SLC6A5 encodes GlyT2, a presynaptic Na+/Cl--dependent glycine transporter that recycles glycine from the synaptic cleft back into presynaptic terminals to sustain glycinergic inhibitory neurotransmission in the caudal CNS (PMID:16751771, PMID:16884688). Its expression is restricted to neurons, predominantly inhibitory neurons of caudal brain regions, distinguishing it from the more broadly distributed glial/neuronal GlyT1 (PMID:41703440). Transport depends on coupled Na+ and Cl- binding: loss-of-function mutations cause defective subcellular localization, reduced glycine uptake, or protein truncation, and operate through disrupted Cl- binding residues, conformational changes in extracellular loop 4, cation-π interactions, and—for the A275T variant—a voltage-sensitive transport defect arising from lowered Na+ affinity (PMID:16751771, PMID:16884688, PMID:22700964). Such loss-of-function mutations cause hereditary hyperekplexia, and genetic ablation of GlyT2 in mice produces handling-induced spasms, accelerated neuromuscular synapse elimination and AChR subunit switching, and early lethality, consistent with loss of glycinergic inhibition and increased motor neuron activity (PMID:16751771, PMID:16884688, PMID:22272310). At the spinal level, selective pharmacological blockade of GlyT2 controls glycine availability for inhibitory neurotransmission relevant to pain processing (PMID:20173309).