title: Ribeye is required for presynaptic Ca(V)1.3a channel localization and afferent innervation of sensory hair cells.
abstract: Ribbon synapses of the ear, eye and pineal gland contain a unique protein component: Ribeye. Ribeye consists of a novel aggregation domain spliced to the transcription factor CtBP2 and is one of the most abundant proteins in synaptic ribbon bodies. Although the importance of Ribeye for the function and physical integrity of ribbon synapses has been shown, a specific role in synaptogenesis has not been described. Here, we have modulated Ribeye expression in zebrafish hair cells and have examined the role of Ribeye in synapse development. Knockdown of ribeye resulted in fewer stimulus-evoked action potentials from afferent neurons and loss of presynaptic Ca(V)1.3a calcium channel clusters in hair cells. Additionally, afferent innervation of hair cells was reduced in ribeye morphants, and the reduction was correlated with depletion of Ribeye punctae. By contrast, transgenic overexpression of Ribeye resulted in Ca(V)1.3a channels colocalized with ectopic aggregates of Ribeye protein. Overexpression of Ribeye, however, was not sufficient to create ectopic synapses. These findings reveal two distinct functions of Ribeye in ribbon synapse formation--clustering Ca(V)1.3a channels at the presynapse and stabilizing contacts with afferent neurons--and suggest that Ribeye plays an organizing role in synaptogenesis.
authors.affiliation: Howard Hughes Medical Institute, Oregon Hearing Research Center, 3181 SW Sam Jackson Park Road, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Lavinia
authors.initials: L
authors.lastname: Sheets
authors.forename: Josef G
authors.initials: JG
authors.lastname: Trapani
authors.forename: Weike
authors.initials: W
authors.lastname: Mo
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Obholzer
authors.forename: Teresa
authors.initials: T
authors.lastname: Nicolson
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000818:Animals; D030801:Animals, Genetically Modified; D020746:Calcium Channels, L-Type / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004594:Electrophysiology; D006198:Hair Cells, Auditory / Q000378:metabolism*; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D020746:Calcium Channels, L-Type; D004268:DNA-Binding Proteins; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D029961:Zebrafish Proteins; C488619:cacna1da protein, zebrafish