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Gap junctions play an important role in differentiation of odontoblasts. Gap junction protein, connexin 43 is expressed in odontoblast. However, the detailed localization in odontoblasts has yet to be fully investigated. We investigated the localization of connexin43 in rat odontoblasts immuno-electron microscopically. The rats were transcardially fixed with 1% paraformaldehyde in 0.1M phosphate buffer, and mandibles were decalcified with 10% ethylene- diamine tetraacetic acid. Pre-embedding method was carried out for immuno-electron micro- scopic analysis. Microscopically, gap junctions were localized between bodies of odontoblasts, and between bodies and processes of odontoblasts. The gap junctions were labeled with gold par- ticles that indicated connexin43. These results suggest that gap junctions between odontoblasts are definitely composed of connexin43 in rats, and our methods used in this study is useful to investigate localization of connexin43 immuno-electron microscopically. Microsc. Res. Tech. 76:988-991, 2013. V C 2013 Wiley Periodicals, Inc.

SUMMARY Plant ‘activators’, such as benzothiadiazole (BTH), protect plants from various diseases by priming the plant salicylic acid (SA) signalling pathway. We have reported previously that a transcription factor identified in rice, WRKY45 (OsWRKY45), plays a pivotal role in BTH-induced disease resistance by mediating SA signalling. Here, we report further functional characterization of WRKY45. Different plant activators vary in their action points, either downstream (BTH and tiadinil) or upstream (probenazole) of SA. Rice resistance to Magnaporthe grisea, induced by both types of plant activator, was markedly reduced in WRKY45-knockdown (WRKY45-kd) rice, indicating a universal role for WRKY45 in chemical-induced resistance. Fungal invasion into rice cells was blocked at most attempted invasion sites (pre-invasive defence) in WRKY45-overexpressing (WRKY45-ox) rice. Hydrogen peroxide accumulated within the cell wall underneath invading fungus appressoria or between the cell wall and the cytoplasm, implying a possible role for H2O2 in pre-invasive defence. Moreover, a hypersensitive reaction-like reaction was observed in rice cells, in which fungal growth was inhibited after invasion (post-invasive defence). The two levels of defence mechanism appear to correspond to Type I and II nonhost resistances. The leaf blast resistance of WRKY45-ox rice plants was much higher than that of other known blast-resistant varieties. WRKY45-ox plants also showed strong panicle blast resistance. BTH-induced resistance to Xanthomonas oryzae pv. oryzae was compromised in WRKY45-kd rice, whereas WRKY45-ox plants were highly resistant to this pathogen. However, WRKY45-ox plants were susceptible to Rhizoctonia solani. These results indicate the versatility and limitations of the application of this gene.

Sugisawa M, Masaoka T, Enokiya Y, Muramatsu T, Hashimoto S, Yamada S, Shimono M. Expression and function of laminin and integrins on adhesion/migration of primary culture cells derived from rat oral epithelium. J Periodont Res 2010; 45: 284–291. © 2010 John Wiley & Sons A/S Background and Objective: It remains controversial whether or not the junctional epithelium cells that are directly attached to teeth migrate on the enamel surface, as those cells are able to adhere firmly to the enamel. The aim of this study was to investigate the expression patterns of laminin γ2, integrin β4 and integrin α3, and to examine their potential function in cell migration. Material and Methods: Oral epithelium cells obtained from Sprague–Dawley rats were established in primary culture. We employed a wound-healing assay to characterize the direction of cell extension at the start of cell migration, and observed different localizations of laminin and integrins using immunofluorescence. For functional analyses of integrins, we employed a phosphatidylinositol-3-kinase (PI3K) activator to promote integrin β4 function and used P1B5 to inhibit integrin α3 function, and we analyzed the percentage of re-epithelialization as the migration function. Results: Marked accumulation of laminin γ2 was detected in the peripheral cytoplasm of cells adjacent to the wound area, as shown by the results of the migration assay. Integrin β4 was detected in the distal cell processes of actively migrating cells, while integrin α3 was found in cell membranes of cells adjacent to the wound area. In the functional analyses, the percentage of re-epithelialization was significantly lower in the PI3K-activator group and in the P1B5-treated group (2.5% and 7.2%, respectively) than in the control group (39.0%) (p

Oral Diseases (2010) 16, 263–268 Objective: The aim of this study was to investigate the proliferation, migration and death of periodontal ligament (PDL) cells after tooth replantation. Materials and methods: Maxillary first molars were extracted from 4-week-old male (n = 28) Sprague–Dawley rats and immediately replanted, after which, proliferation, migration and death of PDL cells were investigated. Results: At 3 days after tooth replantation, many proliferative cell nuclear antigen (PCNA)-positive PDL cells were observed on the alveolar bone side, but fewer on the root side. However, while a gradual decrease was observed in number of PCNA-positive PDL cells on the alveolar bone side until 7 days, an increase was seen on the root side. At 3 weeks, cells labeled with PKH26 (fluorescent dye into plasma membrane) were located in the middle of the PDL space. However, these PKH26-labeled cells did not spread to the surface of the cementum or the alveolar bone. TUNEL-positive cells were observed on both the bone and root sides at 3 days. Number of apoptotic cells increased until 7 days on the bone sides, but decreased on root sides. Conclusion: These results suggest that both cell proliferation and apoptosis occur in different patterns and at different times to maintain regular spacing of the PDL after tooth replantation.

Background and Objective: The 4-META/MMA-TBB [4-(2-methacryloxyethyl)trimellitic anhydride/methyl methacrylate-tributylborane] resin is widely used as a dental adhesive. It has also been applied in the dressing of gingival wound surfaces following periodontal surgery. However, its effect on the regeneration and/or cell attachment of the oral epithelium remains to be clarified. To evaluate the effect of the resin applied as a wound dressing, we investigated expression of laminin 5, integrin β4 and cytokeratin 14 in regenerating oral epithelium treated with this resin following gingivectomy from the viewpoint of cell attachment and differentiation. Material and Methods: The resin was applied to the entire wound surface in rats after gingival surgery, and regenerating epithelium was examined immediately and at 1, 3, 5, 7 and 14 days later. The resin was removed 2 weeks after application in some animals and tissue further examined at 1, 3, 5 and 7 days later. Results: Regenerating epithelium under the resin was not keratinized, but became keratinized immediately after removal of the resin. Laminin 5 and integrin β4 were immunolocalized in the basal lamina, the internal basal lamina, in marginal cells of the regenerating epithelium and at the resin–regenerating epithelium interface. Cytokeratin 14 localized in the regenerating epithelium underneath the resin, as well as in healthy and regenerated junctional epithelial cells. Conclusion: These results suggest that this resin covers the wound surface and that the regenerating epithelium biologically adheres to the resin during the initial process of its regeneration.

Background and objective The junctional epithelium attaches to the enamel surface with hemidesmosomes (of which laminin-5 and integrin-alpha(6)beta(4) are the main components) in the internal basal lamina. Laminin-5 is also involved in cell motility with integrin-alpha(3)beta(1), although their functions have not yet been clarified.The purpose of this study was to determine the functions of those adhesive components between the tooth and the junctional epithelium during cell migration.Because an idea has been proposed that directly attached to tooth cells (DAT cells) may not contribute to cell migration, 5-bromo-2-deoxyuridine staining was performed to confirm cell migration. Material and methods We investigated laminin-gamma(2) (contained only in laminin-5), integrin-beta(4) (involved in cell-extracellular matrix contact) and integrin-alpha(3) (inducing cell migration) in the junctional epithelium, oral gingival epithelium and gingival sulcus epithelium of 6-wk-old ICR mice using laser microdissection, quantitative real-time reverse transcription-polymerase chain reaction, immunofluorescence and 5-bromo-2-deoxyuridine staining. Results Laminin and integrins were clearly immuno-localized in the basal lamina of all epithelium. Quantitative analysis of laminin and integrin mRNAs by laser microdissection showed that they were more highly expressed in DAT cells than in basal cells in the oral gingival epithelium. In particular, a 12-fold higher expression of laminin-5 was observed in the junctional epithelium compared with the oral gingival epithelium. 5-Bromo-2-deoxyuridine staining showed rapid coronal migration of DAT cells. Conclusion These results suggest that the abundant expression of laminin-5 and integrin-alpha(6)beta(4) is involved in the attachment of DAT cells to teeth by hemidesmosomes. Abundant expression of laminin-5 and integrin-alpha(3)beta(1) might assist in DAT cell migration, confirmed by 5-bromo-2-deoxyuridine staining during the turnover of junctional epithelium.

Many morphological and developmental studies have demonstrated the characteristics of tight junctions (TJs) between odontoblasts. However, detailed localization of TJ-associated proteins in odontoblasts and their functions has not yet been clarified. To elucidate the relationship between the establishment of TJ structures and the differentiation of odontoblasts during early dentinogenesis, we studied the expression and localization of constituent proteins of TJs (claudin-1, occludin, ZO-1 and ZO-2) between odontoblasts in rat lower incisors using Western blotting, immunofluorescence and immunoelectron microscopy. When the expression of claudin-1 increases at the distal portion of mature odontoblasts, the TJs form complex networks of strands, and odontoblasts differentiated by developing distal membrane domains and by secreting specific molecules for mineralization. We conclude that the TJs of odontoblasts may play an important role in the differentiation of odontoblasts in rat lower incisors during early dentinogenesis.

Background: Osteopontin (OPN) expression in squamous cell carcinoma (SCC) of the tongue has not been clearly elucidated. Methods: We selected 46 cases of tongue SCC and investigated the expression of OPN by immunohistochemical staining. The immunopositive reaction and score for each case were semiquantitatively evaluated. Results: Scores were significantly higher in carcinoma nests than in neighboring normal epithelium or epithelial dysplasia. The OPN was expressed clearly in the cytoplasm of carcinoma cells. In cases of early invasive carcinoma, in particular, expression of OPN showed a remarkable increase at the invasion front compared with the non-invaded regions. However, there was no significant correlation between expression of OPN in the primary tumor nest and lymphatic metastasis, recurrence, or survival rate. Conclusion: This suggests that OPN is a useful biomarker of early invasion by SCC in tongue.

Background and Objective: It is known that epithelial islands are embedded in the cementum during tooth root formation, but details of this process remain unknown. The purpose of this study was to investigate the dynamic characteristics of Malassez's epithelial rest cells in the cementum during tooth root formation in pigs in vivo. Material and Methods: The first molars of 6-mo-old pigs were used in this study. Specimens were decalcified before being embedded in paraffin. Paraffin sections were investigated using TdT-mediated dUTP-biotin nick end labeling (TUNEL), immunohistochemical, and ultrastructural techniques. Results: Malassez's epithelial rest cells were located close to the root surface at the apical one-third of the periodontal ligament, and epithelial clusters surrounded by distinct lamina cementia were sometimes observed in the cementum. TUNEL-positive cells were detected only in the cementum. Malassez's epithelial rest cells in the periodontal ligament were completely surrounded by basement membranes, but epithelial clusters in the cementum were only intermittently surrounded by such membranes. Cytokeratin-positive cells in the superstratum of the cementum were directly connected by cementocytes and by desmosome-like structures. However, organelles were scarce in the cytokeratin-positive cells in the substratum of the cementum, and the matrix of the cementum was deposited in the cells. Conclusion: These results suggest that the majority of the fragmented Hertwig's root sheath remains in the periodontal ligament and that some cells, which are connected to cementoblasts, are embedded in the cementum and progress to apoptosis.

Objectives The aim of this study was to investigate the differential immunolocalization of laminin gamma(2) and integrin beta(4) in primary cultures of the rat gingival epithelium. Methods The gingival epithelium was obtained from Sprague-Dawley rats and was cultured in serum-free keratinocyte growth medium (DK-SFM). Western blotting analysis, immunofluorescence, confocal laser scanning microscopy (CLSM), and immuno-gold labeling for laminin gamma(2) and integrin beta(4) were employed. CLSM images for laminin and integrin were analyzed in horizontal (x-y axis) and in vertical (x-z axis) sections. Results Both laminin gamma(2) and integrin beta(4) were detected by Western blot analysis in the gingival epithelium. Immunolocalization of laminin gamma(2) was distinct in the cytoplasm to form one or two irregular rings in gingival epithelial cells. By contrast, integrin beta(4) was localized diffusely in the cytoplasm. F-actin (indicating actin filaments) was clearly discernible at the periphery of the cytoplasm to form a cellular fringe. In x-z axis images obtained by CLSM, laminin gamma(2) was recognized as large foci in the most inner portion just above the basal plasma membrane. Integrin beta(4) existed in the area where F-actin was labeled surrounding the membrane. Immuno-electron microscopy showed that 10nm colloidal gold particles indicating laminin gamma(2) were mainly localized at the extracellular portion and in the peripheral cytoplasm, whereas integrin beta(4) was distributed in the cytoplasm close to the basal plasma membrane but not in extracellular regions. Conclusions In primary cultures of the rat gingival epithelium, both laminin gamma(2) and integrin beta(4) may be produced by the epithelium, and irregular rings of laminin gamma(2) are formed in areas where gingival cells adhere to the extracellular matrix.

Objective: It is still an open question whether cells directly attached to the tooth (DAT) cells are migratory or non-migratory cells. The purpose of this study was to examine cytoskeletal and surface structures of DAT cells that might be involved in migration. Methods: We investigated the distribution of stress fibers composed of actin filaments in DAT cells using phallacidin fluorescent dye methods in a confocal laser scanning microscope. To observe the three-dimensional structure of the DAT cell surface, the osmium maceration scanning electron microscope (SEM) method, which removes various soluble materials between DAT cells and the enamel, was employed. Results: Stress fibers were found in the most apically located DAT cells, and were arranged in parallel to the presumable cervical-line, whereas some of the fibers ran parallel to the tooth axis in the more coronally located DAT cells. The parallel arrangement to the tooth axis of the fibers may be involved with migration for turnover, and the parallel accumulation to the presumable cervical-line may be concerned with the cervical contraction of DAT cells. Osmium maceration SEM images at high magnification revealed the existence of microvilli-like structures on the enamel surfaces (facing to the tooth surface) of DAT cells after removal of the soluble matrices. The thicknesses of the microvilli-like structures on the enamel surfaces and cell processes of intercellular bridges were significantly different. Conclusion: DAT cells possess stress fibers arranged in parallel to the tooth axis and to the presumable cervical-line in the cytoplasm, and microvilli-like structures on their enamel surfaces. These results suggest that these structures contribute to DAT cell migration.

Background and objective: It is still an open question why long junctional epithelium can proliferate and occupies the root surface following periodontal surgery or experimentally produced periodontitis, and why the epithelium repopulated once on the root surface is replaced by the connective tissue. The aim of this study is to investigate the proliferative activity of the newly formed regenerative connective tissue and long junctional epithelium during wound healing by staining argyrophilic proteins of the nucleolar organizer regions (AgNORs). Methods: Regenerative connective tissue and long junctional epithelium were experimentally created by insertion of a rubber piece between maxillary molars of rats for 1 week. After removal of the rubber, AgNORs parameters including nuclear area (NA), AgNORs area (AA), AgNORs percentage nuclear area (APNA), AgNORs number (AN) and nuclear number (NN) in regenerative connective tissue and long junctional epithelium were measured and analyzed statistically. Results: APNA in long junctional epithelium after 1 and 4 weeks was over two times greater than that in the regenerative connective tissue. AA in long junctional epithelium was significantly higher than in regenerative connective tissue at 1 and at 4 weeks post-treatment. AN was higher in the central portion than at the root surface except at 20 weeks. APNA and AA decreased remarkably in long junctional epithelium at 12 weeks post-treatment (approximately half at 4 weeks), whereas in regenerative connective tissue, they did not change distinctly. Conclusions: These results imply that long junctional epithelium cannot supply sufficient epithelial cells because of their significantly low rates of proliferation, consequently long junctional epithelium becomes shorter after 12 weeks, whereas the proliferative activity of regenerative connective tissue maintains the same level of proliferation, and ultimately long junctional epithelium is replaced by regenerative connective tissue.

The purpose of this study was to investigate pulp cell responses during hypoxia and reoxygenation. Pulp tissues obtained from beagle dogs were cultured. In the control group, pulp cells were incubated in normoxic conditions (20% O2) for 1-4 d. In the hypoxia group, pulp cells were incubated under hypoxic conditions (2% O2) for 1-4 d. In the reoxygenation group, pulp cells were first incubated under hypoxic conditions for 24 h, and were then incubated in normoxic conditions (20% O2) for one to three additional days. Cell viability, MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction assay, cellular proliferation, and alkaline phosphatase (ALPase) activity were determined. Expression of heat shock protein 70 (HSP70) and vascular endothelial growth factor (VEGF) was analysed by Western blotting. Hypoxia inducible factor-1alpha (HIF-1alpha) in pulp cells was analysed by reverse transcriptase polymerase chain reaction (RT-PCR). The cell growth rate and ALPase activity were significantly higher in the hypoxia group than in the control group. After reoxygenation, cellular proliferation and ALPase activity decreased to the level of the control group while HSP70 expression increased. Hypoxia inducible factor-1alpha expression was detected in pulp cells, and VEGF expression (which is regulated by HIF-1alpha) increased under hypoxic conditions. These results suggest that dynamic responses to hypoxia and reoxygenation occur in pulp cells.

The purpose of this study was to investigate cellular responses of periodontal ligaments during tooth movement. Twenty-eight male Sprague-Dawley rats, weighing 200-250 g each, were used. To create the orthodontic force, elastic rubber blocks (0.65 mm thick) were inserted between the maxillary first and second molars on both sides. On days 3, 7, 10, 14, 21 and 28 after rubber block insertion, histopathological changes in both the tension and the pressure sides were examined by immunohistochemistry using proliferating cell nuclear antigen (PCNA) and by the TUNEL method. The ratios of PCNA-positive cells on the tension side 3 and 7 days after rubber block insertion were higher than those on the pressure side. The ratios of PCNA-positive cells on the tension side were highest at day 3 after insertion and then decreased during the remainder of the experimental period. On the pressure side, the ratios of PCNA-positive cells increased up to day 10 post insertion, then decreased from 14 to 28 days. The ratios of TUNEL-positive cells on both the tension and the pressure sides increased throughout the entire experimental period. These results indicate that the periodontal ligaments on the tension side are able to respond more promptly to orthodontic forces than those on the pressure side. The data also suggest that the ratios of cell proliferation and of cell death are closely related to the regeneration and reconstruction of periodontal ligaments which reflect the orthodontic force.

Effects of experimental warming on shoot developmental growth and biomass production were preliminarily investigated in two evergreen dwarf shrubs Empetrum nigrum and Loiseleuria procumbens, using the International Tundra Experiment’s open-top chamber (OTC) method, in the Tateyama Range, central Japan. An OTC was installed over shrub (E. nigrum and L. procumbens) -dominated vegetation and over shrub-forb (such as Anemone narcissiflora var. nipponica and Solidago virga-aurea ssp. leiocarpa) mixed vegetation, and stem samples of the evergreen shrubs were obtained at 26 months after installing the OTC. The OTC increased the daily mean temperature by 0.1°C to 1.8°C, on average, during the growing season. Shoot developmental growth and biomass production were considerably different between species of different vegetation types. The boreal species E. nigrum generally showed better growth inside the OTC than the arctic and subarctic species L. procumbens. Both species showed significantly larger shoot elongation and biomass production inside the OTC over shrub-dominated vegetation, whereas smaller or reduced growth was detected inside the OTC over shrub-forb mixed vegetation. The variations of growth responses to warming between species of different vegetation types are discussed, especially in relation to interspecific competition under a simulated environmental change.

The purpose of this study was to investigate nerve regeneration and proliferative activity in amputated pulp tissue after the application of 4-META/MMA-TBB adhesive resin (4-META resin). Calcium hydroxide was used as a control material. At 3 days, fibroblast-like cells were positive for proliferating cell nuclear antigen (PCNA) in both 4-META resin- and calcium hydroxide-treated groups and were located mainly within 0.5 mm from the cut surface. Only a few fragmented neurofilament protein (NFP)-positive nerve fibers were observed in this area. At 7 and 14 days, the number of PCNA-positive cells had gradually decreased and regenerated NFP-positive nerve fibers were observed close to the cut surface of the pulp in both groups. At 21 days in the experimental group, several PCNA-positive cells were still found in the area 0.5 mm from the cut surface, and NFP-positive nerve fibers were detected about 0.15-;0.2 mm from the cut surface. In contrast, a dentin bridge was produced under the necrotic layer at 21 days in the control group. PCNA-positive cells were not found underneath the dentin bridge, but NFP-positive nerve fibers had regenerated close to it. These results suggest that although cell differentiation and nerve regeneration are delayed, wound healing occurred even after the application of 4-META resin to exposed pulp surface the same as calcium hydroxide application.

The role played by the internal basal lamina (IBL) and hemidesmosomes between an implant and the peri-implant epithelium (PIE) in the adherence of the epithelium to the implant is controversial. This study used rat maxilla implantation models to clarify the ultrastructure of the PIE-implant interface.Ti-6Al-4V implants were inserted either immediately or 2 weeks after the extraction of the upper left first molar of 6- or 4-week-old rats, respectively. The junctional epithelium (JE) of the upper right molars in the same animals was used as a control. Four weeks after implantation, the animals were sacrificed to prepare specimens for light and immunoelectron microscopy.Under light microscopy, the PIE appeared to attach to the implant surface. Ultrastructurally, IBL, consisting of the lamina densa and lamina lucida, and hemidesmosomes were formed only in the lower region, and rarely in the middle region, of the PIE-implant interface. In control teeth, the IBL and hemidesmosomes formed throughout the dento-JE interface. Laminin-1 was found in the IBL and also in the vesicles and vacuoles of the PIE and JE cells. Statistical analysis showed that there was also a significant difference in the amount of IBL between the PIE-implant and dento-JE interfaces.PIE attached to the implant via hemidesmosomes and IBL in the lower region of the PIE-implant interface. Although PIE cells may secrete laminin-1, which contributes to epidermal cell adhesion, the PIE which attaches to implants only in the lower region of the interface is considered to be the poorly adhered epithelium.

A rare case of clear cell odontogenic carcinoma was investigated using histochemical and immunohistochemical methods. The tumor occurred in the anterior mandible of a 69-year-old Japanese man. Histologically, the tumor was composed mostly of large clear cells and squamous cells. Columnar-shaped cells with basophilic nuclei polarized away from the basement membrane were observed at the periphery of the tumor foci. The tumor cells had aggressively invaded muscle and perineural tissues. The tumor cells were positive for PAS staining. Immunohistochemically, tumor cells reacted positively to keratin, cytokeratin19, epithelial membrane antigen, and S-100 protein. The tumor was diagnosed as a clear cell odontogenic carcinoma. Its characteristics are discussed in term of its histopathological, histochemical and immunohistochemical features.

Lee M-W, Muramatsu T, Uekusa T, Lee J-H, Shimono M. Heat stress induces alkaline phosphatase activity and heatshock protein 25 expression in cultured pulp cells. InternationalEndodontic Journal, 41, 158–162, 2008. Aim To investigate the responses of cultured rat pulpcells to heat stress.Methodology Pulp cells were obtained from ratincisors and cultured at 37 C. The cells were culturedat 42 C for 30 min and then cultured at 37 C again.Morphology, alkaline phosphatase (ALP) activity andexpression of heat shock protein 25 (HSP25) wereinvestigated at 0, 1, 3, 5, 7, 10 and 14 days followingstimulation. As a control, the cells were maintained at37 C.Results Although there were few cells of apoptosisimmediately after heat stress, there were mitotic cellsfrom day 1 after heat stress. ALP activity in the heatstress group significantly increased at days 7 and 14compared with the control group (about 1.7-fold,P < 0.01, Friedman test). HSP25 expression increasedin both groups, with HSP25 in the heat stress groupbeing expressed earlier than in the control group, andnuclear localization of HSP25 was observed at days 0and 1 in heat-stressed cells.Conclusion These results suggest that heat stressnot only induces HSP25 but also enhances ALP activityin pulp cells.Keywords: alkaline phosphatase activity, dentalpulp, heat shock protein 25, heat stress.

To elucidate the biological characteristics of the junctional epithelium (JE) in rats, ultrastructural and morphometric studies of the gingiva in the maxillary molar regions were carried out. Morphometric analysis of three different regions of the JE and the oral epithelium (OE) led to the following results. In the apical JE, intercellular space accounted for about 23% of the total epithelial tissue. At the connective tissue interface, the space accounted for 17% of total epithelial tissue. At the enamel interface, the space accounted for 35% of total JE tissue. A large number of leukocytes were detected in the enlarged intercellular spaces. In the OE, the space accounted for about 11%. In the apical region of the JE. desmosomes occupied approximately 5% of the plasma membrane perimeter; in both the enamel and connective tissue, about 3%; and in the OE, about 5%. However, desmosome densities were approximately 14/100 μm2 in the JE. while being approximately 66/100 μm2 in the OE. From these results, it is suggested that: (a) the volume of intercellular space relative to the entire JE coincides with a dynamic migration of the epithelium; (b) enlargement of the intercellular spaces in the JE causes a low incidence of desmosomes: (c) there is no physiological permeability barrier in the JE; and (d) abundant leukocytes in the intercellular spaces may play an important role in obstructing the passage of external bacteria and toxins into the tissue.