{"gene":"SLC6A5","run_date":"2026-06-10T07:46:34","timeline":{"discoveries":[{"year":2006,"finding":"Missense, nonsense, and frameshift mutations in SLC6A5 (GlyT2) cause hereditary hyperekplexia by resulting in defective subcellular GlyT2 localization, decreased glycine uptake, or both; selected mutations affect predicted glycine and Na+ binding sites, establishing GlyT2 as a presynaptic component of glycinergic transmission whose loss disrupts glycine recycling.","method":"Human patient mutation analysis combined with functional assays of glycine uptake and subcellular localization studies of mutant GlyT2 constructs","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct functional glycine uptake assays and subcellular localization experiments on multiple patient-derived mutations, replicated across two independent papers (PMID:16751771, PMID:16884688)","pmids":["16751771","16884688"],"is_preprint":false},{"year":2012,"finding":"Systematic functional analysis of 16 novel SLC6A5 mutations confirmed all were defective in glycine transport; pathogenic mechanisms include protein truncation, splice site disruption, missense mutations affecting Cl- binding residues, conformational changes mediated by extracellular loop 4, and cation-π interactions. Electrophysiology of mutation A275T specifically showed a voltage-sensitive decrease in glycine transport caused by lower Na+ affinity.","method":"Glycine uptake assays for 16 mutations; detailed electrophysiology (voltage-clamp) of A275T mutant to measure Na+ affinity and transport kinetics","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro functional transport assays combined with electrophysiology and mutagenesis across 16 variants, single lab but multiple orthogonal methods with rigorous controls","pmids":["22700964"],"is_preprint":false},{"year":2012,"finding":"In mice, homozygous loss of GlyT2 (Slc6a5 null allele via MusD retrotransposon insertion) results in undetectable GlyT2 protein, handling-induced spasms by day 5, and death within two weeks; at the neuromuscular junction, synapse elimination and the embryonic-to-adult acetylcholine receptor subunit switch are hastened, consistent with increased motor neuron activity due to loss of glycinergic inhibition.","method":"Mouse knockout model (null allele); immunoblot to confirm absence of GlyT2 protein; neuromuscular junction morphology and AChR subunit expression analysis","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean null allele with confirmed protein loss, defined cellular phenotype at NMJ with molecular readout (AChR subunit switch), single lab but multiple orthogonal analyses","pmids":["22272310"],"is_preprint":false},{"year":2010,"finding":"Intrathecal blockade of GlyT2 (but not GlyT1) with ALX1393 suppresses both dynamic and static mechanical allodynia in herpetic and postherpetic pain in mice, and also suppresses allodynia induced by intrathecal strychnine and NMDA, demonstrating that GlyT2 activity at the spinal level controls glycine availability for inhibitory neurotransmission relevant to pain processing.","method":"Intrathecal pharmacological inhibition with selective GlyT2 inhibitor ALX1393 and GlyT1 inhibitor sarcosine in mouse pain models; behavioral testing for mechanical allodynia","journal":"Journal of pharmacological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — selective pharmacological inhibition with behavioral readout, single lab, single method (pharmacology + behavior)","pmids":["20173309"],"is_preprint":false},{"year":2026,"finding":"Slc6a5 mRNA is restricted to neurons (predominantly inhibitory neurons) in caudal brain regions in mice, whereas Slc6a9 (GlyT1) is distributed in both glial cells and neurons across broader brain regions; this cellular specificity was established using single-cell RNA sequencing, quantitative RT-PCR, and RNAscope.","method":"Single-cell RNA sequencing re-analysis, quantitative RT-PCR, and RNAscope in situ hybridization across mouse brain regions and peripheral organs","journal":"BMC neuroscience","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — three complementary mRNA expression methods converge on the same conclusion about cell-type specificity; localization established but not directly linked to functional consequence in this study","pmids":["41703440"],"is_preprint":false}],"current_model":"SLC6A5 encodes GlyT2, a presynaptic Na+/Cl--dependent glycine transporter expressed specifically in inhibitory neurons of the caudal CNS that recycles glycine from the synaptic cleft back into presynaptic terminals; loss-of-function mutations disrupting glycine uptake (via defective localization, altered Na+ or Cl- binding, or protein truncation) impair glycinergic inhibition, causing hereditary hyperekplexia, and pharmacological blockade of GlyT2 at the spinal level modulates inhibitory tone relevant to pain processing."},"narrative":{"mechanistic_narrative":"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].","teleology":[{"year":2006,"claim":"Established that SLC6A5/GlyT2 is a presynaptic component of glycinergic transmission by showing that patient mutations disrupt glycine recycling, linking the transporter to a human disease.","evidence":"Human patient mutation analysis with glycine uptake assays and subcellular localization of mutant GlyT2 constructs","pmids":["16751771","16884688"],"confidence":"High","gaps":["Did not resolve atomic-level ion-binding mechanism","Genotype-phenotype severity relationships not fully mapped"]},{"year":2010,"claim":"Addressed whether spinal GlyT2 activity gates inhibitory tone in pain, showing selective GlyT2 (not GlyT1) blockade suppresses mechanical allodynia.","evidence":"Intrathecal selective inhibitor ALX1393 in mouse herpetic/postherpetic pain models with behavioral allodynia testing","pmids":["20173309"],"confidence":"Medium","gaps":["Single lab using pharmacology and behavior only","Did not establish molecular site of inhibitor action on GlyT2"]},{"year":2012,"claim":"Defined the in vivo consequence of complete GlyT2 loss, demonstrating motor spasms, lethality, and accelerated NMJ maturation consistent with disinhibition of motor neurons.","evidence":"Slc6a5 null mouse knockout with immunoblot confirmation, NMJ morphology, and AChR subunit expression analysis","pmids":["22272310"],"confidence":"High","gaps":["Mechanism linking glycinergic loss to NMJ remodeling not dissected","No rescue experiment reported"]},{"year":2012,"claim":"Resolved the molecular basis of transport defects across many variants, identifying Cl- binding residues, loop 4 conformation, cation-π interactions, and Na+-affinity-dependent voltage sensitivity as pathogenic mechanisms.","evidence":"Glycine uptake assays for 16 mutations plus voltage-clamp electrophysiology of A275T to measure Na+ affinity","pmids":["22700964"],"confidence":"High","gaps":["No structural model accompanying mutagenesis","Single-lab functional data"]},{"year":2026,"claim":"Clarified the cellular expression boundary of GlyT2, establishing its restriction to neurons (predominantly inhibitory) in caudal brain regions versus the broader glial/neuronal distribution of GlyT1.","evidence":"Single-cell RNA-seq re-analysis, quantitative RT-PCR, and RNAscope across mouse brain regions and peripheral organs","pmids":["41703440"],"confidence":"Medium","gaps":["Expression mapping not linked to a functional consequence in this study","Human cell-type distribution not addressed"]},{"year":null,"claim":"How GlyT2 transport activity is dynamically regulated (trafficking, post-translational modification, partner proteins) at presynaptic terminals remains uncharacterized in this corpus.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No interacting partners or regulatory proteins identified","No structural model of human GlyT2","Regulation of surface localization unresolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005215","term_label":"transporter activity","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[0,3]},{"term_id":"R-HSA-382551","term_label":"Transport of small molecules","supporting_discovery_ids":[0,1]}],"complexes":[],"partners":[],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9Y345","full_name":"Sodium- and chloride-dependent glycine transporter 2","aliases":["Solute carrier family 6 member 5"],"length_aa":797,"mass_kda":87.4,"function":"Sodium- and chloride-dependent glycine transporter (PubMed:10381548, PubMed:10606742, PubMed:16751771, PubMed:31370103, PubMed:9845349). Terminates the action of glycine by its high affinity sodium-dependent reuptake into presynaptic terminals (PubMed:9845349). May be responsible for the termination of neurotransmission at strychnine-sensitive glycinergic synapses (PubMed:9845349) Lacks sodium- and chloride-dependent glycine transporter activity Lacks sodium- and chloride-dependent glycine transporter activity","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q9Y345/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/SLC6A5","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":true,"resolved_as":"NET1","ensg_id":"ENSG00000173848","cell_line_id":"CID000580","localizations":[{"compartment":"nucleoplasm","grade":3}],"interactors":[{"gene":"KPNB1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000580","total_profiled":1310},"omim":[{"mim_id":"614618","title":"HYPEREKPLEXIA 3; HKPX3","url":"https://www.omim.org/entry/614618"},{"mim_id":"604159","title":"SOLUTE CARRIER FAMILY 6 (NEUROTRANSMITTER TRANSPORTER, GLYCINE), MEMBER 5; SLC6A5","url":"https://www.omim.org/entry/604159"},{"mim_id":"163970","title":"SOLUTE CARRIER FAMILY 6 (NEUROTRANSMITTER TRANSPORTER, NORADRENALINE), MEMBER 2; SLC6A2","url":"https://www.omim.org/entry/163970"},{"mim_id":"149400","title":"HYPEREKPLEXIA 1; HKPX1","url":"https://www.omim.org/entry/149400"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Not detected","tissue_distribution":"Not detected","driving_tissues":[],"url":"https://www.proteinatlas.org/search/SLC6A5"},"hgnc":{"alias_symbol":["GLYT2","GlyT-2"],"prev_symbol":["NET1"]},"alphafold":{"accession":"Q9Y345","domains":[{"cath_id":"1.20.1740,1.20.1730","chopping":"215-307_375-766","consensus_level":"medium","plddt":92.1866,"start":215,"end":766}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y345","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y345-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y345-F1-predicted_aligned_error_v6.png","plddt_mean":73.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=SLC6A5","jax_strain_url":"https://www.jax.org/strain/search?query=SLC6A5"},"sequence":{"accession":"Q9Y345","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y345.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y345/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y345"}},"corpus_meta":[{"pmid":"16751771","id":"PMC_16751771","title":"Mutations in the gene encoding GlyT2 (SLC6A5) define a presynaptic component of human startle disease.","date":"2006","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16751771","citation_count":182,"is_preprint":false},{"pmid":"22700964","id":"PMC_22700964","title":"Mutations in the GlyT2 gene (SLC6A5) are a second major cause of startle disease.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22700964","citation_count":70,"is_preprint":false},{"pmid":"16884688","id":"PMC_16884688","title":"Mutations within the human GLYT2 (SLC6A5) gene associated with hyperekplexia.","date":"2006","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/16884688","citation_count":63,"is_preprint":false},{"pmid":"20173309","id":"PMC_20173309","title":"Blockade of glycine transporter (GlyT) 2, but not GlyT1, ameliorates dynamic and static mechanical allodynia in mice with herpetic or postherpetic pain.","date":"2010","source":"Journal of pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/20173309","citation_count":32,"is_preprint":false},{"pmid":"18638388","id":"PMC_18638388","title":"Association study of polymorphisms in the neutral amino acid transporter genes SLC1A4, SLC1A5 and the glycine transporter genes SLC6A5, SLC6A9 with schizophrenia.","date":"2008","source":"BMC psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/18638388","citation_count":22,"is_preprint":false},{"pmid":"22272310","id":"PMC_22272310","title":"A MusD retrotransposon insertion in the mouse Slc6a5 gene causes alterations in neuromuscular junction maturation and behavioral phenotypes.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22272310","citation_count":17,"is_preprint":false},{"pmid":"30847549","id":"PMC_30847549","title":"A glycine transporter SLC6A5 frameshift mutation causes startle disease in Spanish greyhounds.","date":"2019","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30847549","citation_count":8,"is_preprint":false},{"pmid":"33120686","id":"PMC_33120686","title":"Case report of a PRDM5 linked brittle cornea syndrome type 2 in association with a novel SLC6A5 mutation.","date":"2020","source":"Indian journal of ophthalmology","url":"https://pubmed.ncbi.nlm.nih.gov/33120686","citation_count":4,"is_preprint":false},{"pmid":"40012122","id":"PMC_40012122","title":"A frameshift variant in the SLC6A5 gene is associated with startle disease in a family of Old English Sheepdogs.","date":"2025","source":"Animal genetics","url":"https://pubmed.ncbi.nlm.nih.gov/40012122","citation_count":2,"is_preprint":false},{"pmid":"41064056","id":"PMC_41064056","title":"Familial Hyperekplexia Caused by a Novel Homozygous SLC6A5 Variant: A Case Report.","date":"2025","source":"Molecular syndromology","url":"https://pubmed.ncbi.nlm.nih.gov/41064056","citation_count":1,"is_preprint":false},{"pmid":"41703440","id":"PMC_41703440","title":"Slc6a9 is distributed in glial cells and neurons across several nervous system regions, whereas Slc6a5 is more restricted to neurons in the caudal brain.","date":"2026","source":"BMC neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/41703440","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":7377,"output_tokens":1460,"usd":0.022016,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8227,"output_tokens":1995,"usd":0.045505,"stage2_stop_reason":"end_turn"},"total_usd":0.067521,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2006,\n      \"finding\": \"Missense, nonsense, and frameshift mutations in SLC6A5 (GlyT2) cause hereditary hyperekplexia by resulting in defective subcellular GlyT2 localization, decreased glycine uptake, or both; selected mutations affect predicted glycine and Na+ binding sites, establishing GlyT2 as a presynaptic component of glycinergic transmission whose loss disrupts glycine recycling.\",\n      \"method\": \"Human patient mutation analysis combined with functional assays of glycine uptake and subcellular localization studies of mutant GlyT2 constructs\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct functional glycine uptake assays and subcellular localization experiments on multiple patient-derived mutations, replicated across two independent papers (PMID:16751771, PMID:16884688)\",\n      \"pmids\": [\"16751771\", \"16884688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Systematic functional analysis of 16 novel SLC6A5 mutations confirmed all were defective in glycine transport; pathogenic mechanisms include protein truncation, splice site disruption, missense mutations affecting Cl- binding residues, conformational changes mediated by extracellular loop 4, and cation-π interactions. Electrophysiology of mutation A275T specifically showed a voltage-sensitive decrease in glycine transport caused by lower Na+ affinity.\",\n      \"method\": \"Glycine uptake assays for 16 mutations; detailed electrophysiology (voltage-clamp) of A275T mutant to measure Na+ affinity and transport kinetics\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro functional transport assays combined with electrophysiology and mutagenesis across 16 variants, single lab but multiple orthogonal methods with rigorous controls\",\n      \"pmids\": [\"22700964\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"In mice, homozygous loss of GlyT2 (Slc6a5 null allele via MusD retrotransposon insertion) results in undetectable GlyT2 protein, handling-induced spasms by day 5, and death within two weeks; at the neuromuscular junction, synapse elimination and the embryonic-to-adult acetylcholine receptor subunit switch are hastened, consistent with increased motor neuron activity due to loss of glycinergic inhibition.\",\n      \"method\": \"Mouse knockout model (null allele); immunoblot to confirm absence of GlyT2 protein; neuromuscular junction morphology and AChR subunit expression analysis\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean null allele with confirmed protein loss, defined cellular phenotype at NMJ with molecular readout (AChR subunit switch), single lab but multiple orthogonal analyses\",\n      \"pmids\": [\"22272310\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Intrathecal blockade of GlyT2 (but not GlyT1) with ALX1393 suppresses both dynamic and static mechanical allodynia in herpetic and postherpetic pain in mice, and also suppresses allodynia induced by intrathecal strychnine and NMDA, demonstrating that GlyT2 activity at the spinal level controls glycine availability for inhibitory neurotransmission relevant to pain processing.\",\n      \"method\": \"Intrathecal pharmacological inhibition with selective GlyT2 inhibitor ALX1393 and GlyT1 inhibitor sarcosine in mouse pain models; behavioral testing for mechanical allodynia\",\n      \"journal\": \"Journal of pharmacological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — selective pharmacological inhibition with behavioral readout, single lab, single method (pharmacology + behavior)\",\n      \"pmids\": [\"20173309\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Slc6a5 mRNA is restricted to neurons (predominantly inhibitory neurons) in caudal brain regions in mice, whereas Slc6a9 (GlyT1) is distributed in both glial cells and neurons across broader brain regions; this cellular specificity was established using single-cell RNA sequencing, quantitative RT-PCR, and RNAscope.\",\n      \"method\": \"Single-cell RNA sequencing re-analysis, quantitative RT-PCR, and RNAscope in situ hybridization across mouse brain regions and peripheral organs\",\n      \"journal\": \"BMC neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — three complementary mRNA expression methods converge on the same conclusion about cell-type specificity; localization established but not directly linked to functional consequence in this study\",\n      \"pmids\": [\"41703440\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"SLC6A5 encodes GlyT2, a presynaptic Na+/Cl--dependent glycine transporter expressed specifically in inhibitory neurons of the caudal CNS that recycles glycine from the synaptic cleft back into presynaptic terminals; loss-of-function mutations disrupting glycine uptake (via defective localization, altered Na+ or Cl- binding, or protein truncation) impair glycinergic inhibition, causing hereditary hyperekplexia, and pharmacological blockade of GlyT2 at the spinal level modulates inhibitory tone relevant to pain processing.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"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 [#0]. Its expression is restricted to neurons, predominantly inhibitory neurons of caudal brain regions, distinguishing it from the more broadly distributed glial/neuronal GlyT1 [#4]. 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 [#0, #1]. 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 [#0, #2]. At the spinal level, selective pharmacological blockade of GlyT2 controls glycine availability for inhibitory neurotransmission relevant to pain processing [#3].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established that SLC6A5/GlyT2 is a presynaptic component of glycinergic transmission by showing that patient mutations disrupt glycine recycling, linking the transporter to a human disease.\",\n      \"evidence\": \"Human patient mutation analysis with glycine uptake assays and subcellular localization of mutant GlyT2 constructs\",\n      \"pmids\": [\"16751771\", \"16884688\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve atomic-level ion-binding mechanism\", \"Genotype-phenotype severity relationships not fully mapped\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Addressed whether spinal GlyT2 activity gates inhibitory tone in pain, showing selective GlyT2 (not GlyT1) blockade suppresses mechanical allodynia.\",\n      \"evidence\": \"Intrathecal selective inhibitor ALX1393 in mouse herpetic/postherpetic pain models with behavioral allodynia testing\",\n      \"pmids\": [\"20173309\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab using pharmacology and behavior only\", \"Did not establish molecular site of inhibitor action on GlyT2\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined the in vivo consequence of complete GlyT2 loss, demonstrating motor spasms, lethality, and accelerated NMJ maturation consistent with disinhibition of motor neurons.\",\n      \"evidence\": \"Slc6a5 null mouse knockout with immunoblot confirmation, NMJ morphology, and AChR subunit expression analysis\",\n      \"pmids\": [\"22272310\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism linking glycinergic loss to NMJ remodeling not dissected\", \"No rescue experiment reported\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Resolved the molecular basis of transport defects across many variants, identifying Cl- binding residues, loop 4 conformation, cation-π interactions, and Na+-affinity-dependent voltage sensitivity as pathogenic mechanisms.\",\n      \"evidence\": \"Glycine uptake assays for 16 mutations plus voltage-clamp electrophysiology of A275T to measure Na+ affinity\",\n      \"pmids\": [\"22700964\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model accompanying mutagenesis\", \"Single-lab functional data\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Clarified the cellular expression boundary of GlyT2, establishing its restriction to neurons (predominantly inhibitory) in caudal brain regions versus the broader glial/neuronal distribution of GlyT1.\",\n      \"evidence\": \"Single-cell RNA-seq re-analysis, quantitative RT-PCR, and RNAscope across mouse brain regions and peripheral organs\",\n      \"pmids\": [\"41703440\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Expression mapping not linked to a functional consequence in this study\", \"Human cell-type distribution not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How GlyT2 transport activity is dynamically regulated (trafficking, post-translational modification, partner proteins) at presynaptic terminals remains uncharacterized in this corpus.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No interacting partners or regulatory proteins identified\", \"No structural model of human GlyT2\", \"Regulation of surface localization unresolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005215\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"R-HSA-382551\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}