Cepeda, Andreia P.

[["dc.bibliographiccitation.artnumber","13"],["dc.bibliographiccitation.firstpage","1"],["dc.bibliographiccitation.journal","Frontiers in Synaptic Neuroscience"],["dc.bibliographiccitation.lastpage","19"],["dc.bibliographiccitation.volume","9"],["dc.contributor.author","Meese, Sandra"],["dc.contributor.author","Cepeda, Andreia P."],["dc.contributor.author","Gahlen, Felix"],["dc.contributor.author","Adams, Christopher M."],["dc.contributor.author","Ficner, Ralf"],["dc.contributor.author","Ricci, Anthony J."],["dc.contributor.author","Heller, Stefan"],["dc.contributor.author","Reisinger, Ellen"],["dc.contributor.author","Herget, Meike"],["dc.date.accessioned","2018-04-23T11:47:09Z"],["dc.date.available","2018-04-23T11:47:09Z"],["dc.date.issued","2017"],["dc.description.abstract","Otoferlin is essential for fast Ca2+-triggered transmitter release from auditory inner hair cells (IHCs), playing key roles in synaptic vesicle release, replenishment and retrieval. Dysfunction of otoferlin results in profound prelingual deafness. Despite its crucial role in cochlear synaptic processes, mechanisms regulating otoferlin activity have not been studied to date. Here, we identified Ca2+/calmodulin-dependent serine/threonine kinase II delta (CaMKIIδ) as an otoferlin binding partner by pull-downs from chicken utricles and reassured interaction by a co-immunoprecipitation with heterologously expressed proteins in HEK cells. We confirmed the expression of CaMKIIδ in rodent IHCs by immunohistochemistry and real-time PCR. A proximity ligation assay indicates close proximity of the two proteins in rat IHCs, suggesting that otoferlin and CaMKIIδ also interact in mammalian IHCs. In vitro phosphorylation of otoferlin by CaMKIIδ revealed ten phosphorylation sites, five of which are located within C2-domains. Exchange of serines/threonines at phosphorylated sites into phosphomimetic aspartates reduces the Ca2+ affinity of the recombinant C2F domain 10-fold, and increases the Ca2+ affinity of the C2C domain. Concordantly, we show that phosphorylation of otoferlin and/or its interaction partners are enhanced upon hair cell depolarization and blocked by pharmacological CaMKII inhibition. We therefore propose that otoferlin activity is regulated by CaMKIIδ in IHCs."],["dc.identifier.doi","10.3389/fnsyn.2017.00013"],["dc.identifier.gro","3142188"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/13307"],["dc.language.iso","en"],["dc.notes.intern","lifescience updates Crossref Import"],["dc.notes.status","final"],["dc.publisher","Frontiers Media S.A."],["dc.relation.eissn","1663-3563"],["dc.relation.issn","1663-3563"],["dc.rights","http://creativecommons.org/licenses/by/4.0/"],["dc.title","Activity-Dependent Phosphorylation by CaMKIIδ Alters the Ca2+ Affinity of the Multi-C2-Domain Protein Otoferlin"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","EMBO Molecular Medicine"],["dc.bibliographiccitation.volume","11"],["dc.contributor.author","Al‐Moyed, Hanan"],["dc.contributor.author","Cepeda, Andreia P."],["dc.contributor.author","Jung, SangYong"],["dc.contributor.author","Moser, Tobias"],["dc.contributor.author","Kügler, Sebastian"],["dc.contributor.author","Reisinger, Ellen"],["dc.date.accessioned","2020-12-10T18:42:39Z"],["dc.date.available","2020-12-10T18:42:39Z"],["dc.date.issued","2018"],["dc.description.abstract","Normal hearing and synaptic transmission at afferent auditory inner hair cell (IHC) synapses require otoferlin. Deafness DFNB9, caused by mutations in the OTOF gene encoding otoferlin, might be treated by transferring wild-type otoferlin cDNA into IHCs, which is difficult due to the large size of this transgene. In this study, we generated two adeno-associated viruses (AAVs), each containing half of the otoferlin cDNA. Co-injecting these dual-AAV2/6 half-vectors into the cochleae of 6- to 7-day-old otoferlin knock-out (Otof−/−) mice led to the expression of full-length otoferlin in up to 50% of IHCs. In the cochlea, otoferlin was selectively expressed in auditory hair cells. Dual-AAV transduction of Otof−/− IHCs fully restored fast exocytosis, while otoferlin-dependent vesicle replenishment reached 35–50% of wild-type levels. The loss of 40% of synaptic ribbons in these IHCs could not be prevented, indicating a role of otoferlin in early synapse maturation. Acoustic clicks evoked auditory brainstem responses with thresholds of 40–60 dB. Therefore, we propose that gene delivery mediated by dual-AAV vectors might be suitable to treat deafness forms caused by mutations in large genes such as OTOF."],["dc.identifier.doi","10.15252/emmm.201809396"],["dc.identifier.pmid","30509897"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/78034"],["dc.identifier.url","https://mbexc.uni-goettingen.de/literature/publications/207"],["dc.language.iso","en"],["dc.notes.intern","DOI Import GROB-354"],["dc.relation","EXC 2067: Multiscale Bioimaging"],["dc.relation.workinggroup","RG Moser (Molecular Anatomy, Physiology and Pathology of Sound Encoding)"],["dc.rights","CC BY 4.0"],["dc.title","A dual‐AAV approach restores fast exocytosis and partially rescues auditory function in deaf otoferlin knock‐out mice"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.subtype","original_ja"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]