The study of AAV9 expression in cochleae of mice at different ages
10.3760/cma.j.issn.1673-0860.2019.10.012
- VernacularTitle: 腺相关病毒在不同年龄小鼠耳蜗中表达的研究
- Author:
Bei CHEN
1
;
Wenxue TANG
2
;
Jianjun WANG
3
;
Xi LIN
3
Author Information
1. Department of Otology, Otolaryngological Hospital, the First Affiliated Hospitanl of Zhengzhou University, Zhengzhou 450052, China
2. Center for Precision Medicine of Zhengzhou University, Zhengzhou 450052, China
3. Departments of Otolaryngology and Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA
- Publication Type:Journal Article
- Keywords:
Gene therapy;
Adeno-associated virus;
Cochlea;
Mice, inbred C57BL
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2019;54(10):769-775
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To determine the extent of transfection and expression of adeno-associated virus (AAV) serotype 9 (AAV9) in the cochleae of mice at different ages.
Methods:AAV9-green fluorescent protein (GFP) was inoculated into the cochlea of mice via the round window membrane (RWM) or through cochleostomy at different ages. Four groups were divided according to ages and injection sites: P1SM group, AAV9-GFP was delivered to the scala media by cochleostomy at postnatal day 1; P1RW group, AAV9-GFP was delivered to the scala tympani via RWM at postnatal day 1; P9RW group: AAV9-GFP was injected through RWM at postnatal day 9; and P30RW group, adult mice (P30) were injected through RWM. GFP expression in cochlear whole mount was analyzed and auditory brainstem response (ABR) tests were conducted one month after virus injection (for each animal, only left cochlea was injected and the right side was used as a control). GraphPad Prism 5 statistical software was used for data analysis.
Results:All of inner hair cells (IHCs) and most of outer hair cells (OHCs) were transfected via two approaches at P1 injection. There was no significant difference in ABR threshold between injected ears and untreated ears (P>0.05). All of the IHCs and parts of OHCs (69% in apical turn) were transfected via RWM at P9. The strongest GFP expression was observed near the apical turn. Cochlear inoculation via RWM at P30 led to transgene expression in only IHCs. The ABR threshold of injected ears in P9RW group and P30RW group was significantly higher than that of contralateral ears (P<0.01).
Conclusions:AAV9 can be highly expressed in the inner and outer hair cells of the cochlea and hearing sensitivity can be preserved if virus injections are performed in neonatal mice. After AAV9 is transfected into the inner ear of adult mice, it is only expressed in the inner hair cells, which leads to the increase of the ABR response threshold of mice. Transfection efficiency is significant higher in neonate mice than in P9 and adult mice.
