 in the V0 variant, 13 in the V1 variant, 2 in the V2 variant and 0 in the V3 variant. They were deposited in DDBJ. The V1 variant was determined by RT-PCR in the odontoblasts, dental papilla cells, dental follicle cells, periodontal ligament cells, dental pulp cells, and gingival cells of pigs, although a small amount of the V0 valiant was found in the dental papilla cells. The predentin was prepared from developing porcine permanent incisor tooth germs and its soluble proteins were extracted in order to be partially characterized by protein and proteinase profiles. The versican V1 cleavage products were detected in the predentin extract by Western blotting analysis. These results suggested that the versican splice variant V1 implicates both the control of the mineralization and the activities of the predentin metalloproteinases, because it has 13 GAG chains that bind a large amount of calcium.

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“…MMPs are synthesized and secreted as proenzymes and are activated in the extracellular matrix. Their proteolytic activity is regulated by the tissue inhibitors of metalloproteinases (TIMPs) and by the nonspecific inhibitors such as α2-macroglobulin (43,44). MMPs contribute to both physiological and pathological tissue remodelling.…”

Section: Matrix Metalloproteinasesmentioning

confidence: 99%

Bioactive Chitosan Nanoparticles and Photodynamic Therapy Inhibit Collagen Degradation In Vitro

Persadmehr

Torneck

Cvitkovitch

et al. 2014

Journal of Endodontics

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This study evaluated the ability of photodynamic therapy (PDT), chitosan nanoparticles (CSnp), or their combination, to inhibit bacterial collagenase-mediated degradation of collagen. Rat type 1 fibrillar collagen matrices were untreated or treated with 2.5% glutaraldehyde (GD), 2.5% GD followed by 1% CSnp, 1% CSnp, PDT, or 1% CSnp followed by PDT. Samples, except untreated controls, were exposed to Clostridium histolyticum collagenase. The soluble digestion products were assessed by hydroxyproline assay and the remaining adherent collagen was quantified by picrosirius red (PSR) staining. Collagen treated with CSnp, PDT, or a combination of CSnp and PDT, exhibited less degradation than controls. The abundance of post-treatment residual collagen correlated with the extent of degradation. Fourier transform infrared (FTIR) spectroscopy analysis showed that PDT treatment enhanced collagen cross-linking.Immunoblotting of sedimented CSnp indicated that CSnp and collagenase bound with low affinity. However, CSnp-bound collagenase showed a significant reduction in collagenolytic activity compared with controls.III

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Immunohistochemical Localization of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the Forming Rat Incisor

Cited by 44 publications

References 0 publications

Dentin Sialophosphoprotein Is Processed by MMP-2 and MMP-20 in Vitro and in Vivo

Dentin Sialophosphoprotein Is Processed by MMP-2 and MMP-20 in Vitro and in Vivo

A Large Chondroitin Sulfate Proteoglycan, Versican, in Porcine Predentin

Bioactive Chitosan Nanoparticles and Photodynamic Therapy Inhibit Collagen Degradation In Vitro

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