Presynaptic loss of dynamin-related protein 1 impairs synaptic vesicle release and recycling at the mouse calyx of held

Abstract

Impaired mitochondrial biogenesis and function is implicated in many neurodegenerative diseases, and likely affects synaptic neurotransmission prior to cellular loss. Dynamin-related protein 1 (DRP1) is essential for mitochondrial fission and is disrupted in neurodegenerative disease. In this study, we used the mouse calyx of Held synapse as a model to investigate the impact of presynaptic DRP1 loss on synaptic vesicle (SV) recycling and sustained neurotransmission. In vivo viral expression of Cre recombinase in ventral cochlear neurons of floxed-DRP1 mice generated a presynaptic-specific DRP1 knockout (DRP1-preKO), where the innervated postsynaptic cell was unperturbed. Confocal reconstruction of the calyx terminal suggested SV clusters and mitochondrial content were disrupted, and presynaptic terminal volume was decreased. Using postsynaptic voltage-clamp recordings, we found that DRP1-preKO synapses had larger evoked responses at low frequency stimulation. DRP1-preKO synapses also had profoundly altered short-term plasticity, due to defects in SV recycling. Readily releasable pool size, estimated with high-frequency trains, was dramatically reduced in DRP1-preKO synapses, suggesting an important role for DRP1 in maintenance of release-competent SVs at the presynaptic terminal. Presynaptic Ca 2+ accumulation in the terminal was also enhanced in DRP1-preKO synapses. Synaptic transmission defects could be partially rescued with EGTA-AM, indicating close coupling of Ca 2+ channels to SV distance normally found in mature terminals may be compromised by DRP1-preKO. Using paired recordings of the presynaptic and postsynaptic compartments, recycling defects could not be reversed by acute dialysis of ATP into the calyx terminals. Taken together, our results implicate a requirement for mitochondrial fission to coordinate postnatal synapse maturation.