Journal of Otolaryngology and Ophthalmology of Shandong University ›› 2023, Vol. 37 ›› Issue (3): 1-6.doi: 10.6040/j.issn.1673-3770.0.2022.129

• 论著 •    

Effects of a neuron specific AAV2 viral vector on the imaging efficiency of primary cultured spiral ganglion neurons in vitro

LIU Yaoqian1, ZHANG Zhen1,2,3, ZHANG Boya1,2,3, CHEN Zhengnong1,2,3   

  1. 1. Otolaryngology Research Institute, Shanghai Jiaotong University School of Medicine, Shanghai 200233, China;
    2. Department of Otorhinolaryngology, Affiliated Sixth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200233, China;
    3. Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai 200233, China
  • Published:2023-05-24

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Abstract: Objective To explore the effects of AAV2/Anc80L65-hSyn-eGFP virus containing a neuron-specific human synapsin promoter(hSyn)and enhanced green fluorescent protein(eGFP)on the imaging of spiral ganglion neuron(SGN)morphologies in vitro. Methods Dissociated SGNs were transfected with AAV2/Anc80L65-hSyn-eGFP in vitro. Using fluorescence microscopy, the transfection specificity and efficiency of the virus were evaluated by calculating the proportion of fluorescent neurons as observed under low magnification. Elaborate morphologies of single neuron were then observed under high magnification. The imaging application of AAV2/Anc80L65-hSyn-eGFP on living SGNs was further verified by patch clamp studies. Results The AAV2/Anc80L65-hSyn-eGFP virus had both high transfection specificity and efficiency for SGNs in vitro, and did not produce obvious cytotoxicity. Importantly, the virus could reveal the morphology of somas and neurites both rapidly and clearly without causing damage to their structures. Compared with neuron-specific Tuj1 fluorescent staining, AAV2/Anc80L65-hSyn-eGFP was able to mark delicate neurite structures such as filopodia. When utilized for the identification and localization of SGNs in patch clamp studies, AAV2/Anc80L65-hSyn-eGFP significantly promoted the efficiency and accuracy of the experiments. Conclusion AAV2/Anc80L65-hSyn-eGFP is a competent and ideal tool for highlighting the morphology of living SGNs in vitro.

Key words: AAV2/Anc80L65-hSyn-eGFP virus, Spiral ganglion neuron morphology, Primary culture, Imaging effect

CLC Number: 

  • R764.43+1
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