Cochlear expression of a dominant-negative GJB2R75W construct delivered through the round window membrane in mice

Yukihide Maeda; Kunihiro Fukushima; Akihiro Kawasaki; Kazunori Nishizaki; Richard J H Smith

doi:10.1016/j.neures.2007.03.006

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Cochlear expression of a dominant-negative GJB2R75W construct delivered through the round window membrane in mice

Journal article

Peer reviewed

Cochlear expression of a dominant-negative GJB2R75W construct delivered through the round window membrane in mice

Yukihide Maeda, Kunihiro Fukushima, Akihiro Kawasaki, Kazunori Nishizaki and Richard J H Smith

Neuroscience research, Vol.58(3), pp.250-254

07/2007

DOI: 10.1016/j.neures.2007.03.006

PMID: 17462767

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Abstract

Development of a gene-delivery method to the inner ear is an essential step for studies of hearing function and gene therapy. Application of liposomes or adenoviral vectors onto the intact round window membrane (RWM) offers the possibility of atraumatic exogenous gene transfer. GJB2 encodes the gap junction protein Connexin26, which plays a crucial role in potassium recycling in the inner ear. The R75W allele of GJB is a well-characterized mutation that causes deafness at the DFNA3 through a dominant-negative mechanism of action. In this study, a plasmid vector, pGJB2(R75W)-eGFP, was lipocomplexed with N-[1-(2,3-Dioleoloxy)propyl]N,N,N-trimethylammonium methylsulfate: cholesterol and applied onto mouse RWM. At 3 days (3d) post-treatment, immunohistochemistry demonstrated GJB2(R75W)-eGFP transgene expression in the cochlea in: inner and outer pillar cells, outer hair cells, Claudius cells and, in the spiral limbus and ligament. Significant hearing loss was detected by auditory brainstem response testing after 1, 2 and 3d post-treatment; hearing levels returned to control levels at 5d post-treatment. These data confirm that GJB2(R75W) induces functional impairment in the mature cochlea through a dominant negative effect, and importantly, that RWM application of exogenous genes is a feasible method to test their impact on hearing.

Gene Transfer Techniques

Green Fluorescent Proteins - metabolism

Gene Expression

Round Window, Ear - metabolism

Hearing Loss - etiology

Mice, Inbred C57BL

Connexins - genetics

Green Fluorescent Proteins - genetics

Hearing Loss - pathology

Cholesterol

Connexins - metabolism

Feasibility Studies

Connexin 26

Hearing Loss - genetics

Hearing Loss - metabolism

Animals

Cochlea - metabolism

Statistics, Nonparametric

Auditory Threshold - physiology

Female

Liposomes - metabolism

Mice

Evoked Potentials, Auditory, Brain Stem - physiology

Details

Title: Subtitle: Cochlear expression of a dominant-negative GJB2R75W construct delivered through the round window membrane in mice
Creators: Yukihide Maeda - Department of Otolaryngology-Head and Neck Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical science, Okayama 700-8558, Japan
Kunihiro Fukushima
Akihiro Kawasaki
Kazunori Nishizaki
Richard J H Smith
Resource Type: Journal article
Publication Details: Neuroscience research, Vol.58(3), pp.250-254
DOI: 10.1016/j.neures.2007.03.006
PMID: 17462767
NLM abbreviation: Neurosci Res
ISSN: 0168-0102
eISSN: 1872-8111
Publisher: Ireland
Grant note: DC03544 / NIDCD NIH HHS
Language: English
Date published: 07/2007
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Molecular Physiology and Biophysics; Anatomy and Cell Biology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Otolaryngology; Internal Medicine
Record Identifier: 9984007296202771

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