Toriya Naoko, Takuma Taishin, Arakawa Toshiya, Abiko Yoshihiro, Sasano Yasuyuki, Takahashi Ichiro, Sakakura Yasunori, Rahemtulla Firoz, Mizoguchi Itaru
Department of Orthodontics, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-tobetsu, Hokkaido, 061-0293 Japan.
Histochem Cell Biol. 2006 Mar;125(3):205-14. doi: 10.1007/s00418-005-0020-1. Epub 2005 Sep 27.
To analyze the growth-related changes in extracellular matrix components in temporomandibular joint (TMJ) discs, the expression and localization of the core protein of a large chondroitin sulphate proteoglycan, versican, in rat TMJ discs during postnatal development (2-32 weeks) were examined using Western blot analysis, real-time quantitative PCR and immunohistochemistry. Western blot analysis showed that rat TMJ discs predominantly expressed one isoform (V1) and the core protein sharply increased after birth, reached a peak at 8 weeks, and then gradually decreased up to 32 weeks. Real-time quantitative PCR with TaqMan probes indicated that mRNA expression of versican was highest at 2 weeks and gradually decreased with growth. An immunohistochemical study showed that staining for versican was weak and evenly distributed in TMJ discs at 2 weeks. Regional differences in staining for versican became prominent after 8 weeks; staining was intense in the anterior and posterior peripheral attachments, and weak in the central part of the discs. These results demonstrate that growth-related changes and regional differences exist in the expression of versican in the TMJ discs of growing rats, and these probably reflect the changes in the biomechanical environment caused by the development of orofacial functions.
为分析颞下颌关节(TMJ)盘细胞外基质成分与生长相关的变化,采用蛋白质免疫印迹分析、实时定量PCR及免疫组织化学方法,检测了大鼠出生后发育阶段(2 - 32周)颞下颌关节盘中一种大分子硫酸软骨素蛋白聚糖核心蛋白versican的表达及定位。蛋白质免疫印迹分析显示,大鼠颞下颌关节盘主要表达一种异构体(V1),其核心蛋白在出生后急剧增加,于8周时达到峰值,随后直至32周逐渐下降。使用TaqMan探针进行的实时定量PCR表明,versican的mRNA表达在2周时最高,并随生长逐渐降低。免疫组织化学研究显示,2周时颞下颌关节盘中versican染色较弱且分布均匀。8周后versican染色的区域差异变得明显;在前、后周边附着处染色强烈,而在盘的中央部分较弱。这些结果表明,生长中的大鼠颞下颌关节盘中versican的表达存在与生长相关的变化及区域差异,这可能反映了口面部功能发育所引起的生物力学环境的变化。