Yang So-Young, Jeon Soo-Kyung, Kang Jee-Hae, Yoo Hong-Il, Kim Yoo-Seong, Moon Jung-Sun, Kim Min-Seok, Koh Jung-Tae, Oh Won-Mann, Kim Sun-Hun
Dental Science Research Institute, 2nd stage Brain Korea, School of Dentistry, Chonnam National University, Gwangju, South Korea.
Eur J Oral Sci. 2012 Dec;120(6):505-12. doi: 10.1111/j.1600-0722.2012.01001.x. Epub 2012 Oct 4.
The formation of dentin and enamel is initiated by the differentiation of odontogenic precursor cells into odontoblasts and ameloblasts, respectively. This study was performed to identify new molecules involved in the differentiation of odontogenic cells. The genes expressed differentially between the root stage (after the differentiation of odontogenic cells and dental hard-tissue formation) and the cap stage (before the differentiation of odontogenic cells and dental hard-tissue formation) were searched using differential display PCR. For the first time, synaptic vesicle protein (SV) 2b, an important transmembrane transporter of Ca(2+) -stimulated vesicle exocytosis, was identified as a differentially expressed molecule. Real-time PCR and western blotting revealed an increase in the transcriptional and translational levels of SV2b during or after the differentiation of odontogenic cells. Immunofluorescence revealed this molecule to be localized in not only fully differentiated odontoblasts but also in pre-odontoblasts before dentin matrix secretion. The expression pattern of the SV2a isoform was similar to that of the SV2b isoform, whereas the SV2c isoform showed a contrasting pattern of expression. After treatment with alendronate, an inhibitor of protein isoprenylation for the transport of secretory vesicles, the expression of SV2a and SV2b decreased, whereas that of SV2c increased. These results suggest that the SV2 isoforms are functional molecules of (pre)odontoblasts which may be involved in vesicle transport.
牙本质和釉质的形成分别由牙源性前体细胞分化为成牙本质细胞和成釉细胞启动。本研究旨在鉴定参与牙源性细胞分化的新分子。使用差异显示PCR搜索在根阶段(牙源性细胞分化和牙体硬组织形成之后)和帽状期(牙源性细胞分化和牙体硬组织形成之前)之间差异表达的基因。首次鉴定出突触小泡蛋白(SV)2b,它是Ca(2+)刺激的囊泡胞吐作用的重要跨膜转运蛋白,为差异表达分子。实时PCR和蛋白质印迹显示,在牙源性细胞分化期间或之后,SV2b的转录和翻译水平增加。免疫荧光显示该分子不仅定位于完全分化的成牙本质细胞中,也定位于牙本质基质分泌前的前成牙本质细胞中。SV2a亚型的表达模式与SV2b亚型相似,而SV2c亚型显示出相反的表达模式。用阿仑膦酸盐(一种用于分泌小泡运输的蛋白质异戊二烯化抑制剂)处理后,SV2a和SV2b的表达降低,而SV2c的表达增加。这些结果表明,SV2亚型是(前)成牙本质细胞的功能分子,可能参与小泡运输。