Neurotransmitter release depends critically on close spatial coupling of Ca(2+) entry to synaptic vesicles at the nerve terminal; however, the molecular substrates determining their physical proximity are unknown. Using the calyx of Held synapse, where "microdomain" coupling predominates at immature stages and developmentally switches to "nanodomain" coupling, we demonstrate that deletion of the filamentous protein Septin 5 imparts immature synapses with striking morphological and functional features reminiscent of mature synapses. This includes synaptic vesicles tightly localized to active zones, resistance to the slow Ca(2+) buffer EGTA and a reduced number of Ca(2+) channels required to trigger single fusion events. Disrupting Septin 5 organization acutely transforms microdomain to nanodomain coupling and potentiates quantal output in immature wild-type terminals. These observations suggest that Septin 5 is a core molecular substrate that differentiates distinct release modalities at the central synapse.

Non-technical summary Localization of sound sources in the azimuth, which makes use of interaural differences in timing and/or intensity of acoustic signals, is of vital importance for most mammals. Using the small differences in time of arrival and/or intensity at the two ears requires that propagation of electric pulses in the auditory system be temporally precise. In this study, we found that elimination of GluA4, a protein particularly abundant in auditory cells, significantly impairs their ability to faithfully transmit electric signals, leading to profound deficits in auditory responses to sound stimuli in mice. Therefore, we conclude that GluA4 is indispensable for enabling information flow with high fidelity in the auditory circuitry. Our work has identified GluA4 as a potential molecular candidate involved in human hearing deficits and disorders.Abstract Fast excitatory synaptic transmission in central synapses is mediated primarily by AMPA receptors (AMPARs), which are heteromeric assemblies of four subunits, GluA1-4. Among these subunits, rapidly gating GluA3/4 appears to be the most abundantly expressed to enable neurotransmission with submillisecond precision at fast rates in subsets of central synapses. However, neither definitive identification of the molecular substrate for native AMPARs in these neurons, nor their hypothesized functional roles in vivo has been unequivocally demonstrated, largely due to lack of specific antagonists. Using GluA3 or GluA4 knockout (KO) mice, we investigated these issues at the calyx of Held synapse, which is known as a high-fidelity synapse involved in sound localization. Patch-clamp recordings from postsynaptic neurons showed that deletion of GluA4 significantly slowed the time course of both evoked and miniature AMPAR-mediated excitatory postsynaptic currents (AMPAR-EPSCs), reduced their amplitude, and exacerbated AMPAR desensitization and short-term depression (STD). Surprisingly, presynaptic release probability was also elevated, contributing to severe STD at GluA4-KO synapses. In contrast, only marginal changes in AMPAR-EPSCs were found in GluA3-KO mice. Furthermore, independent of changes in intrinsic excitability of postsynaptic neurons, deletion of GluA4 markedly reduced synaptic drive and increased action potential failures during high-frequency activity, leading to profound deficits in specific components of the auditory brainstem responses associated with synchronized spiking in the calyx of Held synapse and other related neurons in vivo. These observations identify GluA4 as the main determinant for fast synaptic response, indispensable for driving high-fidelity neurotransmission and conveying precise temporal information.

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It has been suggested that synchronization of action potentials encodes diverse features of a single image. However, properties of the synchronization, which occurs on a time scale of approximately 1-5 ms, are still poorly understood. We have tested the modulation of synchronization by manipulating the contextual targets introduced in the surround of the receptive field. Experiments were carried out on anaesthetized cats prepared for multiunit and single-cell recordings in area 17. Initially, a patch of sine-wave drifting grating was positioned over the overlapping receptive fields of several neurons. If this coherent motion produced a significant synchronization in cross-correlograms, contextual targets were added. The first contextual stimuli were two sine-wave patches placed above and below the central compound receptive field. Only the contrast of contextual targets changed. Results show that the larger the differential contrast the higher the synchronization. The second contextual stimulus was a lateral shift of a sine-wave patch. Data show that the wider the distance between the central and peripheral patches the better the synchronization. Furthermore, results suggest that the synchrony pattern computed by cross correlating multiunit recordings from two sites differs when the cross correlation is carried out between individual units belonging to each multiunit recording. Together with our previous results it appears that synchronization is stimulus dependent and its strength increases with larger disparities included in the whole stimulating image.