fth1b基因敲除对斑马鱼咽齿早期矿化的影响研究

doi:10.7518/hxkq.2021.01.005

华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (1): 32-37.doi: 10.7518/hxkq.2021.01.005

fth1b基因敲除对斑马鱼咽齿早期矿化的影响研究

周春艳(), 郑雪丹, 杨德琴()

重庆医科大学附属口腔医院牙体牙髓病科口腔疾病与生物医学重庆市重点实验室重庆市高等教育口腔生物医学工程重点实验室，重庆 401147

收稿日期:2019-08-12 修回日期:2020-03-22 出版日期:2021-02-01 发布日期:2021-03-02
通讯作者: 杨德琴 E-mail:2017110871@stu.cqmu.edu.cn;yangdeqin@hospital.cqmu.edu.cn
作者简介:周春艳，硕士，E-mail：2017110871@stu.cqmu.edu.cn
基金资助:
国家自然科学基金(31571508)

Knockout fth1b affects early mineralization of zebrafish pharyngeal teeth

Zhou Chunyan(), Zheng Xuedan, Yang Deqin()

Dept. of Endodontics, Stomatological Hospital of Chongqing Medical University; Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China

Received:2019-08-12 Revised:2020-03-22 Online:2021-02-01 Published:2021-03-02
Contact: Yang Deqin E-mail:2017110871@stu.cqmu.edu.cn;yangdeqin@hospital.cqmu.edu.cn
Supported by:
The National Natural Science Foundation of China(31571508)

摘要/Abstract

摘要： 目的

探索铁蛋白重肽基因（fth1b）在斑马鱼咽齿发育过程中的表达模式及其功能，为后续牙齿相关研究奠定基础。

方法

收集受精后56、72、96及120 h斑马鱼胚胎，运用整胚原位杂交技术检测fth1b基因在斑马鱼咽齿发育过程中的表达；并运用规律成簇间隔短回文重复序列（CRISPR）/Cas9基因编辑技术对fth1b基因特异性敲除，在fth1b^-/-突变体中检测斑马鱼咽齿的发育状况。

结果

fth1b基因与已知斑马鱼咽齿标记基因dlx2b表达模式类似，特异性表达于斑马鱼咽齿发育过程中；靶向性敲除fth1b基因后，早期发育过程中，斑马鱼咽齿标记基因pitx2、dlx2b表达与野生型无较大差别，而咽齿的矿化较野生型斑马鱼弱。

结论

fth1b基因特异性表达于斑马鱼咽齿，对斑马鱼咽齿的早期矿化起作用。

关键词: 铁蛋白重肽基因, 基因敲除, 牙齿发育, 斑马鱼

Abstract: Objective

A study was conducted to explore the expression pattern and function of ferritin heavy polypeptide gene (fth1b) in zebrafish pharyngeal teeth development and lay the foundation for subsequent research on teeth development and mineralization.

Methods

The zebrafish embryos were harvested at 56, 72, 96, and 120 h after ferti-lization. The expression of fth1b in zebrafish pharyngeal teeth development was detected by whole embryo in situ hybri-dization and compared with the known pharyngeal teeth marker dlx2b. The specific knockout of fth1b gene was performed using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing technology. The development of zebrafish pharyngeal teeth was detected in the fth1b^-/- mutant.

Results

The expression pattern of fth1b gene was very similar to that of the known zebrafish pharyngeal teeth marker dlx2b and was specifically expressed in the zebrafish pharyngeal teeth during development. After the specific knockout of the gene fth1b, the earliest gene that can be detect in zebrafish pharyngeal teeth-pitx2 was expressed normally during early development. The dlx2b expression was not significantly different from that of wild type zebrafish, but the mineralization of pharyngeal teeth in the mutant was weaker than that of wild type zebrafish.

Conclusion

The gene fth1b is specifically expressed in zebrafish pharyngeal teeth and acts on their early mineralization.

Key words: ferritin heavy polypeptide gene, gene knockout, teeth development, zebrafish (Danio rerio)

中图分类号:

Q 786

周春艳, 郑雪丹, 杨德琴. fth1b基因敲除对斑马鱼咽齿早期矿化的影响研究[J]. 华西口腔医学杂志, 2021, 39(1): 32-37.

Zhou Chunyan, Zheng Xuedan, Yang Deqin. Knockout fth1b affects early mineralization of zebrafish pharyngeal teeth[J]. West China Journal of Stomatology, 2021, 39(1): 32-37.

图/表 5

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fth1b基因敲除对斑马鱼咽齿早期矿化的影响研究

Knockout fth1b affects early mineralization of zebrafish pharyngeal teeth

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