染色体结构域解旋酶dna结合蛋白3在牙齿发育过程中通过调节尖乳头的早期生长阶段来确定颈宽度的潜在作用。

IF 2.6 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences Pub Date : 2024-12-20 DOI:10.1016/j.job.2024.100604

Kento Shimamura , Toshiki Nojiri , Hisatomo Kondo , Yunosuke Ikeda , Rika Yasuhara , Hiroko Ida-Yonemochi , Keishi Otsu , Hidemitsu Harada , Kenji Mishima , Hayato Ohshima , Takuya Kobayashi , Tarou Irié

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Subsequent histological and immunohistochemical analyses of teeth were performed at each developmental stage. <em>Chd3</em> knockdown in mesenchymal cells from the dental papilla (mDP) and Hertwig's epithelial root sheath (HERS) was performed by <em>Chd3</em> shRNA transduction or a control using an adenoviral vector. These effects were examined using cell proliferation assays and quantitative real-time polymerase chain reaction.</div></div><div><h3>Results</h3><div>Narrowing of tooth cervical width was observed in mandibular first molars of <em>Chd3</em> knockout mice. On postnatal day (PN) 8, the cervical width was narrow before root formation in tooth germs. 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引用次数: 0

摘要

目的：探讨染色体结构域解旋酶dna结合蛋白3 （CHD3）在CHD3基因敲除小鼠牙齿形态发生中的作用。方法：采用CRISPR-Cas9方法构建Chd3基因敲除小鼠。从小鼠及其窝仔身上提取下颌第一磨牙并进行形态计量学分析。随后在每个发育阶段对牙齿进行组织学和免疫组织化学分析。在牙乳头（mDP）和Hertwig上皮根鞘（HERS）的间充质细胞中，通过Chd3 shRNA转导或使用腺病毒载体进行对照，实现Chd3的下调。使用细胞增殖试验和定量实时聚合酶链反应检测这些影响。结果：Chd3基因敲除小鼠下颌第一磨牙牙颈宽度变窄。出生后第8天（PN），牙胚根形成前颈宽较窄。PN1牙间质和PN8牙顶乳头中ki -67阳性细胞数量减少。Chd3对牙间充质细胞增殖有促进作用，但对HERS上皮细胞无明显影响。Chd3在牙间质细胞中维持sonic hedgehog （Shh）的表达，抑制骨形态发生蛋白（Bmp）4的表达，维持Shh和Wnt3a的表达，抑制HERS上皮细胞中Bmp2的表达。结论：Chd3可能通过Shh、Bmp和Wnt信号调控牙颈宽度。

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The potential role of chromodomain helicase DNA-binding protein 3 in defining the cervical width by regulating the early growth stage of the apical papilla during tooth development

Objective

This study aimed to evaluate the role of the chromodomain helicase DNA-binding protein 3 (CHD3) in tooth morphogenesis in Chd3 knockout mice.

Methods

Chd3 knockout mice were generated using the CRISPR-Cas9 method. Mandibular first molars were extracted from the mice and their littermates and morphometrically analyzed. Subsequent histological and immunohistochemical analyses of teeth were performed at each developmental stage. Chd3 knockdown in mesenchymal cells from the dental papilla (mDP) and Hertwig's epithelial root sheath (HERS) was performed by Chd3 shRNA transduction or a control using an adenoviral vector. These effects were examined using cell proliferation assays and quantitative real-time polymerase chain reaction.

Results

Narrowing of tooth cervical width was observed in mandibular first molars of Chd3 knockout mice. On postnatal day (PN) 8, the cervical width was narrow before root formation in tooth germs. The number of Ki-67-positive cells decreased in the dental mesenchyme at PN1 and apical papilla at PN8. Chd3 promoted the proliferation of dental mesenchymal cells, but no significant changes were observed in HERS epithelial cells. Chd3 maintained sonic hedgehog (Shh) expression and inhibited that of bone morphogenetic protein (Bmp)4 in dental mesenchymal cells, maintaining Shh and Wnt3a expression and inhibited that of Bmp2 in HERS epithelial cells.