Contactin-associated protein 4 (Caspr4), also known as contactin-associated protein-like protein (CNTNAP4), is expressed in various regions of the brain. Recent reports suggest that CNTNAP4 is a susceptibility gene of autism spectrum disorders (ASDs). However, the molecular function of Caspr4 in the brain has yet to be identified. In this study, we show an essential role of Caspr4 in neural progenitor cells (NPCs). Caspr4 is expressed in NPCs in the subventricular zone (SVZ), a neurogenic region in the developing cortex. Knocking down of Caspr4 enhances the proliferation of NPCs derived from the SVZ of embryonic day 14 mouse. Neuronal differentiation is increased by overexpression of Caspr4, but decreased by knocking down of Caspr4 in cultured mouse NPCs. Transfection of the intracellular domain of Caspr4 (C4ICD) rescues the abnormal decreased neuronal differentiation of Caspr4-knocking down NPCs. Ligand of Numb protein X2 (LNX2), a binding partner of Numb, interacts with Caspr4 in a PDZ domain-dependent manner and plays a similar role to Caspr4 in NPCs. Moreover, transfection of LNX2 rescues the decreased neuronal differentiation in Caspr4-knocking down NPCs. In contrast, transfection of C4ICD fails to do so in LNX2-knocking down NPCs. These results indicate that Caspr4 inhibits neuronal differentiation in a LNX-dependent manner. Therefore, this study reveals a novel role of Caspr4 through LNX2 in NPCs, which may link to the pathogenesis of ASDs.
Get full access to this article
View all access options for this article.
References
1.
TempleS. (2001). The development of neural stem cells. Nature, 414:112–117.
2.
LiuHK, WangY, BelzT, BockD, TakacsA, RadlwimmerB, BarbusS, ReifenbergerG, LichterP and SchutzG. (2010). The nuclear receptor tailless induces long-term neural stem cell expansion and brain tumor initiation. Genes Dev, 24:683–695.
3.
SwartlingFJ, SavovV, PerssonAI, ChenJ, HackettCS, NorthcottPA, GrimmerMR, LauJ, CheslerL, et al. (2012). Distinct neural stem cell populations give rise to disparate brain tumors in response to N-MYC. Cancer Cell, 21:601–613.
4.
ReifA, FritzenS, FingerM, StrobelA, LauerM, SchmittA and LeschKP. (2006). Neural stem cell proliferation is decreased in schizophrenia, but not in depression. Mol Psychiatry, 11:514–522.
5.
EischAJ and PetrikD. (2012). Depression and hippocampal neurogenesis: a road to remission?. Science, 338:72–75.
6.
SnyderJS, SoumierA, BrewerM, PickelJ and CameronHA. (2011). Adult hippocampal neurogenesis buffers stress responses and depressive behaviour. Nature, 476:458–461.
7.
TaupinP. (2009). Adult neurogenesis, neural stem cells and Alzheimer's disease: developments, limitations, problems and promises. Curr Alzheimer Res, 6:461–470.
8.
MaQH, FutagawaT, YangWL, JiangXD, ZengL, TakedaY, XuRX, BagnardD, SchachnerM, et al. (2008). A TAG1-APP signalling pathway through Fe65 negatively modulates neurogenesis. Nat Cell Biol, 10:283–294.
9.
StateMW and SestanN. (2012). Neuroscience. The emerging biology of autism spectrum disorders. Science, 337:1301–1303.
10.
PenagarikanoO, AbrahamsBS, HermanEI, WindenKD, GdalyahuA, DongH, SonnenblickLI, GruverR, AlmajanoJ, et al. (2011). Absence of CNTNAP2 leads to epilepsy, neuronal migration abnormalities, and core autism-related deficits. Cell, 147:235–246.
11.
LiH, RadfordJC, RagusaMJ, SheaKL, McKercherSR, ZarembaJD, SoussouW, NieZ, KangYJ, et al. (2008). Transcription factor MEF2C influences neural stem/progenitor cell differentiation and maturation in vivo. Proc Natl Acad Sci U S A, 105:9397–9402.
12.
AmiriA, ChoW, ZhouJ, BirnbaumSG, SintonCM, McKayRM and ParadaLF. (2012). Pten deletion in adult hippocampal neural stem/progenitor cells causes cellular abnormalities and alters neurogenesis. J Neurosci, 32:5880–5890.
13.
EinheberS, ZanazziG, ChingW, SchererS, MilnerTA, PelesE and SalzerJL. (1997). The axonal membrane protein Caspr, a homologue of neurexin IV, is a component of the septate-like paranodal junctions that assemble during myelination. J Cell Biol, 139:1495–1506.
14.
CharlesP, TaitS, Faivre-SarrailhC, BarbinG, Gunn-MooreF, Denisenko-NehrbassN, GuennocAM, GiraultJA, BrophyPJ and LubetzkiC. (2002). Neurofascin is a glial receptor for the paranodin/Caspr-contactin axonal complex at the axoglial junction. Curr Biol, 12:217–220.
15.
PoliakS, GollanL, MartinezR, CusterA, EinheberS, SalzerJL, TrimmerJS, ShragerP and PelesE. (1999). Caspr2, a new member of the neurexin superfamily, is localized at the juxtaparanodes of myelinated axons and associates with K+ channels. Neuron, 24:1037–1047.
16.
PoliakS, SalomonD, ElhananyH, SabanayH, KiernanB, PevnyL, StewartCL, XuX, ChiuSY, et al. (2003). Juxtaparanodal clustering of Shaker-like K+ channels in myelinated axons depends on Caspr2 and TAG-1. J Cell Biol, 162:1149–1160.
17.
TrakaM, GoutebrozeL, DenisenkoN, BessaM, NifliA, HavakiS, IwakuraY, FukamauchiF, WatanabeK, et al. (2003). Association of TAG-1 with Caspr2 is essential for the molecular organization of juxtaparanodal regions of myelinated fibers. J Cell Biol, 162:1161–1172.
18.
O'RoakBJ, VivesL, GirirajanS, KarakocE, KrummN, CoeBP, LevyR, KoA, LeeC, et al. (2012). Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature, 485:246–250.
19.
WangLS, HranilovicD, WangK, LindquistIE, YurcabaL, PetkovicZB, GidayaN, JernejB, HakonarsonH and BucanM. (2010). Population-based study of genetic variation in individuals with autism spectrum disorders from Croatia. BMC Med Genet, 11:134
20.
KarayannisT, AuE, PatelJC, KruglikovI, MarkxS, DelormeR, HeronD, SalomonD, GlessnerJ, et al. (2014). Cntnap4 differentially contributes to GABAergic and dopaminergic synaptic transmission. Nature, 511:236–240.
21.
HanS, TaiC, WestenbroekRE, YuFH, CheahCS, PotterGB, RubensteinJL, ScheuerT, de la IglesiaHO and CatterallWA. (2012). Autistic-like behaviour in Scn1a+/− mice and rescue by enhanced GABA-mediated neurotransmission. Nature, 489:385–390.
22.
TrautW, WeichenhanD, HimmelbauerH and WinkingH. (2006). New members of the neurexin superfamily: multiple rodent homologues of the human CASPR5 gene. Mamm Genome, 17:723–731.
23.
RiceDS, NorthcuttGM and KurschnerC. (2001). The Lnx family proteins function as molecular scaffolds for Numb family proteins. Mol Cell Neurosci, 18:525–540.
24.
NieJ, McGillMA, DermerM, DhoSE, WoltingCD and McGladeCJ. (2002). LNX functions as a RING type E3 ubiquitin ligase that targets the cell fate determinant Numb for ubiquitin-dependent degradation. EMBO J, 21:93–102.
25.
CorbinJG, GaianoN, JulianoSL, PoluchS, StancikE and HaydarTF. (2008). Regulation of neural progenitor cell development in the nervous system. J Neurochem, 106:2272–2287.
26.
SpiegelI, SalomonD, ErneB, Schaeren-WiemersN and PelesE. (2002). Caspr3 and caspr4, two novel members of the caspr family are expressed in the nervous system and interact with PDZ domains. Mol Cell Neurosci, 20:283–297.
27.
GarnerCC, NashJ and HuganirRL. (2000). PDZ domains in synapse assembly and signalling. Trends Cell Biol, 10:274–280.
28.
Gonzalez-MariscalL, BetanzosA and Avila-FloresA. (2000). MAGUK proteins: structure and role in the tight junction. Semin Cell Dev Biol, 11:315–324.
29.
ShengM and PakDT. (2000). Ligand-gated ion channel interactions with cytoskeletal and signaling proteins. Annu Rev Physiol, 62:755–778.
30.
ScottK and ZukerCS. (1998). Assembly of the Drosophila phototransduction cascade into a signalling complex shapes elementary responses. Nature, 395:805–808.
31.
DevanathanV, JakovcevskiI, SantuccioneA, LiS, LeeHJ, PelesE, Leshchyns'kaI, SytnykV and SchachnerM. (2010). Cellular form of prion protein inhibits Reelin-mediated shedding of Caspr from the neuronal cell surface to potentiate Caspr-mediated inhibition of neurite outgrowth. J Neurosci, 30:9292–9305.
32.
de la PompaJL, WakehamA, CorreiaKM, SamperE, BrownS, AguileraRJ, NakanoT, HonjoT, MakTW, RossantJ and ConlonRA. (1997). Conservation of the Notch signalling pathway in mammalian neurogenesis. Development, 124:1139–1148.
33.
CampsJ, PittJJ, EmonsG, HummonAB, CaseCM, GradeM, JonesTL, NguyenQT, GhadimiBM, et al. (2013). Genetic amplification of the NOTCH modulator LNX2 upregulates the WNT/beta-catenin pathway in colorectal cancer. Cancer Res, 73:2003–2013.
34.
HirabayashiY, ItohY, TabataH, NakajimaK, AkiyamaT, MasuyamaN and GotohY. (2004). The Wnt/beta-catenin pathway directs neuronal differentiation of cortical neural precursor cells. Development, 131:2791–2801.
35.
KansakuA, HirabayashiS, MoriH, FujiwaraN, KawataA, IkedaM, RokukawaC, KuriharaH and HataY. (2006). Ligand-of-Numb protein X is an endocytic scaffold for junctional adhesion molecule 4. Oncogene, 25:5071–5084.
36.
MirzaM, HreinssonJ, StrandML, HovattaO, SoderO, PhilipsonL, PetterssonRF and SollerbrantK. (2006). Coxsackievirus and adenovirus receptor (CAR) is expressed in male germ cells and forms a complex with the differentiation factor JAM-C in mouse testis. Exp Cell Res, 312:817–830.
37.
D'AgostinoM, TornilloG, CaporasoMG, BaroneMV, GhigoE, BonattiS and MottolaG. (2011). Ligand of Numb proteins LNX1p80 and LNX2 interact with the human glycoprotein CD8alpha and promote its ubiquitylation and endocytosis. J Cell Sci, 124:3545–3556.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
