鸟苷酸交换因子Trio通过调控神经嵴细胞迁徙影响颅颌面发育

口腔生物医学 ›› 2021, Vol. 12 ›› Issue (1): 17-20.

鸟苷酸交换因子Trio通过调控神经嵴细胞迁徙影响颅颌面发育

孟丽，郭舒瑜，刘浩洁，袁礼婵，顾嘉雯，马俊青

南京医科大学口腔疾病研究江苏省重点实验室，南京医科大学附属口腔医院正畸科

收稿日期:2020-09-10 修回日期:2020-12-25 出版日期:2021-03-25 发布日期:2021-04-01
通讯作者: 马俊青 E-mail:jma@njmu.edu.cn
基金资助:
国家自然科学基金;国家自然科学基金;江苏高校优势学科建设工程资助项目(2018-87)

Guanine nucleotide exchange factor Trio effects craniofacial development by regulating neural crest cells migration

Received:2020-09-10 Revised:2020-12-25 Online:2021-03-25 Published:2021-04-01

摘要/Abstract

摘要： 目的：研究鸟苷酸交换因子Trio在胚胎发育过程中对神经嵴细胞的影响。方法：将trio反义寡核苷酸通过显微注射入斑马鱼胚胎内，抑制其trio功能；在斑马鱼胚胎发育96?h时使用体式显微镜观察神经嵴细胞衍生物下颌骨的发育情况，120?h时通过阿尔辛蓝染色观察颅颌面部软骨的发育状况。利用转基因斑马鱼（sox10:EGFP）特异性标记神经嵴细胞，通过延时摄影观察trio敲低后对斑马鱼神经嵴细胞迁徙的影响。体外培养小鼠神经嵴细胞，转染慢病毒敲低Trio的表达，通过划痕实验观测Trio对神经嵴细胞迁徙的影响。结果：斑马鱼体内敲低trio后，下颌发育不足，麦克尔软骨位置靠后，同时神经嵴细胞的迁徙距离减小。在体外实验中，Trio敲低组神经嵴细胞迁移后划痕面积增大。结论：Trio可能通过调节神经嵴细胞的迁徙功能促进颅颌面部神经嵴衍生物的发育。

关键词: Trio, 神经嵴细胞, 迁徙, 颅颌面部发育, 斑马鱼

Abstract: Objective:To investigate the effect of Rho GEF Trio on neural crest cells during embryonic development. Methods:Trio function was inhibited by microinjection of morpholino (MO) into zebrafish embryos，the development of mandibular and neural crest cells derivative was observed at 96 h through stereo microscope, and the development of craniomaxillofacial cartilage was observed at 120 h by Alcian blue staining. The effect of trio on the neural crest cells migration path of zebrafish was observed by using the neural crest cell specific labeling technique of transgenic zebrafish (sox10：EGFP)by means of Timelapse. Trio knockdown neural crest cells were cultured in vitro to observe the effect of Trio on the migration of neural crest cells through measuring the wound area (%) after migration in wound healing assay. Results:When trio function was inhibited，zebrafish mandibular development was insufficient, the Meckel's cartilage became retrusive, and the distance of neural crest cells migration decreased. In vitro，the wound area after neural crest cell migration increased in the Trio knockdown group. Conclusions:Trio may promote craniofacial development of neural crest origin by regulating the neural crest cells migration.

Key words:

Trio;?neural crest cells, ?migration;?craniofacial development;?zebrafish

孟丽郭舒瑜刘浩洁袁礼婵顾嘉雯马俊青. 鸟苷酸交换因子Trio通过调控神经嵴细胞迁徙影响颅颌面发育[J]. 口腔生物医学, 2021, 12(1): 17-20.

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