Kinumatsu T, Hashimoto S, Muramatsu T, Sasaki H, Jung H-S, Yamada S, Shimono M
Department of Periodontics, Tokyo Dental College, Chiba, Japan.
J Periodontal Res. 2009 Feb;44(1):13-20. doi: 10.1111/j.1600-0765.2007.01036.x. Epub 2008 Sep 11.
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.
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.
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.
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.
结合上皮通过半桥粒(其主要成分是层粘连蛋白-5和整合素α(6)β(4))附着于釉质表面的内基膜。层粘连蛋白-5也通过整合素α(3)β(1)参与细胞运动,尽管它们的功能尚未阐明。本研究的目的是确定牙齿与结合上皮之间这些黏附成分在细胞迁移过程中的功能。因为有观点认为直接附着于牙齿的细胞(DAT细胞)可能对细胞迁移没有作用,所以进行了5-溴-2-脱氧尿苷染色以确认细胞迁移。
我们使用激光显微切割、定量实时逆转录-聚合酶链反应、免疫荧光和5-溴-2-脱氧尿苷染色,研究了6周龄ICR小鼠结合上皮、口腔牙龈上皮和龈沟上皮中的层粘连蛋白γ(2)(仅存在于层粘连蛋白-5中)、整合素β(4)(参与细胞与细胞外基质接触)和整合素α(3)(诱导细胞迁移)。
层粘连蛋白和整合素在所有上皮的基膜中均有明显的免疫定位。通过激光显微切割对层粘连蛋白和整合素mRNA进行定量分析表明,它们在口腔牙龈上皮的DAT细胞中的表达高于基底细胞。特别是,与口腔牙龈上皮相比,结合上皮中层粘连蛋白-5的表达高12倍。5-溴-2-脱氧尿苷染色显示DAT细胞向冠方快速迁移。
这些结果表明,层粘连蛋白-5和整合素α(6)β(4)的大量表达通过半桥粒参与DAT细胞与牙齿的附着。层粘连蛋白-5和整合素α(3)β(1)的大量表达可能有助于DAT细胞迁移,这在结合上皮更新过程中的5-溴-2-脱氧尿苷染色中得到了证实。