小型猪乳磨牙胚SHH的时空表达

Spatiotemporal expression of SHH in deciduous molar embryos of miniature pig

目的本研究通过探索SHH在小型猪第三乳磨牙胚的帽状期、钟状期和分泌期的时空表达情况,拟阐明SHH在牙齿发育调控中的作用。方法收集小型孕猪第三乳磨牙胚,通过原位杂交、免疫组化和实时聚合酶链反应检测SHH的m RNA和蛋白表达水平。结果原位杂交及免疫组化可见SHH在帽状期少量表达于成釉器,但在釉结部位表达较强。在钟状期,SHH在间充质中表达明显增强;在分泌期,SHH在间充质和成釉器中都有所表达,尤其在成牙本质细胞和内釉上皮细胞中表达增强。PCR结果显示,在钟状期,SHH在上皮中表达较多,而在帽状期和分泌期,SHH在上皮和间充质中的表达未见明显统计学差异。结论SHH可能在调节上皮形态发育方面具有重要功能,参与调节信号级联,介导上皮和间充质之间的相互作用,本研究进一步完善了小型猪牙齿发育的信号分子网络。

Objective This study aims to elucidate the role of SHH in the regulation of tooth development by exploring the spatiotemporal expression of SHH in the cap stage,bell stage,and secretion phase of the third deciduous molar in miniature pigs.Methods Collect the third molar molar embryos of small pregnant pigs and detect the mRNA and protein expression levels of SHH through in situ hybridization,immunohistochemistry,and real-time polymerase chain reaction.Results Through in situ hybridization and immunohistochemistry,we found that SHH is expressed is small amounts in the cap stage of the enamel,but strongly in the enamel knot.During the bell stage,SHH expression is significantly enhanced in the mesenchyme.During the secretion phase,SHH is expressed in both mesenchymal and enamel cells,particularly in odontoblasts and inner enamel epithelial cell.the PCR results showed that during the bell stage,SHH is more expressed in the epithelium,while in the cap and secretion phases,there was no significant statistical difference in the expression of SHH between the epithelim and mesenchyme.Conclusions SHH may play an important role in regulating epithelial morphogenesis,participating in regulating signal cascades and mediating the interaction between epithelium and mesenchyme.This study further improves the signaling mnolecular network of small pig tooth development.