口腔医学研究

口腔医学研究 ›› 2026, Vol. 42 ›› Issue (10): 853-859.DOI: 10.13701/j.cnki.kqyxyj.2026.10.005

• 口腔生物学研究 • 上一篇    下一篇

G9a在斑马鱼咽齿形态发生过程中的时空表达模式研究

祖帅1, 吴云海1, 彭佳2, 杨菲2, 刘洋1, 胡红梅2, 李伟3*   

  1. 1.井冈山大学医学部中医药学院 江西 吉安 343000;
    2.井冈山大学医学部护理学院 江西 吉安 343000;
    3.井冈山大学临床医学院口腔系 江西 吉安 343000
  • 收稿日期:2025-09-28 发布日期:2026-10-23
  • 通讯作者: * 李伟,E-mail:532184985@qq.com
  • 作者简介:祖帅(1999~ ),男,安徽宿州人,硕士在读,研究方向:牙齿发育。
  • 基金资助:
    江西省教育厅资助项目(编号:GJJ2201614)

Spatiotemporal Expression Mode of G9a in Development Process of Pharyngeal Tooth Morphology of Zebrafish

ZU Shuai1, WU Yunhai1, PENG Jia2, YANG Fei2, LIU Yang1, HU Hongmei2, LI Wei3*   

  1. 1. School of Traditional Chinese Medicine, Medical College, Jinggangshan University, Ji'an 343000, China;
    2. School of Nursing, Medical Division, Jinggangshan University, Ji'an 343000, China;
    3. Department of Stomatology, Clinical Medical College, Jinggangshan University, Ji'an 343000, China
  • Received:2025-09-28 Published:2026-10-23

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摘要： 目的: 本研究旨在探究组蛋白H3K9甲基转移酶G9a在斑马鱼咽齿从发生到早期形态建成关键阶段的时空表达模式。方法: 以野生型AB系斑马鱼为模型,收集36~84 hpf的胚胎。通过逆转录-聚合酶链反应(reverse transcriptase-polymerase chain reaction,RT-PCR)克隆斑马鱼咽齿特异性标记基因配对样同源域2(paired-like homeodomain 2,pitx2)、远端缺失同源盒基因2b(distal-less homeobox 2b,dlx2b)及G9a基因编码区片段,制备地高辛标记的反义RNA探针。采用整体原位杂交(whole-mount in situ hybridization,WISH)技术系统检测G9a在咽齿发育区域的动态表达,并与pitx2和dlx2b的表达模式进行共定位比较。结果: RT-PCR成功克隆得到目的基因片段。原位杂交结果显示,在36~48 hpf,G9a在咽部区域呈现广泛表达。至56~60 hpf,咽齿牙胚开始特化,pitx2与dlx2b在特定区域呈现强阳性信号,此时G9a的表达区域与pitx2、dlx2b的表达区域高度重叠。在72~84 hpf的形态发生阶段,G9a在持续发育的咽齿原基中保持显著表达。结论: 本研究揭示了G9a在斑马鱼咽齿发育关键时期具有特定的时空表达模式,其表达区域与已知的咽齿标记基因高度一致,提示G9a可能通过表观遗传调控机制参与斑马鱼咽齿的形态发生过程。

关键词: 斑马鱼, 咽齿发育, G9a, 表观遗传调控, 全胚原位杂交

Abstract: Objective: To explore the spatiotemporal expression pattern of G9a in zebrafish pharyngeal teeth during the key stage from occurrence to early morphogenesis. Methods: Using wild-type AB zebrafish as the model, embryos of 36-84 hpf (after fertilization within hours) were collected. The zebrafish pharyngoden-specific marker gene paired-like homeodomain 2 (pitx2), distal-less homeobox 2b (dlx2b), and the coding region fragment of the G9a gene were cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) to prepare digoxin-labeled antisense RNA probes. The dynamic expression of G9a in the pharyngeal tooth development region was systematically detected by whole-embryo in situ hybridization whole-mount in situ hybridization (WISH) technology, and a co-localization comparison was made with the expression patterns of pitx2 and dlx2b. Results: The target gene fragment was successfully cloned by RT-PCR. The in situ hybridization showed that G9a was widely expressed in the pharyngeal region at 36-48 hpf. At 56-60 hpf, the pharyngeal tooth germ begins to specialization, and pitx2 and dlx2b show strong positive signals in specific areas. At this time, the expression area of G9a highly overlaps with that of pitx2 and dlx2b. During the morphogenetic stage of 72-84 hpf, G9a remains significantly expressed in the continuously developing pharyngodont primordium. Conclusion: This study, for the first time, revealed that G9a has a specific spatiotemporal expression pattern during the critical period of pharyngeal tooth development in zebrafish. Its expression region is highly consistent with the known pharyngeal tooth marker genes, suggesting that G9a may be involved in the morphogenesis process of zebrafish pharyngeal teeth through epigenetic regulatory mechanisms.

Key words: zebrafish, pharyngeal development, G9a, epigenetic regulation, whole embryonic in situ hybridization