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

doi:10.16098/j.issn.0529-1356.2025.04.003

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Acta Anatomica Sinica ›› 2025, Vol. 56 ›› Issue (4) : 398-403. DOI: 10.16098/j.issn.0529-1356.2025.04.003

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

YONG An-lu WANG Xiao-yu^*

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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.

Key words

Auditory nerve / Neurogenic differentiation factor 1 / GATA binding protein 3 / Synaptogenesis / Immunofluorescence / Chicken embryo

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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

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