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组蛋白去甲基化酶Kdm2a在小鼠磨牙发育过程中的时空表达

Spatial and temporal expression of histone demethylase, Kdm2a, during murine molar development.

作者信息

Yi Q, Cao Y, Liu O S, Lu Y Q, Wang J S, Wang S L, Yao R, Fan Z P

机构信息

a Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology , Beijing , China.

d Xiangya Stomatology Hospital, Central South University , Changsha, Hunan , China.

出版信息

Biotech Histochem. 2016;91(2):137-44. doi: 10.3109/10520295.2015.1106586. Epub 2015 Dec 31.

Abstract

The histone demethylase, lysine (K)-specific demethylase 2A (Kdm2a), is highly conserved and expressed ubiquitously. Kdm2a can regulate cell proliferation and osteo/dentinogenic, adipogenic and chondrogenic differentiation of mesenchymal stem cells (MSCs) derived from dental tissue. We used quantitative real-time RT-PCR analysis and immunohistochemistry to detect Kdm2a expression during development of the murine molar at embryonic days E12, E14, E16 and E17 and postnatal days P3 and P14. Immunohistochemistry results showed no positive staining of Kdm2a at E12. At E14, Kdm2a was expressed weakly in the inner enamel epithelium, stellate reticulum cells and dental sac. At E16, Kdm2a was expressed mainly in the inner and outer enamel epithelium, stratum intermedium and dental sac, but weaker staining was found in cervical loop and dental papilla cells adjacent to the basement membrane. At E17, the strongest Kdm2a staining was detected in the ameloblasts and stronger Kdm2a staining also was detected in the stratum intermedium, outer enamel epithelium and dental papilla cells compared to the expression at E16. Postnatally, we found that Kdm2a was localized in secretory and mature ameloblasts and odontoblasts, and dentin was unstained. Real-time RT-PCR showed that Kdm2a mRNA levels in murine germ cells increased from E12 to E14 and from E14 to E16; no significant change occurred at E16, E17 or P3, then the levels decreased at P14 compared to P3. Kdm2a expression may be closely related to cell proliferation, to ameloblast and odontoblast differentiation and to the secretion of extracellular enamel and dentin during murine tooth development.

摘要

组蛋白去甲基化酶,赖氨酸(K)特异性去甲基化酶2A(Kdm2a)高度保守且广泛表达。Kdm2a可调节源自牙组织的间充质干细胞(MSC)的细胞增殖以及骨/牙本质生成、脂肪生成和成软骨分化。我们使用定量实时逆转录聚合酶链反应(RT-PCR)分析和免疫组织化学方法,检测了小鼠磨牙在胚胎第12天(E12)、第14天(E14)、第16天(E16)和第17天以及出生后第3天(P3)和第14天发育过程中Kdm2a的表达。免疫组织化学结果显示,在E12时Kdm2a无阳性染色。在E14时,Kdm2a在内釉上皮、星网状层细胞和牙囊中有弱表达。在E16时,Kdm2a主要在内釉上皮和外釉上皮、中间层和牙囊中表达,但在与基底膜相邻的颈环和牙乳头细胞中染色较弱。在E17时,成釉细胞中检测到最强的Kdm2a染色,与E16时的表达相比,中间层、外釉上皮和牙乳头细胞中也检测到较强的Kdm2a染色。出生后,我们发现Kdm2a定位于分泌期和成釉细胞以及成牙本质细胞,而牙本质无染色。实时RT-PCR显示,小鼠生殖细胞中Kdm2a mRNA水平从E12到E14以及从E14到E16升高;在E16、E17或P3时无显著变化,然后与P3相比,在P14时水平下降。Kdm2a的表达可能与小鼠牙齿发育过程中的细胞增殖、成釉细胞和成牙本质细胞分化以及细胞外釉质和牙本质的分泌密切相关。

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