{"gene":"AMIGO1","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2003,"finding":"AMIGO1 (AMIGO) is a type I transmembrane protein with six leucine-rich repeats flanked by cysteine-rich LRR N- and C-terminal domains and one immunoglobulin domain; substrate-bound recombinant AMIGO ectodomain promoted neurite extension in hippocampal neurons, while soluble AMIGO ectodomain inhibited neurite fasciculation; AMIGO family members engage in homophilic and heterophilic binding with each other.","method":"Recombinant ectodomain neurite outgrowth assay in hippocampal neurons; homophilic/heterophilic binding assay between AMIGO family members","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct functional assays with recombinant protein and binding assays, single lab with two orthogonal methods","pmids":["12629050"],"is_preprint":false},{"year":2011,"finding":"AMIGO1 is an auxiliary subunit of the Kv2.1 potassium channel complex: it shows extensive spatial and temporal colocalization and co-immunoprecipitation with Kv2.1 in mouse brain, increases Kv2.1 conductance in a voltage-dependent manner in HEK cells, and siRNA-mediated inhibition of endogenous AMIGO suppresses neuronal delayed rectifier current (IK) at negative membrane voltages.","method":"Co-immunoprecipitation, immunofluorescence colocalization, electrophysiology in HEK cells, siRNA knockdown with patch-clamp in neurons","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, electrophysiology in heterologous cells and neurons, siRNA loss-of-function, multiple orthogonal methods in one study","pmids":["22056818"],"is_preprint":false},{"year":2011,"finding":"Crystal structure of AMIGO-1 ectodomain determined at 2.0 Å resolution, revealing a typical LRR domain with N- and C-terminal capping domains containing disulfide bridges followed by a C2-type Ig domain; AMIGO-1 forms a dimer through its LRR-LRR interface, and mutagenesis studies showed that dimerization is necessary for proper cell surface expression, suggesting a role in stable folding in the ER and cell-cell adhesion.","method":"X-ray crystallography (2.0 Å), static light scattering, SAXS, site-directed mutagenesis with cell surface expression assay","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus mutagenesis validating functional consequence, multiple orthogonal structural and biochemical methods","pmids":["21983541"],"is_preprint":false},{"year":2012,"finding":"AMIGO1 localizes predominantly to dendrites in mature primary neurons (while LRR-deleted AMIGO is axonal), and siRNA-mediated silencing of AMIGO reduces dendritic growth of cortical neurons in culture; overexpression of AMIGO in SH-SY5Y cells confers resistance to staurosporine- and H2O2-induced apoptosis.","method":"Immunofluorescence localization, siRNA knockdown with morphometric analysis of dendrites, stable overexpression with apoptosis assay","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA loss-of-function with defined cellular phenotype and localization, single lab with two orthogonal approaches","pmids":["21938721"],"is_preprint":false},{"year":2014,"finding":"In zebrafish, amigo1 is the predominant family member expressed during nervous system development; morpholino knockdown of amigo1 impairs fasciculated tract formation in early fiber scaffolds, and mRNA-mediated overexpression of the Amigo1 ectodomain (which inhibits adhesion by the full-length protein) produces a similar defect, confirming that homophilic interactions underlie fiber tract development; Amigo1 also regulates Kv2.1 potassium channel function to form functional neural circuitry controlling locomotion.","method":"Morpholino antisense knockdown in zebrafish, mRNA overexpression of ectodomain, immunofluorescence imaging of fiber tracts, behavioral analysis","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo loss-of-function and dominant-negative ectodomain rescue in zebrafish, single lab with multiple approaches","pmids":["24904058"],"is_preprint":false},{"year":2015,"finding":"Genetic deletion of AMIGO in mice reduces the amount of Kv2.1 channel protein in brain and alters electrophysiological properties of neurons, demonstrating that AMIGO is required for normal Kv2.1 expression levels and neuronal electrical activity in vivo.","method":"AMIGO knockout mouse, Western blot for Kv2.1 protein, electrophysiology","journal":"Schizophrenia bulletin","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with defined biochemical and electrophysiological phenotypes, single lab","pmids":["26240432"],"is_preprint":false},{"year":2018,"finding":"In adult brain neurons, AMIGO-1 is exclusively colocalized with Kv2.1 and Kv2.2 at plasma membrane sites associated with neuronal ER:PM junctions (hypolemmal subsurface cisternae); Kv2 α subunits are obligatory for AMIGO-1 clustering, PM trafficking, and expression level—coexpression of either Kv2.1 or Kv2.2 is sufficient to drive AMIGO-1 clustering in heterologous cells, and AMIGO-1 expression/localization is lost in Kv2.1 or Kv2.2 knockout mice.","method":"Immunofluorescence colocalization across multiple mammalian species, live-cell imaging in heterologous cells, Kv2.1/Kv2.2 single and double knockout mouse analysis, subcellular fractionation/trafficking assays","journal":"Frontiers in molecular neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal genetic KO and heterologous coexpression with localization readout, replicated across species with multiple orthogonal methods","pmids":["29403353"],"is_preprint":false},{"year":2021,"finding":"All three AMIGO family proteins (AMIGO1, 2, 3) are controlled in surface trafficking and localization by assembly with either Kv2.1 or Kv2.2; AMIGO1 assembly with either Kv2 channel hyperpolarizes the channel activation midpoint by approximately −10 mV; AMIGO1 does not significantly slow inactivation or deactivation (unlike AMIGO2), indicating isoform-specific modulation.","method":"Electrophysiology (voltage-clamp), live-cell imaging of surface trafficking in heterologous cells, co-expression studies","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — electrophysiology and live-cell imaging in heterologous system, single lab with two orthogonal methods","pmids":["34137443"],"is_preprint":false},{"year":2022,"finding":"AMIGO1 modulates the Kv2.1 conductance activation pathway by destabilizing the earliest resting state of Kv2.1 voltage sensors: AMIGO1 speeds early voltage-sensor movements, shifts the gating charge-voltage (Q-V) relationship to more negative voltages, and when voltage sensors are detained at rest by toxins, AMIGO1 exerts a greater effect on the conductance-voltage relationship; fluorescence measurements from voltage-sensor toxins confirm that AMIGO1 makes the earliest resting conformation less stable upon voltage stimulation.","method":"Voltage-clamp recordings, gating current measurements, voltage-sensor toxin fluorescence spectroscopy, heterologous expression in mammalian cell lines","journal":"Biophysical journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstitution in mammalian heterologous system with voltage-clamp plus fluorescence spectroscopy, orthogonal approaches establishing mechanism at voltage-sensor level","pmids":["35314141"],"is_preprint":false},{"year":2022,"finding":"AMIGO1 selectively promotes axon arbor growth of retinal horizontal cells; genetic deletion of Amigo1 in mice reduces horizontal cell axon arbor size without affecting horizontal cell dendrite size or synapse formation, demonstrating a compartment-specific role for AMIGO1 in axon scaling; reduction of horizontal cell axons causes territory matching-mediated shrinkage of rod bipolar cell dendrites, preserving rod bipolar pathway function.","method":"Amigo1 knockout mice, single-cell conditional knockout, morphometric analysis of axon/dendrite arbors, immunofluorescence, electrophysiological recording of retinal output","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO with defined compartment-specific morphological phenotype and functional readout, both global and conditional single-cell KO approaches","pmids":["35169021"],"is_preprint":false},{"year":2025,"finding":"AMIGO-1 clustering and expression in spinal motor neurons is dependent on Kv2 subunits: AMIGO-1 clustering at ER-PM junctions is severely reduced in Kv2.1 knockout mice and moderately reduced in Kv2.2 knockout mice, and is also severely reduced in Kv2.1 S590A mutant mice that cannot bind ER VAP proteins, indicating that Kv2-mediated ER-PM junction formation is required for AMIGO-1 localization in motor neurons.","method":"Kv2.1 and Kv2.2 knockout mice, Kv2.1 S590A knock-in mutant mice, immunofluorescence colocalization in spinal motor neurons","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple genetic mouse models with localization readout, preprint, single lab","pmids":["bio_10.1101_2025.06.04.657913"],"is_preprint":true},{"year":2024,"finding":"Chemogenetic activation of Amigo1-expressing GABAergic neurons within the interpeduncular nucleus (IPN) is critical for anxiety-like behaviors in both naïve mice and those undergoing nicotine withdrawal; stimulation of Amigo1 neurons in nicotine-naïve mice elicits opposite effects on affective versus somatic signs of withdrawal, demonstrating that Amigo1-expressing IPN neurons specifically mediate the affective component of nicotine withdrawal.","method":"Chemogenetic (DREADD) activation of Amigo1-Cre targeted IPN neurons, behavioral assays for anxiety and somatic withdrawal signs","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-type-specific chemogenetics with defined behavioral phenotype, preprint, single lab","pmids":["bio_10.1101_2024.07.05.602259"],"is_preprint":true}],"current_model":"AMIGO1 is a type I transmembrane LRR/Ig-domain cell adhesion molecule that functions as an obligatory auxiliary (β) subunit of Kv2.1 and Kv2.2 voltage-gated potassium channels in brain neurons, where Kv2 α subunits reciprocally control AMIGO1 expression, PM trafficking, and clustering at ER-PM junctions; mechanistically, AMIGO1 destabilizes the earliest resting state of Kv2.1 voltage sensors to hyperpolarize channel activation, while its homophilic LRR-mediated dimerization drives neurite fasciculation and axon arbor expansion (notably in retinal horizontal cells) during neural circuit development."},"narrative":{"mechanistic_narrative":"AMIGO1 is a type I transmembrane leucine-rich-repeat/immunoglobulin-domain cell adhesion molecule that serves dual roles in neural circuit assembly and in shaping neuronal electrical excitability [PMID:12629050, PMID:22056818]. Its ectodomain comprises six leucine-rich repeats with cysteine-rich N- and C-terminal caps followed by a C2-type Ig domain, and crystallographic and biophysical analysis shows it dimerizes through an LRR-LRR interface that is required for stable ER folding and cell-surface expression [PMID:21983541]. Through homophilic LRR-mediated adhesion AMIGO1 promotes neurite extension, fasciculation, and tract formation: the substrate-bound ectodomain drives outgrowth while the soluble ectodomain acts as a dominant-negative that disrupts fasciculation in hippocampal neurons and in zebrafish fiber scaffolds [PMID:12629050, PMID:24904058], and genetic deletion in mice reveals a compartment-specific requirement for AMIGO1 in scaling retinal horizontal cell axon arbors [PMID:35169021]. In parallel, AMIGO1 is an auxiliary subunit of Kv2.1/Kv2.2 voltage-gated potassium channels: it co-immunoprecipitates and colocalizes with Kv2.1, and loss of AMIGO suppresses the neuronal delayed rectifier current and reduces brain Kv2.1 protein in vivo [PMID:22056818, PMID:26240432]. Assembly with Kv2 channels hyperpolarizes the activation midpoint by roughly -10 mV, and mechanistically AMIGO1 destabilizes the earliest resting conformation of the Kv2.1 voltage sensors, speeding early sensor movements and shifting the gating charge-voltage relationship to more negative voltages [PMID:34137443, PMID:35314141]. The relationship is reciprocal: Kv2 α subunits are obligatory for AMIGO1 clustering, plasma-membrane trafficking, and expression, localizing it to ER-PM junctions, a dependency that extends to motor neurons where Kv2-mediated junction formation is required for AMIGO1 localization [PMID:29403353, PMID:bio_10.1101_2025.06.04.657913].","teleology":[{"year":2003,"claim":"Established AMIGO1 as an LRR/Ig adhesion molecule whose ectodomain bidirectionally controls neurite outgrowth and fasciculation, defining its first functional role in neural morphogenesis.","evidence":"Recombinant ectodomain neurite outgrowth assays and homophilic/heterophilic binding assays in hippocampal neurons","pmids":["12629050"],"confidence":"Medium","gaps":["Binding partners beyond the AMIGO family not identified","No in vivo confirmation at this stage","Molecular mechanism of adhesion-to-outgrowth coupling unresolved"]},{"year":2011,"claim":"Revealed an entirely distinct function: AMIGO1 acts as an auxiliary subunit of Kv2.1, coupling the adhesion molecule to control of neuronal potassium currents.","evidence":"Reciprocal Co-IP, immunofluorescence colocalization, HEK electrophysiology, and siRNA knockdown with patch-clamp in neurons","pmids":["22056818"],"confidence":"High","gaps":["Stoichiometry of the AMIGO1-Kv2.1 complex not defined","Biophysical mechanism of conductance modulation not yet resolved","Whether Kv2.2 is also modulated untested here"]},{"year":2011,"claim":"Determined the ectodomain structure and showed dimerization through the LRR interface is required for surface expression, providing the structural basis for both folding and adhesion.","evidence":"2.0 Å X-ray crystallography with SAXS, static light scattering, and mutagenesis-coupled surface expression assays","pmids":["21983541"],"confidence":"High","gaps":["Structure of full-length transmembrane protein and channel-bound complex unknown","Adhesion interface in trans not directly visualized"]},{"year":2012,"claim":"Localized AMIGO1 to dendrites and linked it to dendritic growth and apoptosis resistance, refining its compartment-specific developmental role.","evidence":"Immunofluorescence localization, siRNA knockdown with dendrite morphometry, and overexpression apoptosis assays in SH-SY5Y cells","pmids":["21938721"],"confidence":"Medium","gaps":["Mechanism linking AMIGO1 to apoptosis resistance unknown","Signaling pathways downstream of dendritic AMIGO1 not defined"]},{"year":2014,"claim":"Demonstrated in vivo that homophilic AMIGO1 interactions drive fiber tract fasciculation and that AMIGO1 also regulates Kv2.1 for functional circuitry, unifying its adhesion and channel roles in a whole organism.","evidence":"Morpholino knockdown and dominant-negative ectodomain overexpression in zebrafish with fiber tract imaging and locomotion behavior","pmids":["24904058"],"confidence":"Medium","gaps":["Relative contributions of adhesion vs channel functions to behavior not dissected","Morpholino off-target effects not fully excluded"]},{"year":2015,"claim":"Showed AMIGO is required in vivo for normal Kv2.1 protein levels and neuronal electrical activity, establishing the channel dependency genetically.","evidence":"AMIGO knockout mouse with Western blot for Kv2.1 and electrophysiology","pmids":["26240432"],"confidence":"Medium","gaps":["Whether reduced Kv2.1 reflects trafficking or stability not resolved","Behavioral/circuit consequences not detailed here"]},{"year":2018,"claim":"Defined the reciprocal dependency: Kv2 α subunits are obligatory for AMIGO1 clustering and trafficking to ER-PM junctions, reframing the relationship as bidirectional control.","evidence":"Cross-species immunofluorescence, live-cell imaging in heterologous cells, and Kv2.1/Kv2.2 single and double knockout mouse analysis","pmids":["29403353"],"confidence":"High","gaps":["Molecular interface mediating Kv2-dependent AMIGO1 clustering not mapped","Functional role of ER-PM junction localization for AMIGO1 unclear"]},{"year":2021,"claim":"Distinguished isoform-specific channel modulation, showing AMIGO1 hyperpolarizes Kv2 activation by ~-10 mV without the inactivation/deactivation slowing seen with AMIGO2.","evidence":"Voltage-clamp electrophysiology and live-cell surface trafficking imaging in heterologous cells","pmids":["34137443"],"confidence":"Medium","gaps":["Structural basis of isoform-specific effects unknown","Native neuronal validation of -10 mV shift limited"]},{"year":2022,"claim":"Resolved the biophysical mechanism: AMIGO1 destabilizes the earliest resting state of Kv2.1 voltage sensors, explaining the hyperpolarized activation.","evidence":"Voltage-clamp, gating current measurements, and voltage-sensor toxin fluorescence spectroscopy in mammalian cells","pmids":["35314141"],"confidence":"High","gaps":["Physical contact points between AMIGO1 and the voltage-sensor domain not identified","Whether this mechanism operates identically on Kv2.2 untested"]},{"year":2022,"claim":"Established a precise developmental phenotype: AMIGO1 selectively scales retinal horizontal cell axon arbors, with downstream territory matching of rod bipolar dendrites.","evidence":"Global and single-cell conditional Amigo1 knockout mice with arbor morphometry and retinal electrophysiology","pmids":["35169021"],"confidence":"High","gaps":["Molecular signaling driving compartment-specific axon scaling unknown","Whether channel function contributes to this morphological role not separated"]},{"year":2025,"claim":"Extended the Kv2-dependency to motor neurons and linked it mechanistically to Kv2-VAP-mediated ER-PM junction formation via the Kv2.1 S590A mutant.","evidence":"Kv2.1/Kv2.2 knockout and Kv2.1 S590A knock-in mice with immunofluorescence in spinal motor neurons (preprint)","pmids":["bio_10.1101_2025.06.04.657913"],"confidence":"Medium","gaps":["Preprint, single lab, not peer reviewed","Functional consequence of motor neuron AMIGO1 mislocalization untested"]},{"year":2024,"claim":"Identified an Amigo1-expressing GABAergic neuron population in the interpeduncular nucleus that mediates affective signs of nicotine withdrawal, linking AMIGO1 marker expression to circuit-level behavior.","evidence":"Chemogenetic DREADD activation of Amigo1-Cre IPN neurons with anxiety and somatic withdrawal behavioral assays (preprint)","pmids":["bio_10.1101_2024.07.05.602259"],"confidence":"Medium","gaps":["Preprint, not peer reviewed","Whether AMIGO1 protein itself, rather than as a cell-type marker, contributes to the behavior is unaddressed"]},{"year":null,"claim":"How AMIGO1's adhesion/morphogenic function and its Kv2-channel auxiliary function are coordinated within single neurons, and the physical interface mediating both Kv2 binding and voltage-sensor destabilization, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of the AMIGO1-Kv2 complex","Contact residues for voltage-sensor modulation unmapped","Integration of channel vs adhesion roles in development not dissected"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[0,2,4]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,7,8]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,10]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[2,10]}],"pathway":[{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[1,5,8]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,9]}],"complexes":["Kv2.1 channel complex","Kv2.2 channel complex"],"partners":["KCNB1","KCNB2","AMIGO1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86WK6","full_name":"Amphoterin-induced protein 1","aliases":["AMIGO-1","Alivin-2"],"length_aa":493,"mass_kda":55.2,"function":"Promotes growth and fasciculation of neurites from cultured hippocampal neurons. May be involved in fasciculation as well as myelination of developing neural axons. May have a role in regeneration as well as neural plasticity in the adult nervous system. May mediate homophilic as well as heterophilic cell-cell interaction and contribute to signal transduction through its intracellular domain. Assembled with KCNB1 modulates the gating characteristics of the delayed rectifier voltage-dependent potassium channel KCNB1","subcellular_location":"Cell membrane; Perikaryon; Cell projection, dendrite; Cell projection, axon","url":"https://www.uniprot.org/uniprotkb/Q86WK6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AMIGO1","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/AMIGO1","total_profiled":1310},"omim":[{"mim_id":"615691","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 3; AMIGO3","url":"https://www.omim.org/entry/615691"},{"mim_id":"615690","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 2; AMIGO2","url":"https://www.omim.org/entry/615690"},{"mim_id":"615689","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 1; AMIGO1","url":"https://www.omim.org/entry/615689"},{"mim_id":"612810","title":"LEUCINE-RICH REPEAT AND FIBRONECTIN TYPE III DOMAIN-CONTAINING PROTEIN 4; LRFN4","url":"https://www.omim.org/entry/612810"},{"mim_id":"612809","title":"LEUCINE-RICH REPEAT AND FIBRONECTIN TYPE III DOMAIN-CONTAINING PROTEIN 3; LRFN3","url":"https://www.omim.org/entry/612809"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/AMIGO1"},"hgnc":{"alias_symbol":["AMIGO","KIAA1163"],"prev_symbol":[]},"alphafold":{"accession":"Q86WK6","domains":[{"cath_id":"2.60.40.10","chopping":"277-358","consensus_level":"high","plddt":89.8896,"start":277,"end":358}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86WK6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86WK6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86WK6-F1-predicted_aligned_error_v6.png","plddt_mean":78.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AMIGO1","jax_strain_url":"https://www.jax.org/strain/search?query=AMIGO1"},"sequence":{"accession":"Q86WK6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86WK6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86WK6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86WK6"}},"corpus_meta":[{"pmid":"19033274","id":"PMC_19033274","title":"AmiGO: online access 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Retrieving GO Data Using AmiGO, QuickGO, API, Files, and Tools.","date":"2017","source":"Methods in molecular biology (Clifton, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/27812941","citation_count":21,"is_preprint":false},{"pmid":"24440330","id":"PMC_24440330","title":"Comparison of the safety and feasibility of arrhythmia ablation using the Amigo Robotic Remote Catheter System versus manual ablation.","date":"2013","source":"The American journal of cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/24440330","citation_count":18,"is_preprint":false},{"pmid":"27446084","id":"PMC_27446084","title":"IL-17A and TNF-α Increase the Expression of the Antiapoptotic Adhesion Molecule Amigo-2 in Arthritis Synoviocytes.","date":"2016","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/27446084","citation_count":16,"is_preprint":false},{"pmid":"24178087","id":"PMC_24178087","title":"Chromosome painting of Amigo wheat.","date":"1994","source":"TAG. 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Theoretische und angewandte Genetik","url":"https://pubmed.ncbi.nlm.nih.gov/24178087","citation_count":15,"is_preprint":false},{"pmid":"24904058","id":"PMC_24904058","title":"Amigo adhesion protein regulates development of neural circuits in zebrafish brain.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24904058","citation_count":14,"is_preprint":false},{"pmid":"34137443","id":"PMC_34137443","title":"Kv2 channel-AMIGO β-subunit assembly modulates both channel function and cell adhesion molecule surface trafficking.","date":"2021","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/34137443","citation_count":14,"is_preprint":false},{"pmid":"35314141","id":"PMC_35314141","title":"The AMIGO1 adhesion protein activates Kv2.1 voltage sensors.","date":"2022","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/35314141","citation_count":10,"is_preprint":false},{"pmid":"35169021","id":"PMC_35169021","title":"AMIGO1 Promotes Axon Growth and Territory Matching in the Retina.","date":"2022","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/35169021","citation_count":8,"is_preprint":false},{"pmid":"9491419","id":"PMC_9491419","title":"The pathologic cycle of the infection of the microsporidian Microgemma ovoidea (Thèl., 1895) Amigó et al. 1996 in the liver of the Red Band fish (Cepola macrophthalma L.).","date":"1998","source":"Parasitology research","url":"https://pubmed.ncbi.nlm.nih.gov/9491419","citation_count":5,"is_preprint":false},{"pmid":"40380067","id":"PMC_40380067","title":"A Strategy to Use Artificial MicroRNAs for Gene Silencing and Overcome (AmiGO).","date":"2025","source":"Methods in molecular biology (Clifton, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/40380067","citation_count":0,"is_preprint":false},{"pmid":"41968202","id":"PMC_41968202","title":"AMIGO - Guided assignment of 13C-methyl labelled proteins.","date":"2026","source":"Journal of biomolecular NMR","url":"https://pubmed.ncbi.nlm.nih.gov/41968202","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.11.22.624721","title":"An integrative approach prioritizes the orphan GPR61 genomic region in tissue-specific regulation of chronotype","date":"2024-11-25","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.22.624721","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.07.05.602259","title":"Chemogenetic control of GABAergic activity within the interpeduncular nucleus reveals dissociable behavioral components of the nicotine withdrawal phenotype","date":"2024-07-10","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.05.602259","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.06.04.657913","title":"Kv2/Kv6.4 heteromeric potassium channels are expressed in spinal motor neurons and localized at C-bouton synapses","date":"2025-06-05","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.04.657913","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14325,"output_tokens":3362,"usd":0.046703,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11011,"output_tokens":4124,"usd":0.079077,"stage2_stop_reason":"end_turn"},"total_usd":0.12578,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"AMIGO1 (AMIGO) is a type I transmembrane protein with six leucine-rich repeats flanked by cysteine-rich LRR N- and C-terminal domains and one immunoglobulin domain; substrate-bound recombinant AMIGO ectodomain promoted neurite extension in hippocampal neurons, while soluble AMIGO ectodomain inhibited neurite fasciculation; AMIGO family members engage in homophilic and heterophilic binding with each other.\",\n      \"method\": \"Recombinant ectodomain neurite outgrowth assay in hippocampal neurons; homophilic/heterophilic binding assay between AMIGO family members\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct functional assays with recombinant protein and binding assays, single lab with two orthogonal methods\",\n      \"pmids\": [\"12629050\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AMIGO1 is an auxiliary subunit of the Kv2.1 potassium channel complex: it shows extensive spatial and temporal colocalization and co-immunoprecipitation with Kv2.1 in mouse brain, increases Kv2.1 conductance in a voltage-dependent manner in HEK cells, and siRNA-mediated inhibition of endogenous AMIGO suppresses neuronal delayed rectifier current (IK) at negative membrane voltages.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence colocalization, electrophysiology in HEK cells, siRNA knockdown with patch-clamp in neurons\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, electrophysiology in heterologous cells and neurons, siRNA loss-of-function, multiple orthogonal methods in one study\",\n      \"pmids\": [\"22056818\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Crystal structure of AMIGO-1 ectodomain determined at 2.0 Å resolution, revealing a typical LRR domain with N- and C-terminal capping domains containing disulfide bridges followed by a C2-type Ig domain; AMIGO-1 forms a dimer through its LRR-LRR interface, and mutagenesis studies showed that dimerization is necessary for proper cell surface expression, suggesting a role in stable folding in the ER and cell-cell adhesion.\",\n      \"method\": \"X-ray crystallography (2.0 Å), static light scattering, SAXS, site-directed mutagenesis with cell surface expression assay\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus mutagenesis validating functional consequence, multiple orthogonal structural and biochemical methods\",\n      \"pmids\": [\"21983541\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"AMIGO1 localizes predominantly to dendrites in mature primary neurons (while LRR-deleted AMIGO is axonal), and siRNA-mediated silencing of AMIGO reduces dendritic growth of cortical neurons in culture; overexpression of AMIGO in SH-SY5Y cells confers resistance to staurosporine- and H2O2-induced apoptosis.\",\n      \"method\": \"Immunofluorescence localization, siRNA knockdown with morphometric analysis of dendrites, stable overexpression with apoptosis assay\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA loss-of-function with defined cellular phenotype and localization, single lab with two orthogonal approaches\",\n      \"pmids\": [\"21938721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In zebrafish, amigo1 is the predominant family member expressed during nervous system development; morpholino knockdown of amigo1 impairs fasciculated tract formation in early fiber scaffolds, and mRNA-mediated overexpression of the Amigo1 ectodomain (which inhibits adhesion by the full-length protein) produces a similar defect, confirming that homophilic interactions underlie fiber tract development; Amigo1 also regulates Kv2.1 potassium channel function to form functional neural circuitry controlling locomotion.\",\n      \"method\": \"Morpholino antisense knockdown in zebrafish, mRNA overexpression of ectodomain, immunofluorescence imaging of fiber tracts, behavioral analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo loss-of-function and dominant-negative ectodomain rescue in zebrafish, single lab with multiple approaches\",\n      \"pmids\": [\"24904058\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Genetic deletion of AMIGO in mice reduces the amount of Kv2.1 channel protein in brain and alters electrophysiological properties of neurons, demonstrating that AMIGO is required for normal Kv2.1 expression levels and neuronal electrical activity in vivo.\",\n      \"method\": \"AMIGO knockout mouse, Western blot for Kv2.1 protein, electrophysiology\",\n      \"journal\": \"Schizophrenia bulletin\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with defined biochemical and electrophysiological phenotypes, single lab\",\n      \"pmids\": [\"26240432\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In adult brain neurons, AMIGO-1 is exclusively colocalized with Kv2.1 and Kv2.2 at plasma membrane sites associated with neuronal ER:PM junctions (hypolemmal subsurface cisternae); Kv2 α subunits are obligatory for AMIGO-1 clustering, PM trafficking, and expression level—coexpression of either Kv2.1 or Kv2.2 is sufficient to drive AMIGO-1 clustering in heterologous cells, and AMIGO-1 expression/localization is lost in Kv2.1 or Kv2.2 knockout mice.\",\n      \"method\": \"Immunofluorescence colocalization across multiple mammalian species, live-cell imaging in heterologous cells, Kv2.1/Kv2.2 single and double knockout mouse analysis, subcellular fractionation/trafficking assays\",\n      \"journal\": \"Frontiers in molecular neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal genetic KO and heterologous coexpression with localization readout, replicated across species with multiple orthogonal methods\",\n      \"pmids\": [\"29403353\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"All three AMIGO family proteins (AMIGO1, 2, 3) are controlled in surface trafficking and localization by assembly with either Kv2.1 or Kv2.2; AMIGO1 assembly with either Kv2 channel hyperpolarizes the channel activation midpoint by approximately −10 mV; AMIGO1 does not significantly slow inactivation or deactivation (unlike AMIGO2), indicating isoform-specific modulation.\",\n      \"method\": \"Electrophysiology (voltage-clamp), live-cell imaging of surface trafficking in heterologous cells, co-expression studies\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — electrophysiology and live-cell imaging in heterologous system, single lab with two orthogonal methods\",\n      \"pmids\": [\"34137443\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AMIGO1 modulates the Kv2.1 conductance activation pathway by destabilizing the earliest resting state of Kv2.1 voltage sensors: AMIGO1 speeds early voltage-sensor movements, shifts the gating charge-voltage (Q-V) relationship to more negative voltages, and when voltage sensors are detained at rest by toxins, AMIGO1 exerts a greater effect on the conductance-voltage relationship; fluorescence measurements from voltage-sensor toxins confirm that AMIGO1 makes the earliest resting conformation less stable upon voltage stimulation.\",\n      \"method\": \"Voltage-clamp recordings, gating current measurements, voltage-sensor toxin fluorescence spectroscopy, heterologous expression in mammalian cell lines\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstitution in mammalian heterologous system with voltage-clamp plus fluorescence spectroscopy, orthogonal approaches establishing mechanism at voltage-sensor level\",\n      \"pmids\": [\"35314141\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AMIGO1 selectively promotes axon arbor growth of retinal horizontal cells; genetic deletion of Amigo1 in mice reduces horizontal cell axon arbor size without affecting horizontal cell dendrite size or synapse formation, demonstrating a compartment-specific role for AMIGO1 in axon scaling; reduction of horizontal cell axons causes territory matching-mediated shrinkage of rod bipolar cell dendrites, preserving rod bipolar pathway function.\",\n      \"method\": \"Amigo1 knockout mice, single-cell conditional knockout, morphometric analysis of axon/dendrite arbors, immunofluorescence, electrophysiological recording of retinal output\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO with defined compartment-specific morphological phenotype and functional readout, both global and conditional single-cell KO approaches\",\n      \"pmids\": [\"35169021\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"AMIGO-1 clustering and expression in spinal motor neurons is dependent on Kv2 subunits: AMIGO-1 clustering at ER-PM junctions is severely reduced in Kv2.1 knockout mice and moderately reduced in Kv2.2 knockout mice, and is also severely reduced in Kv2.1 S590A mutant mice that cannot bind ER VAP proteins, indicating that Kv2-mediated ER-PM junction formation is required for AMIGO-1 localization in motor neurons.\",\n      \"method\": \"Kv2.1 and Kv2.2 knockout mice, Kv2.1 S590A knock-in mutant mice, immunofluorescence colocalization in spinal motor neurons\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple genetic mouse models with localization readout, preprint, single lab\",\n      \"pmids\": [\"bio_10.1101_2025.06.04.657913\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Chemogenetic activation of Amigo1-expressing GABAergic neurons within the interpeduncular nucleus (IPN) is critical for anxiety-like behaviors in both naïve mice and those undergoing nicotine withdrawal; stimulation of Amigo1 neurons in nicotine-naïve mice elicits opposite effects on affective versus somatic signs of withdrawal, demonstrating that Amigo1-expressing IPN neurons specifically mediate the affective component of nicotine withdrawal.\",\n      \"method\": \"Chemogenetic (DREADD) activation of Amigo1-Cre targeted IPN neurons, behavioral assays for anxiety and somatic withdrawal signs\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-type-specific chemogenetics with defined behavioral phenotype, preprint, single lab\",\n      \"pmids\": [\"bio_10.1101_2024.07.05.602259\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"AMIGO1 is a type I transmembrane LRR/Ig-domain cell adhesion molecule that functions as an obligatory auxiliary (β) subunit of Kv2.1 and Kv2.2 voltage-gated potassium channels in brain neurons, where Kv2 α subunits reciprocally control AMIGO1 expression, PM trafficking, and clustering at ER-PM junctions; mechanistically, AMIGO1 destabilizes the earliest resting state of Kv2.1 voltage sensors to hyperpolarize channel activation, while its homophilic LRR-mediated dimerization drives neurite fasciculation and axon arbor expansion (notably in retinal horizontal cells) during neural circuit development.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AMIGO1 is a type I transmembrane leucine-rich-repeat/immunoglobulin-domain cell adhesion molecule that serves dual roles in neural circuit assembly and in shaping neuronal electrical excitability [#0, #1]. Its ectodomain comprises six leucine-rich repeats with cysteine-rich N- and C-terminal caps followed by a C2-type Ig domain, and crystallographic and biophysical analysis shows it dimerizes through an LRR-LRR interface that is required for stable ER folding and cell-surface expression [#2]. Through homophilic LRR-mediated adhesion AMIGO1 promotes neurite extension, fasciculation, and tract formation: the substrate-bound ectodomain drives outgrowth while the soluble ectodomain acts as a dominant-negative that disrupts fasciculation in hippocampal neurons and in zebrafish fiber scaffolds [#0, #4], and genetic deletion in mice reveals a compartment-specific requirement for AMIGO1 in scaling retinal horizontal cell axon arbors [#9]. In parallel, AMIGO1 is an auxiliary subunit of Kv2.1/Kv2.2 voltage-gated potassium channels: it co-immunoprecipitates and colocalizes with Kv2.1, and loss of AMIGO suppresses the neuronal delayed rectifier current and reduces brain Kv2.1 protein in vivo [#1, #5]. Assembly with Kv2 channels hyperpolarizes the activation midpoint by roughly -10 mV, and mechanistically AMIGO1 destabilizes the earliest resting conformation of the Kv2.1 voltage sensors, speeding early sensor movements and shifting the gating charge-voltage relationship to more negative voltages [#7, #8]. The relationship is reciprocal: Kv2 \\u03b1 subunits are obligatory for AMIGO1 clustering, plasma-membrane trafficking, and expression, localizing it to ER-PM junctions, a dependency that extends to motor neurons where Kv2-mediated junction formation is required for AMIGO1 localization [#6, #10].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established AMIGO1 as an LRR/Ig adhesion molecule whose ectodomain bidirectionally controls neurite outgrowth and fasciculation, defining its first functional role in neural morphogenesis.\",\n      \"evidence\": \"Recombinant ectodomain neurite outgrowth assays and homophilic/heterophilic binding assays in hippocampal neurons\",\n      \"pmids\": [\"12629050\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Binding partners beyond the AMIGO family not identified\", \"No in vivo confirmation at this stage\", \"Molecular mechanism of adhesion-to-outgrowth coupling unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Revealed an entirely distinct function: AMIGO1 acts as an auxiliary subunit of Kv2.1, coupling the adhesion molecule to control of neuronal potassium currents.\",\n      \"evidence\": \"Reciprocal Co-IP, immunofluorescence colocalization, HEK electrophysiology, and siRNA knockdown with patch-clamp in neurons\",\n      \"pmids\": [\"22056818\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry of the AMIGO1-Kv2.1 complex not defined\", \"Biophysical mechanism of conductance modulation not yet resolved\", \"Whether Kv2.2 is also modulated untested here\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Determined the ectodomain structure and showed dimerization through the LRR interface is required for surface expression, providing the structural basis for both folding and adhesion.\",\n      \"evidence\": \"2.0 \\u00c5 X-ray crystallography with SAXS, static light scattering, and mutagenesis-coupled surface expression assays\",\n      \"pmids\": [\"21983541\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of full-length transmembrane protein and channel-bound complex unknown\", \"Adhesion interface in trans not directly visualized\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Localized AMIGO1 to dendrites and linked it to dendritic growth and apoptosis resistance, refining its compartment-specific developmental role.\",\n      \"evidence\": \"Immunofluorescence localization, siRNA knockdown with dendrite morphometry, and overexpression apoptosis assays in SH-SY5Y cells\",\n      \"pmids\": [\"21938721\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking AMIGO1 to apoptosis resistance unknown\", \"Signaling pathways downstream of dendritic AMIGO1 not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated in vivo that homophilic AMIGO1 interactions drive fiber tract fasciculation and that AMIGO1 also regulates Kv2.1 for functional circuitry, unifying its adhesion and channel roles in a whole organism.\",\n      \"evidence\": \"Morpholino knockdown and dominant-negative ectodomain overexpression in zebrafish with fiber tract imaging and locomotion behavior\",\n      \"pmids\": [\"24904058\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relative contributions of adhesion vs channel functions to behavior not dissected\", \"Morpholino off-target effects not fully excluded\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Showed AMIGO is required in vivo for normal Kv2.1 protein levels and neuronal electrical activity, establishing the channel dependency genetically.\",\n      \"evidence\": \"AMIGO knockout mouse with Western blot for Kv2.1 and electrophysiology\",\n      \"pmids\": [\"26240432\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether reduced Kv2.1 reflects trafficking or stability not resolved\", \"Behavioral/circuit consequences not detailed here\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined the reciprocal dependency: Kv2 \\u03b1 subunits are obligatory for AMIGO1 clustering and trafficking to ER-PM junctions, reframing the relationship as bidirectional control.\",\n      \"evidence\": \"Cross-species immunofluorescence, live-cell imaging in heterologous cells, and Kv2.1/Kv2.2 single and double knockout mouse analysis\",\n      \"pmids\": [\"29403353\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular interface mediating Kv2-dependent AMIGO1 clustering not mapped\", \"Functional role of ER-PM junction localization for AMIGO1 unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Distinguished isoform-specific channel modulation, showing AMIGO1 hyperpolarizes Kv2 activation by ~-10 mV without the inactivation/deactivation slowing seen with AMIGO2.\",\n      \"evidence\": \"Voltage-clamp electrophysiology and live-cell surface trafficking imaging in heterologous cells\",\n      \"pmids\": [\"34137443\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of isoform-specific effects unknown\", \"Native neuronal validation of -10 mV shift limited\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Resolved the biophysical mechanism: AMIGO1 destabilizes the earliest resting state of Kv2.1 voltage sensors, explaining the hyperpolarized activation.\",\n      \"evidence\": \"Voltage-clamp, gating current measurements, and voltage-sensor toxin fluorescence spectroscopy in mammalian cells\",\n      \"pmids\": [\"35314141\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physical contact points between AMIGO1 and the voltage-sensor domain not identified\", \"Whether this mechanism operates identically on Kv2.2 untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established a precise developmental phenotype: AMIGO1 selectively scales retinal horizontal cell axon arbors, with downstream territory matching of rod bipolar dendrites.\",\n      \"evidence\": \"Global and single-cell conditional Amigo1 knockout mice with arbor morphometry and retinal electrophysiology\",\n      \"pmids\": [\"35169021\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular signaling driving compartment-specific axon scaling unknown\", \"Whether channel function contributes to this morphological role not separated\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Extended the Kv2-dependency to motor neurons and linked it mechanistically to Kv2-VAP-mediated ER-PM junction formation via the Kv2.1 S590A mutant.\",\n      \"evidence\": \"Kv2.1/Kv2.2 knockout and Kv2.1 S590A knock-in mice with immunofluorescence in spinal motor neurons (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.06.04.657913\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, single lab, not peer reviewed\", \"Functional consequence of motor neuron AMIGO1 mislocalization untested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified an Amigo1-expressing GABAergic neuron population in the interpeduncular nucleus that mediates affective signs of nicotine withdrawal, linking AMIGO1 marker expression to circuit-level behavior.\",\n      \"evidence\": \"Chemogenetic DREADD activation of Amigo1-Cre IPN neurons with anxiety and somatic withdrawal behavioral assays (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.07.05.602259\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not peer reviewed\", \"Whether AMIGO1 protein itself, rather than as a cell-type marker, contributes to the behavior is unaddressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How AMIGO1's adhesion/morphogenic function and its Kv2-channel auxiliary function are coordinated within single neurons, and the physical interface mediating both Kv2 binding and voltage-sensor destabilization, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of the AMIGO1-Kv2 complex\", \"Contact residues for voltage-sensor modulation unmapped\", \"Integration of channel vs adhesion roles in development not dissected\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [0, 2, 4]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 7, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 10]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [2, 10]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [1, 5, 8]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 9]}\n    ],\n    \"complexes\": [\"Kv2.1 channel complex\", \"Kv2.2 channel complex\"],\n    \"partners\": [\"KCNB1\", \"KCNB2\", \"AMIGO1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}