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

Abstract

Key points This study characterizes the mechanisms underlying defects in synaptic transmission when dynamin‐related protein 1 (DRP1) is genetically eliminated. Viral‐mediated knockout of DRP1 from the presynaptic terminal at the mouse calyx of Held increased initial release probability, reduced the size of the synaptic vesicle recycling pool and impaired synaptic vesicle recycling. Transmission defects could be partially restored by increasing the intracellular calcium buffering capacity with EGTA‐AM, implying close coupling of Ca2+ channels to synaptic vesicles was compromised. Acute restoration of ATP to physiological levels in the presynaptic terminal did not reverse the synaptic defects. Loss of DRP1 impairs mitochondrial morphology in the presynaptic terminal, which in turn seems to arrest synaptic maturation. 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 Ca2+ 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 Ca2+ 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.

Neuronal Glutamatergic Synaptic Clefts Alkalinize Rather Than Acidify during Neurotransmission

“…Targeted viral infection. CNS injection of neonatal mice followed protocols described previously (Singh et al, 2018). Briefly, 1-d-old (P1) mice were chilled on ice for Ͼ5 min until unresponsive, then fixed on a stereotaxic surgery platform.…”

NR4A1 promotes TNF‑α‑induced chondrocyte death and migration injury via activating the�AMPK/Drp1/mitochondrial fission pathway

“…Mitochondrial dysfunction is implicated in the pathological progression of OA via a number of pathways, including oxidative stress, cartilage matrix synthesis and degradation disorder, cytokine mediated inflammatory response activation and chondrocyte death (10,11). Recent studies have highlighted an important role of dynamin-related protein 1 (Drp1)-mediated mitochondrial fission in mitochondrial homeostasis (12,13). Mechanistically, excessive mitochondrial fission contributes to mitochondrial dysfunction as indicated by loss of mitochondrial DNA integrity, reduction in ATP generation, mitochondrial ROS outburst, and loss of mitochondrial membrane potential (13).…”

“…Recent studies have highlighted an important role of dynamin-related protein 1 (Drp1)-mediated mitochondrial fission in mitochondrial homeostasis (12,13). Mechanistically, excessive mitochondrial fission contributes to mitochondrial dysfunction as indicated by loss of mitochondrial DNA integrity, reduction in ATP generation, mitochondrial ROS outburst, and loss of mitochondrial membrane potential (13). Furthermore, excessive fission triggers mitochondrial permeability transition pore (mPTP) opening via regulating voltage-dependent anion channel 1 (VDAC1) and hexokinase 2 (HK2), which plays a critical role in cell death (14).…”

Fenpropathrin induces degeneration of dopaminergic neurons via disruption of the mitochondrial quality control system

“…A previous report indicated that Drp1-dependent division is required for efficient mitochondrial transport and for maintaining synaptic function 31 – 33 . Hence, Drp1 deficiency and reduced ATP production may decrease the capability of mitochondrial movement toward synaptic terminals and lead to mitochondrial depletion in axon terminals as subsequent neurodegeneration 32 , 34 , 35 . Given that we found decreased Drp1 expression in Fen-treated primary neurons, we next investigated mitochondrial morphology and mass in neurites.…”