| 1. |
MA KIDO, T KIYOSHIMA, T KONDO, N AYASAKA, R MOROI, Y TERADA, T TANAKA, DISTRIBUTION OF SUBSTANCE-P AND CALCITONIN GENE-RELATED PEPTIDE-LIKE IMMUNOREACTIVE NERVE-FIBERS IN THE RAT TEMPOROMANDIBULAR-JOINT, JOURNAL OF DENTAL RESEARCH, Vol.72, No.3, pp.592-598, 1993.03, The density and distribution of substance P-like immunoreactive (SP-LI) and calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerve fibers in rat temporomandibular joint (TMJ) were investigated in whole-mount preparations and frozen sections by immunohistochemistry with the avidin-biotin-peroxidase complex method.
Both types of immunoreactive nerves were observed primarily in the joint capsule, the peripheral articular disc, the synovial membrane, and the periosteum. The distribution of CGRP-LI nerves was similar to that of SP-LI nerves. The anterior portion of the joint capsule and disc was most densely innervated, followed by the posterior, lateral, and medial portions. In addition, CGRP-LI nerves were more numerous and more dense in immunointensity than SP-LI nerves. In the synovial membrane, many SP- and CGRP-LI nerves terminated in the subsynovial layer, but some branches extended into the superficial synovial lining layer close to the joint cavity. Immunolabeled nerves were prominently located in the disc attachment and peripheral portion of the disc, and occasional nerves were located in the dense collagenous disc band as an actual disc. However, no fibers were detected in the central disc band. Thus, most of the disc was not innervated by any nerves.
The present study provides a morphological basis for the possible roles of neuropeptides in endocytosis by synoviocytes, regulation of blood flow in the synovial membrane, nociception mechanisms of the TMJ, and modulation of the inflammatory response in the TMJ.. |
| 2. |
T GOTO, T KIYOSHIMA, R MOROI, T TSUKUBA, Y NISHIMURA, M HIMENO, K YAMAMOTO, T TANAKA, LOCALIZATION OF CATHEPSINS-B, CATHEPSINS-D, AND CATHEPSINS-L IN THE RAT OSTEOCLAST BY IMMUNO-LIGHT AND IMMUNOELECTRON MICROSCOPY, HISTOCHEMISTRY, Vol.101, No.1, pp.33-40, 1994.01, The localization of cathepsins B, D, and L was studied in rat osteoclasts by immuno-light and -electron microscopy using the avidin-biotin-peroxidase complex (ABC) method. In cryosections prepared for light microscopy, immunoreactivity for cathepsin D was found in numerous vesicles and vacuoles but was not detected along the resorption lacunae of osteoclasts. However, immunoreactivity for cathepsins B and L occurred strongly along the lacunae, and only weak intracellular immunoreactivity was observed in the vesicles and peripheral part of the vacuoles near the ruffled border. In control sections that were not incubated with the antibody, no cathepsins were found in the osteoclasts or along the resorption lacunae of osteoclasts. At the electron microscopic level, strong intracellular reactivity of cathepsin D was found in numerous vacuoles and vesicles, while extracellular cathepsin D was only slightly detected at the base of the ruffled border but was not found in the eroded bone matrix. Most osteoclasts showed strong extracellular deposition of cathepsins B and L on the collagen fibrils and bone matrix under the ruffled border. The extracellular deposition was stronger for cathepsin L than for cathepsin B. Furthermore cathepsins B and L immunolabled some pits and part of the ampullar extracellular spaces, appearing as vacuoles in the sections. Conversely, the intracellular reactivity for cathepsins B and L was weak: cathepsin-containing vesicles and vacuoles as primary and secondary lysosomes occurred only sparsely. These findings suggest that cathepsins B and L, unlike cathepsin D, are rapidly released into the extracellular matrix and participate in the degradation of organic bone matrix containing collagen fibrils near the tip of the ruffled border. Cathepsin L may be more effective in the degradation of bone matrix than cathepsin B.. |
| 3. |
R Moroi, T Yamaza, T Nishiura, Y Nishimura, Y Terada, K Abe, M Himeno, T Tanaka, Immunocytochemical study of cathepsin L and rat salivary cystatin-3 in rat osteoclasts treated with E-64 in vivo, ARCHIVES OF ORAL BIOLOGY, 10.1016/S0003-9969(97)00003-4, Vol.42, No.4, pp.305-315, 1997.04, The localization of cathepsin L and rat salivary cystatin-3 (RSC-3) in rat osteoclasts (rat femoral and alveolar bones) treated with or without E-64 (control) was examined immunocytochemically. In osteoclasts pretreated with E-64, immunoreactivity for cathepsin L was very weak extracellularly compared to that in the control osteoclasts. However, it was strong intracellularly. The localization of RSC-3 was unclear in the control osteoclasts, while in E-64 treated osteoclasts, both the clear zone and ruffled border areas showed a very strong immunoreaction. At the electron-microscopic level, in normal osteoclasts, numerous immunoreaction products for cathepsin L were found extracellularly in the bone matrix under the ruffled border, while few intracellular products were observed. In contrast, in the E-64-treated osteoclasts, only a few immunoreaction products were found extracellularly, while intracellularly cathepsin L was found in numerous endosome-lysosomal vacuoles. In the immunoreaction for RSC-3, the cytoplasm of the ruffled border was positive, and the tips of the RSC-3-positive ruffled border appeared to enter deeply into the bone matrix. Intracellularly, the granular reaction products of RSC-3 were found in the vacuoles (probably autophagolysosomes). Thus, in E-64-treated osteoclasts, inhibition of the extracellular release of cathepsin L was demonstrated. In addition, intralysosomal accumulation of RSC-3 and deep penetration of the RSC-3-positive ruffled border into the bone matrix were found. These findings suggest that RSC-3 is associated with the inhibition of cathepsin L in both the lysosomes (in the osteoclasts) and bone matrix. (C) 1997 Elsevier Science Ltd.. |
| 4. |
R Moroi, T Yamaza, T Nishiura, Y Nishimura, Y Terada, K Abe, M Himeno, T Tanaka, Immunocytochemical study of cathepsin L and rat salivary cystatin-3 in rat osteoclasts treated with E-64 in vivo, ARCHIVES OF ORAL BIOLOGY, 10.1016/S0003-9969(97)00003-4, Vol.42, No.4, pp.305-315, 1997.04, The localization of cathepsin L and rat salivary cystatin-3 (RSC-3) in rat osteoclasts (rat femoral and alveolar bones) treated with or without E-64 (control) was examined immunocytochemically. In osteoclasts pretreated with E-64, immunoreactivity for cathepsin L was very weak extracellularly compared to that in the control osteoclasts. However, it was strong intracellularly. The localization of RSC-3 was unclear in the control osteoclasts, while in E-64 treated osteoclasts, both the clear zone and ruffled border areas showed a very strong immunoreaction. At the electron-microscopic level, in normal osteoclasts, numerous immunoreaction products for cathepsin L were found extracellularly in the bone matrix under the ruffled border, while few intracellular products were observed. In contrast, in the E-64-treated osteoclasts, only a few immunoreaction products were found extracellularly, while intracellularly cathepsin L was found in numerous endosome-lysosomal vacuoles. In the immunoreaction for RSC-3, the cytoplasm of the ruffled border was positive, and the tips of the RSC-3-positive ruffled border appeared to enter deeply into the bone matrix. Intracellularly, the granular reaction products of RSC-3 were found in the vacuoles (probably autophagolysosomes). Thus, in E-64-treated osteoclasts, inhibition of the extracellular release of cathepsin L was demonstrated. In addition, intralysosomal accumulation of RSC-3 and deep penetration of the RSC-3-positive ruffled border into the bone matrix were found. These findings suggest that RSC-3 is associated with the inhibition of cathepsin L in both the lysosomes (in the osteoclasts) and bone matrix. (C) 1997 Elsevier Science Ltd.. |
| 5. |
T Sakai, T Kiyoshima, Kobayashi, I, R Moroi, T Ibuki, M Nagadome, Y Terada, H Sakai, Age-dependent changes in the distribution of BrdU- and TUNEL-positive cells in the murine gingival tissue, JOURNAL OF PERIODONTOLOGY, 10.1902/jop.1999.70.9.973, Vol.70, No.9, pp.973-981, 1999.09, Background: Age-dependent morphological and cell kinetic changes of the gingival tissue seem to be related to the occurrence of periodontal disease. The purpose of this study was to investigate the age-dependent changes in the distribution of BrdU- and TUNEL-positive cells in murine gingival tissue.
Methods: Gingival tissue of the lower first molar region of 2-, 3-, 5-, 7-, 10-, 15-, 20-, 30-, 40-, 50-, 60-, 70- and 80-week-old mice was used in this study. BrdU- and TUNEL-positive cells were evaluated at the following 4 sites: 1) gingival epithelium (GE); 2) junctional epithelium (JE); 3) submucosal connective tissue of the gingival epithelium (GECT); and 4) submucosal connective tissue of the junctional epithelium (JECT).
Results: No significant differences in the mean number of BrdU-positive cells at each site were demonstrated among the various age groups. No significant change in the mean number of TUNEL-positive cells was demonstrated in either the GE or JE groups among the various age groups. Meanwhile, a significant increase in the TUNEL-positive cells was observed in the GECT of mice 40 weeks or older, and in the JECT of mice 20 weeks or older.
Conclusions: These results indicate that no age-dependent change in the cell proliferation or cell death occurred in the gingival and junctional epithelial layers as well as in the cell proliferation in the submucosal connective tissue. Meanwhile, a significant decrease in the cellular component of the submucosal connective tissue of both gingival and junctional epithelial layers caused by apoptosis occurred with aging. The decreased cellular component in the submucosal connective tissue thus seems to be related to either gingival recession or to the apical migration of the JE with aging. These morphological changes with aging possibly occur in humans and may be related to the susceptibility to periodontal disease in aged individuals.. |
| 6. |
T Sakai, T Kiyoshima, Kobayashi, I, R Moroi, T Ibuki, M Nagadome, Y Terada, H Sakai, Age-dependent changes in the distribution of BrdU- and TUNEL-positive cells in the murine gingival tissue, JOURNAL OF PERIODONTOLOGY, 10.1902/jop.1999.70.9.973, Vol.70, No.9, pp.973-981, 1999.09, Background: Age-dependent morphological and cell kinetic changes of the gingival tissue seem to be related to the occurrence of periodontal disease. The purpose of this study was to investigate the age-dependent changes in the distribution of BrdU- and TUNEL-positive cells in murine gingival tissue.
Methods: Gingival tissue of the lower first molar region of 2-, 3-, 5-, 7-, 10-, 15-, 20-, 30-, 40-, 50-, 60-, 70- and 80-week-old mice was used in this study. BrdU- and TUNEL-positive cells were evaluated at the following 4 sites: 1) gingival epithelium (GE); 2) junctional epithelium (JE); 3) submucosal connective tissue of the gingival epithelium (GECT); and 4) submucosal connective tissue of the junctional epithelium (JECT).
Results: No significant differences in the mean number of BrdU-positive cells at each site were demonstrated among the various age groups. No significant change in the mean number of TUNEL-positive cells was demonstrated in either the GE or JE groups among the various age groups. Meanwhile, a significant increase in the TUNEL-positive cells was observed in the GECT of mice 40 weeks or older, and in the JECT of mice 20 weeks or older.
Conclusions: These results indicate that no age-dependent change in the cell proliferation or cell death occurred in the gingival and junctional epithelial layers as well as in the cell proliferation in the submucosal connective tissue. Meanwhile, a significant decrease in the cellular component of the submucosal connective tissue of both gingival and junctional epithelial layers caused by apoptosis occurred with aging. The decreased cellular component in the submucosal connective tissue thus seems to be related to either gingival recession or to the apical migration of the JE with aging. These morphological changes with aging possibly occur in humans and may be related to the susceptibility to periodontal disease in aged individuals.. |
| 7. |
T Yamaza, Y Tsuji, T Goto, MA Kido, K Nishijma, R Moroi, A Akamine, T Tanaka, Comparison in localization between cystatin C and cathepsin K in osteoclasts and other cells in mouse tibia epiphysis by immunolight and immunoelectron microscopy, BONE, 10.1016/S8756-3282(01)00466-5, Vol.29, No.1, pp.42-53, 2001.07, We compared the distribution of a cysteine proteinase inhibitor, cystatin C, with that of cathepsin K in osteoclasts of the mouse tibia by immunolight and immunoelectron microscopy. Light microscopically, strong immunoreactivity for cystatin C was found extracellularly along the resorption lacuna and intracellularly in the organelles of osteoclasts. In serial sections, various patterns of cystatin C and cathepsin K localization were seen, specifically: (1) some resorption lacuna were positive for both cystatin C and cathepsin K; (2) others were positive for either cystatin C or cathepsin K, but not both; and (3) some lacuna were negative for both. In osteoclasts, the localization of cystatin C was similar to that of cathepsin K. Furthermore, cystatin C immunoreactivity was detected in preosteoclasts and osteoblasts, whereas cathepsin K was seen only in preosteoclasts. Electron microscopically, cystatin C immunoreactive products were found in the rough endoplasmic reticulum (ER), Golgi apparatus, vesicles, granules, and vacuoles of osteoclasts. These cystatin C-positive vesicles had fused or were in the process of fusion with the ampullar vacuoles (extracellular spaces) containing cystatin C-positive, fragmented, fibril-like structures. The extracellular cystatin C was deposited on and between the cytoplasmic processes of ruffled borders, and on and between type I collagen fibrils. In the basolateral region of osteoclasts, cystatin C-positive vesicles and granules also fused with vacuoles that contained cystatin C-positive or negative fibril-like structures. These results indicate that osteoclasts not only synthesize and secrete cathepsin K from the ruffled border into the bone resorption lacunae, but also a cysteine proteinase inhibitor, cystatin C. Therefore, it is suggested that cystatin C regulates the degradation of bone matrix by cathepsin K, both extracellularly and intracellularly. (C) 2001 by Elsevier Science Inc. All rights reserved.. |
| 8. |
T Yamaza, Y Tsuji, T Goto, MA Kido, K Nishijma, R Moroi, A Akamine, T Tanaka, Comparison in localization between cystatin C and cathepsin K in osteoclasts and other cells in mouse tibia epiphysis by immunolight and immunoelectron microscopy, BONE, 10.1016/S8756-3282(01)00466-5, Vol.29, No.1, pp.42-53, 2001.07, We compared the distribution of a cysteine proteinase inhibitor, cystatin C, with that of cathepsin K in osteoclasts of the mouse tibia by immunolight and immunoelectron microscopy. Light microscopically, strong immunoreactivity for cystatin C was found extracellularly along the resorption lacuna and intracellularly in the organelles of osteoclasts. In serial sections, various patterns of cystatin C and cathepsin K localization were seen, specifically: (1) some resorption lacuna were positive for both cystatin C and cathepsin K; (2) others were positive for either cystatin C or cathepsin K, but not both; and (3) some lacuna were negative for both. In osteoclasts, the localization of cystatin C was similar to that of cathepsin K. Furthermore, cystatin C immunoreactivity was detected in preosteoclasts and osteoblasts, whereas cathepsin K was seen only in preosteoclasts. Electron microscopically, cystatin C immunoreactive products were found in the rough endoplasmic reticulum (ER), Golgi apparatus, vesicles, granules, and vacuoles of osteoclasts. These cystatin C-positive vesicles had fused or were in the process of fusion with the ampullar vacuoles (extracellular spaces) containing cystatin C-positive, fragmented, fibril-like structures. The extracellular cystatin C was deposited on and between the cytoplasmic processes of ruffled borders, and on and between type I collagen fibrils. In the basolateral region of osteoclasts, cystatin C-positive vesicles and granules also fused with vacuoles that contained cystatin C-positive or negative fibril-like structures. These results indicate that osteoclasts not only synthesize and secrete cathepsin K from the ruffled border into the bone resorption lacunae, but also a cysteine proteinase inhibitor, cystatin C. Therefore, it is suggested that cystatin C regulates the degradation of bone matrix by cathepsin K, both extracellularly and intracellularly. (C) 2001 by Elsevier Science Inc. All rights reserved.. |
| 9. |
K Shimizu, W Chen, AM Ashique, R Moroi, YP Li, Molecular cloning, developmental expression, promoter analysis and functional characterization of the mouse CNBP gene, GENE, 10.1016/S0378-1119(03)00406-2, Vol.307, pp.51-62, 2003.03, Striking conservation in various organisms suggests that cellular nucleic acid-binding protein (CNBP) plays a fundamental biological role across different species. However, the regulated expression and physiological properties of the CNBP gene are unknown. In this study, we report the molecular cloning, promoter characterization, developmental expression and functional analysis of the mouse CNBP gene. The gene contains five exons and is localized to chromosome 6 in the region corresponding to band 6 D1-D2. Primer extension assay indicates that the transcription start site is located 230 bp upstream of the initiator Met codon. Our promoter analysis indicates that strong transcription enhancer and silencer regions lie within the 1.6 kb proximal region of the promoter and the upstream -3.0 to -1.6 kb region, respectively. The promoter activity is 10 fold higher in embryonic carcinoma cells than that in fibroblast, as determined by CAT assay. Consistent with its function as a transcription factor, CNBP protein is located in the nucleus of cells. During mouse embryogenesis, CNBP is expressed in the anterior region of the early embryo and in the limb, tail and craniofacial region. Overexpression of CNBP strongly stimulates cell proliferation and increases c-myc promoter activity. Our finds suggest that CNBP may play an important role in cell proliferation and tissue patterning during anterior-posterior axis, craniofacial and limb development by targeting c-Myc. (C) 2003 Elsevier Science B.V. All rights reserved.. |
| 10. |
K Shimizu, W Chen, AM Ashique, R Moroi, YP Li, Molecular cloning, developmental expression, promoter analysis and functional characterization of the mouse CNBP gene, GENE, 10.1016/S0378-1119(03)00406-2, Vol.307, pp.51-62, 2003.03, Striking conservation in various organisms suggests that cellular nucleic acid-binding protein (CNBP) plays a fundamental biological role across different species. However, the regulated expression and physiological properties of the CNBP gene are unknown. In this study, we report the molecular cloning, promoter characterization, developmental expression and functional analysis of the mouse CNBP gene. The gene contains five exons and is localized to chromosome 6 in the region corresponding to band 6 D1-D2. Primer extension assay indicates that the transcription start site is located 230 bp upstream of the initiator Met codon. Our promoter analysis indicates that strong transcription enhancer and silencer regions lie within the 1.6 kb proximal region of the promoter and the upstream -3.0 to -1.6 kb region, respectively. The promoter activity is 10 fold higher in embryonic carcinoma cells than that in fibroblast, as determined by CAT assay. Consistent with its function as a transcription factor, CNBP protein is located in the nucleus of cells. During mouse embryogenesis, CNBP is expressed in the anterior region of the early embryo and in the limb, tail and craniofacial region. Overexpression of CNBP strongly stimulates cell proliferation and increases c-myc promoter activity. Our finds suggest that CNBP may play an important role in cell proliferation and tissue patterning during anterior-posterior axis, craniofacial and limb development by targeting c-Myc. (C) 2003 Elsevier Science B.V. All rights reserved.. |
| 11. |
S Kamolmatyakul, W Chen, S Yang, Y Abe, R Moroi, AM Ashique, YP Li, IL-1 alpha stimulates cathepsin K expression in osteoclasts via the tyrosine kinase-NF-kappa B pathway, JOURNAL OF DENTAL RESEARCH, Vol.83, No.10, pp.791-796, 2004.10, Interleukin-1alpha (IL-1alpha) is a powerful activator of osteoclast cells. However, the underlying mechanism for this activation is unknown. In this study, we reveal that IL-1alpha up-regulates the expression of cathepsin K protein, a key protease in bone resorption, by five-fold. Northern blot analysis and promoter analysis show that this induction occurs at the transcriptional level, in a dose-responsive and time-dependent manner. No increase in expression occurs in the presence of either pyrrolidine dithiocarbamate ( PDTC), a selective inhibitor of NF-kappaB, or Genistein, a protein tyrosine kinase inhibitor, suggesting that IL-1alpha up-regulation may be via the tyrosine kinase-NF-kappaB pathway to regulate cathepsin K expression. Antisense oligonucleotides to p65, but not the p50 subunit of NF-kappaB, suppress the IL-1alpha-induced expression of cathepsin K. We therefore conclude that IL-1alpha up-regulates cathepsin K gene expression at the transcription level, and this regulation may be via the tyrosine-kinase-NF-kappaB pathway.. |
| 12. |
YP Li, Y Abe, R Isoda, R Moroi, S Yang, JZ Shao, E Li, W Chen, Cathepsin K-deficient mice exhibit pycnodysostosis geatures due to loss of cathepsin K-mediated osteoclast senescence for bone homeostasis, JOURNAL OF BONE AND MINERAL RESEARCH, Vol.20, No.9, p.S253, 2005.09. |
| 13. |
Yali Cheng, Takako Sakai, Ryoji Moroi, Masaharu Nakagawa, Hidetaka Sakai, Tetsuro Ogata, Yoshihiro Terada, Self-cleaning ability of a photocatalyst-containing denture base material, DENTAL MATERIALS JOURNAL, 10.4012/dmj.27.179, Vol.27, No.2, pp.179-186, 2008.03, This study examined the ability of a photocatalyst mixed in a denture base resin to decompose organic substances which adhered to the denture base resin surface. The photocatalyst was mixed with denture base resin at concentrations of 0, 5, 10, and 15% (w/w). Decomposition test, bending test, and surface roughness measurement were performed at 1, 7, 30, 90, and 180 days after polymerization. Decomposition ability was evaluated based on the residual amount of methylene blue (MB) dissolved in ethanol after UV irradiation for 12 hours. As the mixing ratio increased, the amount of MB in the solution decreased. Meanwhile, no changes in the amount of MB in the immersion solution were observed in the photocatalyst-free resin specimen. Therefore, the results indicated that a denture base resin containing a photocatalyst might have a photocatalytic ability.. |
