| 1991 |
Nr-CAM (NrCAM) was identified as a novel neural glycoprotein with a structure comprising six Ig-like domains, five fibronectin type III repeats, a transmembrane domain, and a short cytoplasmic domain, encoded by a single gene with alternatively spliced mRNAs. Purified protein (Mr 145,000) was confirmed by N-terminal sequencing matching the cDNA-predicted sequence. |
cDNA cloning, protein purification by lentil lectin affinity chromatography/FPLC, N-terminal sequencing, Northern and Southern blotting |
The Journal of cell biology |
High |
2045418
|
| 1992 |
NrCAM (Bravo) mediates both homophilic (divalent cation-independent) and heterophilic (divalent cation-dependent) cell adhesion. Homophilic binding was demonstrated between transfected L cells and between cells and recombinant FGTNr (Ig domains 1-6 + first FNIII repeat fusion protein). Heterophilic binding occurred between NrCAM-transfected and untransfected L cells and between FGTNr and fibroblasts. |
Cell aggregation assay, substrate binding assay, Covasphere aggregation assay, recombinant fusion protein (FGTNr) inhibition, L-cell transfection |
The Journal of cell biology |
High |
1527169
|
| 1992 |
Bravo/NrCAM has a heterodimer structure composed of an alpha chain (Mr 140/130 kD) and a beta chain (60-80 kD) generated by cleavage of an intact polypeptide at a conserved Ser-Arg/Lys-Arg site, analogous to L1 and Ng-CAM. The molecule contains alternatively spliced sequences encoding both extra- and intracellular stretches. |
cDNA cloning, SDS-PAGE, sequence analysis |
The Journal of cell biology |
Medium |
1512296
|
| 1993 |
Neurite extension of tectal cells on immobilized F11 (contactin) is mediated by NrCAM/Bravo on the axonal surface. Direct heterophilic binding between F11 and NrCAM/Bravo was demonstrated, and the interaction was mapped to the second or third Ig-like domain of F11 using domain-specific monoclonal antibodies and deletion mutant proteins expressed on COS cells. |
Neurite outgrowth assay, antibody blocking, COS cell binding assay, deletion mutants |
Neuron |
High |
8274278
|
| 1994 |
The cytoplasmic domains of NrCAM (along with neurofascin, L1, NgCAM, and neuroglian) directly associate with ankyrins. NrCAM and neurofascin together comprise over 0.5% of total membrane protein in adult brain tissue, indicating this ankyrin linkage is a major membrane-cytoskeletal connection. |
Biochemical co-association assay, protein quantification from brain membrane fractions |
The Journal of biological chemistry |
High |
7961622
|
| 1995 |
In vivo perturbation experiments in chick showed that the interaction between axonin-1 on commissural growth cones and Nr-CAM on floor plate cells is required for accurate midline pathfinding. When axonin-1 or Nr-CAM interactions were perturbed (by antibody injection), many commissural axons failed to cross the midline and turned along the ipsilateral floor plate border instead. |
In vivo antibody perturbation in chick embryo spinal cord, histological analysis of axon trajectories |
Neuron |
High |
7541632
|
| 1995 |
Axonin-1 binds directly to NrCAM on peripheral glial cells, mediating neuron-glia contacts. Fluorescent microspheres conjugated with axonin-1 bound to glial cells via NrCAM (identified by antibody blockage). Anti-axonin-1 and anti-NrCAM antibodies both perturbed neurite-glia contact formation in dissociated DRG cultures, implicating this interaction in early axon ensheathment. |
Microsphere binding assay, antibody blocking, purified protein binding assay, dissociated DRG neuron-glia culture perturbation |
The Journal of cell biology |
High |
7490283
|
| 1996 |
NrCAM and a specific isoform of neurofascin (mucin+/third FNIII domain-) are colocalized with ankyrinG and voltage-dependent sodium channels at nodes of Ranvier and axon initial segments. NrCAM was identified by screening a rat brain cDNA expression library with anti-neurofascin antibody; it shares >70% cytoplasmic domain identity with neurofascin. |
cDNA library screening, isoform-specific antibody generation, immunofluorescence co-localization, cDNA sequencing |
The Journal of cell biology |
High |
8947556
|
| 1996 |
Neurofascin promotes neurite extension from tectal cells by heterophilic interaction with NrCAM on axonal surfaces. Conversely, when NrCAM is the substrate, it induces neurite extension through F11 (not neurofascin) as the axonal receptor. Direct binding between neurofascin and NrCAM was demonstrated by transfected COS7 cell binding and immunoprecipitation, and mapped to the Ig domains of neurofascin. Alternative splicing of neurofascin modulates this binding. |
Neurite outgrowth assay on immobilized substrates, antibody blocking, COS7 cell transfection binding assay, immunoprecipitation, deletion mapping |
The Journal of cell biology |
High |
8922386
|
| 1997 |
The extracellular region of glial receptor protein tyrosine phosphatase beta (RPTPbeta) binds to a complex of contactin and NrCAM on neurons. Neurite outgrowth induced by betaCFS (RPTPbeta fusion) was inhibited by antibodies against both NrCAM and contactin, and contactin/NrCAM co-immunoprecipitated with betaCFS. NIH-3T3 cells expressing betaCFS on their surfaces induced neuronal differentiation. |
Antibody blocking of neurite outgrowth, co-immunoprecipitation, recombinant fusion proteins, transfected cell co-culture assay |
The Journal of cell biology |
High |
9049255
|
| 1999 |
NrCAM functions as a substrate ligand for neurite outgrowth from dorsal root ganglion (DRG) and sympathetic ganglion neurons via axonin-1 as the neuronal receptor. A recombinant Nr-CAM-Fc fusion protein (containing all 6 Ig domains and first 2 FNIII repeats) promoted neurite outgrowth from peripheral but not central neurons; anti-axonin-1 antibodies inhibited this outgrowth. In ovo injection of Nr-Fc produced commissural axon guidance errors similar to anti-axonin-1 treatment. |
Recombinant Fc fusion protein substrate assay, antibody blocking of neurite outgrowth, in ovo injection, immunostaining |
Developmental biology |
High |
10328925
|
| 1999 |
NrCAM is the functional receptor on cerebellar granule cells for the neuronal adhesion glycoprotein F3 (contactin). F3Fc-conjugated microspheres bound growth cones via NrCAM (not L1). These beads moved retrogradely at 5.7 µm/min (actin retrograde flow velocity); cytochalasin B (actin disruptor) abolished this movement. NrCAM clustering (induced by cross-linked F3Fc) was required for retrograde mobility. |
Microsphere binding assay, time-lapse video microscopy, cytochalasin B treatment, single particle tracking |
Journal of cell science |
High |
10462518
|
| 2000 |
Axonin-1 interaction with floor-plate NrCAM mediates commissural axon guidance (directionality) without promoting axon elongation. In stripe assays, commissural axons preferred NrCAM+NgCAM stripes; anti-axonin-1 abolished this preference without reducing neurite length. In vivo, axonin-1/NrCAM perturbation selectively caused guidance failure, not elongation defects. |
In vitro stripe assay, antibody blocking, in vivo perturbation with phenotypic measurement of guidance vs. elongation |
The Journal of cell biology |
High |
10811834
|
| 2001 |
Nr-CAM interacts with Nr-CAM-Fc fusion protein and with neurofascin on axonal surfaces to cluster neurofascin and co-precipitate it. Treatment of myelinating DRG-Schwann cell cocultures with Nr-CAM-Fc fusion protein specifically inhibited Na+ channel and ankyrinG accumulation at nodes of Ranvier without affecting myelination extent, demonstrating that NrCAM-neurofascin interactions are required for nodal Na+ channel clustering. |
Myelinating DRG-Schwann cell coculture, Nr-CAM-Fc fusion protein perturbation, immunofluorescence, co-precipitation |
Current biology : CB |
High |
11728309
|
| 2001 |
Nr-CAM-deficient mice have functionally null Nr-CAM; cerebellar granule cells from these mice fail to extend neurites on contactin substrate in vitro, confirming contactin as an Nr-CAM ligand. Combined Nr-CAM/L1 double mutant mice exhibit severe cerebellar folial defects and reduced inner granule cell layer thickness, demonstrating overlapping functions of these related CAMs. |
Nr-CAM knockout mouse, neurite outgrowth assay on contactin substrate, histological analysis of double mutant cerebellum, antibody perturbation in culture |
The Journal of cell biology |
High |
11564762
|
| 2001 |
Targeted ablation of NrCAM in mice causes formation of mature cataracts due to disorganization of lens fiber cells, accompanied by abnormalities in the cytoskeleton and connexin50-containing gap junctions. Ankyrin-B mutant mice display an indistinguishable lens fiber disorganization phenotype, providing genetic evidence that NrCAM and ankyrin-B interact to maintain lens fiber cell contacts. |
NrCAM knockout mouse, ankyrin-B mutant mouse, histology, immunostaining, electron microscopy, genetic epistasis |
The Journal of cell biology |
High |
11449000
|
| 2002 |
Nr-CAM is a direct transcriptional target gene of the beta-catenin/LEF-1 signaling pathway. LEF/TCF binding sites in the Nr-CAM promoter were required for activation by beta-catenin or plakoglobin. Retroviral transduction of Nr-CAM into NIH3T3 cells stimulated cell growth, motility, transformation, and tumor formation in nude mice. Dominant-negative LEF-1 decreased Nr-CAM expression; anti-Nr-CAM antibodies inhibited B16 melanoma motility. |
DNA microarray, promoter-reporter assay with LEF/TCF site mutation, retroviral transduction, cell growth/motility assays, nude mouse xenograft, dominant-negative LEF-1 |
Genes & development |
High |
12183361
|
| 2003 |
NrCAM (ankyrinG-binding protein) precedes Na+ channels at cluster sites adjacent to Schwann cell process tips during node of Ranvier formation. In NrCAM null mutants, both Na+ channel and ankyrinG sequestration at developing nodes are delayed. NrCAM acts locally at individual nodes (not globally on neuronal expression) and its action is linked to glial contact. |
NrCAM null mutant mouse, immunostaining during node development, quantitative analysis of Na+ channel/ankyrinG clustering timing |
The Journal of neuroscience |
High |
14602817
|
| 2004 |
NrCAM coupling to the retrograde actin flow requires both cytoplasmic tail interactions and cis-interactions via FNIII domains (deletion of both is needed to abolish retrograde movement). Additionally, NrCAM-actin coupling requires partitioning into lipid rafts, as cholesterol depletion by methyl-beta-cyclodextrin abolished retrograde movement. TAG-1 bead binding induced coalescence of lipid rafts (caveolin-1 recruitment) at adhesive contact sites. |
Optical tweezers, single particle tracking, deletion mutants of NrCAM, cholesterol depletion with methyl-beta-cyclodextrin, FRAP, immunofluorescence for caveolin-1 |
Molecular biology of the cell |
High |
15254265
|
| 2005 |
NrCAM undergoes metalloprotease-mediated ectodomain shedding. Conditioned medium and purified Nr-CAM-Fc fusion protein both enhanced cell motility, proliferation, and activated ERK and AKT signaling pathways. NrCAM was found in complex with alpha4beta1 integrins in melanoma cells. Stable expression of the ectodomain alone was sufficient to confer cell transformation and tumorigenesis in mice. |
Metalloprotease shedding detection, conditioned medium assays, co-immunoprecipitation with alpha4beta1 integrins, ERK/AKT phosphorylation assays, siRNA knockdown, nude mouse tumor assay |
Cancer research |
High |
16357171
|
| 2006 |
The cytoplasmic carboxy-terminus of NrCAM contains a PDZ-binding motif that specifically interacts with class I PDZ domains of SAP90/PSD95 and SAP97. This interaction is unique among L1 family members (L1, CHL1, Neurofascin do not bind these PDZ domains). In transfected COS-7 cells, NrCAM-mediated recruitment of SAP97 to the plasma membrane required the PDZ-binding motif. NrCAM and SAP97 colocalize in photoreceptor terminals. |
PDZ domain binding assay, transfection of COS-7 cells with PDZ-motif mutants, immunocytochemistry co-localization |
The European journal of neuroscience |
High |
16882004
|
| 2006 |
Nr-CAM expressed on contralateral retinal ganglion cells (RGCs) is critical for guidance of late-born VTC RGCs at the optic chiasm. Blocking Nr-CAM function increases the ipsilateral projection size and reduces neurite outgrowth on chiasm cells in an age- and region-specific manner. EphB1/ephrin-B2-mediated repulsion and Nr-CAM-mediated attraction are distinct parallel molecular programs governing binocular visual pathway formation. |
Nr-CAM null mouse, anterograde axon tracing, in vitro neurite outgrowth on chiasm cell substrate, antibody blocking |
Neuron |
High |
16701205
|
| 2010 |
Initial clustering of Na+ channels at PNS heminodes requires glial NrCAM and gliomedin acting through their axonal receptor neurofascin 186 (NF186). This glial signal is distinct from a second paranodal junction-dependent mechanism. NrCAM and gliomedin cooperate to capture Na+ channels at heminodes before fusion into mature nodes. |
Gliomedin/NrCAM knockout mice, immunostaining during PNS development, electron microscopy |
Neuron |
High |
20188654
|
| 2011 |
NrCAM forms a molecular complex with neuropilin-2 (Npn-2) in brain and neural cells. Genetic deletion of NrCAM causes misprojection of thalamic axons caudally at the ventral telencephalon and striking mistargeting of motor and somatosensory thalamic axons to primary visual cortex. NrCAM is required for Sema3F-induced growth cone collapse in thalamic neuron cultures, consistent with a role in Sema3F-induced axon repulsion via the NrCAM-Npn-2 complex. |
NrCAM null mouse, anterograde axon tracing, co-immunoprecipitation of NrCAM-Npn-2 complex, growth cone collapse assay, visual evoked potentials |
The Journal of neuroscience |
High |
21273439
|
| 2012 |
Sema6D and Nr-CAM are expressed on midline radial glia and Plexin-A1 on chiasm neurons; Plexin-A1 and Nr-CAM are also expressed on contralateral RGCs. Nr-CAM functions as a receptor for Sema6D. Sema6D in combination with Nr-CAM and Plexin-A1 converts Sema6D-mediated repulsion to growth promotion for contralateral RGCs. All three (Sema6D, Plexin-A1, NrCAM) are required for efficient RGC decussation. |
Knockout mouse analysis, in vitro growth assays, receptor identification experiments, genetic interaction analysis |
Neuron |
High |
22632726
|
| 2013 |
NrCAM contributes to mediolateral retinocollicular axon targeting through regulation of RGC axon branch orientation. EphB2 tyrosine kinase (but not kinase-dead EphB2) phosphorylates NrCAM at a conserved tyrosine in the FIGQY ankyrin-binding motif, perturbing ankyrin recruitment in NrCAM-transfected HEK293 cells. In vivo, phospho-FIGQY NrCAM in SC is decreased in EphB1/3 null mice and increased in constitutively active EphB2 mice. |
NrCAM null mouse axon tracing, EphB kinase phosphorylation assay in HEK293 cells, kinase-dead mutant, EphB null mouse analysis, ankyrin recruitment assay |
PloS one |
High |
24023801
|
| 2014 |
NrCAM localizes to dendritic spines of cortical pyramidal neurons and forms a complex with Sema3F receptor subunits Neuropilin-2 (Npn-2) and PlexinA3 (PlexA3) through an Npn-2-binding sequence (TARNER) in the extracellular Ig1 domain. NrCAM deletion elevates spine density on apical dendrites and increases mEPSC frequency. Recombinant Sema3F-Fc induced spine retraction on wild-type but not NrCAM-null neurons; re-expression of NrCAM rescued the response. Trans heterozygous genetic interaction confirmed NrCAM and Sema3F pathways interact in vivo. |
NrCAM null mouse, co-immunoprecipitation, spine density quantification, electron microscopy, whole-cell electrophysiology, rescue experiments, trans heterozygous genetic interaction |
The Journal of neuroscience |
High |
25143608
|
| 2014 |
Long-term maintenance of Na+ channels at PNS nodes of Ranvier requires axoglial contact mediated by both gliomedin and NrCAM together. Mice lacking both molecules (but not either alone) show gradual loss of nodal Na+ channels and other axonal components, formation of binary nodes, dysregulation of nodal gap length, and neurological abnormalities. Node disintegration follows the reverse order of assembly: NF186 disappears first, then Na+ channels and ankyrinG, then βIV spectrin. |
Gliomedin/NrCAM double knockout mouse, immunostaining, electron microscopy, electrophysiology, behavioral analysis |
The Journal of neuroscience |
High |
24719088
|
| 2017 |
NrCAM activates MAPK/Erk and PI3K/Akt signaling pathways via ectodomain shedding and binding to EGFR and α4β1 integrins. These pathways in turn upregulate NrCAM expression through the GSK3β/β-catenin axis, establishing positive feedback loops. NrCAM depletion inhibited thyroid cancer cell growth and invasiveness. |
NrCAM knockdown and overexpression in thyroid cancer cells, phosphorylation assays, co-immunoprecipitation, nude mouse xenograft, transgenic BrafV600E mouse |
The Journal of clinical endocrinology and metabolism |
Medium |
27732334
|
| 2019 |
NrCAM functions as a substrate for ADAM10 metalloprotease cleavage. ADAM10 controls NrCAM surface levels and regulates neurite outgrowth in an NrCAM-dependent manner in vitro. However, ADAM10 cleavage of NrCAM (unlike APP) is not stimulated by the ADAM10 activator acitretin, suggesting substrate-selective ADAM10 activation is feasible and that NrCAM can serve as a biomarker of basal (non-stimulated) ADAM10 activity. |
ADAM10 activity assay, NrCAM surface level quantification, neurite outgrowth assay, human CSF proteomics from clinical trial |
EMBO molecular medicine |
High |
30833305
|
| 2019 |
NrCAM promotes clustering of the Sema3F holoreceptor complex by interfacing with Neuropilin-2 (Npn-2) and the PDZ scaffold protein SAP102. NrCAM-induced receptor clustering stimulates Rap-GAP activity of PlexinA3 (PlexA3) within the holoreceptor complex, which inhibits Rap1-GTPase and inactivates adhesive β1 integrins, mediating Sema3F-induced spine pruning during adolescence. Conditional inducible NrCAM deletion (Nex1Cre-ERT2:NrCAMflox/flox) showed NrCAM acts cell-autonomously in pyramidal neurons. |
Conditional inducible knockout mouse, molecular modeling, holoreceptor complex assembly assay, Rap-GAP activity assay, Rap1-GTPase assay, β1 integrin activity assay, spine density quantification |
Cerebral cortex |
High |
29415226
|
| 2019 |
NrCAM mediates fasciculation of axon fibers in the stria terminalis, regulating amygdala-BNST connectivity. NrCAM null mice show pronounced defasciculation and misprojection of fibers in the stria terminalis and are impaired in context-dependent (but not cued) fear conditioning, linking NrCAM-mediated axon fasciculation to amygdalar-BNST circuit function. |
NrCAM null mouse, neurofilament immunohistochemistry of fiber tracts, contextual vs. cued fear conditioning behavioral testing |
Frontiers in cell and developmental biology |
Medium |
30766872
|
| 2018 |
Neurocan (a chondroitin sulfate proteoglycan) inhibits Sema3F-induced spine elimination through binding to NrCAM. Cell binding and ELISA assays demonstrated association of Neurocan with NrCAM. Neurocan blocked Sema3F-induced morphological retraction in COS-7 cells mediated through NrCAM, Npn-2, and PlexinA3. Glycosaminoglycan chains of Neurocan (but not the C-terminal sushi domain) were required for this inhibition. |
Cell binding assay, ELISA, COS-7 cell morphology assay, cortical neuron culture spine assay, domain deletion analysis |
Frontiers in cellular neuroscience |
Medium |
30356641
|
| 2020 |
Alternative splicing of Nrcam exon 10 in dorsal root ganglia contributes to neuropathic pain. Spinal nerve ligation increases exon 10 insertion (Nrcam+10 variant) at the expense of Nrcam-10 in injured DRG. Antisense oligonucleotides (ASO) targeting exon 10 attenuated mechanical allodynia, thermal hyperalgesia, and cold allodynia in SNL and CCD models in both male and female mice. |
RNA sequencing, ASO treatment (DRG microinjection and intrathecal), behavioral pain assays (von Frey, thermal, cold plate) |
The journal of pain |
Medium |
31917219
|
| 2022 |
NrCAM and Ankyrin B mediate perisomatic synaptic contact between CCK-basket cells (CCK-BCs) and pyramidal neurons (PNs) in mouse medial prefrontal cortex. NrCAM-null mice show significant decreases in CCK-BC (VGLUT3+ and VGAT+) synaptic puncta on PN soma but no decrease in PV-BC puncta or cell loss. Ankyrin B deletion specifically from PNs also reduces VGLUT3+ CCK-BC puncta, establishing that postsynaptic NrCAM-Ankyrin B interaction is required for CCK-BC synapse formation. |
NrCAM null mouse, conditional Ankyrin B deletion (Nex1Cre-ERT2:Ank2flox/flox), CCK-BC reporter mouse (Sncg-tdTomato), immunolabeling, confocal quantification |
Current research in neurobiology |
Medium |
40276719
|
| 2022 |
NrCAM activates the NF-κB signaling pathway by competitively binding to SUMO-1, reducing IκBα SUMOylation and increasing IκBα phosphorylation and degradation, thereby promoting NF-κB-dependent Th17 cell differentiation. NrCAM overexpression increased IL-21 via NF-κB (p65 binding to IL-21 promoter), and NF-κB inhibitor BAY11-7082 partially reversed NrCAM's effects. |
NrCAM overexpression and knockdown in CD4+ T cells, flow cytometry for Th17 markers, co-immunoprecipitation of NrCAM-SUMO-1, p-IκBα western blot, ChIP for p65 on IL-21 promoter, NF-κB inhibitor treatment |
Scandinavian journal of immunology |
Medium |
39155774
|
| 2023 |
In hepatocellular carcinoma (HCC), NrCAM promotes liver cancer stem cell (LCSC) migration, invasion, and metastasis by activating epithelial-mesenchymal transition (EMT) and matrix metalloproteinases (MMPs) through the MACF1-mediated β-catenin signaling pathway in LCSCs. |
MYC-driven LCSC organoids, NrCAM knockdown and overexpression, invasion/migration assays, in vivo tumor allografts (intra-hepatic and lung metastasis), scRNA-seq, MACF1/β-catenin pathway analysis |
Journal of experimental & clinical cancer research |
Medium |
37993901
|
| 2023 |
Rbfox1 (RNA-binding Fox1) regulates alternative splicing of NrCAM exon 10 in DRG after spinal nerve ligation. Downregulation of Rbfox1 following nerve injury amplifies exon 10 insertion (L-Nrcam increase, S-Nrcam decrease). Restoring Rbfox1 mitigates nociceptive hypersensitivity; mimicking Rbfox1 downregulation generates neuropathic pain symptoms. |
Transcriptome profiling (RNA-seq), bioinformatic splicing analysis, Rbfox1 overexpression/knockdown in DRG, splicing isoform quantification, behavioral pain assays |
Neurotherapeutics |
Medium |
38241164
|
| 2025 |
The NRCAM proteoform with microexons 5 and 19 skipped (Δex5Δex19 NRCAM) is uniformly expressed in pediatric high-grade gliomas but not normal brain. This specific splice variant (not full-length NrCAM) is essential for pHGG cell migration, invasion in vitro, and tumor growth in vivo. A monoclonal antibody selective for Δex5Δex19 NRCAM enables T-cell-mediated killing of pHGG cells via an FcRI-based universal immune receptor. |
Bulk and single-nuclei short- and long-read RNA-seq, loss-of-function assays (migration/invasion in vitro), in vivo tumor growth, monoclonal antibody development, T-cell killing assay |
Cell reports |
High |
40782352
|