Adeno-associated viral vector-mediated expression of NT4-ADNF-9 fusion gene protects against aminoglycoside-induced auditory hair cell loss in vitro.
CONCLUSIONS
The present study suggests that adeno-associated viral vector AAV2-mediated expression of activity-dependent neurotrophic factor-9 (ADNF-9) in the cochlea could be a promising approach to protect the cochlea from aminoglycoside-induced impairment, although a further in vivo study is needed.


OBJECTIVE
To construct vectors over-expressing ADNF-9 to overcome its short half-life and investigate the effect of ADNF-9 on aminoglycoside-induced hair cell impairment.


METHODS
We ligated ADNF-9 cDNA to the signal and leader peptides of neurotrophin 4 (NT4) and the fusion gene was named NT4-ADNF-9. NT4-ADNF-9 was subcloned into the prokaryotic expression vector pBV220 to obtain pBV220/NT4-ADNF-9. The induced recombinant ADNF-9 proteins were added into the dorsal root ganglia (DRG) cultures of embryonic day 8 chickens. In addition, we constructed the recombinant vector rAAV-NT4-ADNF-9 and transfected rat neonatal organ of Corti explants in the presence of aminoglycoside G418.


RESULTS
Our data showed that the induced expression of ADNF-9 protein could promote cultured DRG neuronal survival and neurite outgrowth. In addition, transfection of rAAV-NT4-ADNF-9 could prevent hair cell loss induced by G418 treatment in the rat organ of Corti.
Department of Otorhinolaryngology, Second Hospital of Xi'an JiaoTong University, Xi'an, China. zhengguoxi@21cn.com
Guoxi
G
Zheng
Kang
K
Zhu
Junrong
J
Wei
Zhe
Z
Jin
Maoli
M
Duan
D000617:Aminoglycosides / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019677:Artificial Gene Fusion; D018929:Cell Culture Techniques; D002642:Chick Embryo; D000229:Dependovirus; D005727:Ganglia, Spinal / Q000187:drug effects / Q000378:metabolism*; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D006198:Hair Cells, Auditory / Q000187:drug effects / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D014162:Transfection
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D005839:Gentamicins; D009414:Nerve Growth Factors; D009842:Oligopeptides; C410622:activity-dependent neurotrophic factor; C010680:antibiotic G 418; C073922:neurotrophin 4