Yang Guofeng, Jiang Beizhan, Cai Wenping, Liu Shangfeng, Zhao Shouliang
Department of Endodontics, School & Hospital of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai, 200072, People's Republic of China.
Department of Stomatology, Huashan Hospital, Fudan University, Shanghai, 200040, People's Republic of China.
J Mol Histol. 2016 Aug;47(4):413-20. doi: 10.1007/s10735-016-9684-1. Epub 2016 Jun 18.
Hyaluronan (HA) and hyaluronan synthases (HASs) have been shown to play critical roles in embryogenesis and organ development. However, there have not been any studies examining HA and HAS expression and localization during tooth development. The present study was designed to investigate the expression of HA and three isoforms of HASs (HAS1, 2, 3) in embryonic mouse molars. The first mandibular embryonic mouse molars were examined by immunohistochemistry at E11.5, E13.5, E14.5, E16.5, and E18.5. PCR and western blot analyses were performed on RNA and proteins samples from E13.5 to E18.5 tooth germs. At the initial stage (E11.5), HA and HASs were expressed in the dental epithelium but not the underlying dental mesenchyme. HA immunostaining gradually increased in the enamel organ from the bud stage (E13.5) to the late bell stage (E18.5), and HA and HASs were highly expressed in the stellate reticulum and stratum intermedium. HA immunostaining was also enhanced in the dental mesenchyme and its derived tissues, but it was not expressed in the ameloblast and odontoblast regions. The three HAS isoforms had distinct expression patterns, and they were expressed in the dental mesenchyme and odontoblast at various levels. Furthermore, HAS1 and HAS2 expression decreased, while HAS3 expression increased from E13.5 to E18.5. These results suggested that HA synthesized by different HASs is involved in embryonic mouse molar morphogenesis and cytodifferentiation.
透明质酸(HA)和透明质酸合成酶(HASs)已被证明在胚胎发生和器官发育中起关键作用。然而,尚未有任何研究探讨牙齿发育过程中HA和HAS的表达及定位。本研究旨在调查HA和三种HAS亚型(HAS1、2、3)在胚胎小鼠磨牙中的表达。通过免疫组织化学方法,对E11.5、E13.5、E14.5、E16.5和E18.5的第一下颌胚胎小鼠磨牙进行检查。对E13.5至E18.5牙胚的RNA和蛋白质样本进行PCR和蛋白质印迹分析。在初始阶段(E11.5),HA和HASs在牙上皮中表达,但在其下方的牙间充质中不表达。从蕾状期(E13.5)到晚期钟状期(E18.5),釉器中的HA免疫染色逐渐增加,并且HA和HASs在星网状层和中间层中高表达。牙间充质及其衍生组织中的HA免疫染色也增强,但在成釉细胞和成牙本质细胞区域不表达。三种HAS亚型具有不同的表达模式,它们在牙间充质和成牙本质细胞中以不同水平表达。此外,从E13.5到E18.5,HAS1和HAS2的表达下降,而HAS3的表达增加。这些结果表明,由不同HASs合成的HA参与胚胎小鼠磨牙的形态发生和细胞分化。