利用启动子控制的绿色荧光蛋白标记鸡胚听神经

庸安露 王晓钰

doi:10.16098/j.issn.0529-1356.2025.04.003

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解剖学报 ›› 2025, Vol. 56 ›› Issue (4) : 398-403. DOI: 10.16098/j.issn.0529-1356.2025.04.003

神经生物学

利用启动子控制的绿色荧光蛋白标记鸡胚听神经

庸安露王晓钰^*

庸安露王晓钰

作者信息 +

Labeling chicken embryo auditory nerves using promoter-controlled green fluorescent protein

YONG An-lu WANG Xiao-yu^*

Author information +

文章历史 +

摘要

目的利用启动子控制的膜结合绿色荧光蛋白（mGFP）标记鸡胚听神经（AN），以提供更高分辨率的听神经发育图谱。方法在鸡胚2.5 d（E2.5）时将神经源性分化因子1（NeuroD1)和GATA结合蛋白3（GATA3) 启动子控制的mGFP质粒显微注射入耳泡中，并通过鸡胚内电转染将质粒导入听觉神经节前体细胞中。在听神经发育的不同时期追踪听神经轴突到脑干耳蜗核的投射，并对两种启动子标记的特异性进行比较。结果由NeuroD1或GATA3启动子控制的mGFP均可选择性标记听神经，mGFP标记的听神经均投射到听觉脑干的耳蜗核中。GATA3-mGFP可较特异地标记早期（E9~E10）投射的听神经而极少标记前庭神经。而NeuroD1-mGFP可标记至晚期（E15），且显示听神经末梢突触形成，同时，NeuroD1-mGFP也能标记前庭神经并显示投射至脑干前庭核团中。结论利用启动子控制的mGFP标记听神经，可提供更高分辨率的听神经发育图谱。

Abstract

Objective To utilize promoter-controlled membrane-bound (myristoylated) green fluorescent protein (mGFP) to label the auditory nerve (AN) in chicken embryos, providing a higher-resolution map of AN development. Methods At chicken embryonic 2.5 day (E2.5), mGFP plasmids controlled by the neurogenic differentiation factor 1 (NeuroD1) or GATA binding protein 3 (GATA3) promoters were microinjected and electroporated into the epithelium of otocyst in chicken embryos. The projection of AN axons to the cochlear nucleus in the brainstem was tracked at different developmental stages, and synapse formation was observed, and the labeling specificity of the two promoters was compared. Results Both NeuroD1 and GATA3 promoters controlled mGFP selectively labeled the AN, with mGFP-labeled AN axons projecting to the cochlear nucleus in the auditory brainstem. GATA3-mGFP specifically labeled early (E9-E10) projecting AN with significantly fewer labeling in vestibular nerves. NeuroD1-mGFP labeled auditory nerves throughout the development, and showed synaptic formation at E15. A additionally, NeuroD1-mGFP also labeled vestibular nerves projecting to the vestibular nuclei in the brainstem. Conclusion The use of promoter-controlled mGFP to label the AN provides a higher-resolution map of AN development.

导出引用

庸安露王晓钰. 利用启动子控制的绿色荧光蛋白标记鸡胚听神经[J]. 解剖学报. 2025, 56(4): 398-403 https://doi.org/10.16098/j.issn.0529-1356.2025.04.003

YONG An-lu WANG Xiao-yu. Labeling chicken embryo auditory nerves using promoter-controlled green fluorescent protein[J]. Acta Anatomica Sinica. 2025, 56(4): 398-403 https://doi.org/10.16098/j.issn.0529-1356.2025.04.003

中图分类号： R322

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