Impact of glycine transporter 1 knockout on inhibitory neurotransmission in the lateral superior olive
- Glycine constitutes the major neurotransmitter at inhibitory synapses of lower brain regions.
A rapid removal of glycine from the synaptic cleft and consequent recycling is crucial for
synaptic transmission in systems with high effort on temporal precision. This is mainly
achieved by glycine translocation via two glycine transporters (GlyTs), namely GlyT1 and
GlyT2. At inhibitory synapses, GlyT2 was found to be specifically expressed by neurons,
supplying the presynapse with glycine needed for vesicle filling. In contrast, GlyT1 is attributed
to astrocytes and primarily mediates the termination of synaptic transmission by glycine
removal from the synaptic cleft. Employing patch-clamp recordings from principal neurons of
the lateral superior olive (LSO) in acute brainstem slices of GlyT1b/c knockout (KO) mice and
wildtype (WT) littermates at postnatal day 20, I analyzed how postsynaptic responses are
changed in a GlyT1-depleted environment. During spontaneous vesicle release I found no
change of postsynaptic responses, contradicting my initial hypothesis of prolonged decay
times. Electrical stimulation of fibers of the medial nucleus of the trapezoid body (MNTB),
which are known to form fast, reliable and highly precise synapses with LSO principal neurons,
revealed that GlyT1 is involved in proper synaptic function during sustained, high frequent
synaptic transmission. Stimulation with 50 Hz led to a stronger decay time and latency
prolongation in GlyT1b/c KO, accelerating to 60% longer decay times and 30% longer latencies.
Additionally, a more pronounced frequency-dependent depression and fidelity decrease was
observed during stimulation with 200 Hz in GlyT1b/c KO, resulting in 67% smaller amplitudes
and only 25% of WT fidelity at the end of the challenge. Basic properties like readily releasable
pool, release probability, and quantal size (q) were not altered in GlyT1b/c KO, but
interestingly q decreased during 50 Hz and 100 Hz challenges to about 84%, which was not
observed in WT. I conclude that stronger accumulation of extracellular glycine due to GlyT1
loss leads to prolonged activation of postsynaptic glycine receptors (GlyRs). As a further
consequence, activation of presynaptic GlyRs in the vicinity of the synaptic cleft might be
enhanced, accompanied by a stronger occurrence of shunting inhibition. Furthermore, I
assume a GlyT1-dependent glycine shuttle, which is absent at GlyT1b/c KO synapses. This
could result in a diminished glycine supply to GlyT2 located at more distant sites, causing a
disturbed replenishment during periods with excess release of glycine. Conclusively, my study
reveals a contribution of astrocytes in fast and reliable synaptic transmission at the MNTB-LSO
synapse, which in turn is crucial for proper sound source localization.