Inter-spike mitochondrial Ca²⁺ release enhances high frequency synaptic transmission

Presynaptic mitochondria not only absorb but also release Ca²⁺ during high frequency stimulation (HFS) when presynaptic [Ca²⁺ ] is kept low (<500 nm) by high cytosolic Ca²⁺ buffer or strong plasma membrane calcium clearance mechanisms under physiological external [Ca²⁺ ]. Mitochondrial Ca²⁺ release (MCR) does not alter the global presynaptic Ca²⁺ transients. MCR during HFS enhances short-term facilitation and steady state excitatory postsynaptic currents by increasing vesicular release probability. The intra-train MCR may provide residual calcium at interspike intervals, and thus support high frequency neurotransmission at central glutamatergic synapses.

Emerging evidence indicates that mitochondrial Ca²⁺ buffering contributes to local regulation of synaptic transmission. It is unknown, however, whether mitochondrial Ca²⁺ release (MCR) occurs during high frequency synaptic transmission. Confirming the previous notion that 2 μm tetraphenylphosphonium (TPP⁺ ) is a specific inhibitor of the mitochondrial Na⁺ /Ca²⁺ exchanger (mNCX), we studied the role of MCR via mNCX in short-term plasticity during high frequency stimulation (HFS) at the calyx of Held synapse of the rat. TPP⁺ reduced short-term facilitation (STF) and steady state excitatory postsynaptic currents during HFS at mature calyx synapses under physiological extracellular [Ca²⁺ ] ([Ca²⁺ ]_o  = 1.2 mm), but not at immature calyx or at 2 mm [Ca²⁺ ]_o . The inhibitory effects of TPP⁺ were stronger at synapses with morphologically complex calyces harbouring many swellings and at 32°C than at simple calyx synapses and at room temperature. These effects of TPP⁺ on STF were well correlated with those on the presynaptic mitochondrial [Ca²⁺ ] build-up during HFS. Mitochondrial [Ca²⁺ ] during HFS was increased by TPP⁺ at mature calyces under 1.2 mm [Ca²⁺ ]_o , and further enhanced at 32°C, but not under 2 mm [Ca²⁺ ]_o or at immature calyces. The close correlation of the effects of TPP⁺ on mitochondrial [Ca²⁺ ] with those on STF suggests that mNCX contributes to STF at the calyx of Held synapses. The intra-train MCR enhanced vesicular release probability without altering global presynaptic [Ca²⁺ ]. Our results suggest that MCR during HFS elevates local [Ca²⁺ ] near synaptic sites at interspike intervals to enhance STF and to support stable synaptic transmission under physiological [Ca²⁺ ]_o .