Mouse Dspp frameshift model of human dentinogenesis imperfecta.
Mouse Dspp frameshift model of human dentinogenesis imperfecta.
复制标题
人牙本质发育不全的小鼠Dspp移码模型。
DOI：
10.1038/s41598-021-00219-4
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发表时间：
2021-10-19
影响因子：
4.6
通讯作者：
Simmer JP
中科院分区：
文献类型：
--
作者：
Liang T;Hu Y;Zhang H;Xu Q;Smith CE;Zhang C;Kim JW;Wang SK;Saunders TL;Lu Y;Hu JC;Simmer JP
牙本质的非综合征性遗传缺陷是由牙本质涎磷蛋白（DSPP）中的两类显性负性/功能获得性突变引起的：影响N端靶向序列的5′突变和将翻译转移到− 1阅读框架的3′突变。DSPP缺陷导致分类为牙本质发育不良II型和牙本质发生异常II型和III型的表型谱重叠。使用CRISPR/Cas9，我们通过引入FLAG标签，然后在终止前翻译493个外源氨基酸的单核苷酸缺失来生成Dspp−1fs小鼠模型。发育中的切牙和/或磨牙从这只小鼠和DsppP 19 L小鼠的特征在于形态评估，bSEM，纳米硬度测试，组织学分析，原位杂交和免疫组织化学。DsppP 19 L牙本质含有牙本质小管，但生长缓慢，比野生型更柔软，矿化程度更低。DsppP 19 L切牙釉质比正常的更软，而磨牙釉质显示杆/杆间清晰度降低。Dspp-1fs牙本质的形成类似于修复性牙本质：它缺乏牙本质小管，含有细胞碎片，并且比Dspp+/+和DsppP 19 L牙本质明显更软和更薄。Dspp-1fs切牙釉质看起来正常，硬度与野生型相当。我们认为5′和3′ Dspp突变通过不同的病理机制导致牙齿畸形，可以被视为不同的疾病。
Non-syndromic inherited defects of tooth dentin are caused by two classes of dominant negative/gain-of-function mutations in dentin sialophosphoprotein (DSPP): 5′ mutations affecting an N-terminal targeting sequence and 3′ mutations that shift translation into the − 1 reading frame. DSPP defects cause an overlapping spectrum of phenotypes classified as dentin dysplasia type II and dentinogenesis imperfecta types II and III. Using CRISPR/Cas9, we generated a Dspp−1fs mouse model by introducing a FLAG-tag followed by a single nucleotide deletion that translated 493 extraneous amino acids before termination. Developing incisors and/or molars from this mouse and a DsppP19L mouse were characterized by morphological assessment, bSEM, nanohardness testing, histological analysis, in situ hybridization and immunohistochemistry. DsppP19L dentin contained dentinal tubules but grew slowly and was softer and less mineralized than the wild-type. DsppP19L incisor enamel was softer than normal, while molar enamel showed reduced rod/interrod definition. Dspp−1fs dentin formation was analogous to reparative dentin: it lacked dentinal tubules, contained cellular debris, and was significantly softer and thinner than Dspp+/+ and DsppP19L dentin. The Dspp−1fs incisor enamel appeared normal and was comparable to the wild-type in hardness. We conclude that 5′ and 3′ Dspp mutations cause dental malformations through different pathological mechanisms and can be regarded as distinct disorders.
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影响因子：
5.1
作者：
Almushayt, A;Narayanan, K;George, A
通讯作者：
George, A
影响因子：
1.9
作者：
Hu Y;Hu JC;Smith CE;Bartlett JD;Simmer JP
通讯作者：
Simmer JP
影响因子：
--
作者：
Kim J;Lee G;Chang WS;Ki SH;Park JC
通讯作者：
Park JC
影响因子：
2
作者：
Mortier, Geert R.;Cohn, Daniel H.;Warman, Matthew L.
通讯作者：
Warman, Matthew L.
影响因子：
4.2
作者：
Bjorndal, L;Darvann, T
通讯作者：
Darvann, T