Effect of lentiviruses carrying enhanced green fluorescent protein injected into the scala media through a cochleostomy in rats

Abstract Purpose The purposes of the current study were to assess the feasibility of post-auricular microinjection of lentiviruses carrying enhanced green fluorescent protein (EGFP) into the scala media through cochleostomies in rats, determine the expression of viral gene in the cochlea, and record...

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Bibliographic Details
Published in:American journal of otolaryngology 2013-07, Vol.34 (4), p.301-307
Main Authors: Wei, Yan, MD, Fu, Yong, MD, Liu, Shaosheng, MM, Xia, GuiHua, MB, Pan, Song, MD
Format: Article
Language:English
Subjects:
Animals
Cell Survival
Cochlea - surgery
Cochlear Duct - surgery
Cochlear Duct - virology
Disease Models, Animal
Ears & hearing
Efficiency
Electrodes
Evoked Potentials, Auditory, Brain Stem - genetics
Evoked Potentials, Auditory, Brain Stem - physiology
Gene Expression Regulation, Viral
Gene therapy
Genetic Therapy - methods
Green Fluorescent Proteins - pharmacology
Hair Cells, Auditory - physiology
Hearing loss
Injections, Intralesional
Lentivirus
Lentivirus - genetics
Male
Mammals
Otolaryngology
Random Allocation
Rats
Rats, Sprague-Dawley
Rodents
Sensitivity and Specificity
Surgery
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Summary:Abstract Purpose The purposes of the current study were to assess the feasibility of post-auricular microinjection of lentiviruses carrying enhanced green fluorescent protein (EGFP) into the scala media through cochleostomies in rats, determine the expression of viral gene in the cochlea, and record the post-operative changes in the number and auditory function of cochlear hair cells (HCs). Methods Healthy rats were randomly divided into two groups. The left ears of the animals in group I were injected with lentivirus carrying EGFP (n = 10) via scala media lateral wall cochleostomies, and the left ears of the animals in group II were similarly injected with artificial endolymph (n = 10). Prior to and 30 days post-injection, auditory function was assessed with click-auditory brainstem response (ABR) testing, EGFP expression was determined with cochlear frozen sections under fluorescence microscopy, and survival of HCs was estimated based on whole mount preparations. Results Thirty days after surgery, click-ABR testing revealed that there were significant differences in the auditory function, EGFP expression, and survival of HCs in the left ears before and after surgery in the same rats from each group. In group I, EGFP was noted in the strial marginal cells of the scala media, the organ of Corti, spiral nerves, and spiral ganglion cells. Conclusion Lentiviruses were successfully introduced into the scala media through cochleostomies in rats, and the EGFP reporter gene was efficiently expressed in the organ of Corti, spiral nerves, and spiral ganglion cells.
ISSN:0196-0709
1532-818X
DOI:10.1016/j.amjoto.2012.12.011