title: BAAV mediated GJB2 gene transfer restores gap junction coupling in cochlear organotypic cultures from deaf Cx26Sox10Cre mice.
abstract: The deafness locus DFNB1 contains GJB2, the gene encoding connexin26 and GJB6, encoding connexin30, which appear to be coordinately regulated in the inner ear. In this work, we investigated the expression and function of connexin26 and connexin30 from postnatal day 5 to adult age in double transgenic Cx26(Sox10Cre) mice, which we obtained by crossing connexin26 floxed mice with a deleter Sox10-Cre line. Cx26(Sox10Cre) mice presented with complete connexin26 ablation in the epithelial gap junction network of the cochlea, whereas connexin30 expression was developmentally delayed; immunolabeling patterns for both connexins were normal in the cochlear lateral wall. In vivo electrophysiological measurements in Cx26(Sox10Cre) mice revealed profound hearing loss accompanied by reduction of endocochlear potential, and functional experiments performed in postnatal cochlear organotypic cultures showed impaired gap junction coupling. Transduction of these cultures with a bovine adeno associated virus vector restored connexin26 protein expression and rescued gap junction coupling. These results suggest that restoration of normal connexin levels by gene delivery via recombinant adeno associated virus could be a way to rescue hearing function in DFNB1 mouse models and, in future, lead to the development of therapeutic interventions in humans.
authors.affiliation: Fondazione per la Ricerca Biomedica Avanzata, Istituto Veneto di Medicina Molecolare, Padova, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Crispino
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Pasquale
authors.forename: Pietro
authors.initials: P
authors.lastname: Scimemi
authors.forename: Laura
authors.initials: L
authors.lastname: Rodriguez
authors.forename: Fabian
authors.initials: F
authors.lastname: Galindo Ramirez
authors.forename: Romolo Daniele
authors.initials: RD
authors.lastname: De Siati
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Santarelli
authors.forename: Edoardo
authors.initials: E
authors.lastname: Arslan
authors.forename: Mario
authors.initials: M
authors.lastname: Bortolozzi
authors.forename: John A
authors.initials: JA
authors.lastname: Chiorini
authors.forename: Fabio
authors.initials: F
authors.lastname: Mammano
mesh_terms: D000818:Animals; D002417:Cattle; D003051:Cochlea / Q000473:pathology* / Q000503:physiopathology; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics* / Q000627:therapeutic use*; D003638:Deafness / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D000229:Dependovirus / Q000378:metabolism*; D055724:Electrophysiological Phenomena; D036681:Fluorescence Recovery After Photobleaching; D017629:Gap Junctions / Q000378:metabolism*; D015316:Genetic Therapy; D006198:Hair Cells, Auditory / Q000378:metabolism / Q000473:pathology; D019426:Integrases / Q000378:metabolism; D051379:Mice; D009924:Organ Culture Techniques; D009925:Organ of Corti / Q000473:pathology; D010539:Permeability; D011993:Recombinant Fusion Proteins; D055757:SOXE Transcription Factors / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic
chemical_list: D017630:Connexins; C000607042:GJB2 protein, human; C000607043:Gjb2 protein, mouse; D011993:Recombinant Fusion Proteins; D055757:SOXE Transcription Factors; C527924:Sox10 protein, mouse; D000072259:Connexin 26; C045073:Cre recombinase; D019426:Integrases