Individual synaptic vesicles mediate stimulated exocytosis from cochlear inner hair cells

Significance

Synaptic transmission is codetermined by presynaptic and postsynaptic neurons. Therefore, to understand how the inner hair cell (IHC) signals to spiral ganglion neurons at the first synapse in the auditory pathway, here we directly studied individual membrane fusion events by making cell-attached membrane capacitance recordings from IHCs, for which the quantal size is debated. The observed fusion steps in membrane capacitance are consistent with the quantal hypothesis of synaptic transmission in which individual synaptic vesicles undergo exocytosis independently from each other. This finding, in conjunction with previous work, raises the exciting possibility that action potential generation can be triggered by the release of a single vesicle at the IHC synapse.

Abstract

Spontaneous excitatory postsynaptic currents (sEPSCs) measured from the first synapse in the mammalian auditory pathway reach a large mean amplitude with a high level of variance (CV between 0.3 and 1). This has led some to propose that each inner hair cell (IHC) ribbon-type active zone (AZ), on average, releases ∼6 synaptic vesicles (SVs) per sEPSC in a coordinated manner. If true, then the predicted change in membrane capacitance (C _m) for such multivesicular fusion events would equate to ∼300 attofarads (aF). Here, we performed cell-attached C _m measurements to directly examine the size of fusion events at the basolateral membrane of IHCs where the AZs are located. The frequency of events depended on the membrane potential and the expression of Ca _v1.3, the principal Ca ²⁺-channel type of IHCs. Fusion events averaged 40 aF, which equates to a normal-sized SV with an estimated diameter of 37 nm. The calculated SV volumes showed a high degree of variance (CV > 0.6). These results indicate that SVs fused individually with the plasma membrane during spontaneous and evoked release and SV volume may contribute more variability in EPSC amplitude than previously assumed.