Ca²⁺-permeable AMPARs mediate glutamatergic transmission and excitotoxic damage at the hair cell ribbon synapse

We report functional and structural evidence for GluA2-lacking Ca²⁺-permeable AMPARs (CP-AMPARs) at the mature hair cell ribbon synapse. By using the methodological advantages of three species (of either sex), we demonstrate that CP-AMPARs are present at the hair cell synapse in an evolutionarily conserved manner. Via a combination of in vivo electrophysiological and Ca²⁺ imaging approaches in the larval zebrafish, we show that hair cell stimulation leads to robust Ca²⁺ influx into afferent terminals. Prolonged application of AMPA caused loss of afferent terminal responsiveness, whereas blocking CP-AMPARs protects terminals from excitotoxic swelling. Immuno his to chemical analysis of AMPAR subunits in mature rat cochlea show regions within synapses lacking the GluA2 subunit. Paired recordings from adult bullfrog auditory synapses demonstrate that CP-AMPARs mediate a major component of glutamatergic transmission. Together, our results support the importance of CP-AMPARs in mediating transmission at the hair cell ribbon synapse. Further, excess Ca²⁺ entry via CP-AMPARs may underlie afferent terminal damage following excitotoxic challenge, suggesting that limiting Ca²⁺ levels in the afferent terminal may protect against cochlear synaptopathy associated with hearing loss.