Developmental Acquisition of a Rapid Calcium-Regulated Vesicle Supply Allows Sustained High Rates of Exocytosis in Auditory Hair Cells 英文参考文献.docVIP

DevelopmentalAcquisitionofaRapid Calcium-RegulatedVesicleSupplyAllowsSustainedHigh RatesofExocytosisinAuditoryHairCells SnezanaLevic*,YohanBouleau,DidierDulon* Equipe Neurophysiologie de la Synapse Auditive, Unite′ Mixte de Recherche, Inserm U587 et Universite′ Victor Segalen, Institut des Neurosciences de Bordeaux, Centre Hospitalier Universitaire Pellegrin, Bordeaux, France Abstract Auditory hair cells (HCs) have the remarkable property to indefinitely sustain high rates of synaptic vesicle release during ongoing sound stimulation. The mechanisms of vesicle supply that allow such indefatigable exocytosis at the ribbon active zone remain largely unknown. To address this issue, we characterized the kinetics of vesicle recruitment and release in developing chick auditory HCs. Experiments were done using the intact chick basilar papilla from E10 (embryonic day 10) to P2 (two days post-hatch) by monitoring changes in membrane capacitance and Ca2+ currents during various voltage stimulations. Compared to immature pre-hearing HCs (E10-E12), mature post-hearing HCs (E18-P2) can steadily mobilize a larger readily releasable pool (RRP) of vesicles with faster kinetics and higher Ca2+ efficiency. As assessed by varying the inter-pulse interval of a 100 ms paired-pulse depolarization protocol, the kinetics of RRP replenishment were found much faster in mature HCs. Unlike mature HCs, exocytosis in immature HCs showed large depression during repetitive stimulations. Remarkably, when the intracellular concentration of EGTA was raised from 0.5 to 2 mM, the paired-pulse 2+ depression level remained unchanged in immature HCs but was drastically increased in mature HCs, indicating that the Ca sensitivity of the vesicle replenishment process increases during maturation. Concomitantly, the immunoreactivity of the calcium sensor otoferli