敲除 fth1b 会影响斑马鱼咽齿的早期矿化。
Knockout fth1b affects early mineralization of zebrafish pharyngeal teeth.
机构信息
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.
出版信息
Hua Xi Kou Qiang Yi Xue Za Zhi. 2021 Feb 1;39(1):32-37. doi: 10.7518/hxkq.2021.01.005.
OBJECTIVES
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 fertilization. The expression of fth1b in zebrafish pharyngeal teeth development was detected by whole embryo hybridization 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.
CONCLUSIONS
The gene fth1b is specifically expressed in zebrafish pharyngeal teeth and acts on their early mineralization.
目的
本研究旨在探讨铁蛋白重链多肽基因（fth1b）在斑马鱼咽齿发育过程中的表达模式和功能，为后续牙齿发育和矿化研究奠定基础。
方法
收集受精后 56、72、96 和 120 h 的斑马鱼胚胎，通过全胚胎杂交检测 fth1b 在斑马鱼咽齿发育中的表达，并与已知的咽齿标记基因 dlx2b 进行比较。利用成簇规律间隔短回文重复序列（CRISPR）/Cas9 基因编辑技术特异性敲除 fth1b 基因。检测 fth1b 突变体中斑马鱼咽齿的发育情况。
结果
fth1b 基因的表达模式与已知的斑马鱼咽齿标记基因 dlx2b 非常相似，在发育过程中特异性表达于斑马鱼咽齿中。基因 fth1b 特异性敲除后，最早可检测到的基因 pitx2 在早期发育过程中表达正常。dlx2b 的表达与野生型斑马鱼无显著差异，但突变体中咽齿的矿化程度弱于野生型斑马鱼。
结论
fth1b 基因在斑马鱼咽齿中特异性表达，并作用于其早期矿化。
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